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Cristallini C, Rossin D, Vanni R, Barbani N, Bulgheresi C, Labardi M, Perveen S, Burchielli S, Terlizzi D, Kusmic C, Del Ry S, Cabiati M, Trouki C, Rossino D, Sergi F, Villano A, Aquaro GD, Scarpellino G, Ruffinatti FA, Amorim S, Pires RA, Reis RL, Rastaldo R, Giachino C. A biodegradable, microstructured, electroconductive and nano-integrated drug eluting patch (MENDEP) for myocardial tissue engineering. Bioact Mater 2025; 50:246-272. [PMID: 40270551 PMCID: PMC12017858 DOI: 10.1016/j.bioactmat.2025.04.008] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/04/2024] [Revised: 03/24/2025] [Accepted: 04/05/2025] [Indexed: 04/25/2025] Open
Abstract
We produced a microstructured, electroconductive and nano-functionalized drug eluting cardiac patch (MENDEP) designed to attract endogenous precursor cells, favor their differentiation and counteract adverse ventricular remodeling in situ. MENDEP showed mechanical anisotropy and biaxial strength comparable to porcine myocardium, reduced impedance, controlled biodegradability, molecular recognition ability and controlled drug release activity. In vitro, cytocompatibility and cardioinductivity were demonstrated. Migration tests showed the chemoattractive capacity of the patches and conductivity assays showed unaltered cell-cell interactions and cell beating synchronicity. MENDEP was then epicardially implanted in a rat model of ischemia/reperfusion (I/R). Histological, immunofluorescence and biomarker analysis indicated that implantation did not cause damage to the healthy myocardium. After I/R, MENDEP recruited precursor cells into the damaged myocardium and triggered their differentiation towards the vascular lineage. Under the patch, the myocardial tissue appeared well preserved and cardiac gap junctions were correctly distributed at the level of the intercalated discs. The fibrotic area measured in the I/R group was partially reduced in the patch group. Overall, these results demonstrate that MENDEP was fully retained on the epicardial surface of the left ventricle over 4-week implantation period, underwent progressive vascularization, did not perturb the healthy myocardium and showed great potential in repairing the infarcted area.
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Affiliation(s)
- Caterina Cristallini
- Institute for Chemical and Physical Processes, CNR-IPCF, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
- Department of Civil and Industrial Engineering, DICI, University of Pisa, Largo Lucio Lazzarino, 56126, Pisa, Italy
| | - Daniela Rossin
- Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, 10043, Orbassano, Italy
| | - Roberto Vanni
- Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, 10043, Orbassano, Italy
| | - Niccoletta Barbani
- Institute for Chemical and Physical Processes, CNR-IPCF, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
- Department of Civil and Industrial Engineering, DICI, University of Pisa, Largo Lucio Lazzarino, 56126, Pisa, Italy
| | - Chiara Bulgheresi
- Department of Civil and Industrial Engineering, DICI, University of Pisa, Largo Lucio Lazzarino, 56126, Pisa, Italy
| | - Massimiliano Labardi
- Institute for Chemical and Physical Processes, CNR-IPCF, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
| | - Sadia Perveen
- Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, 10043, Orbassano, Italy
| | | | | | - Claudia Kusmic
- Clinical Physiology Institute, CNR-IFC, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
| | - Silvia Del Ry
- Clinical Physiology Institute, CNR-IFC, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
| | - Manuela Cabiati
- Clinical Physiology Institute, CNR-IFC, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
| | - Cheherazade Trouki
- Institute for Chemical and Physical Processes, CNR-IPCF, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
- Department of Pharmacy, University of Pisa, Via Bonanno 6, 56126, Pisa, Italy
| | - Dawid Rossino
- Institute for Chemical and Physical Processes, CNR-IPCF, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
- Department of Civil and Industrial Engineering, DICI, University of Pisa, Largo Lucio Lazzarino, 56126, Pisa, Italy
| | - Francesca Sergi
- Department of Civil and Industrial Engineering, DICI, University of Pisa, Largo Lucio Lazzarino, 56126, Pisa, Italy
| | - Anthea Villano
- Institute for Chemical and Physical Processes, CNR-IPCF, Via Giuseppe Moruzzi 1, 56124, Pisa, Italy
| | - Giovanni D. Aquaro
- Academic Radiology Unit, Department of Surgical, Medical and Molecular Pathology and of the Critical Area, University of Pisa, Via Paradisa 2, 56124, Pisa, Italy
| | - Giorgia Scarpellino
- Department of Life Sciences and Systems Biology, University of Turin, Via Accademia Albertina 13, 10123, Torino, Italy
- Department of Biology and Biotechnology “L. Spallanzani”, University of Pavia, Via Adolfo Ferrata 9, 27100, Pavia, Italy
| | - Federico A. Ruffinatti
- Department of Life Sciences and Systems Biology, University of Turin, Via Accademia Albertina 13, 10123, Torino, Italy
| | - Sara Amorim
- 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, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal
| | - Ricardo A. Pires
- 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, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal
| | - Rui L. Reis
- 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, AvePark, Parque de Ciência e Tecnologia, Zona Industrial da Gandra, 4805-017, Barco, Guimarães, Portugal
- ICVS/3B's-PT Government Associate Laboratory, Braga, Guimarães, Portugal
| | - Raffaella Rastaldo
- Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, 10043, Orbassano, Italy
| | - Claudia Giachino
- Department of Clinical and Biological Sciences, University of Turin, Regione Gonzole 10, 10043, Orbassano, Italy
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2
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Wongsirichot P. Pilot scale polyhydroxyalkanoates biopolymer production using pure cultures: current status and future opportunities. Crit Rev Biotechnol 2025; 45:887-903. [PMID: 39428339 DOI: 10.1080/07388551.2024.2409112] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/31/2024] [Revised: 07/08/2024] [Accepted: 08/08/2024] [Indexed: 10/22/2024]
Abstract
The development and commercialization of bio-based and biodegradable polyhydroxyalkanoates (PHAs) biopolymers could be crucial for the transition toward a sustainable circular economy. However, despite potential traditional and novel applications in the packaging, textiles, agriculture, automotive, electronics, and biomedical industries, the commercialization of PHAs is limited by their current market competitiveness. This review provides the first critical assessment of the current pure culture pilot-scale PHA literature, which could be crucial in translating promising laboratory-scale developments into industrial-scale commercial PHA production. It will also complement reviews of mixed microbial cultures currently dominating pilot-scale PHA literature. Pure culture fermentations could provide advantages, such as ease of characterizing microbial producers' behavior, higher PHA productivities, and better alignment with existing PHA commercialization and industrial biotechnology approaches. Key aspects, including producer organisms, fermentation volumes and schemes, control schemes, optimization, and properties of the polymers produced, are discussed in-depth, to elucidate important trends, achievements, and knowledge gaps. Furthermore, specific ways for future pilot-scale studies to help address current PHA commercialization challenges are also identified. The insights, and recommendations provided will be extremely beneficial for the future development of PHA production, at both pilot and commercial scales, whilst also being beneficial to the production of other microbial polymers and industrial biotechnology as a whole.
