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Orash Mahmoudsalehi A, Soleimani M, Stalin Catzim Rios K, Ortega-Lara W, Mamidi N. Advanced 3D scaffolds for corneal stroma regeneration: a preclinical progress. J Mater Chem B 2025; 13:5980-6020. [PMID: 40105794 DOI: 10.1039/d5tb00090d] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/20/2025]
Abstract
Corneal stromal defects represent a significant global cause of blindness, necessitating innovative therapeutic strategies to address the limitations of conventional treatments, such as corneal transplantation. Tissue engineering, a cornerstone of regenerative medicine, offers a transformative approach by leveraging biomaterial-based solutions to restore damaged tissues. Among these, three-dimensional (3D) scaffolds fabricated using advanced techniques like 3D printing have emerged as a promising platform for corneal regeneration. These scaffolds replicate the native extracellular matrix (ECM) architecture, providing a biomimetic microenvironment that supports cell proliferation, differentiation, and tissue integration. This review highlights recent advances in the design and fabrication of 3D scaffolds for corneal stroma engineering (CSE), emphasizing the critical interplay between scaffold architecture, mechanical properties, and bioactive signaling in directing cellular behavior and tissue regeneration. Likewise, we emphasize the diverse range of biomaterials utilized in scaffold fabrication, highlighting their influence on cellular interactions and tissue reconstruction. By elucidating the complex relationship between scaffold design and biologics, this review aims to illuminate the evolution of next-generation strategies for engineering functional corneal tissue. Eventually, this review will provide a comprehensive synthesis of the current state-of-the-art in 3D scaffold-based corneal tissue engineering (CTE), offering insights that could advance progress toward effective vision restoration therapies.
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Affiliation(s)
- Amin Orash Mahmoudsalehi
- Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Monterrey 64849, Mexico.
| | - Maryam Soleimani
- Silesian University of Technology, Faculty of Mechanical Engineering, Department of Didactic Laboratory of Nanotechnology and Material Technologies, 18a Konareskiego Str, 44-100 Gliwice, Poland
| | - Kevin Stalin Catzim Rios
- Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Monterrey 64849, Mexico.
| | - Wendy Ortega-Lara
- Tecnologico de Monterrey, Escuela de Ingeniería y Ciencias, Av. Eugenio Garza Sada 2501 Sur, Monterrey 64849, Mexico.
| | - Narsimha Mamidi
- School of Pharmacy, Wisconsin Center for NanoBioSystems, University of Wisconsin-Madison, Madison, Wisconsin, USA.
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Ghosh A, Bera AK, Singh V, Basu S, Pati F. Bioprinting of anisotropic functional corneal stroma using mechanically robust multi-material bioink based on decellularized cornea matrix. BIOMATERIALS ADVANCES 2024; 165:214007. [PMID: 39216318 DOI: 10.1016/j.bioadv.2024.214007] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 04/27/2024] [Revised: 07/08/2024] [Accepted: 08/23/2024] [Indexed: 09/04/2024]
Abstract
Corneal scarring is a common cause of blindness, affecting millions globally each year. A huge gap between the demand and supply of donor tissue currently limits corneal transplantation, the only definitive therapy for patients with corneal scarring. To overcome this challenge, researchers have harnessed the efficacy of 3D bioprinting to fabricate artificial corneal stromal constructs. With all the different bioinks available, the decellularized corneal matrix-based bioprinted construct can fulfill the required biological functionality but is limited by the lack of mechanical stiffness. Additionally, from a biophysical standpoint, it is necessary for an ideal corneal substitute to mimic the anisotropy of the cornea from the central optic zone to the surrounding periphery. In this study, we enhanced the mechanical robustness of decellularized cornea matrix (DCM) hydrogel by blending it with another natural polymer, sonicated silk fibroin solution in a defined ratio. Although hybrid hydrogel has an increased complex modulus than DCM hydrogel, it has a lower in vitro degradation rate and increased opaqueness due to the presence of crystalline beta-sheet conformation within the hydrogel. Therefore, we used this multi-material bioink-based approach to fabricate a corneal stromal equivalent where the outer peripheral corneal rim was printed with a mechanically robust polymeric blend of DCM and sonicated silk fibroin and the central optic zone was printed with only DCM. The bioprinted corneal stroma thus maintained its structural integrity and did not break when lifted with forceps. The two different bioinks were encapsulated with human limbus-derived mesenchymal stem cells (hLMSC) individually and 3D bioprinted in different patterns (concentric and parallel) to attain a native-like structure in terms of architecture and transparency. Thus, the bilayer cornea constructs maintained high cell viability and expressed keratocyte core proteins indicating optimal functionality. This approach helped to gain insight into bioprinting corneas with heterogeneous mechanical property without disturbing the structural clarity of the central optic zone.
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Affiliation(s)
- Anwesha Ghosh
- Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy 502284, Telangana, India
| | - Ashis Kumar Bera
- Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy 502284, Telangana, India
| | - Vivek Singh
- Centre Ocular Regeneration, Prof. Brien Holden Eye Research Centre L.V. Prasad Eye Institute, Hyderabad 500034, Telangana, India
| | - Sayan Basu
- Centre Ocular Regeneration, Prof. Brien Holden Eye Research Centre L.V. Prasad Eye Institute, Hyderabad 500034, Telangana, India
| | - Falguni Pati
- Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy 502284, Telangana, India.
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Tabatabai TS, Salehi M, Rezakhani L, Arabpour Z, Djalilian AR, Alizadeh M. Decellularization of various tissues and organs through chemical methods. Tissue Cell 2024; 91:102573. [PMID: 39393204 PMCID: PMC11993266 DOI: 10.1016/j.tice.2024.102573] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/01/2024] [Revised: 09/20/2024] [Accepted: 09/23/2024] [Indexed: 10/13/2024]
Abstract
Due to the increase in demand for donor organs and tissues during the past 20 years, new approaches have been created. These methods include, for example, tissue engineering in vitro and the production of regenerative biomaterials for transplantation. Applying the natural extracellular matrix (ECM) as a bioactive biomaterial for clinical applications is a unique approach known as decellularization technology. Decellularization is the process of eliminating cells from an extracellular matrix while preserving its natural components including its structural and functional proteins and glycosaminoglycan. This can be achieved by physical, chemical, or biological processes. A naturally formed three-dimensional structure with a biocompatible and regenerative structure is the result of the decellularization process. Decreasing the biological factors and antigens at the transplant site reduces the risk of adverse effects including inflammatory responses and immunological rejection. Regenerative medicine and tissue engineering applications can benefit from the use of decellularization, a promising approach that provides a biomaterial that preserves its extracellular matrix.
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Affiliation(s)
- Tayebeh Sadat Tabatabai
- Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
| | - Majid Salehi
- Tissue Engineering and Stem Cells Research Center, Shahroud University of Medical Sciences, Shahroud, Iran; Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
| | - Leila Rezakhani
- Fertility and Infertility Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran; Department of Tissue Engineering, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran
| | - Zohreh Arabpour
- Department of Ophthalmology and Visual Sciences, University of Illinois, Chicago, IL 60612, USA
| | - Ali R Djalilian
- Department of Ophthalmology and Visual Sciences, University of Illinois, Chicago, IL 60612, USA
| | - Morteza Alizadeh
- Department of Tissue Engineering and Biomaterials, School of Advanced Medical Sciences and Technologies, Hamadan University of Medical Sciences, Hamadan, Iran.
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Al Monla R, Daien V, Michon F. Advanced bioengineering strategies broaden the therapeutic landscape for corneal failure. Front Bioeng Biotechnol 2024; 12:1480772. [PMID: 39605752 PMCID: PMC11598527 DOI: 10.3389/fbioe.2024.1480772] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2024] [Accepted: 10/30/2024] [Indexed: 11/29/2024] Open
Abstract
The cornea acts as the eye foremost protective layer and is essential for its focusing power. Corneal blindness may arise from physical trauma or conditions like dystrophies, keratitis, keratoconus, or ulceration. While conventional treatments involve medical therapies and donor allografts-sometimes supplemented with keratoprostheses-these options are not suitable for all corneal defects. Consequently, the development of bioartificial corneal tissue has emerged as a critical research area, aiming to address the global shortage of human cornea donors. Bioengineered corneas hold considerable promise as substitutes, with the potential to replace either specific layers or the entire thickness of damaged corneas. This review first delves into the structural anatomy of the human cornea, identifying key attributes necessary for successful corneal tissue bioengineering. It then examines various corneal pathologies, current treatments, and their limitations. Finally, the review outlines the primary approaches in corneal tissue engineering, exploring cell-free, cell-based, and scaffold-based options as three emerging strategies to address corneal failure.
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Affiliation(s)
- Reem Al Monla
- Institute for Neurosciences of Montpellier, INSERM, University of Montpellier, Montpellier, France
| | - Vincent Daien
- Department of Ophthalmology, Gui de Chauliac Hospital, Montpellier, France
- Sydney Medical School, The Save Sight Institute, The University of Sydney, Sydney, NSW, Australia
| | - Frederic Michon
- Institute for Neurosciences of Montpellier, INSERM, University of Montpellier, Montpellier, France
- Department of Ophthalmology, Gui de Chauliac Hospital, Montpellier, France
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Pal P, Sambhakar S, Paliwal S, Kumar S, Kalsi V. Biofabrication paradigms in corneal regeneration: bridging bioprinting techniques, natural bioinks, and stem cell therapeutics. JOURNAL OF BIOMATERIALS SCIENCE. POLYMER EDITION 2024; 35:717-755. [PMID: 38214998 DOI: 10.1080/09205063.2024.2301817] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/21/2023] [Accepted: 12/29/2023] [Indexed: 01/14/2024]
Abstract
Corneal diseases are a major cause of vision loss worldwide. Traditional methods like corneal transplants from donors are effective but face challenges like limited donor availability and the risk of graft rejection. Therefore, new treatment methods are essential. This review examines the growing field of bioprinting and biofabrication in corneal tissue engineering. We begin by discussing various bioprinting methods such as stereolithography, inkjet, and extrusion printing, highlighting their strengths and weaknesses for eye-related uses. We also explore how biological tissues are made suitable for bioprinting through a process called decellularization, which can be achieved using chemical, physical, or biological methods. The review then looks at natural materials, known as bioinks, used in bioprinting. We focus on materials like gelatin, collagen, fibrin, chitin, chitosan, silk fibroin, and alginate, examining their mechanical and biological properties. The importance of hydrogel scaffolds, particularly those based on collagen and other materials, is also discussed in the context of repairing corneal tissue. Another key area we cover is the use of stem cells in corneal regeneration. We pay special attention to limbal epithelial stem cells and mesenchymal stromal cells, highlighting their roles in this process. The review concludes with an overview of the latest advancements in corneal tissue bioprinting, from early techniques to advanced methods of delivering stem cells using bioengineered materials. In summary, this review presents the current state and future potential of bioprinting and biofabrication in creating functional corneal tissues, highlighting new developments and ongoing challenges with a view towards restoring vision.
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Affiliation(s)
- Pankaj Pal
- Department of Pharmacy, Banasthali Vidyapith, Radha Kishnpura, Rajasthan, India
| | - Sharda Sambhakar
- Department of Pharmacy, Banasthali Vidyapith, Radha Kishnpura, Rajasthan, India
| | - Shailendra Paliwal
- Department of Pharmacy, L.L.R.M Medical College, Meerut, Uttar Pradesh, India
| | - Shobhit Kumar
- Department of Pharmaceutical Technology, Meerut Institute of Engineering and Technology, Meerut, Uttar Pradesh, India
| | - Vandna Kalsi
- School of Pharmaceutical Sciences, Lovely Professional University, Phagwara, Punjab, India
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Procházková A, Poláchová M, Dítě J, Netuková M, Studený P. Chemical, Physical, and Biological Corneal Decellularization Methods: A Review of Literature. J Ophthalmol 2024; 2024:1191462. [PMID: 38567029 PMCID: PMC10985644 DOI: 10.1155/2024/1191462] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/16/2023] [Revised: 02/28/2024] [Accepted: 03/08/2024] [Indexed: 04/04/2024] Open
Abstract
The cornea is one of the most commonly transplanted tissues worldwide. It is used to restore vision when severe visual impairment or blindness occurs in patients with corneal diseases or after trauma. Due to the global shortage of healthy donor corneas, decellularized corneal tissue has significant potential as an alternative to corneal transplantation. It preserves the native and biological ultrastructure of the cornea and, therefore, represents the most promising scaffold. This article discusses different methods of corneal decellularization based on the current literature. We searched PubMed.gov for articles from January 2009 to December 2023 using the following keywords: corneal decellularization, decellularization methods, and corneal transplantation. Although several methods of decellularization of corneal tissue have been reported, a universal standardised protocol of corneal decellularization has not yet been introduced. In general, a combination of decellularization methods has been used for efficient decellularization while preserving the optimal properties of the corneal tissue.
