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Radi S, EzEldeen M, Végvári Á, Coates D, Jacobs R, Bostanci N, Bao K. The proteome of osteoblasts in a 3D culture perfusion bioreactor model compared with static conditions. Sci Rep 2025; 15:12120. [PMID: 40204872 PMCID: PMC11982442 DOI: 10.1038/s41598-025-96632-0] [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: 12/10/2024] [Accepted: 03/31/2025] [Indexed: 04/11/2025] Open
Abstract
Bone disorders represent a significant global burden. Currently, animal models are used to develop and screen novel treatments. However, interspecies variations and ethical concerns highlight the need for a more complex 3D bone model. In this study, we developed a simplified in vitro bone-like model using a U-CUP perfusion-based bioreactor system, designed to provide continuous nutrient flow and mechanostimulation through 3D cultures. An immortalized human fetal osteoblastic cell line was seeded on collagen scaffolds and cultured for 21 days in both a perfusion bioreactor system and in static cultures. PrestoBlue™ assay, scanning electron microscopy, and proteomics allowed monitoring of metabolic activity and compared morphological and proteome differences between both conditions. Results indicated an altered cellular morphology in the bioreactor compared to the static cultures and identified a total of 3494 proteins. Of these, 105 proteins exhibited significant upregulation in the static culture, while 86 proteins displayed significant downregulation. Enrichment analyses of these proteins revealed ten significant pathways including epithelial-mesenchymal transition, TNF-alpha signaling via NF-kB, and KRAS pathway. The current data indicated of osteogenic differentiation enhancement within the bioreactor on day 21 compared to static cultures. In conclusion, the U-CUP perfusion bioreactor is beneficial for facilitating osteogenic differentiation in 3D cultures.
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Affiliation(s)
- Sonya Radi
- OMFS-IMPATH Research Group, Faculty of Medicine, Department of Imaging and Pathology, KU Leuven and Oral and Maxillofacial Surgery, University Hospitals Leuven, Leuven, Belgium
| | - Mostafa EzEldeen
- OMFS-IMPATH Research Group, Faculty of Medicine, Department of Imaging and Pathology, KU Leuven and Oral and Maxillofacial Surgery, University Hospitals Leuven, Leuven, Belgium
- Department of Oral Health Sciences, KU Leuven and Paediatric Dentistry and Special Dental Care, University Hospitals Leuven, Leuven, Belgium
| | - Ákos Végvári
- Proteomics Biomedicum, Division of Chemistry I, Department of Medical Biochemistry and Biophysics (MBB), Karolinska Institutet, Solna, Stockholm, Sweden
| | - Dawn Coates
- Sir John Walsh Research Institute, Faculty of Dentistry, University of Otago, Dunedin, New Zealand
| | - Reinhilde Jacobs
- OMFS-IMPATH Research Group, Faculty of Medicine, Department of Imaging and Pathology, KU Leuven and Oral and Maxillofacial Surgery, University Hospitals Leuven, Leuven, Belgium
- Department of Dental Medicine (DENTMED), Karolinska Institutet, Huddinge, Stockholm, Sweden
| | - Nagihan Bostanci
- Division of Oral Health and Periodontology, Department of Dental Medicine, Karolinska Institutet, Huddinge, Stockholm, Sweden
| | - Kai Bao
- Division of Oral Health and Periodontology, Department of Dental Medicine, Karolinska Institutet, Huddinge, Stockholm, Sweden.
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Inchingolo AM, Inchingolo AD, Nardelli P, Latini G, Trilli I, Ferrante L, Malcangi G, Palermo A, Inchingolo F, Dipalma G. Stem Cells: Present Understanding and Prospects for Regenerative Dentistry. J Funct Biomater 2024; 15:308. [PMID: 39452606 PMCID: PMC11508604 DOI: 10.3390/jfb15100308] [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: 08/05/2024] [Revised: 10/02/2024] [Accepted: 10/11/2024] [Indexed: 10/26/2024] Open
Abstract
Regenerative medicine in dentistry focuses on repairing damaged oral tissues using advanced tools like stem cells, biomaterials, and tissue engineering (TE). Mesenchymal stem cells (MSCs) from dental sources, such as dental pulp and periodontal ligament, show significant potential for tissue regeneration due to their proliferative and differentiative abilities. This systematic review, following PRISMA guidelines, evaluated fifteen studies and identified effective strategies for improving dental, periodontal, and bone tissue regeneration through scaffolds, secretomes, and bioengineering methods. Key advancements include the use of dental pulp stem cells (DPSCs) and periodontal ligament stem cells (PDLSCs) to boost cell viability and manage inflammation. Additionally, pharmacological agents like matrine and surface modifications on biomaterials improve stem cell adhesion and promote osteogenic differentiation. By integrating these approaches, regenerative medicine and TE can optimize dental therapies and enhance patient outcomes. This review highlights the potential and challenges in this field, providing a critical assessment of current research and future directions.
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Affiliation(s)
- Angelo Michele Inchingolo
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Alessio Danilo Inchingolo
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Paola Nardelli
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Giulia Latini
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Irma Trilli
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Laura Ferrante
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Giuseppina Malcangi
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Andrea Palermo
- College of Medicine and Dentistry, Birmingham B4 6BN, UK;
| | - Francesco Inchingolo
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
| | - Gianna Dipalma
- Department of Interdisciplinary Medicine, University of Bari “Aldo Moro”, 70124 Bari, Italy; (A.M.I.); (A.D.I.); (P.N.); (G.L.); (I.T.); (L.F.); (G.D.)
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Li X, Li Y, Zhang X, Xu J, Kang J, Li B, Zhao B, Wang L. Cross-Linking Methods of the Silk Protein Hydrogel in Oral and Craniomaxillofacial Tissue Regeneration. Tissue Eng Regen Med 2024; 21:529-544. [PMID: 38294593 PMCID: PMC11087422 DOI: 10.1007/s13770-023-00624-y] [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: 09/04/2023] [Revised: 12/16/2023] [Accepted: 12/17/2023] [Indexed: 02/01/2024] Open
Abstract
BACKGROUND Craniomaxillofacial tissue defects are clinical defects involving craniomaxillofacial and oral soft and hard tissues. They are characterized by defect-shaped irregularities, bacterial and inflammatory environments, and the need for functional recovery. Conventional clinical treatments are currently unable to achieve regeneration of high-quality oral craniomaxillofacial tissue. As a natural biomaterial, silk fibroin (SF) has been widely studied in biomedicine and has broad prospects for use in tissue regeneration. Hydrogels made of SF showed excellent water retention, biocompatibility, safety and the ability to combine with other materials. METHODS To gain an in-depth understanding of the current development of SF, this article reviews the structure, preparation and application prospects in oral and craniomaxillofacial tissue regenerative medicine. It first briefly introduces the structure of SF and then summarizes the principles, advantages and disadvantages of the different cross-linking methods (physical cross-linking, chemical cross-linking and double network structure) of SF. Finally, the existing research on the use of SF in tissue engineering and the prospects of using SF with different cross-linking methods in oral and craniomaxillofacial tissue regeneration are also discussed. CONCLUSIONS This review is intended to show the advantages of SF hydrogels in tissue engineering and provides theoretical support for establishing novel and viable silk protein hydrogels for regeneration.
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Affiliation(s)
- Xiujuan Li
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China
| | - Yuanjiao Li
- School of Medical Sciences, Shanxi Medical University, Taiyuan, Shanxi, China
| | - Xinsong Zhang
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China
| | - Jie Xu
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China
| | - Jie Kang
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China
| | - Bing Li
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China
| | - Bin Zhao
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China.
| | - Lu Wang
- Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials, Shanxi Medical University School and Hospital of Stomatology, Taiyuan, 030001, Shanxi, China.
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Bryniarska-Kubiak N, Basta-Kaim A, Kubiak A. Mechanobiology of Dental Pulp Cells. Cells 2024; 13:375. [PMID: 38474339 PMCID: PMC10931140 DOI: 10.3390/cells13050375] [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: 12/23/2023] [Revised: 02/06/2024] [Accepted: 02/11/2024] [Indexed: 03/14/2024] Open
Abstract
The dental pulp is the inner part of the tooth responsible for properly functioning during its lifespan. Apart from the very big biological heterogeneity of dental cells, tooth microenvironments differ a lot in the context of mechanical properties-ranging from 5.5 kPa for dental pulp to around 100 GPa for dentin and enamel. This physical heterogeneity and complexity plays a key role in tooth physiology and in turn, is a great target for a variety of therapeutic approaches. First of all, physical mechanisms are crucial for the pain propagation process from the tooth surface to the nerves inside the dental pulp. On the other hand, the modulation of the physical environment affects the functioning of dental pulp cells and thus is important for regenerative medicine. In the present review, we describe the physiological significance of biomechanical processes in the physiology and pathology of dental pulp. Moreover, we couple those phenomena with recent advances in the fields of bioengineering and pharmacology aiming to control the functioning of dental pulp cells, reduce pain, and enhance the differentiation of dental cells into desired lineages. The reviewed literature shows great progress in the topic of bioengineering of dental pulp-although mainly in vitro. Apart from a few positions, it leaves a gap for necessary filling with studies providing the mechanisms of the mechanical control of dental pulp functioning in vivo.