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Affiliation(s)
- Phavit Wongsirichot
- Department of Chemical Engineering, The University of Manchester, Manchester, United Kingdom
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3
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Bianchi E, Ruggeri M, Vigani B, Aguzzi C, Rossi S, Sandri G. Synthesis and use of thermoplastic polymers for tissue engineering purposes. Int J Pharm X 2025; 9:100313. [PMID: 39807177 PMCID: PMC11729033 DOI: 10.1016/j.ijpx.2024.100313] [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: 07/25/2024] [Revised: 11/27/2024] [Accepted: 12/12/2024] [Indexed: 01/16/2025] Open
Abstract
Thermoplastic polymers provide a versatile platform to mimic various aspects of physiological extracellular matrix properties such as chemical composition, stiffness, and topography for use in cell and tissue engineering applications. In this review, we provide a brief overview of the most promising thermoplastic polymers, and in particular the thermoplastic polyesters, such as poly(lactic acid), poly(glycolic acid), and polycaprolactone, and the thermoplastic elastomers, such as polyurethanes, polyhydroxyalkanoates, and poly(butyl cyanoacrylate). A particular focus has been made on the synthesis processes, the processability and the biocompatibility. We also discuss how these materials can be applied in tissue engineering, mimicking tissues' structure and function, and stimulate mesenchymal stem cells differentiation and mechanotransduction.
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Affiliation(s)
- Eleonora Bianchi
- Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
| | - Marco Ruggeri
- Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
| | - Barbara Vigani
- Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
| | - Carola Aguzzi
- Department of Pharmacy and Pharmaceutical Technology, University of Granada, Cartuja Campus, Granada 18071, Spain
| | - Silvia Rossi
- Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
| | - Giuseppina Sandri
- Department of Drug Sciences, University of Pavia, Viale Taramelli 12, 27100 Pavia, Italy
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4
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Bolla M, Pettinato M, Ferrari PF, Fabiano B, Perego P. Polyhydroxyalkanoates production from laboratory to industrial scale: A review. Int J Biol Macromol 2025; 310:143255. [PMID: 40250686 DOI: 10.1016/j.ijbiomac.2025.143255] [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: 02/12/2025] [Revised: 04/06/2025] [Accepted: 04/15/2025] [Indexed: 04/20/2025]
Abstract
Environmental issues related to fossil-based plastics are getting the attention of the media and legislative authorities, addressing the need to improve the plastics' design, collection, and circular economy. In this regard, polyhydroxyalkanoates (PHAs) represent a promising alternative to the conventional polymers, given their biological origin, biodegradability, and biocompatibility. To date, their commercialization covers only a little percentage of the biodegradable plastic application, mainly due to their high cost. However, new production strategies are being investigated and patented, enhancing the PHA market competitiveness. This review tries to fill the gap about the critical investigation on innovative and up-to-date process strategies in PHA production field, deeply evaluating them from a plant-engineering point of view. Several aspects are considered regarding the reduction of the production costs and the increase in the overall PHA productivity and recovery. Among them, the feeding of pre-treated carbon sources derived from food and agro-industrial wastes, the use of mixed microbial cultures as convenient substitutes to the pure ones, and optimized downstream processes are widely discussed. The overlook of the topic is completed by evaluating the innovative technologies existing at pilot and industrial scale, able to achieve improved production yields. Finally, PHA economic and market current conditions are investigated.
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Affiliation(s)
- Maria Bolla
- Department of Civil, Chemical and Environmental Engineering, University of Genoa, via Opera Pia, 15, 16145 Genoa, Italy.
| | - Margherita Pettinato
- Department of Civil, Chemical and Environmental Engineering, University of Genoa, via Opera Pia, 15, 16145 Genoa, Italy; Research Center for Biologically Inspired Engineering in Vascular Medicine and Longevity, University of Genoa, via Montallegro, 1, 16145 Genoa, Italy.
| | - Pier Francesco Ferrari
- Department of Civil, Chemical and Environmental Engineering, University of Genoa, via Opera Pia, 15, 16145 Genoa, Italy; Research Center for Biologically Inspired Engineering in Vascular Medicine and Longevity, University of Genoa, via Montallegro, 1, 16145 Genoa, Italy; IRCCS Ospedale Policlinico San Martino, largo Rosanna Benzi, 10, 16132, Genoa, Italy.
| | - Bruno Fabiano
- Department of Civil, Chemical and Environmental Engineering, University of Genoa, via Opera Pia, 15, 16145 Genoa, Italy; Research Center for Biologically Inspired Engineering in Vascular Medicine and Longevity, University of Genoa, via Montallegro, 1, 16145 Genoa, Italy.
| | - Patrizia Perego
- Department of Civil, Chemical and Environmental Engineering, University of Genoa, via Opera Pia, 15, 16145 Genoa, Italy; Research Center for Biologically Inspired Engineering in Vascular Medicine and Longevity, University of Genoa, via Montallegro, 1, 16145 Genoa, Italy; IRCCS Ospedale Policlinico San Martino, largo Rosanna Benzi, 10, 16132, Genoa, Italy.
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5
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Chen Y, Liu X, Zhang L, Yang J, Guo W, Zheng S, Wang J, Wu F, Yan X, Wu Q, Chen G. Cell Sizes Matter for Industrial Bioproduction, a Case of Polyhydroxybutyrate. ADVANCED SCIENCE (WEINHEIM, BADEN-WURTTEMBERG, GERMANY) 2025; 12:e2412256. [PMID: 39965051 PMCID: PMC11984891 DOI: 10.1002/advs.202412256] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/02/2024] [Revised: 01/16/2025] [Indexed: 02/20/2025]
Abstract
Most bacterial cells are 1-2 microns in size, limiting intracellular products like polyhydroxyalkanoates (PHA) accumulation. Cell size is regulated by key genes such as mreB and minCD, which encode cellular skeleton protein and control cell fission ring location, respectively. Their expression changes significantly affect microbial growth. This study successfully redesigns the ClpXP protein degradation system by deleting the sspB gene and using mutated SsrA tags with different degradation rates to control MreB degradation. Dynamic degradation of MreB allows non-model bacterium Halomonas bluephagenesis to grow normally and increase cell size simultaneously. Combined with overexpression of minCD, H. bluephagenesis with progressive MreB degradation increases the cell size further, albeit with a reduced growth rate. H. bluephagenesis CYL0307, with the PHB granule-associated protein PhaP1 deleted and phaABRe overexpressed in the MreB-degraded strain, increases cell volume more than nine times compared to the original strain. CYL0307 produces 149 g L-1 cell dry weight containing 82% PHB after 44 h in a 5000 L bioreactor, with cells containing single large PHB granules, simplifying recovery and purification. These results provide a post-translational gene regulation method in H. bluephagenesis and a strategy for enhancing PHB production via morphological engineering.