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Affiliation(s)
- Alexandra Procházková
- Department of Ophthalmology, Kralovske Vinohrady University Hospital and 3rd Medical Faculty, Srobarova 1150/50, Prague 10 100 34, Czech Republic
| | - Martina Poláchová
- Department of Ophthalmology, Kralovske Vinohrady University Hospital and 3rd Medical Faculty, Srobarova 1150/50, Prague 10 100 34, Czech Republic
| | - Jakub Dítě
- Department of Ophthalmology, Kralovske Vinohrady University Hospital and 3rd Medical Faculty, Srobarova 1150/50, Prague 10 100 34, Czech Republic
| | - Magdaléna Netuková
- Department of Ophthalmology, Kralovske Vinohrady University Hospital and 3rd Medical Faculty, Srobarova 1150/50, Prague 10 100 34, Czech Republic
| | - Pavel Studený
- Department of Ophthalmology, Kralovske Vinohrady University Hospital and 3rd Medical Faculty, Srobarova 1150/50, Prague 10 100 34, Czech Republic
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El Zarif M, Abdul Jawad K, Alió JL, Makdissy N, De Miguel MP. In vivo confocal microscopy evaluation of infiltrated immune cells in corneal stroma treated with cell therapy in advanced keratoconus. J Ophthalmic Inflamm Infect 2024; 14:5. [PMID: 38277094 PMCID: PMC10817874 DOI: 10.1186/s12348-024-00385-2] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/25/2023] [Accepted: 01/06/2024] [Indexed: 01/27/2024] Open
Abstract
PURPOSE This study investigates immune cell (ICs) infiltration in advanced keratoconus patients undergoing autologous adipose-derived adult stem cell (ADASC) therapy with recellularized human donor corneal laminas (CL). METHODS A prospective clinical trial included fourteen patients divided into three groups: G-1, ADASCs; G-2, decellularized CL (dCL); and G-3, dCL recellularized with ADASCs (ADASCs-rCL). Infiltrated ICs were assessed using in vivo confocal microscopy (IVCM) at 1,3,6, and12 months post-transplant. RESULTS Infiltrated ICs, encompassing granulocytes and agranulocytes, were observed across all groups, categorized by luminosity, structure, and area. Stromal ICs infiltration ranged from 1.19% to 6.62%, with a consistent increase in group-related cell density (F = 10.68, P < .0001), independent of post-op time (F = 0.77, P = 0.511); the most substantial variations were observed in G-3 at 6 and 12 months (2.0 and 1.87-fold, respectively). Similarly, significant size increases were more group-dependent (F = 5.76, P < .005) rather than time-dependent (F = 2.84, P < .05); G-3 exhibited significant increases at 6 and 12 months (3.70-fold and 2.52-fold, respectively). A lamina-induced shift in IC size occurred (F = 110.23, P < .0001), primarily with 50-100 μm2 sizes and up to larger cells > 300μm2, presumably macrophages, notably in G-3, indicating a potential role in tissue repair and remodeling, explaining reductions in cells remnants < 50μm2. CONCLUSIONS ADASCs-rCL therapy may lead to increased IC infiltration compared to ADASCs alone, impacting cell distribution and size due to the presence of the lamina. The findings reveal intricate immune patterns shaped by the corneal microenvironment and highlight the importance of understanding immune responses for the development of future therapeutic strategies.
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Affiliation(s)
- Mona El Zarif
- Optica General, Saida, Lebanon
- Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain
- Doctoral School of Sciences and Technology, Lebanese University, Hadath, Lebanon
| | | | - Jorge L Alió
- Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain
- Cornea, Cataract and Refractive Surgery Unit, Vissum (Miranza Group), Alicante, Spain
| | - Nehman Makdissy
- Genomic Surveillance and Biotherapy GSBT, Faculty of Sciences, Lebanese University, RasMaska, Lebanon.
| | - María P De Miguel
- Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Health Research Institute, Madrid, Spain.
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Chameettachal S, Venuganti A, Parekh Y, Prasad D, Joshi VP, Vashishtha A, Basu S, Singh V, Bokara KK, Pati F. Human cornea-derived extracellular matrix hydrogel for prevention of post-traumatic corneal scarring: A translational approach. Acta Biomater 2023; 171:289-307. [PMID: 37683964 DOI: 10.1016/j.actbio.2023.09.002] [Citation(s) in RCA: 12] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/24/2023] [Revised: 08/24/2023] [Accepted: 09/01/2023] [Indexed: 09/10/2023]
Abstract
Corneal scarring and opacification are a significant cause of blindness affecting millions worldwide. The current standard of care for corneal blindness is corneal transplantation, which suffers from several drawbacks. One alternative approach that has shown promise is the use of xenogeneic corneal extracellular matrix (ECM), but its clinical applicability is challenging due to safety concerns. This study reports the innovative use of human cornea-derived ECM to prevent post-traumatic corneal scarring. About 30 - 40% of corneas donated to the eye banks do not meet the standards defined for clinical use and are generally discarded, although they are completely screened for their safety. In this study, human cornea-derived decellularized ECM hydrogel was prepared from the non-transplantation grade human cadaveric corneas obtained from an accredited eye-bank. The prepared hydrogel was screened for its efficacy against corneal opacification following an injury in an animal model. Our in vivo study revealed that, the control collagen-treated group developed corneal opacification, while the prophylactic application of human cornea-derived hydrogel effectively prevented corneal scarring and opacification. The human hydrogel-treated corneas were indistinguishable from healthy corneas and comparable to those treated with the xenogeneic bovine corneal hydrogel. We also demonstrated that the application of the hydrogel retained the biological milieu including cell behavior, protein components, optical properties, curvature, and nerve regeneration by remodeling the corneal wound after injury. The hydrogel application is also sutureless, resulting in faster corneal healing. We envision that this human cornea-derived ECM-based hydrogel has potential clinical application in preventing scarring from corneal wounding. STATEMENT OF SIGNIFICANCE: There are significant challenges surrounding corneal regeneration after injury due to extensive scarring. Although there is substantial research on corneal regeneration, much of it uses synthetic materials with chemical cross-linking methods or xenogeneic tissue-based material devices which have to undergo exhaustive safety analysis before clinical trials. Herein, we demonstrate the potential application of a human corneal extracellular matrix hydrogel without any additional materials for scarless corneal tissue regeneration, and a method to reduce the wasting of donated allogenic corneal tissue from eye banks. We found no difference in efficacy between the usage of human tissues compared to xenogeneic sources. This may help ease clinical translation and can be used topically without sutures as an outpatient procedure.
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Affiliation(s)
- Shibu Chameettachal
- Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy 502285, Telangana, India
| | - Animith Venuganti
- Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, L V Prasad Eye Institute, Hyderabad, Telangana, India
| | - Yash Parekh
- CSIR-Center for Cellular and Molecular Biology, ANNEXE II, Medical Biotechnology Complex, Uppal Road, Hyderabad, Telangana, India
| | - Deeksha Prasad
- Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, L V Prasad Eye Institute, Hyderabad, Telangana, India
| | - Vineet P Joshi
- Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy 502285, Telangana, India; Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, L V Prasad Eye Institute, Hyderabad, Telangana, India
| | - Anviti Vashishtha
- CSIR-Center for Cellular and Molecular Biology, ANNEXE II, Medical Biotechnology Complex, Uppal Road, Hyderabad, Telangana, India
| | - Sayan Basu
- Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, L V Prasad Eye Institute, Hyderabad, Telangana, India.
| | - Vivek Singh
- Centre for Ocular Regeneration, Prof. Brien Holden Eye Research Centre, L V Prasad Eye Institute, Hyderabad, Telangana, India.
| | - Kiran Kumar Bokara
- CSIR-Center for Cellular and Molecular Biology, ANNEXE II, Medical Biotechnology Complex, Uppal Road, Hyderabad, Telangana, India.
| | - Falguni Pati
- Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy 502285, Telangana, India.
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Wang X, Elbahrawi RT, Abdukadir AM, Ali ZM, Chan V, Corridon PR. A proposed model of xeno-keratoplasty using 3D printing and decellularization. Front Pharmacol 2023; 14:1193606. [PMID: 37799970 PMCID: PMC10548234 DOI: 10.3389/fphar.2023.1193606] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2023] [Accepted: 09/06/2023] [Indexed: 10/07/2023] Open
Abstract
Corneal opacity is a leading cause of vision impairment and suffering worldwide. Transplantation can effectively restore vision and reduce chronic discomfort. However, there is a considerable shortage of viable corneal graft tissues. Tissue engineering may address this issue by advancing xeno-keratoplasty as a viable alternative to conventional keratoplasty. In particular, livestock decellularization strategies offer the potential to generate bioartificial ocular prosthetics in sufficient supply to match existing and projected needs. To this end, we have examined the best practices and characterizations that have supported the current state-of-the-art driving preclinical and clinical applications. Identifying the challenges that delimit activities to supplement the donor corneal pool derived from acellular scaffolds allowed us to hypothesize a model for keratoprosthesis applications derived from livestock combining 3D printing and decellularization.
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Affiliation(s)
- Xinyu Wang
- Biomedical Engineering and Healthcare Engineering Innovation Center, Khalifa University, Abu Dhabi, United Arab Emirates
- Department of Immunology and Physiology, College of Medicine and Health Sciences, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Rawdah Taha Elbahrawi
- Department of Immunology and Physiology, College of Medicine and Health Sciences, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Azhar Mohamud Abdukadir
- Department of Immunology and Physiology, College of Medicine and Health Sciences, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Zehara Mohammed Ali
- Department of Immunology and Physiology, College of Medicine and Health Sciences, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Vincent Chan
- Biomedical Engineering and Healthcare Engineering Innovation Center, Khalifa University, Abu Dhabi, United Arab Emirates
| | - Peter R. Corridon
- Biomedical Engineering and Healthcare Engineering Innovation Center, Khalifa University, Abu Dhabi, United Arab Emirates
- Department of Immunology and Physiology, College of Medicine and Health Sciences, Khalifa University, Abu Dhabi, United Arab Emirates
- Center for Biotechnology, Khalifa University, Abu Dhabi, United Arab Emirates
- Hleathcare, Engineering and Innovation Center, Khalifa University, Abu Dhabi, United Arab Emirates
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Corneal Stromal Densitometry Evolution in a Clinical Model of Cellular Therapy for Advanced Keratoconus. Cornea 2023; 42:332-343. [PMID: 36256440 DOI: 10.1097/ico.0000000000003152] [Citation(s) in RCA: 8] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2022] [Accepted: 08/12/2022] [Indexed: 11/26/2022]
Abstract
PURPOSE The aim of this study was to report the corneal densitometry (CD) evolution studied by Scheimpflug tomography, anterior segment optical coherence tomography (AS-OCT), and confocal microscopy changes, in patients with advanced keratoconus included in a clinical experience of advanced cell therapy using autologous humans adipose-derived adult stem cells (ADASCs) and corneal decellularized and ADASCs-recellularized human donor corneal laminas. METHODS This study is an interventional prospective, consecutive, randomized, comparative series of cases. Fourteen patients with keratoconus were randomly distributed into 3 groups for 3 types of surgical interventions: group 1 (G-1), autologous ADASC implantation (n = 5); group 2 (G-2), decellularized human corneal stroma (n = 5); and group 3 (G-3), autologous ADASCs + decellularized human corneal stroma (n = 4). Participants were assessed with Scheimpflug-based Oculus Pentacam CD module, AS-OCT (Visante; Carl Zeiss), and confocal microscopy (HRT3 RCM Heidelberg). RESULTS A significant improvement of 1 to 2 logMAR lines in all visual parameters in the 3 groups was obtained. The central and total CD were statistically significantly higher in G-2 compared with G-1 and G-3 compared with G-1 at the studied annular zones centered on the corneal apex (0-2, 2-6, and 6-10 mm). There was statistical significance higher in G-3 compared with G-2 at the central corneal stroma at 0-2 and 2-6 mm. The confocal microscopy findings and the AS-OCT reflected the densitometry changes. CONCLUSIONS The intrastromal implantation of ADASCs produced very subtle changes in CD at the level of the central corneal stroma. However, the intrastromal implantation of decellularized corneal laminas increases it slightly, but with lower values than the implantation of recellularized laminas with ADASCs.
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Hazra S, Akepogu J, Krishna S, Pulipaka S, Bagga B, Ramachandran C. Use of Decellularized SMILE (Small-Incision Lenticule Extraction) Lenticules for Engineering the Corneal Endothelial Layer: A Proof-of-Concept. Curr Eye Res 2023; 48:251-262. [PMID: 36458563 DOI: 10.1080/02713683.2022.2151018] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/05/2022]
Abstract
PURPOSE To demonstrate the suitability of using decellularized SMILE (Small-incision Lenticule Extraction) lenticules for culturing and transplanting the corneal endothelium (CE). METHODS The SMILE lenticules, obtained during refractive surgery, were decellularized by incubating in CE culture medium and fetal bovine serum. Decellularization was confirmed by hematoxylin and eosin staining, DAPI staining, and gel electrophoresis. The amount of DNA per milligram of dry tissue weight was calculated to quantify the residual nuclear content. The transparency of the decellularized lenticules was determined by calculating the modulation transfer function. Immunostaining for stromal collagens and glycosaminoglycan was performed using specific antibodies. Engineered tissue was constructed by culturing the CE cells on lenticules and staining for ZO-1, Na/K ATPase, and N-cadherin. The functionality of the engineered tissues was assessed by transplanting them onto edematous human donor corneas and perfusing for 10 days ex-vivo. RESULTS The residual DNA per milligram of dry tissue weight was found to be significantly reduced (p < 0.0001) in serum (0.255 µg/mg) and Opti-MEM (0.140 µg/mg) when compared to fresh lenticules (3.9 µg/mg). Decellularization did not alter the arrangement of the collagen fibers or the transparency of the lenticules. CE cells attached and matured to express ZO-1, Na/K ATPase, and N-cadherin at two weeks after seeding. The engineered tissue upon transplantation significantly reduced the corneal edema (p < 0.05) and the transplanted cells remained intact on the SMILE lenticule post-transplantation. CONCLUSION This study demonstrates the suitability of using SMILE lenticules decellularized using a simple, chemical-free method for engineering the corneal endothelium for transplantation.