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Affiliation(s)
- Natalia Bryniarska-Kubiak
- Laboratory of Immunoendocrinology, Department of Experimental Neuroendocrinology, Maj Institute of Pharmacology, Polish Academy of Sciences, 12 Smętna St., 31-343 Kraków, Poland;
- Laboratory of Stem Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St., 30-387 Kraków, Poland
| | - Agnieszka Basta-Kaim
- Laboratory of Immunoendocrinology, Department of Experimental Neuroendocrinology, Maj Institute of Pharmacology, Polish Academy of Sciences, 12 Smętna St., 31-343 Kraków, Poland;
| | - Andrzej Kubiak
- Laboratory of Stem Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 7 Gronostajowa St., 30-387 Kraków, Poland
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Astudillo-Ortiz E, Babo PS, Sunde PT, Galler KM, Gomez-Florit M, Gomes ME. Endodontic Tissue Regeneration: A Review for Tissue Engineers and Dentists. TISSUE ENGINEERING. PART B, REVIEWS 2023; 29:491-513. [PMID: 37051704 DOI: 10.1089/ten.teb.2022.0211] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 04/14/2023]
Abstract
The paradigm shift in the endodontic field from replacement toward regenerative therapies has witnessed the ever-growing research in tissue engineering and regenerative medicine targeting pulp-dentin complex in the past few years. Abundant literature on the subject that has been produced, however, is scattered over diverse areas of knowledge. Moreover, the terminology and concepts are not always consensual, reflecting the range of research fields addressing this subject, from endodontics to biology, genetics, and engineering, among others. This fact triggered some misinterpretations, mainly when the denominations of different approaches were used as synonyms. The evaluation of results is not precise, leading to biased conjectures. Therefore, this literature review aims to conceptualize the commonly used terminology, summarize the main research areas on pulp regeneration, identify future trends, and ultimately clarify whether we are really on the edge of a paradigm shift in contemporary endodontics toward pulp regeneration.
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Affiliation(s)
- Esteban Astudillo-Ortiz
- 3B's Research Group, I3Bs-Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Braga, Portugal
- ICVS/3B's-PT Government Associate Laboratory, Braga, Portugal
- Department of Endodontics, School of Dentistry, University of Cuenca, Cuenca, Ecuador
| | - Pedro S Babo
- 3B's Research Group, I3Bs-Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Braga, Portugal
- ICVS/3B's-PT Government Associate Laboratory, Braga, Portugal
| | - Pia T Sunde
- Department of Endodontics, Institute of Clinical Dentistry, Faculty of Dentistry, University of Oslo, Oslo, Norway
| | - Kerstin M Galler
- Department of Operative Dentistry and Periodontology, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany
| | | | - Manuela E Gomes
- 3B's Research Group, I3Bs-Research Institute on Biomaterials, Biodegradables and Biomimetics, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine, Braga, Portugal
- ICVS/3B's-PT Government Associate Laboratory, Braga, Portugal
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6
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Gross T, Dieterle MP, Vach K, Altenburger MJ, Hellwig E, Proksch S. Biomechanical Modulation of Dental Pulp Stem Cell (DPSC) Properties for Soft Tissue Engineering. Bioengineering (Basel) 2023; 10:bioengineering10030323. [PMID: 36978714 PMCID: PMC10045720 DOI: 10.3390/bioengineering10030323] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/23/2023] [Revised: 02/14/2023] [Accepted: 02/28/2023] [Indexed: 03/08/2023] Open
Abstract
Dental pulp regeneration strategies frequently result in hard tissue formation and pulp obliteration. The aim of this study was to investigate whether dental pulp stem cells (DPSCs) can be directed toward soft tissue differentiation by extracellular elasticity. STRO-1-positive human dental pulp cells were magnetically enriched and cultured on substrates with elasticities of 1.5, 15, and 28 kPa. The morphology of DPSCs was assessed visually. Proteins relevant in mechanobiology ACTB, ITGB1, FAK, p-FAK, TALIN, VINCULIN, PAXILLIN, ERK 1/2, and p-ERK 1/2 were detected by immunofluorescence imaging. Transcription of the pulp marker genes BMP2, BMP4, MMP2, MMP3, MMP13, FN1, and IGF2 as well as the cytokines ANGPT1, VEGF, CCL2, TGFB1, IL2, ANG, and CSF1 was determined using qPCR. A low stiffness, i.e., 1.5 kPa, resulted in a soft tissue-like phenotype and gene expression, whereas DPSCs on 28 kPa substrates exhibited a differentiation signature resembling hard tissues with a low cytokine expression. Conversely, the highest cytokine expression was observed in cells cultured on intermediate elasticity, i.e., 15 kPa, substrates possibly allowing the cells to act as “trophic mediators”. Our observations highlight the impact of biophysical cues for DPSC fate and enable the design of scaffold materials for clinical pulp regeneration that prevent hard tissue formation.
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Affiliation(s)
- Tara Gross
- Department of Operative Dentistry and Periodontology, Center for Dental Medicine, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany
- G.E.R.N. Research Center for Tissue Replacement, Regeneration and Neogenesis, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Engesserstr. 4, 79108 Freiburg, Germany
- Correspondence: ; Tel.: +49-(0)761-270-48850; Fax: +49-(0)761-270-47620
| | - Martin Philipp Dieterle
- Division of Oral Biotechnology, Center for Dental Medicine, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Hugstetter Str. 55, 79106 Freiburg, Germany
| | - Kirstin Vach
- Institute of Medical Biometry and Statistics, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs—University of Freiburg, Stefan-Meier-Str. 26, 79104 Freiburg, Germany
| | - Markus Joerg Altenburger
- Department of Operative Dentistry and Periodontology, Center for Dental Medicine, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany
- G.E.R.N. Research Center for Tissue Replacement, Regeneration and Neogenesis, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Engesserstr. 4, 79108 Freiburg, Germany
| | - Elmar Hellwig
- Department of Operative Dentistry and Periodontology, Center for Dental Medicine, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany
| | - Susanne Proksch
- Department of Operative Dentistry and Periodontology, Center for Dental Medicine, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Hugstetter Straße 55, 79106 Freiburg, Germany
- G.E.R.N. Research Center for Tissue Replacement, Regeneration and Neogenesis, Medical Center—University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Engesserstr. 4, 79108 Freiburg, Germany
- Dental Clinic 1–Operative Dentistry and Periodontology, University Hospital Erlangen, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Glückstr. 11, 91054 Erlangen, Germany
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7
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Choi B, Kim JE, Park SO, Kim EY, Oh S, Choi H, Yoon D, Min HJ, Kim HR, Chang EJ. Sphingosine-1-phosphate hinders the osteogenic differentiation of dental pulp stem cells in association with AKT signaling pathways. Int J Oral Sci 2022; 14:21. [PMID: 35459199 PMCID: PMC9033766 DOI: 10.1038/s41368-022-00173-5] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2021] [Revised: 02/24/2022] [Accepted: 02/24/2022] [Indexed: 11/16/2022] Open
Abstract
Sphingosine-1-phosphate (S1P) is an important lipid mediator that regulates a diverse range of intracellular cell signaling pathways that are relevant to tissue engineering and regenerative medicine. However, the precise function of S1P in dental pulp stem cells (DPSCs) and its osteogenic differentiation remains unclear. We here investigated the function of S1P/S1P receptor (S1PR)-mediated cellular signaling in the osteogenic differentiation of DPSCs and clarified the fundamental signaling pathway. Our results showed that S1P-treated DPSCs exhibited a low rate of differentiation toward the osteogenic phenotype in association with a marked reduction in osteogenesis-related gene expression and AKT activation. Of note, both S1PR1/S1PR3 and S1PR2 agonists significantly downregulated the expression of osteogenic genes and suppressed AKT activation, resulting in an attenuated osteogenic capacity of DPSCs. Most importantly, an AKT activator completely abrogated the S1P-mediated downregulation of osteoblastic markers and partially prevented S1P-mediated attenuation effects during osteogenesis. Intriguingly, the pro-inflammatory TNF-α cytokine promoted the infiltration of macrophages toward DPSCs and induced S1P production in both DPSCs and macrophages. Our findings indicate that the elevation of S1P under inflammatory conditions suppresses the osteogenic capacity of the DPSCs responsible for regenerative endodontics.
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Affiliation(s)
- Bongkun Choi
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Ji-Eun Kim
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Si-On Park
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Eun-Young Kim
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Soyoon Oh
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Hyuksu Choi
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Dohee Yoon
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Hyo-Jin Min
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea.,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea
| | - Hyung-Ryong Kim
- Department of Pharmacology, College of Dentistry, Jeonbuk National University, Jeonju, Korea.
| | - Eun-Ju Chang
- Department of Biomedical Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul, 05505, Korea. .,Stem Cell Immunomodulation Research Center, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea. .,Department of Biochemistry and Molecular Biology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea.
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8
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Effects of mechanical force on proliferation and apoptosis of stem cells from human exfoliated deciduous teeth. Clin Oral Investig 2022; 26:5205-5213. [PMID: 35441898 DOI: 10.1007/s00784-022-04488-9] [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: 12/20/2021] [Accepted: 04/08/2022] [Indexed: 11/03/2022]
Abstract
OBJECTIVES This study was designed to explore the effects of mechanical force on the proliferation, apoptosis, and morphology of stem cells from human exfoliated deciduous tooth pulp (SHEDs). MATERIALS AND METHODS Caries-free stranded deciduous teeth were extracted, and SHEDs were isolated through enzymatic digestion. The cultured SHEDs were subjected to different levels of mechanical stimuli (0, 100, 200, and 300 g) for 7 days (30 min/day) using external centrifugal force. Cell proliferation was evaluated with the CCK-8 assay, and the cell cycle and apoptosis were assessed by flow cytometry. The cell morphology was examined using transmission electron microscopy. RESULTS Cell proliferation assay showed no differences between the three stimulation groups and the control group in day 1 to day 3. From the 4th day, cell proliferation was significantly lower in the mechanical force groups than in the control group, but no significant difference was observed among the three mechanical force groups. Besides, there was no significant difference in cell apoptosis among the four groups for 7 days. On day 7 after stimulation, the SHEDs were shrunken, with significantly increased isochromosome in the nucleus and an increase in lysosomes. CONCLUSIONS Mechanical force can inhibit the proliferation and affect morphology of SHEDs, but it has no effect on cell apoptosis. CLINICAL RELEVANCE Mechanical force stimulation significantly inhibited cell proliferation of SHEDs. Mechanical force stimulation had no significant effect on cell apoptosis of SHEDs. The morphology and ultrastructure of SHEDs changed after mechanical force stimulation.