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Affiliation(s)
- Yi‐Ling Chen
- School of Life SciencesTsinghua UniversityBeijing100084China
- Tsinghua‐Peking Center for Life SciencesTsinghua UniversityBeijing100084China
| | - Xu Liu
- PhaBuilder Biotechnology Co. Ltd.Shunyi DistrictBeijing101309China
- State Key Laboratory of Animal NutritionInstitute of Animal ScienceChinese Academy of Agricultural SciencesBeijing100193China
| | - Li‐Zhan Zhang
- PhaBuilder Biotechnology Co. Ltd.Shunyi DistrictBeijing101309China
| | - Ji‐Shuai Yang
- PhaBuilder Biotechnology Co. Ltd.Shunyi DistrictBeijing101309China
| | - Wei‐Ke Guo
- School of Life SciencesTsinghua UniversityBeijing100084China
| | - Shuang Zheng
- School of Life SciencesTsinghua UniversityBeijing100084China
| | - Jia‐Le Wang
- School of Life SciencesTsinghua UniversityBeijing100084China
| | - Fu‐Qing Wu
- School of Life SciencesTsinghua UniversityBeijing100084China
- Tsinghua‐Peking Center for Life SciencesTsinghua UniversityBeijing100084China
| | - Xu Yan
- School of Life SciencesTsinghua UniversityBeijing100084China
| | - Qiong Wu
- School of Life SciencesTsinghua UniversityBeijing100084China
| | - Guo‐Qiang Chen
- School of Life SciencesTsinghua UniversityBeijing100084China
- Tsinghua‐Peking Center for Life SciencesTsinghua UniversityBeijing100084China
- Key Lab of Industrial Biocatalysts of the Ministry of EducationDepartment of Chemical EngineeringTsinghua UniversityBeijing100084China
- Center for Synthetic and Systems BiologyTsinghua UniversityBeijing100084China
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6
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Naik TJ, Salgaonkar BB. Unlocking the potential of microbes: Concomitant production of polyhydroxyalkanoates and carotenoids. Int J Biol Macromol 2025; 303:140654. [PMID: 39909243 DOI: 10.1016/j.ijbiomac.2025.140654] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2024] [Revised: 01/21/2025] [Accepted: 02/02/2025] [Indexed: 02/07/2025]
Abstract
The escalating environmental concerns and depletion of crude oil resources have catalyzed interest in biologically derived polymers, particularly biodegradable ones such as polyhydroxyalkanoates. However, the high production costs associated with polyhydroxyalkanoates, driven by raw material expenses, stringent production conditions and low yields, hinder their widespread adoption. A potential strategy to mitigate these costs involves the production of PHAs and other high-value bioproducts, such as carotenoids simultaneously in microbial systems, utilizing shared metabolic pathways. Carotenoids, known for their antioxidant properties and applications in the food, cosmetics and pharmaceutical industries, offer substantial market potential. This review presents a comprehensive overview of the current progress in polyhydroxyalkanoate and carotenoid co-production, explores the co-synthesis pathways, addresses the challenges involved and explores the future prospects of this integrated bioprocess. By diversifying the product portfolio and optimizing microbial production systems, the co-production strategy could pave the way for more sustainable and economically viable bioplastics.
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Affiliation(s)
- Tejas Jagannath Naik
- Microbiology Programme, School of Biological Sciences and Biotechnology (SBSB), Goa University, Taleigao Plateau, Goa 403 206, India.
| | - Bhakti Balkrishna Salgaonkar
- Microbiology Programme, School of Biological Sciences and Biotechnology (SBSB), Goa University, Taleigao Plateau, Goa 403 206, India.
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7
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Kalia VC, Patel SKS, Krishnamurthi P, Singh RV, Lee JK. Exploiting latent microbial potentials for producing polyhydroxyalkanoates: A holistic approach. ENVIRONMENTAL RESEARCH 2025; 269:120895. [PMID: 39832546 DOI: 10.1016/j.envres.2025.120895] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/30/2024] [Revised: 01/14/2025] [Accepted: 01/16/2025] [Indexed: 01/22/2025]
Abstract
Plastics are versatile, however, nonbiodegradable polymers that are primarily derived from fossil fuels and pose notable environmental challenges. However, biopolymers, such as polyhydroxyalkanoates (PHAs), poly(lactic acid), starch, and cellulose have emerged as sustainable alternatives to conventional plastics. Among these, PHAs stand out as strong contenders as they are completely bio-based and biodegradable and are synthesized by microbes as an energy reserve under stress conditions. Despite their limitations, including low mechanical strength, susceptibility to degradation, a restricted scope of application, and high production costs, biopolymers have promising potential. This review explores strategies for enhancing PHA production to address these challenges, emphasizing the need for sustainable PHA production. These strategies include selecting robust microbial strains and feedstock combinations, optimizing cell biomass and biopolymer yields, genetically engineering biosynthetic pathways, and improving downstream processing techniques. Additives such as plasticizers, thermal stabilizers, and antioxidants are crucial for modifying PHA characteristics, and its processing for achieving the desired balance between processability and end-use performance. By overcoming these complications, biopolymers have become more viable, versatile, and eco-friendly alternatives to conventional plastics, offering hope for a more sustainable future.
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Affiliation(s)
- Vipin Chandra Kalia
- Department of Chemical Engineering, Konkuk University, Gwangjin-Gu, Seoul, 05029, Republic of Korea
| | - Sanjay K S Patel
- Department of Biotechnology, Hemvati Nandan Bahuguna Garhwal University (A Central University), Srinagar, 246174, Uttarakhand, India
| | | | - Rahul Vikram Singh
- Department of Chemical Engineering, Konkuk University, Gwangjin-Gu, Seoul, 05029, Republic of Korea
| | - Jung-Kul Lee
- Department of Chemical Engineering, Konkuk University, Gwangjin-Gu, Seoul, 05029, Republic of Korea.