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Affiliation(s)
- Swatilekha Hazra
- Professor Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
- Manipal Academy of Higher Education, Manipal University, Manipal, India
| | - Jacquelyn Akepogu
- Professor Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
- Manipal Academy of Higher Education, Manipal University, Manipal, India
| | - Supriya Krishna
- Professor Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
- Ludwig-Maximilians University, Munich, Germany
| | - SriRavali Pulipaka
- Professor Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
- Department of Applied Biology, CSIR-Indian Institute of Chemical Technology, Hyderabad, Telangana, India
| | - Bhupesh Bagga
- Department of Cornea and Anterior Segment, LV Prasad Eye Institute, Hyderabad, Telangana, India
| | - Charanya Ramachandran
- Professor Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, Telangana, India
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12
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Wang B, Qinglai T, Yang Q, Li M, Zeng S, Yang X, Xiao Z, Tong X, Lei L, Li S. Functional acellular matrix for tissue repair. Mater Today Bio 2023; 18:100530. [PMID: 36601535 PMCID: PMC9806685 DOI: 10.1016/j.mtbio.2022.100530] [Citation(s) in RCA: 25] [Impact Index Per Article: 12.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/14/2022] [Revised: 12/23/2022] [Accepted: 12/26/2022] [Indexed: 12/29/2022] Open
Abstract
In view of their low immunogenicity, biomimetic internal environment, tissue- and organ-like physicochemical properties, and functionalization potential, decellularized extracellular matrix (dECM) materials attract considerable attention and are widely used in tissue engineering. This review describes the composition of extracellular matrices and their role in stem-cell differentiation, discusses the advantages and disadvantages of existing decellularization techniques, and presents methods for the functionalization and characterization of decellularized scaffolds. In addition, we discuss progress in the use of dECMs for cartilage, skin, nerve, and muscle repair and the transplantation or regeneration of different whole organs (e.g., kidneys, liver, uterus, lungs, and heart), summarize the shortcomings of using dECMs for tissue and organ repair after refunctionalization, and examine the corresponding future prospects. Thus, the present review helps to further systematize the application of functionalized dECMs in tissue/organ transplantation and keep researchers up to date on recent progress in dECM usage.
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Affiliation(s)
- Bin Wang
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Tang Qinglai
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Qian Yang
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Mengmeng Li
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Shiying Zeng
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Xinming Yang
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Zian Xiao
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
| | - Xinying Tong
- Department of Hemodialysis, The Second Xiangya Hospital, Central South University, Changsha 410011, Hunan, China
| | - Lanjie Lei
- State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, China
| | - Shisheng Li
- Department of Otorhinolaryngology Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha 410011, China
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13
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Crouch DJ, Sheridan CM, Behnsen JG, Bosworth LA. An Optimized Method to Decellularize Human Trabecular Meshwork. Bioengineering (Basel) 2022; 9:194. [PMID: 35621472 PMCID: PMC9137515 DOI: 10.3390/bioengineering9050194] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/31/2022] [Revised: 04/27/2022] [Accepted: 04/28/2022] [Indexed: 12/12/2022] Open
Abstract
Glaucoma is linked to raised intraocular pressure (IOP). The trabecular meshwork (TM) plays a major role in regulating IOP by enabling outflow of aqueous humor from the eye through its complex 3D structure. A lack of therapies targeting the dysfunctional TM highlights the need to develop biomimetic scaffolds that provide 3D in vitro models for glaucoma research or as implantable devices to regenerate TM tissue. To artificially mimic the TM's structure, we assessed methods for its decellularization and outline an optimized protocol for cell removal and structural retention. Using bovine TM, we trialed 2 lysing agents-Trypsin (0.05% v/v) and Ammonium Hydroxide (NH4OH; 2% v/v). Twenty-four hours in Trypsin caused significant structural changes. Shorter exposure (2 h) reduced this disruption whilst decellularizing the tissue (dsDNA 26 ± 14 ng/mL (control 1970 ± 146 ng/mL)). In contrast, NH4OH lysed all cells (dsDNA 25 ± 21 ng/mL), and the TM structure remained intact. For human TM, 2% v/v NH4OH similarly removed cells (dsDNA 52 ± 4 ng/mL (control 1965 ± 233 ng/mL)), and light microscopy and SEM presented no structural damage. X-ray computed tomography enabled a novel 3D reconstruction of decellularized human TM and observation of the tissue's intricate architecture. This study provides a new, validated method using NH4OH to decellularize delicate human TM without compromising tissue structure.
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Affiliation(s)
- Devon J. Crouch
- Department of Eye and Vision Science, Institute of Life Course and Medical Sciences, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L7 8TX, UK;
| | - Carl M. Sheridan
- Department of Eye and Vision Science, Institute of Life Course and Medical Sciences, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L7 8TX, UK;
| | - Julia G. Behnsen
- Department of Mechanical, Materials, and Aerospace Engineering, University of Liverpool, Liverpool L69 6GB, UK;
| | - Lucy A. Bosworth
- Department of Eye and Vision Science, Institute of Life Course and Medical Sciences, Faculty of Health and Life Sciences, University of Liverpool, Liverpool L7 8TX, UK;
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Neishabouri A, Soltani Khaboushan A, Daghigh F, Kajbafzadeh AM, Majidi Zolbin M. Decellularization in Tissue Engineering and Regenerative Medicine: Evaluation, Modification, and Application Methods. Front Bioeng Biotechnol 2022; 10:805299. [PMID: 35547166 PMCID: PMC9081537 DOI: 10.3389/fbioe.2022.805299] [Citation(s) in RCA: 104] [Impact Index Per Article: 34.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/30/2021] [Accepted: 04/04/2022] [Indexed: 12/14/2022] Open
Abstract
Reproduction of different tissues using scaffolds and materials is a major element in regenerative medicine. The regeneration of whole organs with decellularized extracellular matrix (dECM) has remained a goal despite the use of these materials for different purposes. Recently, decellularization techniques have been widely used in producing scaffolds that are appropriate for regenerating damaged organs and may be able to overcome the shortage of donor organs. Decellularized ECM offers several advantages over synthetic compounds, including the preserved natural microenvironment features. Different decellularization methods have been developed, each of which is appropriate for removing cells from specific tissues under certain conditions. A variety of methods have been advanced for evaluating the decellularization process in terms of cell removal efficiency, tissue ultrastructure preservation, toxicity, biocompatibility, biodegradability, and mechanical resistance in order to enhance the efficacy of decellularization methods. Modification techniques improve the characteristics of decellularized scaffolds, making them available for the regeneration of damaged tissues. Moreover, modification of scaffolds makes them appropriate options for drug delivery, disease modeling, and improving stem cells growth and proliferation. However, considering different challenges in the way of decellularization methods and application of decellularized scaffolds, this field is constantly developing and progressively moving forward. This review has outlined recent decellularization and sterilization strategies, evaluation tests for efficient decellularization, materials processing, application, and challenges and future outlooks of decellularization in regenerative medicine and tissue engineering.
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Affiliation(s)
- Afarin Neishabouri
- Pediatric Urology and Regenerative Medicine Research Center, Children’s Medical Center, Pediatric Center of Excellence, Tehran University of Medical Science, Tehran, Iran
| | - Alireza Soltani Khaboushan
- Pediatric Urology and Regenerative Medicine Research Center, Children’s Medical Center, Pediatric Center of Excellence, Tehran University of Medical Science, Tehran, Iran
- Students’ Scientific Research Center, Tehran University of Medical Sciences, Tehran, Iran
| | - Faezeh Daghigh
- Department of Physiology, Faculty of Medicine, Tabriz Medical Sciences, Islamic Azad University, Tabriz, Iran
| | - Abdol-Mohammad Kajbafzadeh
- Pediatric Urology and Regenerative Medicine Research Center, Children’s Medical Center, Pediatric Center of Excellence, Tehran University of Medical Science, Tehran, Iran
- *Correspondence: Masoumeh Majidi Zolbin, ; Abdol-Mohammad Kajbafzadeh,
| | - Masoumeh Majidi Zolbin
- Pediatric Urology and Regenerative Medicine Research Center, Children’s Medical Center, Pediatric Center of Excellence, Tehran University of Medical Science, Tehran, Iran
- *Correspondence: Masoumeh Majidi Zolbin, ; Abdol-Mohammad Kajbafzadeh,
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15
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Himmler M, Schubert DW, Fuchsluger TA. Examining the Transmission of Visible Light through Electrospun Nanofibrous PCL Scaffolds for Corneal Tissue Engineering. NANOMATERIALS 2021; 11:nano11123191. [PMID: 34947541 PMCID: PMC8705195 DOI: 10.3390/nano11123191] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/29/2021] [Revised: 11/21/2021] [Accepted: 11/23/2021] [Indexed: 12/28/2022]
Abstract
The transparency of nanofibrous scaffolds is of highest interest for potential applications like corneal wound dressings in corneal tissue engineering. In this study, we provide a detailed analysis of light transmission through electrospun polycaprolactone (PCL) scaffolds. PCL scaffolds were produced via electrospinning, with fiber diameters in the range from (35 ± 13) nm to (167 ± 35) nm. Light transmission measurements were conducted using UV-vis spectroscopy in the range of visible light and analyzed with respect to the influence of scaffold thickness, fiber diameter, and surrounding medium. Contour plots were compiled for a straightforward access to light transmission values for arbitrary scaffold thicknesses. Depending on the fiber diameter, transmission values between 15% and 75% were observed for scaffold thicknesses of 10 µm. With a decreasing fiber diameter, light transmission could be improved, as well as with matching refractive indices of fiber material and medium. For corneal tissue engineering, scaffolds should be designed as thin as possible and fabricated from polymers with a matching refractive index to that of the human cornea. Concerning fiber diameter, smaller fiber diameters should be favored for maximizing graft transparency. Finally, a novel, semi-empirical formulation of light transmission through nanofibrous scaffolds is presented.
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Affiliation(s)
- Marcus Himmler
- Department of Ophthalmology, University Medical Center Rostock, Doberaner Straße 140, 18057 Rostock, Germany
- Institute of Polymer Materials, Friedrich-Alexander University Erlangen-Nuremberg, Martenstraße 7, 91058 Erlangen, Germany;
- Correspondence: (M.H.); (T.A.F.)
| | - Dirk W. Schubert
- Institute of Polymer Materials, Friedrich-Alexander University Erlangen-Nuremberg, Martenstraße 7, 91058 Erlangen, Germany;
| | - Thomas A. Fuchsluger
- Department of Ophthalmology, University Medical Center Rostock, Doberaner Straße 140, 18057 Rostock, Germany
- Correspondence: (M.H.); (T.A.F.)
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16
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Kim DK, Lee S, Choi JH, Jung BS, Kim KS, Song JE, Reis RL, Khang G. Enhanced Silk Fibroin-Based Film Scaffold Using Curcumin for Corneal Endothelial Cell Regeneration. Macromol Res 2021. [DOI: 10.1007/s13233-021-9081-9] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/24/2022]
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17
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Maqueda M, Mosquera JL, García-Arumí J, Veiga A, Duarri A. Repopulation of decellularized retinas with hiPSC-derived retinal pigment epithelial and ocular progenitor cells shows cell engraftment, organization and differentiation. Biomaterials 2021; 276:121049. [PMID: 34332373 DOI: 10.1016/j.biomaterials.2021.121049] [Citation(s) in RCA: 16] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2021] [Revised: 07/22/2021] [Accepted: 07/23/2021] [Indexed: 11/18/2022]
Abstract
The retinal extracellular matrix (ECM) provides architectural support, adhesion and signal guidance that controls retinal development. Decellularization of the ECM affords great potential to tissue engineering; however, how structural retinal ECM affects in vitro development, differentiation and maturation of ocular cells remains to be elucidated. Here, mouse and porcine retinas were decellularized and the protein profile analyzed. Acellular retinal ECM (arECM) scaffolds were then repopulated with human iPSC-derived retinal pigment epithelial (RPE) cells or ocular progenitor cells (OPC) to assess their integration, proliferation and organization. 3837 and 2612 unique proteins were identified in mouse and porcine arECM, respectively, of which 93 and 116 proteins belong to the matrisome. GO analysis shows that matrisome-related proteins were associated with the extracellular region and cell junction and KEGG pathways related to signalling transduction, nervous and endocrine systems and cell junctions were enriched. Interestingly, mouse and porcine arECMs were successfully repopulated with both RPE and OPC, the latter exhibiting cell lineage-specific clusters. Retinal cells organized into different layers containing well-defined areas with pigmented cells, photoreceptors, Müller glia, astrocytes, and ganglion cells, whereas in other areas, conjunctival/limbal, corneal and lens cells re-arranged in cell-specific self-organized areas. In conclusion, our results demonstrated that decellularization of both mouse and porcine retinas retains common native ECM components that upon cell repopulation could guide similar ocular cell adhesion, migration and organization.