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Atila D, Hasirci V, Tezcaner A. Coaxial electrospinning of composite mats comprised of core/shell poly(methyl methacrylate)/silk fibroin fibers for tissue engineering applications. J Mech Behav Biomed Mater 2022; 128:105105. [DOI: 10.1016/j.jmbbm.2022.105105] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/23/2021] [Revised: 01/17/2022] [Accepted: 01/23/2022] [Indexed: 01/01/2023]
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10
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Three-Dimensional Culture Systems for Dissecting Notch Signalling in Health and Disease. Int J Mol Sci 2021; 22:ijms222212473. [PMID: 34830355 PMCID: PMC8618738 DOI: 10.3390/ijms222212473] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/12/2021] [Revised: 11/16/2021] [Accepted: 11/17/2021] [Indexed: 12/12/2022] Open
Abstract
Three-dimensional (3D) culture systems opened up new horizons in studying the biology of tissues and organs, modelling various diseases, and screening drugs. Producing accurate in vitro models increases the possibilities for studying molecular control of cell–cell and cell–microenvironment interactions in detail. The Notch signalling is linked to cell fate determination, tissue definition, and maintenance in both physiological and pathological conditions. Hence, 3D cultures provide new accessible platforms for studying activation and modulation of the Notch pathway. In this review, we provide an overview of the recent advances in different 3D culture systems, including spheroids, organoids, and “organ-on-a-chip” models, and their use in analysing the crucial role of Notch signalling in the maintenance of tissue homeostasis, pathology, and regeneration.
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11
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Rubert M, Vetsch JR, Lehtoviita I, Sommer M, Zhao F, Studart AR, Müller R, Hofmann S. Scaffold Pore Geometry Guides Gene Regulation and Bone-like Tissue Formation in Dynamic Cultures. Tissue Eng Part A 2021; 27:1192-1204. [PMID: 33297842 DOI: 10.1089/ten.tea.2020.0121] [Citation(s) in RCA: 12] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022] Open
Abstract
Cells sense and respond to scaffold pore geometry and mechanical stimuli. Many fabrication methods used in bone tissue engineering render structures with poorly controlled pore geometries. Given that cell-scaffold interactions are complex, drawing a conclusion on how cells sense and respond to uncontrolled scaffold features under mechanical loading is difficult. In this study, monodisperse templated scaffolds (MTSC) were fabricated and used as well-defined porous scaffolds to study the effect of dynamic culture conditions on bone-like tissue formation. Human bone marrow-derived stromal cells were cultured on MTSC or conventional salt-leached scaffolds (SLSC) for up to 7 weeks, either under static or dynamic conditions (wall shear stress [WSS] using spinner flask bioreactors). The influence of controlled spherical pore geometry of MTSC subjected to static or dynamic conditions on osteoblast cell differentiation, bone-like tissue formation, structure, and distribution was investigated. WSS generated within the two idealized geometrical scaffold features was assessed. Distinct response to fluid flow in osteoblast cell differentiation were shown to be dependent on scaffold pore geometry. As revealed by collagen staining and microcomputed tomography images, dynamic conditions promoted a more regular extracellular matrix (ECM) formation and mineral distribution in both scaffold types compared with static conditions. The results showed that regulation of bone-related genes and the amount and the structure of mineralized ECM were dependent on scaffold pore geometry and the mechanical cues provided by the two different culture conditions. Under dynamic conditions, SLSC favored osteoblast cell differentiation and ECM formation, whereas MTSC enhanced ECM mineralization. The spherical pore shape in MTSC supported a more trabecular bone-like structure under dynamic conditions compared with MTSC statically cultured or to SLSC under either static or dynamic conditions. These results suggest that cell activity and bone-like tissue formation is driven not only by the pore geometry but also by the mechanical environment. This should be taken into account in the future design of complex scaffolds, which should favor cell differentiation while guiding the formation, structure, and distribution of the engineered bone tissue. This could help to mimic the anatomical complexity of the bone tissue structure and to adapt to each bone defect needs. Impact statement Aging of the human population leads to an increasing need for medical implants with high success rate. We provide evidence that cell activity and the amount and structure of bone-like tissue formation is dependent on the scaffold pore geometry and on the mechanical environment. Fabrication of complex scaffolds comprising concave and planar pore geometries might represent a promising direction toward the tunability and mimicry the structural complexity of the bone tissue. Moreover, the use of fabrication methods that allow a systematic fabrication of reproducible and geometrically controlled structures would simplify scaffold design optimization.
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Affiliation(s)
- Marina Rubert
- Department of Health Sciences and Technology, Institute for Biomechanics, ETH Zurich, Zurich, Switzerland
| | - Jolanda Rita Vetsch
- Department of Health Sciences and Technology, Institute for Biomechanics, ETH Zurich, Zurich, Switzerland
| | - Iina Lehtoviita
- Department of Health Sciences and Technology, Institute for Biomechanics, ETH Zurich, Zurich, Switzerland
| | - Marianne Sommer
- Complex Materials, Department of Materials, ETH Zurich, Zurich, Switzerland
| | - Feihu Zhao
- Zienkiewicz Centre for Computational Engineering, College of Engineering, Swansea University, Swansea, United Kingdom
| | - André R Studart
- Complex Materials, Department of Materials, ETH Zurich, Zurich, Switzerland
| | - Ralph Müller
- Department of Health Sciences and Technology, Institute for Biomechanics, ETH Zurich, Zurich, Switzerland
| | - Sandra Hofmann
- Department of Health Sciences and Technology, Institute for Biomechanics, ETH Zurich, Zurich, Switzerland.,Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands.,Institute for Complex Molecular Systems, Eindhoven University of Technology Eindhoven, The Netherlands
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12
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Diana R, Ardhani R, Kristanti Y, Santosa P. Dental pulp stem cells response on the nanotopography of scaffold to regenerate dentin-pulp complex tissue. Regen Ther 2020; 15:243-250. [PMID: 33426225 PMCID: PMC7770425 DOI: 10.1016/j.reth.2020.09.007] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2020] [Revised: 08/05/2020] [Accepted: 09/16/2020] [Indexed: 12/13/2022] Open
Abstract
The study of regenerative dentistry receives a fast growing interest. The potential ability of the dentin-pulp complex to regenerate is both promising and perplexing. To answer the challenging nature of the dental environment, scientists have developed various combinations of biomaterial scaffolds, stem cells, and incorporation of several growth factors. One of the crucial elements of this tissue engineering plan is the selection and fabrication of scaffolds. However, further findings suggest that cell behavior hugely depends on mechanical signaling. Nanotopography modifies scaffolds to alter cell migration and differentiation. However, to the best of the author's knowledge, there are very few studies addressing the correlation between nanotopography and dentin-pulp complex regeneration. Therefore, this article presents a comprehensive review of these studies and suggests a direction for future developments, particularly in the incorporation of nanotopography design for dentin-pulp complex regeneration.
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Key Words
- BDNF, brain-derived neurotrophic factor
- BMP, bone morphogenetic protein
- DPSC, dental pulp stem cell
- Dental pulp stem cell
- Dentin-pulp complex tissue
- ECM, extracellular matrix
- FGF2, fibroblast growth factor-2
- GDNF, glial cell line-derived neurotrophic factor
- GO, graphene oxide
- GelMA, methacrylated gelatin
- IGF, insulin-like growth factor
- ION-CPC, iron oxide nanoparticle-incorporating calcium phosphate cement
- LPS, lipopolysaccharide
- NGF, nerve growth factor
- Nanotopography
- PCL, polycaprolactone
- PDGF, platelet-derived growth factor
- PEGMA, poly(ethylene glycol) dimethacrylate
- PGA, polyglycolic acid
- PHMS, polyhydroxymethylsiloxane
- PLGA, poly-dl-lactic-co-glycolic acid
- PLLA, poly-l-lactic acid
- RGO, reduced graphene oxide
- Regenerative dentistry
- SACP, stem cells from apical papilla
- SDF-1, stromal cell-derived factor-1
- SHED, stem cells from human exfoliated deciduous teeth
- Scaffold
- TGF-β, transforming growth factor-β
- TNF-α, t umour necrosis factor-alpha
- VEGF, vascular endothelial growth factor
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Affiliation(s)
- Rasda Diana
- Department of Conservative Dentistry, Faculty of Dentistry Universitas Gadjah Mada, Jl Denta Sekip Utara, Yogyakarta, 55281, Indonesia
| | - Retno Ardhani
- Department of Dental Biomedical Sciences, Faculty of Dentistry Universitas Gadjah Mada, Jl Denta Sekip Utara, Yogyakarta, 55281, Indonesia
- Corresponding author. Fax: +62274 515307.
| | - Yulita Kristanti
- Department of Conservative Dentistry, Faculty of Dentistry Universitas Gadjah Mada, Jl Denta Sekip Utara, Yogyakarta, 55281, Indonesia
| | - Pribadi Santosa
- Department of Conservative Dentistry, Faculty of Dentistry Universitas Gadjah Mada, Jl Denta Sekip Utara, Yogyakarta, 55281, Indonesia
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13
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Zhao F, Lacroix D, Ito K, van Rietbergen B, Hofmann S. Changes in scaffold porosity during bone tissue engineering in perfusion bioreactors considerably affect cellular mechanical stimulation for mineralization. Bone Rep 2020; 12:100265. [PMID: 32613033 PMCID: PMC7315008 DOI: 10.1016/j.bonr.2020.100265] [Citation(s) in RCA: 18] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/27/2019] [Revised: 03/24/2020] [Accepted: 04/02/2020] [Indexed: 11/24/2022] Open
Abstract
Bone tissue engineering (BTE) experiments in vitro have shown that fluid-induced wall shear stress (WSS) can stimulate cells to produce mineralized extracellular matrix (ECM). The application of WSS on seeded cells can be achieved through bioreactors that perfuse medium through porous scaffolds. In BTE experiments in vitro, commonly a constant flow rate is used. Previous studies have found that tissue growth within the scaffold will result in an increase of the WSS over time. To keep the WSS in a reported optimal range of 10–30 mPa, the applied external flow rate can be decreased over time. To investigate what reduction of the external flow rate during culturing is needed to keep the WSS in the optimal range, we here conducted a computational study, which simulated the formation of ECM, and in which we investigated the effect of constant fluid flow and different fluid flow reduction scenarios on the WSS. It was found that for both constant and reduced fluid flow scenarios, the WSS did not exceed a critical value, which was set to 60 mPa. However, the constant flow velocity resulted in a reduction of the cell/ECM surface being exposed to a WSS in the optimal range from 50% at the start of culture to 18.6% at day 21. Reducing the fluid flow over time could avoid much of this effect, leaving the WSS in the optimal range for 40.9% of the surface at 21 days. Therefore, for achieving more mineralized tissue, the conventional manner of loading the perfusion bioreactors (i.e. constant flow rate/velocity) should be changed to a decreasing flow over time in BTE experiments. This study provides an in silico tool for finding the best fluid flow reduction strategy.