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8
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Zennifer A, Praveenn Kumar S, Bagewadi S, Unnamalai S, Chellappan D, Abdulmalik S, Yu X, Sethuraman S, Sundaramurthi D, Kumbar SG. Innovative spiral nerve conduits: Addressing nutrient transport and cellular activity for critical-sized nerve defects. Bioact Mater 2025; 44:544-557. [PMID: 39584067 PMCID: PMC11583721 DOI: 10.1016/j.bioactmat.2024.10.028] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/27/2024] [Accepted: 10/31/2024] [Indexed: 11/26/2024] Open
Abstract
Large-gap nerve defects require nerve guide conduits (NGCs) for complete regeneration and muscle innervation. Many NGCs have been developed using various scaffold designs and tissue engineering strategies to promote axon regeneration. Still, most are tubular with inadequate pore sizes and lack surface cues for nutrient transport, cell attachment, and tissue infiltration. This study developed a porous spiral NGC to address these issues using a 3D-printed thermoplastic polyurethane (TPU) fiber lattice. The lattice was functionalized with poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) electrospun aligned (aPHBV) and randomly (rPHBV) oriented nanofibers to enhance cellular activity. TPU lattices were made with 25 %, 35 %, and 50 % infill densities to create scaffolds with varied mechanical compliance. The fabricated TPU/PHBV spiral conduits had significantly higher surface areas (25 % TPU/PHBV: 698.97 mm2, 35 % TPU/PHBV: 500.06 mm2, 50 % TPU/PHBV: 327.61 mm2) compared to commercially available nerve conduits like Neurolac™ (205.26 mm2). Aligned PHBV nanofibers showed excellent Schwann cell (RSC96) adhesion, proliferation, and neurogenic gene expression for all infill densities. Spiral TPU/PHBV conduits with 25 % and 35 % infill densities exhibited Young's modulus values comparable to Neurotube® and ultimate tensile strength like acellular cadaveric human nerves. A 10 mm sciatic nerve defect in Wistar rats treated with TPU/aPHBV NGCs demonstrated muscle innervation and axon healing comparable to autografts over 4 months, as evaluated by gait analysis, functional recovery, and histology. The TPU/PHBV NGC developed in this study shows promise as a treatment for large-gap nerve defects.
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Affiliation(s)
- Allen Zennifer
- Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials, School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
- Department of Orthopedic Surgery, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030-4037, USA
| | - S.K. Praveenn Kumar
- Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials, School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
| | - Shambhavi Bagewadi
- Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials, School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
| | - Swathi Unnamalai
- Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials, School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
| | - Davidraj Chellappan
- Central Animal Facility (CAF), School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
| | - Sama Abdulmalik
- Department of Orthopedic Surgery, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030-4037, USA
| | - Xiaojun Yu
- Department of Biomedical Engineering, Stevens Institute of Technology, 1 Castle Point on Hudson Hoboken, New Jersey, 07030, USA
| | - Swaminathan Sethuraman
- Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials, School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
| | - Dhakshinamoorthy Sundaramurthi
- Tissue Engineering & Additive Manufacturing (TEAM) Lab, Centre for Nanotechnology & Advanced Biomaterials, School of Chemical & Biotechnology, SASTRA Deemed University, Tamil Nadu, India
| | - Sangamesh G. Kumbar
- Department of Orthopedic Surgery, University of Connecticut Health Center, 263 Farmington Avenue, Farmington, CT, 06030-4037, USA
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9
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Lalonde JN, Pilania G, Marrone BL. Materials designed to degrade: structure, properties, processing, and performance relationships in polyhydroxyalkanoate biopolymers. Polym Chem 2025; 16:235-265. [PMID: 39464417 PMCID: PMC11498330 DOI: 10.1039/d4py00623b] [Citation(s) in RCA: 3] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/07/2024] [Accepted: 10/05/2024] [Indexed: 10/29/2024]
Abstract
Conventional plastics pose significant environmental and health risks across their life cycle, driving intense interest in sustainable alternatives. Among these, polyhydroxyalkanoates (PHAs) stand out for their biocompatibility, degradation characteristics, and diverse applications. Yet, challenges like production cost, scalability, and limited chemical variety hinder their widespread adoption, impacting material selection and design. This review examines PHA research through the lens of the classical materials tetrahedron, exploring property-structure-processing-performance (PSPP) relationships. By analyzing recent literature and addressing current limitations, we gain valuable insights into PHA development. Despite challenges, we remain optimistic about the role of PHAs in transitioning towards a circular plastic economy, emphasizing the need for further research to unlock their full potential.
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Affiliation(s)
- Jessica N Lalonde
- Department of Mechanical Engineering and Materials Science, Duke University Durham NC 27708 USA
- Bioscience Division, Los Alamos National Laboratory Los Alamos NM 87545 USA
| | | | - Babetta L Marrone
- Bioscience Division, Los Alamos National Laboratory Los Alamos NM 87545 USA
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10
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Kolpek DJ, Kim J, Mohammed H, Gensel JC, Park J. Physicochemical Property Effects on Immune Modulating Polymeric Nanoparticles: Potential Applications in Spinal Cord Injury. Int J Nanomedicine 2024; 19:13357-13374. [PMID: 39691455 PMCID: PMC11649979 DOI: 10.2147/ijn.s497859] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/25/2024] [Accepted: 11/26/2024] [Indexed: 12/19/2024] Open
Abstract
Nanoparticles (NPs) offer promising potential as therapeutic agents for inflammation-related diseases, owing to their capabilities in drug delivery and immune modulation. In preclinical studies focusing on spinal cord injury (SCI), polymeric NPs have demonstrated the ability to reprogram innate immune cells. This reprogramming results in redirecting immune cells away from the injury site, downregulating pro-inflammatory signaling, and promoting a regenerative environment post-injury. However, to fully understand the mechanisms driving these effects and maximize therapeutic efficacy, it is crucial to assess NP interactions with innate immune cells. This review examines how the physicochemical properties of polymeric NPs influence their modulation of the immune system. To achieve this, the review delves into the roles played by innate immune cells in SCI and investigates how various NP properties influence cellular interactions and subsequent immune modulation. Key NP properties such as size, surface charge, molecular weight, shape/morphology, surface functionalization, and polymer composition are thoroughly examined. Furthermore, the review establishes connections between these properties and their effects on the immunomodulatory functions of NPs. Ultimately, this review suggests that leveraging NPs and their physicochemical properties could serve as a promising therapeutic strategy for treating SCI and potentially other inflammatory diseases.