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Affiliation(s)
- Maria Maqueda
- Bioinformatics Unit, Institut d'Investigació Biomèdica de Bellvitge - IDIBELL, L'Hospitalet de Llobregat, Barcelona, Spain
| | - Jose Luis Mosquera
- Bioinformatics Unit, Institut d'Investigació Biomèdica de Bellvitge - IDIBELL, L'Hospitalet de Llobregat, Barcelona, Spain
| | - José García-Arumí
- Ophthalmology Research Group, Vall d'Hebron Institut de Recerca - VHIR, Vall d'Hebron Hospital Universitari, Barcelona, Spain
| | - Anna Veiga
- Pluripotent Stem Cell Therapy Group, Regenerative Medicine Program, Institut d'Investigació Biomèdica de Bellvitge - IDIBELL, L'Hospitalet de Llobregat, Barcelona, Spain; National Stem Cell Bank-Barcelona Node, Biomolecular and Bioinformatics Resources Platform (PRB2), ISCIII, Madrid, Spain
| | - Anna Duarri
- Ophthalmology Research Group, Vall d'Hebron Institut de Recerca - VHIR, Vall d'Hebron Hospital Universitari, Barcelona, Spain; National Stem Cell Bank-Barcelona Node, Biomolecular and Bioinformatics Resources Platform (PRB2), ISCIII, Madrid, Spain.
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18
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El Zarif M, Alió JL, Alió Del Barrio JL, Abdul Jawad K, Palazón-Bru A, Abdul Jawad Z, De Miguel MP, Makdissy N. Corneal Stromal Regeneration Therapy for Advanced Keratoconus: Long-term Outcomes at 3 Years. Cornea 2021; 40:741-754. [PMID: 33591032 DOI: 10.1097/ico.0000000000002646] [Citation(s) in RCA: 35] [Impact Index Per Article: 8.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2020] [Accepted: 11/20/2020] [Indexed: 01/02/2023]
Abstract
PURPOSE To report the 3-year clinical outcomes of corneal stromal cell therapy consisting of the intrastromal implantation with autologous adipose-derived adult stem cells (ADASCs), and decellularized or ADASC-recellularized human donor corneal laminas in advanced keratoconus. METHODS Fourteen patients were enrolled in 3 experimental groups. Group 1 (G-1) patients underwent implantation of ADASCs alone (3 × 10⁶ cells/1 mL) (n = 5). Group 2 (G-2) patients received a 120-μm decellularized corneal stroma lamina (n = 5). Group 3 (G-3) patients received a 120-μm lamina recellularized with ADASCs (1 × 10⁶ cells/1 mL) (n = 4). ADASCs were obtained by elective liposuction. Implantation was performed into a femtosecond pocket under topical anesthesia. RESULTS At 3 years, a significant improvement of 1 to 2 logMAR lines in uncorrected distance visual acuity was observed in all groups. A statistically significant decrease in corrected distance visual acuity was obtained in G-2 and G-3 (P < 0.001) when compared with that of G-1. Rigid contact lens distance visual acuity showed a statistically significant worsening in G-2 (P < 0.001) compared with that of G-1. A statistically significant increase in central corneal thickness was observed in G-2 (P = 0.012) and G-3 (P < 0.001); in the Scheimpflug corneal topography, the thinnest point was observed in G-2 (P = 0.007) and G-3 (P = 0.001) when compared with that of G-1. CONCLUSIONS Intrastromal implantation of ADASCs and decellularized or ADASC-recellularized human corneal stroma laminas did not have complications at 3 years. The technique showed a moderate improvement in (uncorrected distance visual acuity) and (corrected distance visual acuity) in advanced keratoconus.
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Affiliation(s)
- Mona El Zarif
- Lebaneese university Hadath: EDST of Biotechnology, Optica General, Saida, Lebanon
- Division of Ophthalmology, Universidad Miguel Hernández University, Alicante, Spain
- Lebaneese university Hadath: EDST of Biotechnology, Doctoral School of Sciences and Technology, Lebanese University, Hadath, Lebanon
- GSBT Genomic Surveillance and Biotherapy Team, Faculty of Sciences, Lebanese University, Hadath, Lebanon
| | - Jorge L Alió
- Division of Ophthalmology, Universidad Miguel Hernández University, Alicante, Spain
- Cornea, Cataract and Refractive Surgery Unit, Vissum Instituto Oftalmologico de Alicante, Grupo Miranza, Alicante, Spain
| | - Jorge L Alió Del Barrio
- Division of Ophthalmology, Universidad Miguel Hernández University, Alicante, Spain
- Cornea, Cataract and Refractive Surgery Unit, Vissum Instituto Oftalmologico de Alicante, Grupo Miranza, Alicante, Spain
| | - Karim Abdul Jawad
- Lebaneese university Hadath: EDST of Biotechnology, Optica General, Saida, Lebanon
| | - Antonio Palazón-Bru
- Department of Clinical Medicine, Miguel Hernández University, Alicante, Spain; and
| | - Ziad Abdul Jawad
- Lebaneese university Hadath: EDST of Biotechnology, Optica General, Saida, Lebanon
| | - María P De Miguel
- Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Health Research Institute, Madrid, Spain
| | - Nehman Makdissy
- GSBT Genomic Surveillance and Biotherapy Team, Faculty of Sciences, Lebanese University, Hadath, Lebanon
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Öztürk-Öncel MÖ, Erkoc-Biradli FZ, Rasier R, Marcali M, Elbuken C, Garipcan B. Rose petal topography mimicked poly(dimethylsiloxane) substrates for enhanced corneal endothelial cell behavior. MATERIALS SCIENCE & ENGINEERING. C, MATERIALS FOR BIOLOGICAL APPLICATIONS 2021; 126:112147. [PMID: 34082958 DOI: 10.1016/j.msec.2021.112147] [Citation(s) in RCA: 11] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 12/03/2020] [Revised: 04/05/2021] [Accepted: 04/26/2021] [Indexed: 12/29/2022]
Abstract
Low proliferation capacity of corneal endothelial cells (CECs) and worldwide limitations in transplantable donor tissues reveal the critical need of a robust approach for in vitro CEC growth. However, preservation of CEC-specific phenotype with increased proliferation has been a great challenge. Here we offer a biomimetic cell substrate design, by optimizing mechanical, topographical and biochemical characteristics of materials with CEC microenvironment. We showed the surprising similarity between topographical features of white rose petals and corneal endothelium due to hexagonal cell shapes and physiologically relevant cell density (≈ 2000 cells/mm2). Polydimethylsiloxane (PDMS) substrates with replica of white rose petal topography and cornea-friendly Young's modulus (211.85 ± 74.9 kPa) were functionalized with two of the important corneal extracellular matrix (ECM) components, collagen IV (COL 4) and hyaluronic acid (HA). White rose petal patterned and COL 4 modified PDMS with optimized stiffness provided enhanced bovine CEC response with higher density monolayers and increased phenotypic marker expression. This biomimetic approach demonstrates a successful platform to improve in vitro cell substrate properties of PDMS for corneal applications, suggesting an alternative environment for CEC-based therapies, drug toxicity investigations, microfluidics and organ-on-chip applications.
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Affiliation(s)
| | | | - Rıfat Rasier
- Department of Ophthalmology, Demiroglu Bilim University, Istanbul, Turkey
| | - Merve Marcali
- UNAM-National Nanotechnology Research Center, Institute of Materials Science and Nanotechnology, Bilkent University, Ankara, Turkey
| | - Caglar Elbuken
- UNAM-National Nanotechnology Research Center, Institute of Materials Science and Nanotechnology, Bilkent University, Ankara, Turkey; Faculty of Biochemistry and Molecular Medicine, Faculty of Medicine, University of Oulu, 90014 Oulu, Finland
| | - Bora Garipcan
- Institute of Biomedical Engineering, Boğaziçi University, Istanbul, Turkey.
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20
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Asgari F, Asgari HR, Najafi M, Eftekhari BS, Vardiani M, Gholipourmalekabadi M, Koruji M. Optimization of decellularized human placental macroporous scaffolds for spermatogonial stem cells homing. JOURNAL OF MATERIALS SCIENCE. MATERIALS IN MEDICINE 2021; 32:47. [PMID: 33891169 PMCID: PMC8065005 DOI: 10.1007/s10856-021-06517-7] [Citation(s) in RCA: 20] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/08/2020] [Accepted: 03/19/2021] [Indexed: 06/08/2023]
Abstract
Decellularized scaffolds have been found to be excellent platforms for tissue engineering applications. The attempts are still being made to optimize a decellularization protocol with successful removal of the cells with minimal damages to extracellular matrix components. We examined twelve decellularization procedures using different concentrations of Sodium dodecyl sulfate and Triton X-100 (alone or in combination), and incubation time points of 15 or 30 min. Then, the potential of the decellularized scaffold as a three-dimensional substrate for colony formation capacity of mouse spermatogonial stem cells was determined. The morphological, degradation, biocompatibility, and swelling properties of the samples were fully characterized. The 0.5%/30 SDS/Triton showed optimal decellularization with minimal negative effects on ECM (P ≤ 0.05). The swelling ratios increased with the increase of SDS and Triton concentration and incubation time. Only 0.5%/15 and 30 SDS showed a significant decrease in the SSCs viability compared with other groups (P < 0.05). The SSCs colony formation was clearly observed under SEM and H&E stained slides. The cells infiltrated into the subcutaneously implanted scaffold at days 7 and 30 post-implantation with no sign of graft rejection. Our data suggest the %0.5/30 SDS/Triton as an excellent platform for tissue engineering and reproductive biology applications.
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Affiliation(s)
- Fatemeh Asgari
- Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran
- Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Hamid Reza Asgari
- Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran
- Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Mohammad Najafi
- Biochemistry Department, Iran University of Medical Sciences, Tehran, Iran
- Cellular and Molecular Research Centre, Iran University of Medicine Sciences, Tehran, Iran
| | - Behnaz Sadat Eftekhari
- Biomaterials Group, Faculty of Biomedical Engineering, Amirkabir University of Technology, Tehran, Iran
- Department of Physiology and Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, USA
| | - Mina Vardiani
- Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran, Tehran, Iran
| | - Mazaher Gholipourmalekabadi
- Cellular and Molecular Research Centre, Iran University of Medicine Sciences, Tehran, Iran.
- Department of Tissue Engineering & Regenerative Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran.
- Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran.
| | - Morteza Koruji
- Stem Cell and Regenerative Medicine Research Center, Iran University of Medical Sciences, Tehran, Iran.
- Department of Anatomical Sciences, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
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21
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Ulag S, Uysal E, Bedir T, Sengor M, Ekren N, Ustundag CB, Midha S, Kalaskar DM, Gunduz O. Recent developments and characterization techniques in
3D
printing of corneal stroma tissue. POLYM ADVAN TECHNOL 2021. [DOI: 10.1002/pat.5340] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
Affiliation(s)
- Songul Ulag
- Center for Nanotechnology & Biomaterials Application and Research (NBUAM) Marmara University Istanbul Turkey
| | - Ebru Uysal
- Department of Bioengineering, Faculty of Chemistry and Metallurgy Yildiz Technical University Istanbul Turkey
| | - Tuba Bedir
- Center for Nanotechnology & Biomaterials Application and Research (NBUAM) Marmara University Istanbul Turkey
| | - Mustafa Sengor
- Center for Nanotechnology & Biomaterials Application and Research (NBUAM) Marmara University Istanbul Turkey
| | - Nazmi Ekren
- Center for Nanotechnology & Biomaterials Application and Research (NBUAM) Marmara University Istanbul Turkey
| | - Cem Bulent Ustundag
- Center for Nanotechnology & Biomaterials Application and Research (NBUAM) Marmara University Istanbul Turkey
- Department of Bioengineering, Faculty of Chemistry and Metallurgy Yildiz Technical University Istanbul Turkey
| | - Swati Midha
- UCL Division of Surgery & Interventional Science University College London (UCL) London UK
| | - Deepak M. Kalaskar
- UCL Division of Surgery & Interventional Science University College London (UCL) London UK
| | - Oguzhan Gunduz
- Center for Nanotechnology & Biomaterials Application and Research (NBUAM) Marmara University Istanbul Turkey
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22
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Chandru A, Agrawal P, Ojha SK, Selvakumar K, Shiva VK, Gharat T, Selvam S, Thomas MB, Damala M, Prasad D, Basu S, Bhowmick T, Sangwan VS, Singh V. Human Cadaveric Donor Cornea Derived Extra Cellular Matrix Microparticles for Minimally Invasive Healing/Regeneration of Corneal Wounds. Biomolecules 2021; 11:532. [PMID: 33918484 PMCID: PMC8066719 DOI: 10.3390/biom11040532] [Citation(s) in RCA: 19] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/02/2021] [Revised: 03/24/2021] [Accepted: 03/25/2021] [Indexed: 01/10/2023] Open
Abstract
Biological materials derived from extracellular matrix (ECM) proteins have garnered interest as their composition is very similar to that of native tissue. Herein, we report the use of human cornea derived decellularized ECM (dECM) microparticles dispersed in human fibrin sealant as an accessible therapeutic alternative for corneal anterior stromal reconstruction. dECM microparticles had good particle size distribution (≤10 µm) and retained the majority of corneal ECM components found in native tissue. Fibrin-dECM hydrogels exhibited compressive modulus of 70.83 ± 9.17 kPa matching that of native tissue, maximum burst pressure of 34.3 ± 3.7 kPa, and demonstrated a short crosslinking time of ~17 min. The fibrin-dECM hydrogels were found to be biodegradable, cytocompatible, non-mutagenic, non-sensitive, non-irritant, and supported the growth and maintained the phenotype of encapsulated human corneal stem cells (hCSCs) in vitro. In a rabbit model of anterior lamellar keratectomy, fibrin-dECM bio-adhesives promoted corneal re-epithelialization within 14 days, induced stromal tissue repair, and displayed integration with corneal tissues in vivo. Overall, our results suggest that the incorporation of cornea tissue-derived ECM microparticles in fibrin hydrogels is non-toxic, safe, and shows tremendous promise as a minimally invasive therapeutic approach for the treatment of superficial corneal epithelial wounds and anterior stromal injuries.