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Affiliation(s)
- Feihu Zhao
- Orthopaedic Biomechanics, Department of Biomedical Engineering, Eindhoven University of Technology, 5600 MB, Eindhoven, the Netherlands
- Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, 5600 MB Eindhoven, the Netherlands
- Zienkiewicz Centre for Computational Engineering (ZCCE), College of Engineering, Swansea University, SA1 8EN Swansea, United Kingdom
| | - Damien Lacroix
- INSIGNEO Institute for in silico Medicine, Department of Mechanical Engineering, University of Sheffield, S1 3JD Sheffield, United Kingdom
| | - Keita Ito
- Orthopaedic Biomechanics, Department of Biomedical Engineering, Eindhoven University of Technology, 5600 MB, Eindhoven, the Netherlands
- Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, 5600 MB Eindhoven, the Netherlands
| | - Bert van Rietbergen
- Orthopaedic Biomechanics, Department of Biomedical Engineering, Eindhoven University of Technology, 5600 MB, Eindhoven, the Netherlands
- Corresponding authors at: Orthopaedic Biomechanics, Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, the Netherlands.
| | - Sandra Hofmann
- Orthopaedic Biomechanics, Department of Biomedical Engineering, Eindhoven University of Technology, 5600 MB, Eindhoven, the Netherlands
- Institute for Complex Molecular Systems (ICMS), Eindhoven University of Technology, 5600 MB Eindhoven, the Netherlands
- Corresponding authors at: Orthopaedic Biomechanics, Department of Biomedical Engineering, Eindhoven University of Technology, PO Box 513, 5600 MB Eindhoven, the Netherlands.
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14
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Salgado CL, Barrias CC, Monteiro FJM. Clarifying the Tooth-Derived Stem Cells Behavior in a 3D Biomimetic Scaffold for Bone Tissue Engineering Applications. Front Bioeng Biotechnol 2020; 8:724. [PMID: 32671055 PMCID: PMC7333018 DOI: 10.3389/fbioe.2020.00724] [Citation(s) in RCA: 26] [Impact Index Per Article: 5.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/20/2020] [Accepted: 06/09/2020] [Indexed: 12/24/2022] Open
Abstract
Massive amounts of cell are needed for creating tissue engineered 3D constructs, which often requires culture on scaffolds under dynamic conditions to facilitate nutrients and oxygen diffusion. Dynamic cultures are expected to improve cell viability and proliferation rate, when compared to static conditions. However, cells from distinct types and/or tissues sources may respond differently to external stimuli and be incompatible with culture under mechanical shear stress. The first aim of this work was to show that dental stem cells are a valuable source for improving bone regeneration potential of artificial grafts. Mesenchymal stem/stromal cells (MSCs) were isolated from human dental follicle (hDFMSC) and pulp tissues (hDPMSC) and shown to express prototypical stem cell markers. The follicle and pulp dental MSCs capacity to differentiate into osteoblast lineage was evaluated after seeding on 3D porous scaffolds of collagen-nanohydroxyapatite/phosphoserine biocomposite cryogel with osteogenic factors in the culture medium. Both tooth-derived MSCs were able to show high ALP activity, express osteogenic gene markers and secrete osteopontin (OPN). Thereafter, designed multicompartment holder adaptable to spinner flasks was used for dynamic culture (50 rpm) of both dental MSCs types within the porous 3D scaffolds. Standard static culture conditions were used as control. Culture under dynamic conditions promoted follicle MSCs proliferation, while improving their spatial distribution within the scaffold. Under dynamic conditions, the biocomposite scaffold promoted MSCs osteogenic differentiation, as suggested by increased alkaline phosphatase (ALP) activity, higher osteogenic gene expression and OPN deposition. In a similar manner, under dynamic conditions, dental pulp MSCs also showed higher ALP activity and proliferation rate, but lower amounts of osteopontin secretion, when compared to static conditions. After implantation, dental follicle MSCs-loaded 3D scaffolds cultured under dynamic conditions showed higher tissue ingrowth and osteogenic differentiation (higher human OPN secretion) than dental pulp cells. Overall, this study explored the use of tooth-derived stem cells as a clinical alternative source for bone tissue engineering, together with an innovative device for dynamic culture of cell-laden 3D scaffolds. Results showed that human MSCs response upon culture on 3D scaffolds, depends on the cells source and the culture regimen. This suggests that both the type of cells and their culture conditions should be carefully adjusted according to the final clinical application.
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Affiliation(s)
- Christiane L Salgado
- i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.,INEB, Instituto Nacional de Engenharia Biomédica, Universidade do Porto, Porto, Portugal
| | - Cristina C Barrias
- i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.,INEB, Instituto Nacional de Engenharia Biomédica, Universidade do Porto, Porto, Portugal.,ICBAS, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto, Portugal
| | - Fernando J M Monteiro
- i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal.,INEB, Instituto Nacional de Engenharia Biomédica, Universidade do Porto, Porto, Portugal.,FEUP, Faculdade de Engenharia da Universidade do Porto, Departamento de Engenharia Metalúrgica e Materiais, Porto, Portugal
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15
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Zhang W, Liu H, Yang W, Liu C, Xie M, Guo R, Liang J, Ye Z, Xu H. Hydroxyapatite/silk fibroin composite biomimetic scaffold for dental pulp repair. BIOINSPIRED BIOMIMETIC AND NANOBIOMATERIALS 2019. [DOI: 10.1680/jbibn.18.00050] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/27/2022]
Abstract
Dental pulp repair is a difficult clinical problem. In the present study, the authors aimed to mimic the extracellular matrix of dental pulp tissue structurally and compositionally. Nanofibrous silk fibroin (SF) scaffolds containing hydroxyapatite (HAp) nanoparticles were fabricated by using the freeze-drying approach. Rod-shaped HAp was successfully embedded in the composite scaffold, the diameter of which was about 100–200 nm as shown by transmission electron microscopy analysis. The three-dimensional microstructure of the composite scaffold prepared in various ratios of HAp to SF was observed by scanning electron microscopy and the pore size of the optimal scaffold was about 30–120 μm. Meanwhile, the hemocompatibility of the composite scaffolds was evaluated based on their impact on the clotting function by way of activated partial thromboplastin time, prothrombin time and thromboelastographic assays. The scaffolds possessed a low hemolysis rate of red blood cells. Furthermore, cell culture tests using dental pulp stem cells found that the scaffolds had good biocompatibility. There biomimetic HAp/SF composite scaffolds may serve as a promising biomaterial for dental pulp repair.
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Affiliation(s)
- Wu Zhang
- The First Affiliated Hospital of Jinan University, Guangzhou, China; School of Stomatology, Jinan University, Guangzhou, China
| | - Haixia Liu
- Guangzhou Nansha District Maternal and Child Health Care Hospital, Guangzhou, China
| | - Wei Yang
- Key Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University, Guangzhou, China
| | - Chang Liu
- School of Stomatology, Jinan University, Guangzhou, China
| | - Miaomiao Xie
- Department of Stomatology, People’s Hospital of Baoan District, Shenzhen, China
| | - Rui Guo
- Key Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University, Guangzhou, China
| | - Jianqiang Liang
- Guangzhou Haizhu District Stomatological Hospital, Guangzhou, China
| | - Zhongtai Ye
- Department of Stomatology, People’s Hospital of Baoan District, Shenzhen, China
| | - Hao Xu
- The First Affiliated Hospital of Jinan University, Guangzhou, China
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16
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Marolt Presen D, Traweger A, Gimona M, Redl H. Mesenchymal Stromal Cell-Based Bone Regeneration Therapies: From Cell Transplantation and Tissue Engineering to Therapeutic Secretomes and Extracellular Vesicles. Front Bioeng Biotechnol 2019; 7:352. [PMID: 31828066 PMCID: PMC6890555 DOI: 10.3389/fbioe.2019.00352] [Citation(s) in RCA: 81] [Impact Index Per Article: 13.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/06/2019] [Accepted: 11/06/2019] [Indexed: 12/12/2022] Open
Abstract
Effective regeneration of bone defects often presents significant challenges, particularly in patients with decreased tissue regeneration capacity due to extensive trauma, disease, and/or advanced age. A number of studies have focused on enhancing bone regeneration by applying mesenchymal stromal cells (MSCs) or MSC-based bone tissue engineering strategies. However, translation of these approaches from basic research findings to clinical use has been hampered by the limited understanding of MSC therapeutic actions and complexities, as well as costs related to the manufacturing, regulatory approval, and clinical use of living cells and engineered tissues. More recently, a shift from the view of MSCs directly contributing to tissue regeneration toward appreciating MSCs as "cell factories" that secrete a variety of bioactive molecules and extracellular vesicles with trophic and immunomodulatory activities has steered research into new MSC-based, "cell-free" therapeutic modalities. The current review recapitulates recent developments, challenges, and future perspectives of these various MSC-based bone tissue engineering and regeneration strategies.