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Affiliation(s)
- Daniel J Kolpek
- Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY, USA
| | - Jaechang Kim
- Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY, USA
| | - Hisham Mohammed
- Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY, USA
| | - John C Gensel
- Spinal Cord and Brain Injury Research Center, Department of Physiology, College of Medicine, University of Kentucky, Lexington, KY, USA
| | - Jonghyuck Park
- Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY, USA
- Spinal Cord and Brain Injury Research Center, Department of Physiology, College of Medicine, University of Kentucky, Lexington, KY, USA
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11
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Martinka Maksymiak M, Andrä-Żmuda S, Sikorska W, Janeczek H, Chaber P, Musioł M, Godzierz M, Kowalczuk M, Adamus G. Structural and Thermal Characterization of Bluepha ® Biopolyesters: Insights into Molecular Architecture and Potential Applications. MATERIALS (BASEL, SWITZERLAND) 2024; 17:5863. [PMID: 39685298 DOI: 10.3390/ma17235863] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/14/2024] [Revised: 11/21/2024] [Accepted: 11/26/2024] [Indexed: 12/18/2024]
Abstract
This study presents an in-depth molecular and structural characterization of novel biopolyesters developed under the trademark Bluepha®. The primary aim was to elucidate the relationship between chemical structure, chain architecture, and material properties of these biopolyesters to define their potential applications across various sectors. Proton nuclear magnetic resonance (1H NMR) analysis identified the biopolyesters as poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] (PHBH) copolymers, containing 4% and 10% molar content of hydroxyhexanoate (HH) units, respectively. Mass spectrometry analysis of PHBH oligomers, produced via controlled thermal degradation, further confirmed the chemical structure and molecular architecture of the PHBH samples. Additionally, multistage electrospray ionization mass spectrometry (ESI-MS/MS) provided insights into the chemical homogeneity and arrangement of comonomer units within the copolyester chains, revealing a random distribution of hydroxyhexanoate (HH) and hydroxybutyrate (HB) units along the PHBH chains. X-ray diffraction (XRD) patterns demonstrated partial crystallinity in the PHBH samples. The thermal properties, including glass transition temperature (Tg), melting temperature (Tm), and melting enthalpy (ΔHm), were found to be lower in PHBH than in poly(R)-3-hydroxybutyrate (PHB), suggesting a broader application potential for the tested PHBH biopolyesters.
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Affiliation(s)
- Magdalena Martinka Maksymiak
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Silke Andrä-Żmuda
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Wanda Sikorska
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Henryk Janeczek
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Paweł Chaber
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Marta Musioł
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Marcin Godzierz
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Marek Kowalczuk
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
| | - Grazyna Adamus
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, 34. M. Curie-Skłodowska St., 41-819 Zabrze, Poland
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12
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Chaber P, Andrä-Żmuda S, Śmigiel-Gac N, Zięba M, Dawid K, Martinka Maksymiak M, Adamus G. Enhancing the Potential of PHAs in Tissue Engineering Applications: A Review of Chemical Modification Methods. MATERIALS (BASEL, SWITZERLAND) 2024; 17:5829. [PMID: 39685265 DOI: 10.3390/ma17235829] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 10/18/2024] [Revised: 11/22/2024] [Accepted: 11/25/2024] [Indexed: 12/18/2024]
Abstract
Polyhydroxyalkanoates (PHAs) are a family of polyesters produced by many microbial species. These naturally occurring polymers are widely used in tissue engineering because of their in vivo degradability and excellent biocompatibility. The best studied among them is poly(3-hydroxybutyrate) (PHB) and its copolymer with 3-hydroxyvaleric acid (PHBV). Despite their superior properties, PHB and PHBV suffer from high crystallinity, poor mechanical properties, a slow resorption rate, and inherent hydrophobicity. Not only are PHB and PHBV hydrophobic, but almost all members of the PHA family struggle because of this characteristic. One can overcome the limitations of microbial polyesters by modifying their bulk or surface chemical composition. Therefore, researchers have put much effort into developing methods for the chemical modification of PHAs. This paper explores a rarely addressed topic in review articles-chemical methods for modifying the structure of PHB and PHBV to enhance their suitability as biomaterials for tissue engineering applications. Different chemical strategies for improving the wettability and mechanical properties of PHA scaffolds are discussed in this review. The properties of PHAs that are important for their applications in tissue engineering are also discussed.
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Affiliation(s)
- Paweł Chaber
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowska 34, 41-819 Zabrze, Poland
| | - Silke Andrä-Żmuda
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowska 34, 41-819 Zabrze, Poland
| | - Natalia Śmigiel-Gac
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowska 34, 41-819 Zabrze, Poland
| | - Magdalena Zięba
- Department of Optoelectronics, Silesian University of Technology, ul. B. Krzywoustego 2, 44-100 Gliwice, Poland
| | - Kamil Dawid
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowska 34, 41-819 Zabrze, Poland
| | - Magdalena Martinka Maksymiak
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowska 34, 41-819 Zabrze, Poland
| | - Grażyna Adamus
- Centre of Polymer and Carbon Materials, Polish Academy of Sciences, ul. M. Curie-Skłodowska 34, 41-819 Zabrze, Poland
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13
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Ben Abdeladhim R, Reis JA, Vieira AM, de Almeida CD. Polyhydroxyalkanoates: Medical Applications and Potential for Use in Dentistry. MATERIALS (BASEL, SWITZERLAND) 2024; 17:5415. [PMID: 39597239 PMCID: PMC11595952 DOI: 10.3390/ma17225415] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/18/2024] [Revised: 10/29/2024] [Accepted: 10/31/2024] [Indexed: 11/29/2024]
Abstract
Polyhydroxyalkanoates (PHAs) are promising biopolymers as an alternative to traditional synthetic polymers due to their biodegradability and biocompatibility. The PHA market is blooming in response to the growing demand for biodegradable and environmentally friendly plastics. These biopolyesters are produced and degraded by a variety of microorganisms, making them environmentally friendly, while offering benefits such as biocompatibility (when adequately processed) and biodegradability. Their versatility extends to various areas, from biomedicine to agriculture and composite materials, where they pave the way for significative innovations. In the field of regenerative medicine, some PHAs have key applications, namely in vascular grafts, oral tissue regeneration, and development of self-healing polymers. In addition, PHAs have the potential to be used in the creation of dental implant materials and dental medical devices. PHAs can also be used to encapsulate hydrophobic drugs, providing an approach for more targeted and effective treatments. To summarize, PHAs open new perspectives in the field of medicine by improving drug delivery and offering ecologically biocompatible solutions for medical devices. The aim of this review is to present the medical and dental applications of PHA, their advantages, disadvantages, and indications.
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Affiliation(s)
- Rim Ben Abdeladhim
- Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal; (R.B.A.); (J.A.R.); (A.M.V.)
| | - José Alexandre Reis
- Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal; (R.B.A.); (J.A.R.); (A.M.V.)
- Egas Moniz Center for Interdisciplinary Research (CiiEM), Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal
| | - Ana Maria Vieira
- Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal; (R.B.A.); (J.A.R.); (A.M.V.)
- Egas Moniz Center for Interdisciplinary Research (CiiEM), Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal
| | - Catarina Dias de Almeida
- Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal; (R.B.A.); (J.A.R.); (A.M.V.)