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Affiliation(s)
- Arun Chandru
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Parinita Agrawal
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Sanjay Kumar Ojha
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Kamalnath Selvakumar
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Vaishnavi K. Shiva
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Tanmay Gharat
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Shivaram Selvam
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Midhun Ben Thomas
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Mukesh Damala
- Brien Holden Eye Research Center, LV Prasad Eye Institute, Hyderabad, Telangana 500034, India; (M.D.); (D.P.); (S.B.); (V.S.S.)
| | - Deeksha Prasad
- Brien Holden Eye Research Center, LV Prasad Eye Institute, Hyderabad, Telangana 500034, India; (M.D.); (D.P.); (S.B.); (V.S.S.)
| | - Sayan Basu
- Brien Holden Eye Research Center, LV Prasad Eye Institute, Hyderabad, Telangana 500034, India; (M.D.); (D.P.); (S.B.); (V.S.S.)
- Center for Ocular Regeneration (CORE), LV Prasad Eye Institute, Hyderabad, Telangana 500034, India
| | - Tuhin Bhowmick
- Pandorum Technologies Private Limited, Bangalore, Karnataka 560100, India; (P.A.); (S.K.O.); (K.S.); (V.K.S.); (T.G.); (S.S.); (M.B.T.)
| | - Virender Singh Sangwan
- Brien Holden Eye Research Center, LV Prasad Eye Institute, Hyderabad, Telangana 500034, India; (M.D.); (D.P.); (S.B.); (V.S.S.)
- Center for Ocular Regeneration (CORE), LV Prasad Eye Institute, Hyderabad, Telangana 500034, India
| | - Vivek Singh
- Brien Holden Eye Research Center, LV Prasad Eye Institute, Hyderabad, Telangana 500034, India; (M.D.); (D.P.); (S.B.); (V.S.S.)
- Center for Ocular Regeneration (CORE), LV Prasad Eye Institute, Hyderabad, Telangana 500034, India
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23
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Capella-Monsonís H, Zeugolis DI. Decellularized xenografts in regenerative medicine: From processing to clinical application. Xenotransplantation 2021; 28:e12683. [PMID: 33709410 DOI: 10.1111/xen.12683] [Citation(s) in RCA: 18] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/25/2020] [Revised: 01/28/2021] [Accepted: 02/25/2021] [Indexed: 12/13/2022]
Abstract
Decellularized xenografts are an inherent component of regenerative medicine. Their preserved structure, mechanical integrity and biofunctional composition have well established them in reparative medicine for a diverse range of clinical indications. Nonetheless, their performance is highly influenced by their source (ie species, age, tissue) and processing (ie decellularization, crosslinking, sterilization and preservation), which govern their final characteristics and determine their success or failure for a specific clinical target. In this review, we provide an overview of the different sources and processing methods used in decellularized xenografts fabrication and discuss their effect on the clinical performance of commercially available decellularized xenografts.
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Affiliation(s)
- Héctor Capella-Monsonís
- 1Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.,Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CÚRAM), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland
| | - Dimitrios I Zeugolis
- 1Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.,Science Foundation Ireland (SFI) Centre for Research in Medical Devices (CÚRAM), Biomedical Sciences Building, National University of Ireland Galway (NUI Galway), Galway, Ireland.,Regenerative, Modular & Developmental Engineering Laboratory (REMODEL), Faculty of Biomedical Sciences, Università della Svizzera Italiana (USI), Lugano, Switzerland
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24
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El Zarif M, Alió JL, Alió Del Barrio JL, De Miguel MP, Abdul Jawad K, Makdissy N. Corneal Stromal Regeneration: A Review of Human Clinical Studies in Keratoconus Treatment. Front Med (Lausanne) 2021; 8:650724. [PMID: 33708786 PMCID: PMC7940685 DOI: 10.3389/fmed.2021.650724] [Citation(s) in RCA: 26] [Impact Index Per Article: 6.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2021] [Accepted: 01/28/2021] [Indexed: 12/18/2022] Open
Abstract
The use of advanced therapies with stem cells to reconstruct the complex tissue of corneal stroma has gained interest in recent years. Besides, collagen-based scaffolds bioengineering has been offered as another alternative over the last decade. The outcomes of the first clinical experience with stem cells therapy on corneal stroma regeneration in patients with advanced keratoconus were recently reported. Patients were distributed into three experimental groups: Group 1 (G-1) patients underwent implantation of autologous adipose-derived adult stem cells (ADASCs) alone, Group 2 (G-2) received a 120 μm decellularized donor corneal stromal laminas, and Group 3 (G-3) received a 120 μm recellularized donor laminas with ADASCs. A follow up of 36 months of clinical data, and 12 months of confocal microscopy study was performed, the authors found significant clinical improvement in almost all studied mean values of primary and secondary outcomes. Corneal confocal microscopy demonstrated an increase in cell density in the host stroma, as well as in the implanted tissue. Using different approaches, allogenic small incision lenticule extraction (SMILE) implantation was applied in cases with advanced keratoconus. Some authors reported the implantation of SMILE intrastromal lenticules combined with accelerated collagen cross-linking. Others performed intrastromal implantation of negative meniscus-shaped corneal stroma lenticules. Others have compared the outcomes of penetrating keratoplasty (PKP) vs. small-incision Intralase femtosecond (IFS) intracorneal concave lenticule implantation (SFII). Femtosecond laser-assisted small incision sutureless intrasotromal lamellar keratoplasty (SILK) has been also investigated. The published evidence shows that the implantation of autologous ADASCs, decellularized or recellularized human corneal stroma, allogenic SMILE lenticules corneal inlay, and recombinant cross-linked collagen have shown initially to be potentially effective for the treatment of advanced keratoconus. In light of the present evidence available, it can be said that the era of corneal stromal regeneration therapy has been already started.
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Affiliation(s)
- Mona El Zarif
- Optica General, Saida, Lebanon.,Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain.,Faculty of Sciences, GSBT Genomic Surveillance and Biotherapy Team, Mont Michel Campus, Lebanese University, Beirut, Lebanon.,Doctoral School of Sciences and Technology, Lebanese University, Hadath, Lebanon
| | - Jorge L Alió
- Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain.,Cornea, Cataract and Refractive Surgery Unit, Vissum (Miranza Group), Alicante, Spain
| | - Jorge L Alió Del Barrio
- Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain.,Cornea, Cataract and Refractive Surgery Unit, Vissum (Miranza Group), Alicante, Spain
| | - Maria P De Miguel
- Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain
| | | | - Nehman Makdissy
- Faculty of Sciences, GSBT Genomic Surveillance and Biotherapy Team, Mont Michel Campus, Lebanese University, Beirut, Lebanon
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25
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Hancox Z, Heidari Keshel S, Yousaf S, Saeinasab M, Shahbazi MA, Sefat F. The progress in corneal translational medicine. Biomater Sci 2020; 8:6469-6504. [PMID: 33174878 DOI: 10.1039/d0bm01209b] [Citation(s) in RCA: 16] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Cornea tissue is in high demand by tissue donation centres globally, and thus tissue engineering cornea, which is the main topic of corneal translational medicine, can serve as a limitless alternative to a donated human cornea tissue. Tissue engineering aims to produce solutions to the challenges associated with conventional cornea tissue, including transplantation and use of human amniotic membrane (HAM), which have issues with storage and immune rejection in patients. Accordingly, by carefully selecting biomaterials and fabrication methods to produce these therapeutic tissues, the demand for cornea tissue can be met, with an improved healing outcome for recipients with less associated harmful risks. In this review paper, we aim to present the recent advancements in the research and clinical applications of cornea tissue, applications including biomaterial selection, fabrication methods, scaffold structure, cellular response to these scaffolds, and future advancements of these techniques.
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Affiliation(s)
- Zoe Hancox
- Department of Biomedical and Electronics Engineering, School of Engineering, University of Bradford, Bradford, UK.
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26
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Mahdavi SS, Abdekhodaie MJ, Mashayekhan S, Baradaran-Rafii A, Djalilian AR. Bioengineering Approaches for Corneal Regenerative Medicine. Tissue Eng Regen Med 2020; 17:567-593. [PMID: 32696417 PMCID: PMC7373337 DOI: 10.1007/s13770-020-00262-8] [Citation(s) in RCA: 57] [Impact Index Per Article: 11.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/22/2020] [Accepted: 04/06/2020] [Indexed: 12/13/2022] Open
Abstract
BACKGROUND Since the cornea is responsible for transmitting and focusing light into the eye, injury or pathology affecting any layer of the cornea can cause a detrimental effect on visual acuity. Aging is also a reason for corneal degeneration. Depending on the level of the injury, conservative therapies and donor tissue transplantation are the most common treatments for corneal diseases. Not only is there a lack of donor tissue and risk of infection/rejection, but the inherent ability of corneal cells and layers to regenerate has led to research in regenerative approaches and treatments. METHODS In this review, we first discussed the anatomy of the cornea and the required properties for reconstructing layers of the cornea. Regenerative approaches are divided into two main categories; using direct cell/growth factor delivery or using scaffold-based cell delivery. It is expected delivered cells migrate and integrate into the host tissue and restore its structure and function to restore vision. Growth factor delivery also has shown promising results for corneal surface regeneration. Scaffold-based approaches are categorized based on the type of scaffold, since it has a significant impact on the efficiency of regeneration, into the hydrogel and non-hydrogel based scaffolds. Various types of cells, biomaterials, and techniques are well covered. RESULTS The most important characteristics to be considered for biomaterials in corneal regeneration are suitable mechanical properties, biocompatibility, biodegradability, and transparency. Moreover, a curved shape structure and spatial arrangement of the fibrils have been shown to mimic the corneal extracellular matrix for cells and enhance cell differentiation. CONCLUSION Tissue engineering and regenerative medicine approaches showed to have promising outcomes for corneal regeneration. However, besides proper mechanical and optical properties, other factors such as appropriate sterilization method, storage, shelf life and etc. should be taken into account in order to develop an engineered cornea for clinical trials.