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Affiliation(s)
- Darja Marolt Presen
- Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, Vienna, Austria.,Austrian Cluster for Tissue Regeneration, Vienna, Austria
| | - Andreas Traweger
- Austrian Cluster for Tissue Regeneration, Vienna, Austria.,Spinal Cord Injury & Tissue Regeneration Center Salzburg, Institute of Tendon and Bone Regeneration, Paracelsus Medical University, Salzburg, Austria
| | - Mario Gimona
- GMP Unit, Spinal Cord Injury & Tissue Regeneration Center Salzburg, Paracelsus Medical University, Salzburg, Austria
| | - Heinz Redl
- Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, Vienna, Austria.,Austrian Cluster for Tissue Regeneration, Vienna, Austria
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17
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Mirzaeian L, Eivazkhani F, Hezavehei M, Moini A, Esfandiari F, Valojerdi MR, Fathi R. Optimizing The Cell Seeding Protocol to Human Decellularized Ovarian Scaffold: Application of Dynamic System for Bio-Engineering. CELL JOURNAL 2019; 22:227-235. [PMID: 31721538 PMCID: PMC6874796 DOI: 10.22074/cellj.2020.6604] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/11/2018] [Accepted: 02/26/2019] [Indexed: 12/22/2022]
Abstract
Objective Decellularized tissue scaffolds provide an extracellular matrix to control stem cells differentiation toward
specific lineages. The application of mesenchymal stem cells for artificial ovary production may enhance ex vivo functions
of the ovary. On the other hand, the scaffold needs interaction and integration with cells. Thus, the development of
ovarian engineered constructs (OVECs) requires the use of efficient methods for seeding of the cells into the ovarian
and other types of scaffolds. The main goal of the present study was to develop an optimized culture system for efficient
seeding of peritoneum mesenchymal stem cells (PMSCs) into human decellularized ovarian scaffold.
Materials and Methods In this experimental study, three methods were used for cellular seeding including rotational
(spinner flask) and static (conventional and injection) seeding cultures. OVECs were evaluated with Hematoxylin and
Eosin staining and viability analyses for the seeded PMSCs. Then, immunohistochemistry analysis was performed
using the best method of cellular seeding for primordial germ cell-like cells, mesenchymal stem cells and proliferation
markers. Stereology analysis was also performed for the number of penetrated cells into the OVECs.
Results Our results showed that rotational seeding increases the permeability of PMSCs into the scaffold and survival
rate of the seeded PMSCs, comparing to the other methods. On the other hand, rotationally seeded PMSCs had a more
favorable capability of proliferation with Ki67 expression and differentiation to ovarian specific cells with expression
of primordial germ cell line markers without mesenchymal stem cells markers production. Furthermore, stereology
showed a more favorable distribution of PMSCs along the outer surfaces of the OVEC with further distribution at the
central part of the scaffold. The average total cell values were determined 2142187 cells/mm3 on each OVEC.
Conclusion The rotational seeding method is a more favorable approach to cell seeding into ovarian decellularized
tissue than static seeding.
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Affiliation(s)
- Leila Mirzaeian
- Department of Developmental Biology, University of Science and Culture, Tehran, Iran.,Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Farideh Eivazkhani
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR,Tehran, Iran
| | - Maryam Hezavehei
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran
| | - Ashraf Moini
- Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.,Department of Gynecology and Obstetrics, Arash Women's Hospital, Tehran University of Medical Sciences, Tehran, Iran.,Vali-e-Asr Reproductive Health Research Center, Tehran University of Medical Sciences, Tehran, Iran
| | - Fereshteh Esfandiari
- Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
| | - Mojtaba Rezazadeh Valojerdi
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.,Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran
| | - Rouhollah Fathi
- Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
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18
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Marrelli M, Codispoti B, Shelton RM, Scheven BA, Cooper PR, Tatullo M, Paduano F. Dental Pulp Stem Cell Mechanoresponsiveness: Effects of Mechanical Stimuli on Dental Pulp Stem Cell Behavior. Front Physiol 2018; 9:1685. [PMID: 30534086 PMCID: PMC6275199 DOI: 10.3389/fphys.2018.01685] [Citation(s) in RCA: 90] [Impact Index Per Article: 12.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/12/2018] [Accepted: 11/08/2018] [Indexed: 12/28/2022] Open
Abstract
Dental pulp is known to be an accessible and important source of multipotent mesenchymal progenitor cells termed dental pulp stem cells (DPSCs). DPSCs can differentiate into odontoblast-like cells and maintain pulp homeostasis by the formation of new dentin which protects the underlying pulp. DPSCs similar to other mesenchymal stem cells (MSCs) reside in a niche, a complex microenvironment consisting of an extracellular matrix, other local cell types and biochemical stimuli that influence the decision between stem cell (SC) self-renewal and differentiation. In addition to biochemical factors, mechanical factors are increasingly recognized as key regulators in DPSC behavior and function. Thus, microenvironments can significantly influence the role and differentiation of DPSCs through a combination of factors which are biochemical, biomechanical and biophysical in nature. Under in vitro conditions, it has been shown that DPSCs are sensitive to different types of force, such as uniaxial mechanical stretch, cyclic tensile strain, pulsating fluid flow, low-intensity pulsed ultrasound as well as being responsive to biomechanical cues presented in the form of micro- and nano-scale surface topographies. To understand how DPSCs sense and respond to the mechanics of their microenvironments, it is essential to determine how these cells convert mechanical and physical stimuli into function, including lineage specification. This review therefore covers some aspects of DPSC mechanoresponsivity with an emphasis on the factors that influence their behavior. An in-depth understanding of the physical environment that influence DPSC fate is necessary to improve the outcome of their therapeutic application for tissue regeneration.
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Affiliation(s)
- Massimo Marrelli
- Stem Cells Unit, Biomedical Section, Tecnologica Research Institute and Marrelli Health, Crotone, Italy
| | - Bruna Codispoti
- Stem Cells Unit, Biomedical Section, Tecnologica Research Institute and Marrelli Health, Crotone, Italy
| | - Richard M. Shelton
- School of Dentistry, Institute of Clinical Sciences, College of Medical and Dental Sciences, The University of Birmingham, Birmingham, United Kingdom
| | - Ben A. Scheven
- School of Dentistry, Institute of Clinical Sciences, College of Medical and Dental Sciences, The University of Birmingham, Birmingham, United Kingdom
| | - Paul R. Cooper
- School of Dentistry, Institute of Clinical Sciences, College of Medical and Dental Sciences, The University of Birmingham, Birmingham, United Kingdom
| | - Marco Tatullo
- Stem Cells Unit, Biomedical Section, Tecnologica Research Institute and Marrelli Health, Crotone, Italy
| | - Francesco Paduano
- Stem Cells Unit, Biomedical Section, Tecnologica Research Institute and Marrelli Health, Crotone, Italy
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19
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Raspini G, Wolff J, Helminen M, Raspini G, Raspini M, Sándor GK. Dental Stem Cells Harvested from Third Molars Combined with Bioactive Glass Can Induce Signs of Bone Formation In Vitro. EJOURNAL OF ORAL MAXILLOFACIAL RESEARCH 2018; 9:e2. [PMID: 29707181 PMCID: PMC5913415 DOI: 10.5037/jomr.2018.9102] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 11/24/2017] [Accepted: 02/20/2018] [Indexed: 12/23/2022]
Abstract
Objectives The aim of this study was to assess the interaction of a bioactive glass scaffold with cells derived from dental pulp, dental follicle and periodontal ligament. Material and Methods Impacted third molars were surgically removed from three young donors. Cells from the dental pulp, follicle and periodontal ligament tissues were isolated and expanded. Different cell populations were characterised using specific CD markers. Expanded pulp, follicle and periodontal cells were then seeded onto bioactive glass scaffolds and cultured in osteogenic medium or basic medium. Cell attachment, viability, proliferation and alkaline phosphatase activity were assessed. Results This study revealed good biocompatibility of the specific bioactive glass configuration tested and the osteogenic induction of cells derived from dental pulp, dental follicle and periodontal ligament. Osteogenic medium seemed to increase the differentiation pattern and dental pulp stem cells showed the most positive results compared to periodontal ligament and dental follicle stem cells. Conclusions Dental pulp stem cells combined with a bioactive glass scaffold and exposed to osteogenic medium in vitro represent a promising combination for future study of hard tissue regeneration in the cranio-maxillofacial skeleton.