- Egas Moniz Center for Interdisciplinary Research (CiiEM), Egas Moniz School of Health & Science, Campus Universitário, Quinta da Granja, 2829-511 Caparica, Portugal
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14
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Bhatia SK, Patel AK, Yang YH. The green revolution of food waste upcycling to produce polyhydroxyalkanoates. Trends Biotechnol 2024; 42:1273-1287. [PMID: 38582658 DOI: 10.1016/j.tibtech.2024.03.002] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/05/2024] [Revised: 02/16/2024] [Accepted: 03/07/2024] [Indexed: 04/08/2024]
Abstract
This review emphasizes the urgent need for food waste upcycling as a response to the mounting global food waste crisis. Focusing on polyhydroxyalkanoates (PHAs) as an alternative to traditional plastics, it examines the potential of various food wastes as feedstock for microbial fermentation and PHA production. The upcycling of food waste including cheese whey, waste cooking oil, coffee waste, and animal fat is an innovative practice for food waste management. This approach not only mitigates environmental impacts but also contributes to sustainable development and economic growth. Downstream processing techniques for PHAs are discussed, highlighting their role in obtaining high-quality materials. The study also addresses sustainability considerations, emphasizing biodegradability and recycling, while acknowledging the challenges associated with this path.
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Affiliation(s)
- Shashi Kant Bhatia
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea; Institute for Ubiquitous Information Technology and Applications, Seoul 05029, Republic of Korea
| | - Anil Kumar Patel
- Institute of Aquatic Science and Technology, National Kaohsiung University of Science and Technology, Kaohsiung City, Taiwan
| | - Yung-Hun Yang
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea; Institute for Ubiquitous Information Technology and Applications, Seoul 05029, Republic of Korea.
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15
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Cruz-Romero CL, Chávez-Ramírez AU, Flores-Juárez CR, Arjona N, Álvarez-López A, del Bosque Plata L, Vallejo-Becerra V, Galindo-de-la-Rosa JDD. Biosynthesis of Polyhydroalkanoates Doped with Silver Nanoparticles Using Pseudomonas putida and Pseudomonas aeruginosa for Antibacterial Polymer Applications. Int J Mol Sci 2024; 25:8996. [PMID: 39201681 PMCID: PMC11354355 DOI: 10.3390/ijms25168996] [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: 07/19/2024] [Revised: 07/26/2024] [Accepted: 07/29/2024] [Indexed: 09/03/2024] Open
Abstract
In this study, the biosynthesis of polyhydroxyalkanoates (PHAs) was carried out using Pseudomonas putida and Pseudomonas aeruginosa. These PHAs were produced using reagent-grade glycerol and crude glycerol as the carbon sources. The objective was to compare the production of PHAs and to functionalize these polymers with silver nanoparticles to provide antibacterial properties for potential biomedical applications. The findings from the physical and chemical analyses confirmed the successful synthesis and extraction of PHAs, achieving comparable yields using both crude glycerol and reagent-grade glycerol as carbon sources across both strains. Approximately 16% higher PHAs production was obtained using Pseudomonas putida compared to Pseudomonas aeruginosa, and no significant difference was observed in the production rate of PHAs between the two carbon sources used, which means that crude glycerol could be utilized even though it has more impurities. Notably, PHAs functionalized with silver nanoparticles showed improved antibacterial effectiveness, especially those derived from reagent-grade glycerol and the Pseudomonas aeruginosa strain.
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Affiliation(s)
- Carmen Liliana Cruz-Romero
- Facultad de Ingeniería, División de Investigación y Posgrado, Centro Universitario Cerro de las Campanas, Universidad Autónoma de Querétaro, Querétaro, Qro. C.P. 76010, Mexico; (C.L.C.-R.); (A.Á.-L.)
| | - Abraham Ulises Chávez-Ramírez
- Centro de Investigación y Desarrollo Tecnológico en Electroquímica, Pedro Escobedo, Qro. C.P. 76703, Mexico; (A.U.C.-R.); (N.A.)
| | - Cyntia R. Flores-Juárez
- División Industrial Área de Nanotecnología, Universidad Tecnológica de Querétaro, Querétaro, Qro. C.P. 76148, Mexico;
| | - Noé Arjona
- Centro de Investigación y Desarrollo Tecnológico en Electroquímica, Pedro Escobedo, Qro. C.P. 76703, Mexico; (A.U.C.-R.); (N.A.)
| | - Alejandra Álvarez-López
- Facultad de Ingeniería, División de Investigación y Posgrado, Centro Universitario Cerro de las Campanas, Universidad Autónoma de Querétaro, Querétaro, Qro. C.P. 76010, Mexico; (C.L.C.-R.); (A.Á.-L.)
| | | | - Vanessa Vallejo-Becerra
- Facultad de Ingeniería, División de Investigación y Posgrado, Centro Universitario Cerro de las Campanas, Universidad Autónoma de Querétaro, Querétaro, Qro. C.P. 76010, Mexico; (C.L.C.-R.); (A.Á.-L.)
| | - Juan de Dios Galindo-de-la-Rosa
- Facultad de Ingeniería, División de Investigación y Posgrado, Centro Universitario Cerro de las Campanas, Universidad Autónoma de Querétaro, Querétaro, Qro. C.P. 76010, Mexico; (C.L.C.-R.); (A.Á.-L.)
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16
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Heidarian P, Aziz S, Halley PJ, McNally T, Peijs T, Vandi LJ, Varley RJ. Poly(3-Hydroxybutyrate- co-3-Hydroxyvalerate) Self-Reinforced Composites via Solvent-Induced Interfiber Welding of Nanofibers. Biomacromolecules 2024; 25:5039-5047. [PMID: 39041249 DOI: 10.1021/acs.biomac.4c00441] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 07/24/2024]
Abstract
In this study, we explore an approach to enhance the mechanical performance of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) by utilizing the self-reinforcing effect of β-phase-induced PHBV electrospun nanofiber mats. This involves electrospinning combined with low-temperature postspun vapor solvent interfiber welding. Scanning electron microscopy imaging confirmed fiber alignment, while XRD diffraction revealed the presence of both α and β crystalline phases under optimized electrospinning conditions. The resulting composite exhibited significant improvements in mechanical properties attributed to the formation of more perfectly structured α and β polymorphs and enhanced interfacial adhesion of electrospun nanofibers after vapor solvent treatment. This approach offers entirely recyclable and biodegradable materials, presenting the potential for a new family of sustainable bioplastics.