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Affiliation(s)
- S Sharareh Mahdavi
- Department of Chemical and Petroleum Engineering, Sharif University of Technology, 1393 Azadi Ave., Tehran, 11365-11155, Iran
| | - Mohammad J Abdekhodaie
- Department of Chemical and Petroleum Engineering, Sharif University of Technology, 1393 Azadi Ave., Tehran, 11365-11155, Iran.
| | - Shohreh Mashayekhan
- Department of Chemical and Petroleum Engineering, Sharif University of Technology, 1393 Azadi Ave., Tehran, 11365-11155, Iran
| | - Alireza Baradaran-Rafii
- Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, SBUMS, Arabi Ave, Daneshjoo Blvd, Velenjak, Tehran, 19839-63113, Iran
| | - Ali R Djalilian
- Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, 1200 W Harrison St, Chicago, IL, 60607, USA
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27
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Li J, Rao Z, Zhao Y, Xu Y, Chen L, Shen Z, Bai Y, Lin Z, Huang Q. A Decellularized Matrix Hydrogel Derived from Human Dental Pulp Promotes Dental Pulp Stem Cell Proliferation, Migration, and Induced Multidirectional Differentiation In Vitro. J Endod 2020; 46:1438-1447.e5. [DOI: 10.1016/j.joen.2020.07.008] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/06/2020] [Revised: 07/04/2020] [Accepted: 07/04/2020] [Indexed: 01/17/2023]
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28
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Biomimetic corneal stroma using electro-compacted collagen. Acta Biomater 2020; 113:360-371. [PMID: 32652228 DOI: 10.1016/j.actbio.2020.07.004] [Citation(s) in RCA: 17] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/06/2020] [Revised: 07/01/2020] [Accepted: 07/02/2020] [Indexed: 12/14/2022]
Abstract
Engineering substantia propria (or stroma of cornea) that mimics the function and anatomy of natural tissue is vital for in vitro modelling and in vivo regeneration. There are, however, few examples of bioengineered biomimetic corneal stroma. Here we describe the construction of an orthogonally oriented 3D corneal stroma model (3D-CSM) using pure electro-compacted collagen (EC). EC films comprise aligned collagen fibrils and support primary human corneal stromal cells (hCSCs). Cell-laden constructs are analogous to the anatomical structure of native human cornea. The hCSCs are guided by the topographical cues provided by the aligned collagen fibrils of the EC films. Importantly, the 3D-CSM are biodegradable, highly transparent, glucose-permeable and comprise quiescent hCSCs. Gene expression analysis indicated the presence of aligned collagen fibrils is strongly coupled to downregulation of active fibroblast/myofibroblast markers α-SMA and Thy-1, with a concomitant upregulation of the dormant keratocyte marker ALDH3. The 3D-CSM represents the first example of an optimally robust biomimetic engineered corneal stroma that is constructed from pure electro-compacted collagen for cell and tissue support. The 3D-CSM is a significant advance for synthetic corneal stroma engineering, with the potential to be used for full-thickness and functional cornea replacement, as well as informing in vivo tissue regeneration. STATEMENT OF SIGNIFICANCE: This manuscript represents the first example of a robust, transparent, glucose permeable and pure collagen-based biomimetic 3D corneal stromal model (3D-CSM) constructed from pure electro-compacted collagen. The collagen fibrils of 3D-CSM are aligned and orthogonally arranged, mimicking native human corneal stroma. The alignment of collagen fibrils correlates with the direction of current applied for electro-compaction and influences human corneal stromal cell (hCSC) orientation. Moreover, 3D-CSM constructs support a corneal keratocyte phenotype; an essential requirement for modelling healthy corneal stroma. As-prepared 3D-CSM hold great promise as corneal stromal substitutes for research and translation, with the potential to be used for full-thickness cornea replacement.
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29
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Riau AK, Liu YC, Yam GH, Mehta JS. Stromal keratophakia: Corneal inlay implantation. Prog Retin Eye Res 2020; 75:100780. [DOI: 10.1016/j.preteyeres.2019.100780] [Citation(s) in RCA: 22] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2019] [Revised: 07/28/2019] [Accepted: 09/02/2019] [Indexed: 12/31/2022]
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30
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Kilic Bektas C, Hasirci V. Cell Loaded GelMA:HEMA IPN hydrogels for corneal stroma engineering. JOURNAL OF MATERIALS SCIENCE. MATERIALS IN MEDICINE 2019; 31:2. [PMID: 31811387 DOI: 10.1007/s10856-019-6345-4] [Citation(s) in RCA: 20] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/15/2019] [Accepted: 11/16/2019] [Indexed: 06/10/2023]
Abstract
Stroma is the main refractive element of the cornea and damage to it is one of the main causes of blindness. In this study, cell loaded hydrogels of methacrylated gelatin (GelMA) and poly(2-hydroxyethyl methacrylate) (pHEMA) (8:2) interpenetrating network (IPN) hydrogels were prepared as the corneal stroma substitute and tested in situ and in vitro. Compressive modulus of the GelMA hydrogels was significantly enhanced with the addition of pHEMA in the structure (6.53 vs 155.49 kPa, respectively). More than 90% of the stromal keratocytes were viable in the GelMA and GelMA-HEMA hydrogels as calculated by Live-Dead Assay and NIH Image-J program. Cells synthesized representative collagens and proteoglycans in the hydrogels indicating that they preserved their keratocyte functions. Transparency of the cell loaded GelMA-HEMA hydrogels was increased significantly up to 90% at 700 nm during three weeks of incubation and was comparable with the transparency of native cornea. Cell loaded GelMA-HEMA corneal stroma model is novel and reported for the first time in the literature in terms of introduction of cells during the preparation phase of the hydrogels. The appropriate mechanical strength and high transparency of the cell loaded constructs indicates a viable alternative to the current devices used in the treatment of corneal blindness.
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Affiliation(s)
- Cemile Kilic Bektas
- Department of Biological Sciences, Middle East Technical University (METU), Ankara, Turkey
- Department of Biotechnology, METU, Ankara, Turkey
- BIOMATEN, METU Center of Excellence in Biomaterials and Tissue Engineering, Ankara, Turkey
| | - Vasif Hasirci
- Department of Biological Sciences, Middle East Technical University (METU), Ankara, Turkey.
- Department of Biotechnology, METU, Ankara, Turkey.
- BIOMATEN, METU Center of Excellence in Biomaterials and Tissue Engineering, Ankara, Turkey.
- Department of Medical Engineering, Acıbadem Mehmet Ali Aydınlar University, Istanbul, Turkey.
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31
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Uyanıklar M, Günal G, Tevlek A, Hosseinian P, Aydin HM. Hybrid Cornea: Cell Laden Hydrogel Incorporated Decellularized Matrix. ACS Biomater Sci Eng 2019; 6:122-133. [DOI: 10.1021/acsbiomaterials.9b01286] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
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32
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Isidan A, Liu S, Li P, Lashmet M, Smith LJ, Hara H, Cooper DK, Ekser B. Decellularization methods for developing porcine corneal xenografts and future perspectives. Xenotransplantation 2019; 26:e12564. [PMID: 31659811 PMCID: PMC6908750 DOI: 10.1111/xen.12564] [Citation(s) in RCA: 49] [Impact Index Per Article: 8.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2019] [Revised: 09/11/2019] [Accepted: 10/13/2019] [Indexed: 12/23/2022]
Abstract
Corneal transplantation is the only option to cure corneal opacities. However, there is an imbalance between supply and demand of corneal tissues in the world. To solve the problem of corneal shortage, corneal xenotransplantation studies have been implemented in the past years using porcine corneas. The corneal xenografts could come from (a) wild-type pigs, (b) genetically engineered pigs, (c) decellularized porcine corneas, and (d) decellularized porcine corneas that are recellularized with human corneal cells, eventually with patients' own cells, as in all type of xenografts. All approaches except, the former would reduce or mitigate recipient immune responses. Although several techniques in decellularization have been reported, there is still no standardized protocol for the complete decellularization of corneal tissue. Herein, we reviewed different decellularization methods for porcine corneas based on the mechanism of action, decellularization efficacy, biocompatibility, and the undesirable effects on corneal ultrastructure. We compared 9 decellularization methods including: (a) sodium dodecyl sulfate, (b) triton x-100, (c) hypertonic saline, (d) human serum with electrophoresis, (e) high hydrostatic pressure, (f) freeze-thaw, (h) nitrogen gas, (h) phospholipase A2 , and (i) glycerol with chemical crosslinking methods. It appears that combined methods could be more useful to perform efficient corneal decellularization.
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Affiliation(s)
- Abdulkadir Isidan
- Transplant Division, Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA
| | - Shaohui Liu
- Department of Ophthalmology, Glick Eye Institute, Indiana University School of Medicine, Indianapolis, IN, USA
| | - Ping Li
- Transplant Division, Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA
| | - Matthew Lashmet
- Transplant Division, Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA
| | - Lester J. Smith
- Radiology and Imaging Sciences, Indiana University School of Medicine, Indianapolis, IN, USA
- 3D Bioprinting Core, Indiana University School of Medicine, Indianapolis, IN, USA
| | - Hidetaka Hara
- Xenotransplantation Program, Department of Surgery, University of Birmingham at Alabama, Birmingham, AL, USA
| | - David K.C. Cooper
- Xenotransplantation Program, Department of Surgery, University of Birmingham at Alabama, Birmingham, AL, USA
| | - Burcin Ekser
- Transplant Division, Department of Surgery, Indiana University School of Medicine, Indianapolis, IN, USA
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Alió JL, Alió Del Barrio JL, El Zarif M, Azaar A, Makdissy N, Khalil C, Harb W, El Achkar I, Jawad ZA, De Miguel MP. Regenerative Surgery of the Corneal Stroma for Advanced Keratoconus: 1-Year Outcomes. Am J Ophthalmol 2019; 203:53-68. [PMID: 30772348 DOI: 10.1016/j.ajo.2019.02.009] [Citation(s) in RCA: 55] [Impact Index Per Article: 9.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/13/2018] [Revised: 02/06/2019] [Accepted: 02/06/2019] [Indexed: 12/13/2022]
Abstract
PURPOSE This study evaluated 1-year safety and efficacy outcomes of corneal stroma cell therapy. Therapy consisted of implanting autologous adipose-derived adult stem cells (ADASc) with or without sheets of decellularized donor human corneal stroma within the stroma of patients with advanced keratoconus. DESIGN This was a prospective interventional non-randomized series of cases. METHODS Fourteen consecutive patients were selected and divided into 3 experimental groups. Group A patients underwent implantation of autologous ADASc alone (3 × 106 cells/1 mL) (n = 5). Group B patients received decellularized donor 120-μm-thick corneal stroma lamina alone (n = 5). Group C patients had implantation of recellularized donor lamina with 1 × 106 autologous ADASc plus another 1 × 106 cells/1 mL at the time of the surgery (n = 4). Autologous ADASc were obtained by elective liposuction. Implantation was performed in the corneal stroma through a femtosecond-assisted 9.5-mm diameter lamellar dissection with the patient under topical anesthesia. Twelve months of follow-up data are presented. RESULTS No complications were observed during the 1-year follow-up, and full corneal transparency was recovered within 3 months in all patients. No patient lost lines of visual acuity. Corrected distance visual acuity improved 0.231, 0.264, and 0.094 Snellen lines in groups 1, 2, and 3, respectively. In group 1, refractive parameters showed an overall stability, whereas in groups 2 and 3, sphere improved 2.35 diopter (D) and 0.625 D, respectively. Anterior keratometry remained stable (group 1) and improved in groups 2 and 3 (mean improvement of 2D). Corneal aberrometry improved significantly. In optical coherence tomography scans, corneal thickness showed a mean improvement of 14.5 μm (group 1) and 116.4 μm (groups 2 and 3) in the central thickness, and new collagen production was observed at the surgical plane (group 1). Confocal biomicroscopy confirmed the host recellularization of the implanted laminas. CONCLUSIONS Intrastromal implantation of autologous ADASc and decellularized human corneal stroma did not show complications at 1 year of follow-up and were moderately effective for the treatment of advanced keratoconus. NOTE: Publication of this article is sponsored by the American Ophthalmological Society.
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34
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Capella-Monsonís H, Kelly J, Kearns S, Zeugolis DI. Decellularised porcine peritoneum as a tendon protector sheet. Biomed Mater 2019; 14:044102. [DOI: 10.1088/1748-605x/ab2301] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022]
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35
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Wang YH, Young TH, Wang TJ. Investigating the effect of chitosan/ polycaprolactone blends in differentiation of corneal endothelial cells and extracellular matrix compositions. Exp Eye Res 2019; 185:107679. [PMID: 31129253 DOI: 10.1016/j.exer.2019.05.019] [Citation(s) in RCA: 25] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2018] [Revised: 04/10/2019] [Accepted: 05/22/2019] [Indexed: 12/14/2022]
Abstract
This study aimed to investigate the underlying mechanisms of corneal endothelial cells (CECs) differentiation and identify the extracellular matrix (ECM) compositions using chitosan/polycaprolactone (PCL) blended membrane, hence exploring the potential use of chitosan/PCL blends in tissue engineering of CECs. We utilized the chitosan/PCL blends named as PCL25 consisting of PCL at 25% by weight. The surface characteristics of PCL25 were confirmed by using Fourier Transform Infrared Spectroscopy (FTIR) and Atomic Force Microscope (AFM). Bovine CECs were cultured on the blends, compared with TCPS and pure chitosan membrane. Cell behaviors in terms of cell attachment, proliferation, differentiation phenotype and expression of differentiation proteins were examined. Furthermore, ECM protein productions were also analyzed. From the experiments, we found the topography (roughness) of PCL25 membrane examined by AFM was greater than pure chitosan membrane. FTIR results confirmed the functional groups of C=O bond of PCL. The CECs displayed hexagonal morphology and similar proliferation rate on both PCL25 membrane and TCPS. In addition, the immunofluorescence evidence showed well-localized ZO-1 and Na+/K+ ATPase expression of membrane proteins. ECM protein productions of CECs on PCL were no inferior to TCPS. Moreover, western blot results verified the higher amount of collagen type IV, and reduced TGF-β2 expression on PCL25 membrane compared to TCPS substrate. In conclusions, chitosan/PCL blends membrane provided a favorable environment for CECs in terms of ECM compositions, therefore enhancing the growth and differentiation. Accordingly, for CEC tissue engineering applications, PCL 25 might be a suitable alternative for cadaveric cornea transplantation in the near future.