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Affiliation(s)
- Gregorio Raspini
- Department of Oral and Maxillofacial Surgery, University of Oulu, OuluFinland
| | - Jan Wolff
- Department of Oral and Maxillofacial Surgery/Pathology & 3D Innovation Lab, VU University Medical Center, AmsterdamThe Netherlands
| | - Mika Helminen
- Faculty of Social Sciences, University of Tampere and Science Center, Pirkanmaa Hospital District, TampereFinland
| | | | | | - George K Sándor
- Department of Oral and Maxillofacial Surgery, University of Oulu, OuluFinland.,Tissue Engineering and Oral and Maxillofacial Surgery, Medical Research Center, University of Oulu and Oulu University Hospital, OuluFinland
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20
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Gnanasegaran N, Govindasamy V, Kathirvaloo P, Musa S, Abu Kasim NH. Effects of cell cycle phases on the induction of dental pulp stem cells toward dopaminergic-like cells. J Tissue Eng Regen Med 2017; 12:e881-e893. [PMID: 28079995 DOI: 10.1002/term.2401] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/13/2016] [Revised: 11/16/2016] [Accepted: 01/09/2017] [Indexed: 12/22/2022]
Abstract
Parkinson's disease (PD) is characterized by tremors and cognitive issues, and is due to the death of dopaminergic (DA-ergic) neurons in brain circuits that are responsible for producing neurotransmitter dopamine (DA). Currently, cell replacement therapies are underway to improve upon existing therapeutic approaches such as drug treatments and electrical stimulation. Among the widely available sources, dental pulp stem cells (DPSCs) from deciduous teeth have gained popularity because of their neural crest origin and inherent propensity toward neuronal lineage. Despite the various pre-clinical studies conducted, an important factor yet to be elucidated is the influence of growth phases in a typical trans-differentiation process. This study selected DPSCs at three distinct time points with variable growth phase proportions (G0/G1, S and G2/M) for in vitro trans-differentiation into DA-ergic-like cells. Using commercially available PCR arrays, we identified distinct gene profiles pertaining to cell cycles in these phases. The differentiation outcomes were assessed in terms of morphology and gene and protein expression, as well as with functional assays. It was noted that DPSCs with the highest G0/G1 phase were comparatively the best, representing at least a 2-fold up regulation (p < 0.05) of DA-ergic molecular cues compared to those from the remaining time points. Further investigations in terms of protein expression and DA-release assays also revealed a similar phenomenon (p < 0.05). These findings are expected to provide vital information for consideration in improving standard operating procedures in future cell transplantation work. Copyright © 2017 John Wiley & Sons, Ltd.
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Affiliation(s)
- Nareshwaran Gnanasegaran
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Vijayendran Govindasamy
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Premasangery Kathirvaloo
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Sabri Musa
- Department of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Noor Hayaty Abu Kasim
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
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In Vitro and In Vivo Dentinogenic Efficacy of Human Dental Pulp-Derived Cells Induced by Demineralized Dentin Matrix and HA-TCP. Stem Cells Int 2017; 2017:2416254. [PMID: 28761445 PMCID: PMC5518496 DOI: 10.1155/2017/2416254] [Citation(s) in RCA: 13] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/11/2017] [Revised: 04/26/2017] [Accepted: 05/03/2017] [Indexed: 01/09/2023] Open
Abstract
Human dental pulp cells have been known to have the stem cell features such as self-renewal and multipotency. These cells are differentiated into hard tissue by addition of proper cytokines and biomaterials. Hydroxyapatite-tricalcium phosphates (HA-TCPs) are essential components of hard tissue and generally used as a biocompatible material in tissue engineering of bone. Demineralized dentin matrix (DDM) has been reported to increase efficiency of bone induction. We compared the efficiencies of osteogenic differentiation and in vivo bone formation of HA-TCP and DDM on human dental pulp stem cells (hDPSCs). DDM contains inorganic components as with HA-TCP, and organic components such as collagen type-1. Due to these components, osteoinduction potential of DDM on hDPSCs was remarkably higher than that of HA-TCP. However, the efficiencies of in vivo bone formation are similar in HA-TCP and DDM. Although osteogenic gene expression and bone formation in immunocompromised nude mice were similar levels in both cases, dentinogenic gene expression level was slightly higher in DDM transplantation than in HA-TCP. All these results suggested that in vivo osteogenic potentials in hDPSCs are induced with both HA-TCP and DDM by osteoconduction and osteoinduction, respectively. In addition, transplantation of hDPSCs/DDM might be more effective for differentiation into dentin.
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22
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Collado-González M, Pecci-Lloret MP, García-Bernal D, Aznar-Cervantes S, Oñate-Sánchez RE, Moraleda JM, Cenis JL, Rodríguez-Lozano FJ. Biological effects of silk fibroin 3D scaffolds on stem cells from human exfoliated deciduous teeth (SHEDs). Odontology 2017; 106:125-134. [PMID: 28616672 DOI: 10.1007/s10266-017-0310-9] [Citation(s) in RCA: 14] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/11/2017] [Accepted: 05/22/2017] [Indexed: 10/25/2022]
Abstract
The aim is to investigate in vitro biological effects of silk fibroin 3D scaffolds on stem cells from human exfoliated deciduous teeth (SHEDs) in terms of proliferation, morphological appearance, cell viability, and expression of mesenchymal stem cell markers. Silk fibroin 3D scaffolding materials may represent promising suitable scaffolds for their application in regenerative endodontic therapy approaches. SHEDs were cultured in silk fibroin 3D scaffolds. Then, cell numbers were counted and the Alamar blue colorimetric assay was used to analyse cell proliferation after 24, 48, 72, and 168 h of culture. The morphological features of SHEDs cultured on silk fibroin scaffolds were evaluated by scanning electron microscopy (SEM). Finally, cell viability and the expression of mesenchymal stem cell markers were analysed by flow cytometry. One-way analysis of variance (ANOVA) followed by a Bonferroni post-test was performed (P < 0.05). At 24 and 48 h of culture, SHED proliferation on scaffolds was modest compared to the control although still significant (p < 0.05). However, cell proliferation progressively increased from 72 to 168 h compared with the control (p < 0.001; p < 0.01). In addition, flow cytometry analysis showed that the culture of SHEDs on silk fibroin scaffolds did not significantly alter the level of expression of the mesenchymal markers CD73, CD90, or CD105 up to 168 h; in addition, cell viability in silk fibroin was similar to than obtained in plastic. Moreover, SEM studies revealed a suitable degree of proliferation, cell spreading, and attachment, especially after 168 h of culture. The findings from the current study suggest that silk fibroin 3D scaffolds had a favourable effect on the biological responses of SHEDs. Further in vivo investigations are required to confirm these results.
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Affiliation(s)
- M Collado-González
- Cellular Therapy and Hematopoietic Transplant Unit, Haematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain.,School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain
| | - M P Pecci-Lloret
- School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain
| | - D García-Bernal
- Cellular Therapy and Hematopoietic Transplant Unit, Haematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain
| | - S Aznar-Cervantes
- Biotechnology Department, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), Murcia, Spain
| | - R E Oñate-Sánchez
- School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain
| | - J M Moraleda
- Cellular Therapy and Hematopoietic Transplant Unit, Haematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain
| | - J L Cenis
- Biotechnology Department, Instituto Murciano de Investigación y Desarrollo Agrario y Alimentario (IMIDA), Murcia, Spain
| | - F J Rodríguez-Lozano
- Cellular Therapy and Hematopoietic Transplant Unit, Haematology Department, Virgen de la Arrixaca Clinical University Hospital, IMIB, University of Murcia, Murcia, Spain. .,School of Dentistry, Faculty of Medicine, University of Murcia, Murcia, Spain. .,Special Patients and Gerodontology Unit, School of Dentistry, University of Murcia, IMIB-Arrixaca, Morales Meseguer Hospital, Avda, Marqués de los Vélez s/n, 30007, Murcia, Spain.
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23
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Gnanasegaran N, Govindasamy V, Simon C, Gan QF, Vincent-Chong VK, Mani V, Krishnan Selvarajan K, Subramaniam V, Musa S, Abu Kasim NH. Effect of dental pulp stem cells in MPTP-induced old-aged mice model. Eur J Clin Invest 2017; 47:403-414. [PMID: 28369799 DOI: 10.1111/eci.12753] [Citation(s) in RCA: 19] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/05/2017] [Accepted: 03/24/2017] [Indexed: 12/13/2022]
Abstract
BACKGROUND Parkinson's disease (PD) is a neurodegenerative disease caused by the loss of dopaminergic (DA-ergic) neurons in the substantia nigra (SN) and represented as a huge threat to the geriatric population. Cell replacement therapies (CRTs) have been proposed as a promising strategy to slow down or replace neuronal loss. Among the widely available cell sources, dental pulp stem cells (DPSCs) portray as an attractive source primarily due to their neural crest origin, ease of tissue procurement and less ethical hurdles. MATERIALS AND METHODS We first demonstrated the in vitro differentiation ability of DPSCs towards DA-ergic-like cells before evaluating their neuro-protection/neuro-restoration capacities in MPTP-induced mice. Transplantation via intrathecal was performed with behavioural assessments being evaluated every fortnight. Subsequent analysis investigating their immuno-modulatory behaviour was conducted using neuronal and microglial cell lines. RESULTS It was apparent that the behavioural parameters began to improve corresponding to tyrosine hydroxylase (TH), dopamine transporter (DAT) and dopamine decarboxylase (AADC) immunostaining in SN and striatum as early as 8-week post-transplantation (P < 0·05). About 60% restoration of DA-ergic neurons was observed at SN in MPTP-treated mice after 12-week post-transplantation. Similarly, their ability to reduce toxic effects of MPTP (DNA damages, reactive oxygen species and nitric oxide release) and regulate cytokine levels was distinctly noted (P < 0·05) upon exposure in in vitro model. CONCLUSIONS Our results suggest that DPSCs may provide a therapeutic benefit in the old-aged PD mice model and may be explored in stem cell-based CRTs especially in geriatric population as an attempt towards 'personalized medicine'.