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Affiliation(s)
- Pejman Heidarian
- Carbon Nexus at the Institute for Frontier Materials, Deakin University, Geelong, Victoria 3216, Australia
| | - Shazed Aziz
- School of Chemical Engineering, University of Queensland, St Lucia 4072, Australia
| | - Peter J Halley
- School of Chemical Engineering, University of Queensland, St Lucia 4072, Australia
- Centre for Advanced Materials Processing and Manufacturing AMPAM, The University of Queensland, St Lucia 4072, Australia
| | - Tony McNally
- International Institute for Nanocomposite Manufacturing (IINM), University of Warwick, Coventry CV4 74L, U.K
| | - Ton Peijs
- Materials Engineering Centre, WMG, University of Warwick, Coventry CV4 74L, U.K
| | - Luigi-Jules Vandi
- School of Mechanical and Mining Engineering, University of Queensland, St Lucia 4072, Australia
| | - Russell J Varley
- Carbon Nexus at the Institute for Frontier Materials, Deakin University, Geelong, Victoria 3216, Australia
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17
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Mi CH, Qi XY, Zhou YW, Ding YW, Wei DX, Wang Y. Advances in medical polyesters for vascular tissue engineering. DISCOVER NANO 2024; 19:125. [PMID: 39115796 PMCID: PMC11310390 DOI: 10.1186/s11671-024-04073-x] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/03/2024] [Accepted: 07/25/2024] [Indexed: 08/11/2024]
Abstract
Blood vessels are highly dynamic and complex structures with a variety of physiological functions, including the transport of oxygen, nutrients, and metabolic wastes. Their normal functioning involves the close and coordinated cooperation of a variety of cells. However, adverse internal and external environmental factors can lead to vascular damage and the induction of various vascular diseases, including atherosclerosis and thrombosis. This can have serious consequences for patients, and there is an urgent need for innovative techniques to repair damaged blood vessels. Polyesters have been extensively researched and used in the treatment of vascular disease and repair of blood vessels due to their excellent mechanical properties, adjustable biodegradation time, and excellent biocompatibility. Given the high complexity of vascular tissues, it is still challenging to optimize the utilization of polyesters for repairing damaged blood vessels. Nevertheless, they have considerable potential for vascular tissue engineering in a range of applications. This summary reviews the physicochemical properties of polyhydroxyalkanoate (PHA), polycaprolactone (PCL), poly-lactic acid (PLA), and poly(lactide-co-glycolide) (PLGA), focusing on their unique applications in vascular tissue engineering. Polyesters can be prepared not only as 3D scaffolds to repair damage as an alternative to vascular grafts, but also in various forms such as microspheres, fibrous membranes, and nanoparticles to deliver drugs or bioactive ingredients to damaged vessels. Finally, it is anticipated that further developments in polyesters will occur in the near future, with the potential to facilitate the wider application of these materials in vascular tissue engineering.
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Affiliation(s)
- Chen-Hui Mi
- Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Medicine, Department of Life Sciences and Medicine, Northwest University, Xi'an, 710069, China
| | - Xin-Ya Qi
- Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Medicine, Department of Life Sciences and Medicine, Northwest University, Xi'an, 710069, China
| | - Yan-Wen Zhou
- Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Medicine, Department of Life Sciences and Medicine, Northwest University, Xi'an, 710069, China
| | - Yan-Wen Ding
- Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Medicine, Department of Life Sciences and Medicine, Northwest University, Xi'an, 710069, China
| | - Dai-Xu Wei
- Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, School of Medicine, Department of Life Sciences and Medicine, Northwest University, Xi'an, 710069, China.
- School of Clinical Medicine, Chengdu University, Chengdu, China.
- Shaanxi Key Laboratory for Carbon-Neutral Technology, Xi'an, 710069, China.
| | - Yong Wang
- Department of Interventional Radiology and Vascular Surgery, Second Affiliated Hospital of Hainan Medical University, Haikou, China.
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18
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Narayanasamy A, Patel SKS, Singh N, Rohit MV, Lee JK. Valorization of Algal Biomass to Produce Microbial Polyhydroxyalkanoates: Recent Updates, Challenges, and Perspectives. Polymers (Basel) 2024; 16:2227. [PMID: 39125253 PMCID: PMC11314723 DOI: 10.3390/polym16152227] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/22/2024] [Revised: 07/28/2024] [Accepted: 07/30/2024] [Indexed: 08/12/2024] Open
Abstract
Biopolymers are highly desirable alternatives to petrochemical-based plastics owing to their biodegradable nature. The production of bioplastics, such as polyhydroxyalkanoates (PHAs), has been widely reported using various bacterial cultures with substrates ranging from pure to biowaste-derived sugars. However, large-scale production and economic feasibility are major limiting factors. Now, using algal biomass for PHA production offers a potential solution to these challenges with a significant environmental benefit. Algae, with their unique ability to utilize carbon dioxide as a greenhouse gas (GHG) and wastewater as feed for growth, can produce value-added products in the process and, thereby, play a crucial role in promoting environmental sustainability. The sugar recovery efficiency from algal biomass is highly variable depending on pretreatment procedures due to inherent compositional variability among their cell walls. Additionally, the yields, composition, and properties of synthesized PHA vary significantly among various microbial PHA producers from algal-derived sugars. Therefore, the microalgal biomass pretreatments and synthesis of PHA copolymers still require considerable investigation to develop an efficient commercial-scale process. This review provides an overview of the microbial potential for PHA production from algal biomass and discusses strategies to enhance PHA production and its properties, focusing on managing GHGs and promoting a sustainable future.
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Affiliation(s)
- Anand Narayanasamy
- Bioconversion Technology Division, Sardar Patel Renewable Energy Research Institute, Vallabh Vidyanagar, Anand 388120, Gujarat, India; (A.N.); (N.S.); (M.V.R.)
| | - Sanjay K. S. Patel
- Department of Biotechnology, Hemvati Nandan Bahuguna Garhwal University (A Central University), Srinagar 246174, Uttarakhand, India;
- Department of Chemical Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
| | - Neha Singh
- Bioconversion Technology Division, Sardar Patel Renewable Energy Research Institute, Vallabh Vidyanagar, Anand 388120, Gujarat, India; (A.N.); (N.S.); (M.V.R.)
| | - M. V. Rohit
- Bioconversion Technology Division, Sardar Patel Renewable Energy Research Institute, Vallabh Vidyanagar, Anand 388120, Gujarat, India; (A.N.); (N.S.); (M.V.R.)