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Affiliation(s)
- Yu-Hsin Wang
- Department of Biomedical Engineering, National Taiwan University, Taipei, Taiwan
| | - Tai-Horng Young
- Department of Biomedical Engineering, National Taiwan University, Taipei, Taiwan
| | - Tsung-Jen Wang
- Department of Ophthalmology, Taipei Medical University Hospital, Taipei, Taiwan; Department of Ophthalmology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
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Fernández-Pérez J, Ahearne M. Decellularization and recellularization of cornea: Progress towards a donor alternative. Methods 2019; 171:86-96. [PMID: 31128238 DOI: 10.1016/j.ymeth.2019.05.009] [Citation(s) in RCA: 42] [Impact Index Per Article: 7.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/12/2019] [Revised: 05/09/2019] [Accepted: 05/10/2019] [Indexed: 12/20/2022] Open
Abstract
The global shortage of donor corneas for transplantation has led to corneal bioengineering being investigated as a method to generate transplantable tissues. Decellularized corneas are among the most promising materials for engineering corneal tissue since they replicate the complex structure and composition of real corneas. Decellularization is a process that aims to remove cells from organs or tissues resulting in a cell-free scaffold consisting of the tissues extracellular matrix. Here different decellularization techniques are described, including physical, chemical and biological methods. Analytical techniques to confirm decellularization efficiency are also discussed. Different cell sources for the recellularization of the three layers of the cornea, recellularization methods used in the literature and techniques used to assess the outcome of the implantation of such scaffolds are examined. Studies involving the application of decellularized corneas in animal models and human clinical studies are discussed. Finally, challenges for this technology are explored involving scalability, automatization and regulatory affairs.
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Affiliation(s)
- Julia Fernández-Pérez
- Dept of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, University of Dublin, Ireland; Trinity Centre for Biomedical Engineering, Trinity Biomedical Science Institute, Trinity College Dublin, University of Dublin, Ireland
| | - Mark Ahearne
- Dept of Mechanical and Manufacturing Engineering, School of Engineering, Trinity College Dublin, University of Dublin, Ireland; Trinity Centre for Biomedical Engineering, Trinity Biomedical Science Institute, Trinity College Dublin, University of Dublin, Ireland.
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Development and characterization of Lyophilized Transparized Decellularized stroma as a replacement for living cornea in deep anterior lamellar keratoplasty. Cell Tissue Bank 2019; 20:49-59. [DOI: 10.1007/s10561-018-9742-x] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2018] [Accepted: 12/01/2018] [Indexed: 11/27/2022]
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Dorgau B, Felemban M, Hilgen G, Kiening M, Zerti D, Hunt NC, Doherty M, Whitfield P, Hallam D, White K, Ding Y, Krasnogor N, Al-Aama J, Asfour HZ, Sernagor E, Lako M. Decellularised extracellular matrix-derived peptides from neural retina and retinal pigment epithelium enhance the expression of synaptic markers and light responsiveness of human pluripotent stem cell derived retinal organoids. Biomaterials 2019; 199:63-75. [PMID: 30738336 DOI: 10.1016/j.biomaterials.2019.01.028] [Citation(s) in RCA: 58] [Impact Index Per Article: 9.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2018] [Revised: 01/11/2019] [Accepted: 01/20/2019] [Indexed: 12/13/2022]
Abstract
Tissue specific extracellular matrices (ECM) provide structural support and enable access to molecular signals and metabolites, which are essential for directing stem cell renewal and differentiation. To mimic this phenomenon in vitro, tissue decellularisation approaches have been developed, resulting in the generation of natural ECM scaffolds that have comparable physical and biochemical properties of the natural tissues and are currently gaining traction in tissue engineering and regenerative therapies due to the ease of standardised production, and constant availability. In this manuscript we report the successful generation of decellularised ECM-derived peptides from neural retina (decel NR) and retinal pigment epithelium (decel RPE), and their impact on differentiation of human pluripotent stem cells (hPSCs) to retinal organoids. We show that culture media supplementation with decel RPE and RPE-conditioned media (CM RPE) significantly increases the generation of rod photoreceptors, whilst addition of decel NR and decel RPE significantly enhances ribbon synapse marker expression and the light responsiveness of retinal organoids. Photoreceptor maturation, formation of correct synapses between retinal cells and recording of robust light responses from hPSC-derived retinal organoids remain unresolved challenges for the field of regenerative medicine. Enhanced rod photoreceptor differentiation, synaptogenesis and light response in response to addition of decellularised matrices from RPE and neural retina as shown herein provide a novel and substantial advance in generation of retinal organoids for drug screening, tissue engineering and regenerative medicine.
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Affiliation(s)
- Birthe Dorgau
- Institute of Genetic Medicine, Newcastle University, UK
| | | | | | | | - Darin Zerti
- Institute of Genetic Medicine, Newcastle University, UK
| | | | | | | | - Dean Hallam
- Institute of Genetic Medicine, Newcastle University, UK
| | | | - Yuchun Ding
- Interdisciplinary Computing and Complex Biosystems (ICOS) Research Group, Newcastle University, UK
| | - Natalio Krasnogor
- Interdisciplinary Computing and Complex Biosystems (ICOS) Research Group, Newcastle University, UK
| | - Jumana Al-Aama
- Department of Genetic Medicine and Princess Al-Jawhara Center of Excellence in Research of Hereditary Disorders, Faculty of Medicine, King Abdulaziz University, Saudi Arabia
| | - Hani Z Asfour
- Department of Medical Microbiology and Parasitology, Faculty of Medicine, Princess Al-Jawhara Center of Excellence in Research o Hereditary Disorders, King Abdulaziz University, Jeddah, Saudi Arabia
| | | | - Majlinda Lako
- Institute of Genetic Medicine, Newcastle University, UK.
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Kim H, Park MN, Kim J, Jang J, Kim HK, Cho DW. Characterization of cornea-specific bioink: high transparency, improved in vivo safety. J Tissue Eng 2019; 10:2041731418823382. [PMID: 30719272 PMCID: PMC6348552 DOI: 10.1177/2041731418823382] [Citation(s) in RCA: 75] [Impact Index Per Article: 12.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/01/2018] [Accepted: 12/16/2018] [Indexed: 12/22/2022] Open
Abstract
Corneal transplantation is a typical surgical procedure for severe corneal diseases. However, the waiting time for a donor cornea has gradually increased due to a decrease in supply caused by an aging population and increased cases of laser-based surgeries. Artificial corneas were developed to meet the increase in demand; however, these approaches have suffered from material deterioration resulted by the limited tissue integration. Here, we introduce a cornea-derived decellularized extracellular matrix (Co-dECM) as a bioink for corneal regeneration. The developed Co-dECM bioink had similar quantitative measurement results for collagen and GAGs compared with that of the native cornea and also had the proper transparency for vision. The differentiation potential of human turbinate-derived mesenchymal stem cells (hTMSCs) to a keratocyte lineage was only observed in the Co-dECM group. Moreover, the developed bioink did not have any cytotoxic effect on encapsulated cells for three-dimensional (3D) culture and has great biocompatibility evident by the xeno-implantation of the Co-dECM gel into mice and rabbits for two and one month, respectively. An in vivo safety similar to clinical-grade collagen was seen with the Co-dECM, which helped to maintain the keratocyte-specific characteristics in vivo, compared with collagen. Taken together, the Co-dECM bioink has the potential to be used in various types of corneal diseases based on its corneal-specific ability and design flexibility through 3D cell printing technology.
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Affiliation(s)
- Hyeonji Kim
- Department of Mechanical Engineering and Center for Rapid Prototyping-based 3D Tissue/Organ Printing, Pohang University of Science and Technology, Pohang, Republic of Korea
| | - Moon-Nyeo Park
- College of Korean Medicines, Kyung Hee University, Seoul, Republic of Korea
| | - Jisoo Kim
- School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Republic of Korea
| | - Jinah Jang
- School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Republic of Korea
- Department of Creative IT Engineering, Pohang University of Science and Technology, Pohang, Republic of Korea
| | - Hong-Kyun Kim
- Department of Ophthalmology, Kyungpook National University School of Medicine, Daegu, Republic of Korea
| | - Dong-Woo Cho
- Department of Mechanical Engineering and Center for Rapid Prototyping-based 3D Tissue/Organ Printing, Pohang University of Science and Technology, Pohang, Republic of Korea
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Dorazehi F, Nabiuni M, Jalali H. Potential Use of Amniotic Membrane - Derived Scaffold for Cerebrospinal Fluid Applications. INTERNATIONAL JOURNAL OF MOLECULAR AND CELLULAR MEDICINE 2018; 7:91-101. [PMID: 30276164 PMCID: PMC6148501 DOI: 10.22088/ijmcm.bums.7.2.91] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 04/08/2018] [Accepted: 08/14/2018] [Indexed: 01/23/2023]
Abstract
Scaffolds derived from decellularized tissues provide a natural microenvironment for cell culture. Embryonic cerebrospinal fluid (e-CSF) contains factors which play vital roles in the development of the nervous system. This research was aimed to survey the effect of Wistar rat e-CSF on neural differentiation of bone marrow derived mesenchymal stem cells (BM-MSCs) cultured on the human amniotic membrane (AM). BM-MSCs were collected from femurs and tibias, and were cultured in Dulbecco's Modified Eagle's Medium. The placenta was harvested from healthy women during cesarean section and AM was acellularized using EDTA and physical scrubbing. e- CSF was harvested from rat fetuses at E17. Adequate numbers of BM-MSCs were cultured on acellularized membrane, and were treated with E17 CSF for 7 days. MTT (3-(4, 5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide) assay confirmed the survival and proliferation of BM-MSCs cultured on AM derived scaffold. Hematoxylin/eosin staining and scanning electron microscopy showed the morphological and the structural changes of BM-MSCs throughout the culture and treatment with e-CSF. The results of immunocytochemistry showed that microtubule associated protein 2 and beta-III tubulin were expressed in BM-MSCs cultured on acellular amnion scaffold and treated with e-CSF. Our results showed for the first time that the combination of acellular AM as a natural scaffold and e-CSF as a source of neurological factors could effectively improve the BM-MSCs cultivation and differentiation.
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Affiliation(s)
- Fereshteh Dorazehi
- Department of Cell and Molecular Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran
| | - Mohammad Nabiuni
- Department of Cell and Molecular Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran
| | - Hanieh Jalali
- Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran
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Toprakhisar B, Nadernezhad A, Bakirci E, Khani N, Skvortsov GA, Koc B. Development of Bioink from Decellularized Tendon Extracellular Matrix for 3D Bioprinting. Macromol Biosci 2018; 18:e1800024. [DOI: 10.1002/mabi.201800024] [Citation(s) in RCA: 51] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/17/2018] [Revised: 06/09/2018] [Indexed: 01/12/2023]
Affiliation(s)
- Burak Toprakhisar
- Faculty of Engineering and Natural Sciences; Sabanci University; Orhanli-Tuzla Istanbul 34956 Turkey
- 3D Bioprinting Laboratory; Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
| | - Ali Nadernezhad
- Faculty of Engineering and Natural Sciences; Sabanci University; Orhanli-Tuzla Istanbul 34956 Turkey
- 3D Bioprinting Laboratory; Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
| | - Ezgi Bakirci
- Faculty of Engineering and Natural Sciences; Sabanci University; Orhanli-Tuzla Istanbul 34956 Turkey
- 3D Bioprinting Laboratory; Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
| | - Navid Khani
- Faculty of Engineering and Natural Sciences; Sabanci University; Orhanli-Tuzla Istanbul 34956 Turkey
- 3D Bioprinting Laboratory; Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
| | - Gozde Akdeniz Skvortsov
- 3D Bioprinting Laboratory; Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
- Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
| | - Bahattin Koc
- Faculty of Engineering and Natural Sciences; Sabanci University; Orhanli-Tuzla Istanbul 34956 Turkey
- 3D Bioprinting Laboratory; Sabanci University Nanotechnology Research and Application Center; Orhanli-Tuzla Istanbul 34956 Turkey
- Faculty of Engineering and Natural Sciences; Sabanci University; Orhanli-Tuzla Istanbul 34956 Turkey
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Lin CH, Kao YC, Ma H, Tsay RY. An investigation on the correlation between the mechanical property change and the alterations in composition and microstructure of a porcine vascular tissue underwent trypsin-based decellularization treatment. J Mech Behav Biomed Mater 2018; 86:199-207. [PMID: 29986294 DOI: 10.1016/j.jmbbm.2018.06.029] [Citation(s) in RCA: 18] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/18/2018] [Revised: 06/14/2018] [Accepted: 06/19/2018] [Indexed: 01/15/2023]
Abstract
PURPOSE The nonlinear pseudoelastic behavior of a native/decellularized vascular tissue is closely related to the detailed composition and microstructure of the extracellular matrix and is important in maintaining the patency of a small-caliber vascular graft. A commonly used enzyme-detergent based decellularization protocol is effective in cell component removal but it also changes the microstructure and composition of the decellularized tissues. Previous studies provide limited information to correlate the mechanical property change with the alterations in composition and microstructure in a decellularization process. In this study, the correlations were studied by implementing a previously established fiber-progressive-engagement model to describe the nonlinear pseudoelastic behavior of a vascular tissue and to evaluate the effects of trypsin concentration and exposure duration on porcine coronary artery decellularization RESULTS: Results showed that tissue length and width increased and thickness and wet weight decreased with the exposure of trypsin. The effects of trypsin exposure times on the four mechanical parameters, i.e. initial strain, turning strain, initial modulus and stiffness modulus, in the longitudinal and circumferential directions were similar, but stronger in the circumferential direction. Major components of the extracellular matrix were vulnerable to the trypsin-based decellularization process. The decreases in initial and turning strain and the increase in initial modulus in circumferential direction were correlated with the significant decrease of collagen and glycosaminoglycans in the media layer. CONCLUSIONS Although trypsin-based decellularization achieved cell component removal and preservation of ultimate tensile stress, the microstructure and composition changed with alterations in the pseudoelastic behavior of the porcine coronary artery. Taken together, the current observations suggested less waviness, early engagement, or re-alignment of insoluble collagen fibers in the media layer, which resulted in turning from anisotropic into isotropic uniaxial mechanical property of porcine vascular tissue. Selecting the proper trypsin concentration (< 0.03-0.5%) and duration (< 12 h) of trypsin exposure in combination with other methods will achieve optimal porcine coronary artery decellularization.