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Affiliation(s)
- Nareshwaran Gnanasegaran
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Vijayendran Govindasamy
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Christopher Simon
- Faculty of Applied Sciences, AIMST University, Semeling, Bedong, Kedah, Malaysia
| | - Quan Fu Gan
- Faculty of Applied Sciences, AIMST University, Semeling, Bedong, Kedah, Malaysia
| | - Vui King Vincent-Chong
- Oral Cancer Research and Coordinating Center (OCRCC), Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Vasudevan Mani
- Department of Pharmacology and Toxicology, College of Pharmacy, Qassim University, Buraidah, Saudi Arabia
| | | | - Vellayan Subramaniam
- Laboratory Animal Facility and Management (LAFAM), Faculty of Pharmacy, UiTM Puncak, Alam Selangor, Malaysia
| | - Sabri Musa
- Department of Paediatric Dentistry and Orthodontics, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
| | - Noor Hayaty Abu Kasim
- Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, Kuala Lumpur, Malaysia
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24
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Analysis of the Adherence of Dental Pulp Stem Cells on Two-Dimensional and Three-Dimensional Silk Fibroin-Based Biomaterials. J Craniofac Surg 2017; 28:939-943. [DOI: 10.1097/scs.0000000000003596] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
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25
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Dou Q, Low ZWK, Zhang K, Loh XJ. A new light triggered approach to develop a micro porous tough hydrogel. RSC Adv 2017. [DOI: 10.1039/c7ra03214e] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/03/2023] Open
Abstract
A porous tough hydrogel (alginate–polyacrylamide) is successfully synthesized using a photo-gelling polymer as a templating agent.
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Affiliation(s)
- Qingqing Dou
- Institute of Materials Research and Engineering (IMRE)
- Singapore 138634
| | - Zhi Wei Kenny Low
- Institute of Materials Research and Engineering (IMRE)
- Singapore 138634
- Department of Materials Science and Engineering
- National University of Singapore
- Singapore 117576
| | - Kangyi Zhang
- Institute of Materials Research and Engineering (IMRE)
- Singapore 138634
| | - Xian Jun Loh
- Institute of Materials Research and Engineering (IMRE)
- Singapore 138634
- Department of Materials Science and Engineering
- National University of Singapore
- Singapore 117576
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26
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Woloszyk A, Buschmann J, Waschkies C, Stadlinger B, Mitsiadis TA. Human Dental Pulp Stem Cells and Gingival Fibroblasts Seeded into Silk Fibroin Scaffolds Have the Same Ability in Attracting Vessels. Front Physiol 2016; 7:140. [PMID: 27148078 PMCID: PMC4835714 DOI: 10.3389/fphys.2016.00140] [Citation(s) in RCA: 29] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/10/2016] [Accepted: 03/30/2016] [Indexed: 12/20/2022] Open
Abstract
Neovascularization is one of the most important processes during tissue repair and regeneration. Current healing approaches based on the use of biomaterials combined with stem cells in critical-size bone defects fail due to the insufficient implant vascularization and integration into the host tissues. Therefore, here we studied the attraction, ingrowth, and distribution of blood vessels from the chicken embryo chorioallantoic membrane into implanted silk fibroin scaffolds seeded with either human dental pulp stem cells or human gingival fibroblasts. Perfusion capacity was evaluated by non-invasive in vivo Magnetic Resonance Imaging while the number and density of blood vessels were measured by histomorphometry. Our results demonstrate that human dental pulp stem cells and gingival fibroblasts possess equal abilities in attracting vessels within silk fibroin scaffolds. Additionally, the prolonged in vitro pre-incubation period of these two cell populations favors the homogeneous distribution of vessels within silk fibroin scaffolds, which further improves implant survival and guarantees successful healing and regeneration.
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Affiliation(s)
- Anna Woloszyk
- Orofacial Development and Regeneration, Center of Dental Medicine, Institute of Oral Biology, University of Zurich Zurich, Switzerland
| | - Johanna Buschmann
- Plastic Surgery and Hand Surgery, University Hospital Zurich Zurich, Switzerland
| | - Conny Waschkies
- Institute for Biomedical Engineering, ETH and University of ZurichZurich, Switzerland; Visceral and Transplant Surgery, University Hospital ZurichZurich, Switzerland
| | - Bernd Stadlinger
- Clinic of Cranio-Maxillofacial and Oral Surgery, University of Zurich, University Hospital Zurich Zurich, Switzerland
| | - Thimios A Mitsiadis
- Orofacial Development and Regeneration, Center of Dental Medicine, Institute of Oral Biology, University of Zurich Zurich, Switzerland
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27
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Kołbuk D, Guimond-Lischer S, Sajkiewicz P, Maniura-Weber K, Fortunato G. Morphology and surface chemistry of bicomponent scaffolds in terms of mesenchymal stromal cell viability. J BIOACT COMPAT POL 2016. [DOI: 10.1177/0883911515621571] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
Abstract
Biological interaction between cells and scaffolds is mediated through events at surfaces. Proteins present in the culture medium adsorb on substrates, generating a protein adlayer that triggers further downstream events governing cell adhesion. Polymer blends often combine the properties of the individual components, for example, can provide mechanical as well as surface properties in one fibre. Therefore, mixtures of synthetic polycaprolactone and gelatin as a denatured form of collagen were electrospun at selected conditions and polymer weight ratios. Fibre morphologies and chemical properties of the surfaces were analysed. These scaffolds were seeded with human mesenchymal stromal cells and their viability was studied. Gelatin addition to polycaprolactone leads to a reduction in fibre diameter. A linear increase in gelatin at the fibre surface was observed in function of the weighed polymers, except for polycaprolactone/gelatin fibres incorporating equal weight ratios. Thereby, a depletion of gelatin at the fibre surface is stated for equally mixed polymers. The depletion of gelatin at the fibre surface is most probably due to hydrophobic interactions between hydrophobic segments of polycaprolactone and gelatin, affecting the spinning mechanism and thus fibre structure. Furthermore, polycaprolactone/gelatin blends show enhanced wettability properties compared to pure gelatin, at least partly due to molecular segregation. Results of in vitro studies reveal an increase in cellular viability and proliferation for cells cultivated on nanofibres containing gelatin, caused by the cell-attractive surface composition as well as the hydrophilic nature of the scaffolds. Contact guidance of cells seeded on parallelised fibres is observed, and DNA tests show evidently enhanced cell numbers on nanofibres containing 20 wt% of gelatin.
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Affiliation(s)
- Dorota Kołbuk
- Institute of Fundamental Technological Research, Polish Academy of Sciences, Warsaw, Poland
- Advanced Fibres, Swiss Federal Laboratories for Materials Science and Technology (EMPA), St. Gallen, Switzerland
| | - Stefanie Guimond-Lischer
- Biointerfaces, Swiss Federal Laboratories for Materials Science and Technology (EMPA), St. Gallen, Switzerland
| | - Paweł Sajkiewicz
- Institute of Fundamental Technological Research, Polish Academy of Sciences, Warsaw, Poland
| | - Katharina Maniura-Weber
- Biointerfaces, Swiss Federal Laboratories for Materials Science and Technology (EMPA), St. Gallen, Switzerland
| | - Giuseppino Fortunato
- Protection and Physiology, Swiss Federal Laboratories for Materials Science and Technology (EMPA), St. Gallen, Switzerland
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28
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Maioli M, Basoli V, Santaniello S, Cruciani S, Delitala AP, Pinna R, Milia E, Grillari-Voglauer R, Fontani V, Rinaldi S, Muggironi R, Pigliaru G, Ventura C. Osteogenesis from Dental Pulp Derived Stem Cells: A Novel Conditioned Medium Including Melatonin within a Mixture of Hyaluronic, Butyric, and Retinoic Acids. Stem Cells Int 2016; 2016:2056416. [PMID: 26880937 PMCID: PMC4736970 DOI: 10.1155/2016/2056416] [Citation(s) in RCA: 34] [Impact Index Per Article: 3.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/11/2015] [Revised: 11/30/2015] [Accepted: 12/14/2015] [Indexed: 01/09/2023] Open
Abstract
Human dental pulp stem cells (hDPSCs) have shown relevant potential for cell therapy in the orthopedic and odontoiatric fields. The optimization of their osteogenic potential is currently a major challenge. Vascular endothelial growth factor A (VEGF A) has been recently reported to act as a major conductor of osteogenesis in vitro and in vivo. Here, we attempted to prime endogenous VEGF A expression without the need for viral vector mediated gene transfer technologies. We show that hDPSCs exposure to a mixture of hyaluronic, butyric, and retinoic acids (HA + BU + RA) induced the transcription of a gene program of osteogenesis and the acquirement of an osteogenic lineage. Such response was also elicited by cell exposure to melatonin, a pleiotropic agent that recently emerged as a remarkable osteogenic inducer. Interestingly, the commitment to the osteogenic fate was synergistically enhanced by the combinatorial exposure to a conditioned medium containing both melatonin and HA + BU + RA. These in vitro results suggest that in vivo osteogenesis might be improved and further studies are needed.
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Affiliation(s)
- Margherita Maioli
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
- Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Via Massarenti, 40138 Bologna, Italy
| | - Valentina Basoli
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
- Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Via Massarenti, 40138 Bologna, Italy
- Research Department, Rinaldi Fontani Foundation, Viale Belfiore 43, 50144 Florence, Italy
- Department of Biotechnology, University of Natural Resources and Life Sciences Vienna, Muthgasse 18, A-1190 Vienna, Austria
| | - Sara Santaniello
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
- Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Via Massarenti, 40138 Bologna, Italy
| | - Sara Cruciani
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
| | | | - Roberto Pinna
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
| | - Egle Milia
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
| | - Regina Grillari-Voglauer
- Department of Biotechnology, University of Natural Resources and Life Sciences Vienna, Muthgasse 18, A-1190 Vienna, Austria
| | - Vania Fontani
- Research Department, Rinaldi Fontani Foundation, Viale Belfiore 43, 50144 Florence, Italy
- Department of Regenerative Medicine, Rinaldi Fontani Institute, Viale Belfiore 43, 50144 Florence, Italy
| | - Salvatore Rinaldi
- Research Department, Rinaldi Fontani Foundation, Viale Belfiore 43, 50144 Florence, Italy
- Department of Regenerative Medicine, Rinaldi Fontani Institute, Viale Belfiore 43, 50144 Florence, Italy
| | - Roberta Muggironi
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
- Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Via Massarenti, 40138 Bologna, Italy
| | - Gianfranco Pigliaru
- Department of Biomedical Sciences, University of Sassari, Viale San Pietro 43/B, 07100 Sassari, Italy
- Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Via Massarenti, 40138 Bologna, Italy
| | - Carlo Ventura
- Laboratory of Molecular Biology and Stem Cell Engineering, National Institute of Biostructures and Biosystems, Via Massarenti, 40138 Bologna, Italy
- Stem Wave Institute for Tissue Healing (SWITH), Gruppo Villa Maria and Ettore Sansavini Health Science Foundation, Via Provinciale per Cotignola 9, 48022 Lugo, Ravenna, Italy
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Liu G, Xu G, Gao Z, Liu Z, Xu J, Wang J, Zhang C, Wang S. Demineralized Dentin Matrix Induces Odontoblastic Differentiation of Dental Pulp Stem Cells. Cells Tissues Organs 2015; 201:65-76. [DOI: 10.1159/000440952] [Citation(s) in RCA: 28] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 09/07/2015] [Indexed: 11/19/2022] Open
Abstract
The aim of this study was to investigate the effect of demineralized dentin matrix (DDM) on dental pulp stem cells (DPSCs) and the potential of complexes with DPSCs and DDM for mineralized tissue formation. Stem cells derived from the dental pulp of healthy pigs aged 18 months were isolated and cultured. DPSCs were incubated with alpha-minimum essential medium treated with DDM extract at 1 mg/ml (DDM1) or 10 mg/ml (DDM10). The concentrations of 3 growth factors in DDM extract was measured by enzyme-linked immunosorbent assay. Adhesion of DPSCs on DDM and hydroxyapatite-tricalcium phosphate (HA-TCP) surfaces was observed using scanning electron microscopy. Cell proliferation was evaluated with cell counting kit-8 and migration by Transwell migration assays. Odontoblastic differentiation was assessed by alkaline phosphatase (ALP) and alizarin red staining, ALP activity and real-time polymerase chain reaction analysis of markers of ALP, runt-related transcription factor 2, type I collagen, dentin matrix acidic phosphoprotein-1, osteonectin and dentin sialophosphoprotein (DSPP). Finally, DPSCs were combined with DDM and placed subcutaneously in nude mice for 12 weeks; DPSCs combined with HA-TCP and DDM alone served as controls. DDM could promote DPSC adhesion, migration and odontoblastic differentiation. Mineralized tissue formation was observed with the DPSC and DDM combination and the DPSC and HA-TCP combination. The mineralized tissue of the DPSC + DDM combination stained positive for DSPP, similar to the dentin tissue. These results indicate that DDM induces DPSC odontoblastic differentiation, suggesting applications for dentin regeneration.
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30
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Ducret M, Fabre H, Degoul O, Atzeni G, McGuckin C, Forraz N, Alliot-Licht B, Mallein-Gerin F, Perrier-Groult E, Farges JC. Manufacturing of dental pulp cell-based products from human third molars: current strategies and future investigations. Front Physiol 2015; 6:213. [PMID: 26300779 PMCID: PMC4526817 DOI: 10.3389/fphys.2015.00213] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/03/2015] [Accepted: 07/16/2015] [Indexed: 01/01/2023] Open
Abstract
In recent years, mesenchymal cell-based products have been developed to improve surgical therapies aimed at repairing human tissues. In this context, the tooth has recently emerged as a valuable source of stem/progenitor cells for regenerating orofacial tissues, with easy access to pulp tissue and high differentiation potential of dental pulp mesenchymal cells. International guidelines now recommend the use of standardized procedures for cell isolation, storage and expansion in culture to ensure optimal reproducibility, efficacy and safety when cells are used for clinical application. However, most dental pulp cell-based medicinal products manufacturing procedures may not be fully satisfactory since they could alter the cells biological properties and the quality of derived products. Cell isolation, enrichment and cryopreservation procedures combined to long-term expansion in culture media containing xeno- and allogeneic components are known to affect cell phenotype, viability, proliferation and differentiation capacities. This article focuses on current manufacturing strategies of dental pulp cell-based medicinal products and proposes a new protocol to improve efficiency, reproducibility and safety of these strategies.
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Affiliation(s)
- Maxime Ducret
- Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Claude Bernard Lyon 1 Lyon, France ; Faculté d'Odontologie, Université de Lyon, Université Claude Bernard Lyon 1 Lyon, France ; Hospices Civils de Lyon, Service de Consultations et Traitements Dentaires Lyon, France
| | - Hugo Fabre
- Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Claude Bernard Lyon 1 Lyon, France
| | - Olivier Degoul
- CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France
| | | | - Colin McGuckin
- CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France
| | - Nico Forraz
- CTI-BIOTECH, Cell Therapy Research Institute Meyzieu, France
| | - Brigitte Alliot-Licht
- Faculté d'Odontologie, Institut National de la Santé et de la Recherche Médicale UMR1064, Centre de Recherche en Transplantation et Immunologie, Université de Nantes Nantes, France
| | - Frédéric Mallein-Gerin
- Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Claude Bernard Lyon 1 Lyon, France
| | - Emeline Perrier-Groult
- Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Claude Bernard Lyon 1 Lyon, France
| | - Jean-Christophe Farges
- Laboratoire de Biologie Tissulaire et Ingénierie thérapeutique, UMR5305 Centre National de la Recherche Scientifique/Université Claude Bernard Lyon 1 Lyon, France ; Faculté d'Odontologie, Université de Lyon, Université Claude Bernard Lyon 1 Lyon, France ; Hospices Civils de Lyon, Service de Consultations et Traitements Dentaires Lyon, France
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31
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Osteogenic Potential of Dental Mesenchymal Stem Cells in Preclinical Studies: A Systematic Review Using Modified ARRIVE and CONSORT Guidelines. Stem Cells Int 2015; 2015:378368. [PMID: 26106427 PMCID: PMC4464683 DOI: 10.1155/2015/378368] [Citation(s) in RCA: 35] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/27/2014] [Accepted: 02/01/2015] [Indexed: 12/22/2022] Open
Abstract
Background and Objective. Dental stem cell-based tissue engineered constructs are emerging as a promising alternative to autologous bone transfer for treating bone defects. The purpose of this review is to systematically assess the preclinical in vivo and in vitro studies which have evaluated the efficacy of dental stem cells on bone regeneration. Methods. A literature search was conducted in Ovid Medline, Embase, PubMed, and Web of Science up to October 2014. Implantation of dental stem cells in animal models for evaluating bone regeneration and/or in vitro studies demonstrating osteogenic potential of dental stem cells were included. The preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were used to ensure the quality of the search. Modified ARRIVE (Animal research: reporting in invivo experiments) and CONSORT (Consolidated reporting of trials) were used to critically analyze the selected studies. Results. From 1914 citations, 207 full-text articles were screened and 137 studies were included in this review. Because of the heterogeneity observed in the studies selected, meta-analysis was not possible. Conclusion. Both in vivo and in vitro studies indicate the potential use of dental stem cells in bone regeneration. However well-designed randomized animal trials are needed before moving into clinical trials.
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Shen G, Hu X, Guan G, Wang L. Surface Modification and Characterisation of Silk Fibroin Fabric Produced by the Layer-by-Layer Self-Assembly of Multilayer Alginate/Regenerated Silk Fibroin. PLoS One 2015; 10:e0124811. [PMID: 25919690 PMCID: PMC4412632 DOI: 10.1371/journal.pone.0124811] [Citation(s) in RCA: 29] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2014] [Accepted: 03/17/2015] [Indexed: 02/05/2023] Open
Abstract
Silk-based medical products have a long history of use as a material for surgical sutures because of their desirable mechanical properties. However, silk fibroin fabric has been reported to be haemolytic when in direct contact with blood. The layer-by-layer self-assembly technique provides a method for surface modification to improve the biocompatibility of silk fibroin fabrics. Regenerated silk fibroin and alginate, which have excellent biocompatibility and low immunogenicity, are outstanding candidates for polyelectrolyte deposition. In this study, silk fabric was degummed and positively charged to create a silk fibroin fabric that could undergo self-assembly. The multilayer self-assembly of the silk fibroin fabric was achieved by alternating the polyelectrolyte deposition of a negatively charged alginate solution (pH = 8) and a positively charged regenerated silk fibroin solution (pH = 2). Finally, the negatively charged regenerated silk fibroin solution (pH = 8) was used to assemble the outermost layer of the fabric so that the surface would be negatively charged. A stable structural transition was induced using 75% ethanol. The thickness and morphology were characterised using atomic force microscopy. The properties of the self-assembled silk fibroin fabric, such as the bursting strength, thermal stability and flushing stability, indicated that the fabric was stable. In addition, the cytocompatibility and haemocompatibility of the self-assembled silk fibroin fabrics were evaluated. The results indicated that the biocompatibility of the self-assembled multilayers was acceptable and that it improved markedly. In particular, after the self-assembly, the fabric was able to prevent platelet adhesion. Furthermore, other non-haemolytic biomaterials can be created through self-assembly of more than 1.5 bilayers, and we propose that self-assembled silk fibroin fabric may be an attractive candidate for anticoagulation applications and for promoting endothelial cell adhesion for vascular prostheses.
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Affiliation(s)
- Gaotian Shen
- Key Laboratory of Textile Science and Technology, Ministry of Education, College of Textiles, Donghua University, Songjiang District, Shanghai 201620, China
| | - Xingyou Hu
- Key Laboratory of Textile Science and Technology, Ministry of Education, College of Textiles, Donghua University, Songjiang District, Shanghai 201620, China
| | - Guoping Guan
- Key Laboratory of Textile Science and Technology, Ministry of Education, College of Textiles, Donghua University, Songjiang District, Shanghai 201620, China
- Engineering Research Center of Technical Textiles, Ministry of Education, Shanghai 201620, China
| | - Lu Wang
- Key Laboratory of Textile Science and Technology, Ministry of Education, College of Textiles, Donghua University, Songjiang District, Shanghai 201620, China
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