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Affiliation(s)
- Chih-Hsun Lin
- Division of Plastic Surgery, Department of Surgery, Taipei Veterans General Hospital, No. 201, Section 2, Shipai Rd., Beitou Dist., Taipei City 112, Taiwan, ROC; Department of Surgery, School of Medicine, National Yang-Ming University, No. 155, Section 2, Linong St., Beitou Dist., Taipei City 112, Taiwan, ROC
| | - Yun-Chu Kao
- Institute of Biomedical Engineering, National Yang-Ming University, No. 155, Section 2, Linong St., Beitou Dist., Taipei City 112, Taiwan, ROC
| | - Hsu Ma
- Division of Plastic Surgery, Department of Surgery, Taipei Veterans General Hospital, No. 201, Section 2, Shipai Rd., Beitou Dist., Taipei City 112, Taiwan, ROC; Department of Surgery, School of Medicine, National Yang-Ming University, No. 155, Section 2, Linong St., Beitou Dist., Taipei City 112, Taiwan, ROC
| | - Ruey-Yug Tsay
- Institute of Biomedical Engineering, National Yang-Ming University, No. 155, Section 2, Linong St., Beitou Dist., Taipei City 112, Taiwan, ROC; Center for Advanced Pharmaceutics and Drug Delivery Research, National Yang-Ming University, No. 155, Section 2, Linong St., Beitou Dist., Taipei City 112, Taiwan, ROC.
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Zhang Y, Iwata T, Nam K, Kimura T, Wu P, Nakamura N, Hashimoto Y, Kishida A. Water absorption by decellularized dermis. Heliyon 2018; 4:e00600. [PMID: 29862362 PMCID: PMC5968173 DOI: 10.1016/j.heliyon.2018.e00600] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/04/2017] [Revised: 02/09/2018] [Accepted: 04/04/2018] [Indexed: 11/20/2022] Open
Abstract
Water absorption by decellularized dermis was investigated and compared with biopolymer and synthetic polymer hydrogels (glutaraldehyde-crosslinked gelatin and crosslinked poly(acrylamide) hydrogel, respectively). Porcine dermis was decellularized in an aqueous sodium dodecyl sulfate (SDS) solution. Histological evaluation revealed that the SDS-treated dermis has much larger gaps between collagen fibrils than non-treated dermis, and that water absorption depends on these gaps. Decellularized dermis has low water absorptivity and the absorption obeys Fick's second law. During absorption, the water diffusion rate decreases with time and occurs in two steps. The first is rapid absorption into the large gaps, followed by slow absorption by the collagen fiber layer. Because of the gaps, decellularized dermis can absorb more water than native dermis and shows different water absorption behavior to glutaraldehyde-crosslinked gelatin and crosslinked poly(acrylamide) hydrogels.
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Chen Z, You J, Liu X, Cooper S, Hodge C, Sutton G, Crook JM, Wallace GG. Biomaterials for corneal bioengineering. ACTA ACUST UNITED AC 2018; 13:032002. [PMID: 29021411 DOI: 10.1088/1748-605x/aa92d2] [Citation(s) in RCA: 81] [Impact Index Per Article: 11.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/11/2022]
Abstract
Corneal transplantation is an important surgical treatment for many common corneal diseases. However, a worldwide shortage of tissue from suitable corneal donors has meant that many people are not able to receive sight-restoring operations. In addition, rejection is a major cause of corneal transplant failure. Bioengineering corneal tissue has recently gained widespread attention. In order to facilitate corneal regeneration, a range of materials is currently being investigated. The ideal substrate requires sufficient tectonic durability, biocompatibility with cultured cellular elements, transparency, and perhaps biodegradability and clinical compliance. This review considers the anatomy and function of the native cornea as a precursor to evaluating a variety of biomaterials for corneal regeneration including key characteristics for optimal material form and function. The integration of appropriate cells with the most appropriate biomaterials is also discussed. Taken together, the information provided offers insight into the requirements for fabricating synthetic and semisynthetic corneas for in vitro modeling of tissue development and disease, pharmaceutical screening, and in vivo application for regenerative medicine.
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Affiliation(s)
- Zhi Chen
- ARC Centre of Excellence for Electromaterials Science, Intelligent Polymer Research Institute, AIIM Facility, Innovation Campus, University of Wollongong, Squires Way, Fairy Meadow, New South Wales 2519, Australia
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45
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Sivandzade F, Mashayekhan S. Design and fabrication of injectable microcarriers composed of acellular cartilage matrix and chitosan. JOURNAL OF BIOMATERIALS SCIENCE-POLYMER EDITION 2018; 29:683-700. [DOI: 10.1080/09205063.2018.1433422] [Citation(s) in RCA: 14] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/31/2022]
Affiliation(s)
- Farzane Sivandzade
- Department of Chemical and Petroleum Engineering, Sharif University of Technology, Tehran, Iran
| | - Shohreh Mashayekhan
- Department of Chemical and Petroleum Engineering, Sharif University of Technology, Tehran, Iran
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Alió Del Barrio JL, El Zarif M, Azaar A, Makdissy N, Khalil C, Harb W, El Achkar I, Jawad ZA, de Miguel MP, Alió JL. Corneal Stroma Enhancement With Decellularized Stromal Laminas With or Without Stem Cell Recellularization for Advanced Keratoconus. Am J Ophthalmol 2018; 186:47-58. [PMID: 29103962 DOI: 10.1016/j.ajo.2017.10.026] [Citation(s) in RCA: 76] [Impact Index Per Article: 10.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/21/2017] [Revised: 10/25/2017] [Accepted: 10/26/2017] [Indexed: 12/13/2022]
Abstract
PURPOSE This phase 1 study seeks to preliminarily evaluate the safety and efficacy of decellularized human corneal stromal lamina transplantation with or without autologous adipose-derived adult stem cell recellularization within the corneal stroma of patients with advanced keratoconus. DESIGN Phase 1 clinical trial. METHODS Femtosecond-assisted 120-μm thickness and 9-mm diameter laminas were obtained from the anterior stroma of human donor corneas and decellularized with a sodium dodecyl sulfate solution. Autologous adipose-derived adult stem cells were obtained by elective liposuction and cultured onto both sides of the lamina. Five patients received the decellularized lamina alone and 4 patients the recellularized lamina into a femtosecond-assisted 9.5-mm diameter lamellar pocket under topical anesthesia. The total duration of follow-up was 6 months. RESULTS No case showed clinical haze or scarring by month 3. Six months after surgery, patients showed a general improvement of all visual parameters, with a mean unaided visual acuity from 0.109 to 0.232 (P = .05) and corrected distance visual acuity from 0.22 to 0.356 (P = .068). Refractive sphere improved in all patients (from -4.55 diopters [D] to -2.69 D; P = .017), but refractive cylinder remained stable (from -2.83 to -2.61; P = .34). An improvement tendency of all anterior keratometric values was observed. A mean improvement of 120 μm in all thickness parameters was confirmed (P = .008), as well as an improvement in the spherical aberration (P = .018), coma (P = .23) and total higher order aberrations (P = .31). No significant differences among groups were detected. CONCLUSIONS Decellularized human corneal stromal laminas transplantation seems safe and moderately effective for advanced keratoconus. Potential benefits of its recellularization with autologous adipose-derived adult stem cells remains unclear.
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Affiliation(s)
- Jorge L Alió Del Barrio
- Cornea, Cataract and Refractive Surgery Unit, Vissum Corporación, Alicante, Spain; Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain
| | | | - Albert Azaar
- Reviva Regenerative Medicine Center, Middle East Hospital, Beirut, Lebanon
| | - Nehman Makdissy
- Reviva Regenerative Medicine Center, Middle East Hospital, Beirut, Lebanon; Lebanese University, Beirut, Lebanon
| | - Charbel Khalil
- Reviva Regenerative Medicine Center, Middle East Hospital, Beirut, Lebanon
| | - Walid Harb
- Reviva Regenerative Medicine Center, Middle East Hospital, Beirut, Lebanon
| | | | | | - María P de Miguel
- Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain
| | - Jorge L Alió
- Cornea, Cataract and Refractive Surgery Unit, Vissum Corporación, Alicante, Spain; Division of Ophthalmology, Universidad Miguel Hernández, Alicante, Spain.
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Kilic Bektas C, Hasirci V. Mimicking corneal stroma using keratocyte-loaded photopolymerizable methacrylated gelatin hydrogels. J Tissue Eng Regen Med 2018; 12:e1899-e1910. [PMID: 29193831 DOI: 10.1002/term.2621] [Citation(s) in RCA: 55] [Impact Index Per Article: 7.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2017] [Revised: 10/31/2017] [Accepted: 11/22/2017] [Indexed: 11/06/2022]
Abstract
Cell-laden methacrylated gelatin (GelMA) hydrogels with high (approximately 90%) transparency were prepared to mimic the natural form and function of corneal stroma. They were synthesized from GelMA with a methacrylation degree of 70% as determined by nuclear magnetic resonance. Hydrogels were strong enough to withstand handling. Stability studies showed that 87% of the GelMA hydrogels remained after 21 days in phosphate buffered saline (PBS). Cell viability in the first 2 days was over 90% for the human keratocytes loaded in the gels as determined with the live-dead analysis. Cells in the hydrogel elongated and connected to each other as observed by confocal laser scanning microscopy (CLSM) images and scanning electron microscope analysis after 3 weeks in the culture medium and cells were seen to be distributed throughout the hydrogel bulk. Cells were found to synthesize collagen Types I and V, decorin, and biglycan (representative collagens and proteoglycans of human corneal stroma, respectively) showing that keratocytes maintained their functions and preserved their phenotypes in the hydrogels. Transparency of cell-loaded and cell-free hydrogels after 21 days was found to be over 90% at all time points in the visible light range and was comparable to the transparency of the native cornea. The corneal stroma equivalent produced in this study that has cells entrapped in it leads to a product with homogenous distribution of cells. It was transparent at the very beginning and is expected to allow better vision than nontransparent substrates. It, therefore, has a significant potential to be used as an alternative to the current products used to treat corneal blindness.
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Affiliation(s)
- Cemile Kilic Bektas
- Department of Biological Sciences, METU, Ankara, Turkey.,Department of Biotechnology, METU, Ankara, Turkey.,BIOMATEN, METU Center of Excellence in Biomaterials and Tissue Engineering, Ankara, Turkey
| | - Vasif Hasirci
- Department of Biological Sciences, METU, Ankara, Turkey.,Department of Biotechnology, METU, Ankara, Turkey.,BIOMATEN, METU Center of Excellence in Biomaterials and Tissue Engineering, Ankara, Turkey
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Functionalized silk fibroin film scaffold using β-Carotene for cornea endothelial cell regeneration. Colloids Surf B Biointerfaces 2017; 164:340-346. [PMID: 29413615 DOI: 10.1016/j.colsurfb.2017.11.052] [Citation(s) in RCA: 34] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2017] [Revised: 11/13/2017] [Accepted: 11/21/2017] [Indexed: 11/21/2022]
Abstract
Design of corneal endothelium substitute is important for replacement of cadaveric cornea tissue. Our previous study has shown the suitability of silk fibroin (SF) as a biomaterial for cornea scaffold. In this study, we used β-Carotene (β-C) to enhance the regeneration of corneal endothelial cells (CEnCs) and maintain CEnC specific function. The fabricated film scaffolds showed desired transparency and hydrophilic properties which are crucial factors for vision recovery. The cell viability, phenotype and gene expression was examined using MTT assay, immunofluorescence and reverse transcription polymerase chain reactions. Compared with pristine SF scaffold, proper amount of β-C incorporated with SF scaffolds showed higher initial cell attachment, cell viability and mRNA expression. The results indicate that the fabricated SF film scaffold incorporated with small amount of β-C might be the suitable alternative corneal endothelium substitute for transplantation.
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Brunette I, Roberts CJ, Vidal F, Harissi-Dagher M, Lachaine J, Sheardown H, Durr GM, Proulx S, Griffith M. Alternatives to eye bank native tissue for corneal stromal replacement. Prog Retin Eye Res 2017; 59:97-130. [DOI: 10.1016/j.preteyeres.2017.04.002] [Citation(s) in RCA: 41] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/17/2016] [Revised: 04/15/2017] [Accepted: 04/21/2017] [Indexed: 12/13/2022]
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