| | - Jung-Kul Lee
- Department of Chemical Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
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Sharifulden NSAN, Barrios Silva LV, Mandakhbayar NE, Shin SJ, Kim HW, Knowles JC, Nguyen LTB, Chau DYS. The biological and therapeutic assessment of a P(3HB-co-4HB)-bioactive glass-graphene composite biomaterial for tissue regeneration. J Biomed Mater Res B Appl Biomater 2024; 112:e35441. [PMID: 38923274 DOI: 10.1002/jbm.b.35441] [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: 02/29/2024] [Revised: 05/04/2024] [Accepted: 06/09/2024] [Indexed: 06/28/2024]
Abstract
An ideal wound dressing should create a healing environment that relieves pain, protects against infections, maintains moisture, removes debris, and speeds up wound closure and repair. However, conventional options like gauze often fall short in fulfilling these requirements, especially for chronic or nonhealing wounds. Hence there is a critical need for inventive formulations that offer efficient, cost-effective, and eco-friendly alternatives. This study focuses on assessing the innovative formulation based on a microbial-derived copolymer known as poly(3-hydroxybutyrate-co-4-hydroxybutyrate), P(3HB-co-4HB) bioactive glass and graphene particles, and exploring their biological response in vitro and in vivo-to find the best combination that promotes cell adhesion and enhances wound healing. The formulation optimized at concentration of bioactive glass (1 w/w%) and graphene (0.01 w/w%) showed accelerated degradation and enhanced blood vessel formation. Meanwhile biocompatibility was evaluated using murine osteoblasts, human dermal fibroblasts, and standard cell culture assays, demonstrating no adverse effects after 7 days of culture and well-regulated inflammatory kinetics. Whole thickness skin defect using mice indicated the feasibility of the biocomposites for a faster wound closure and reduced inflammation. Overall, this biocomposite appears promising as an ideal wound dressing material and positively influencing wound healing rates.
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Affiliation(s)
- Nik S A N Sharifulden
- Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, Royal Free Hospital, London, UK
| | - Lady V Barrios Silva
- Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, Royal Free Hospital, London, UK
| | - Nandin-Erdene Mandakhbayar
- Institute of Tissue Regeneration Engineering, Dankook University, Cheonan, South Korea
- Department of Biochemistry, School of Biomedicine, Mongolian National University of Medical Sciences, Ulaanbaatar, Mongolia
| | - Seong-Jin Shin
- Institute of Tissue Regeneration Engineering, Dankook University, Cheonan, South Korea
| | - Hae-Won Kim
- Institute of Tissue Regeneration Engineering, Dankook University, Cheonan, South Korea
- UCL Eastman-Korea Dental Medicine Innovation Centre, Dankook University, Cheonan, South Korea
- Department of Nanobiomedical Science, Dankook University, Cheonan, South Korea
- BK21 NBM Global Research Centre for Regenerative Medicine, Dankook University, Cheonan, South Korea
| | - Jonathan C Knowles
- Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, Royal Free Hospital, London, UK
- UCL Eastman-Korea Dental Medicine Innovation Centre, Dankook University, Cheonan, South Korea
- Department of Nanobiomedical Science, Dankook University, Cheonan, South Korea
- BK21 NBM Global Research Centre for Regenerative Medicine, Dankook University, Cheonan, South Korea
| | - Linh T B Nguyen
- Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, Royal Free Hospital, London, UK
| | - David Y S Chau
- Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, Royal Free Hospital, London, UK
- UCL Eastman-Korea Dental Medicine Innovation Centre, Dankook University, Cheonan, South Korea
- Department of Nanobiomedical Science, Dankook University, Cheonan, South Korea
- BK21 NBM Global Research Centre for Regenerative Medicine, Dankook University, Cheonan, South Korea
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20
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Kesharwani P, Alexander A, Shukla R, Jain S, Bisht A, Kumari K, Verma K, Sharma S. Tissue regeneration properties of hydrogels derived from biological macromolecules: A review. Int J Biol Macromol 2024; 271:132280. [PMID: 38744364 DOI: 10.1016/j.ijbiomac.2024.132280] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/13/2024] [Revised: 05/02/2024] [Accepted: 05/08/2024] [Indexed: 05/16/2024]
Abstract
The successful tissue engineering depends on the development of biologically active scaffolds that possess optimal characteristics to effectively support cellular functions, maintain structural integrity and aid in tissue regeneration. Hydrogels have emerged as promising candidates in tissue regeneration due to their resemblance to the natural extracellular matrix and their ability to support cell survival and proliferation. The integration of hydrogel scaffold into the polymer has a variable impact on the pseudo extracellular environment, fostering cell growth/repair. The modification in size, shape, surface morphology and porosity of hydrogel scaffolds has consequently paved the way for addressing diverse challenges in the tissue engineering process such as tissue architecture, vascularization and simultaneous seeding of multiple cells. The present review provides a comprehensive update on hydrogel production using natural and synthetic biomaterials and their underlying mechanisms. Furthermore, it delves into the application of hydrogel scaffolds in tissue engineering for cardiac tissues, cartilage tissue, adipose tissue, nerve tissue and bone tissue. Besides, the present article also highlights various clinical studies, patents, and the limitations associated with hydrogel-based scaffolds in recent times.
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Affiliation(s)
- Payal Kesharwani
- Department of Pharmacy, Banasthali Vidyapith, Banasthali, Rajasthan, India; Institute of Pharmacy, Ram-Eesh Institute of Vocational and Technical Education Greater Noida, India
| | - Amit Alexander
- Department of Pharmaceuticals, National Institute of Pharmaceutical Education and Research, Guwahati, Assam, India
| | - Rahul Shukla
- Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER)-Raebareli, Lucknow, Uttar Pradesh, India
| | - Smita Jain
- Department of Pharmacy, Banasthali Vidyapith, Banasthali, Rajasthan, India
| | - Akansha Bisht
- Department of Pharmacy, Banasthali Vidyapith, Banasthali, Rajasthan, India
| | - Kajal Kumari
- Department of Pharmacy, Banasthali Vidyapith, Banasthali, Rajasthan, India
| | - Kanika Verma
- Department of Pharmacy, Banasthali Vidyapith, Banasthali, Rajasthan, India
| | - Swapnil Sharma
- Department of Pharmacy, Banasthali Vidyapith, Banasthali, Rajasthan, India.
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21
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Park H, He H, Yan X, Liu X, Scrutton NS, Chen GQ. PHA is not just a bioplastic! Biotechnol Adv 2024; 71:108320. [PMID: 38272380 DOI: 10.1016/j.biotechadv.2024.108320] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/11/2023] [Revised: 01/21/2024] [Accepted: 01/22/2024] [Indexed: 01/27/2024]
Abstract
Polyhydroxyalkanoates (PHA) have evolved into versatile biopolymers, transcending their origins as mere bioplastics. This extensive review delves into the multifaceted landscape of PHA applications, shedding light on the diverse industries that have harnessed their potential. PHA has proven to be an invaluable eco-conscious option for packaging materials, finding use in films foams, paper coatings and even straws. In the textile industry, PHA offers a sustainable alternative, while its application as a carbon source for denitrification in wastewater treatment showcases its versatility in environmental remediation. In addition, PHA has made notable contributions to the medical and consumer sectors, with various roles ranging from 3D printing, tissue engineering implants, and cell growth matrices to drug delivery carriers, and cosmetic products. Through metabolic engineering efforts, PHA can be fine-tuned to align with the specific requirements of each industry, enabling the customization of material properties such as ductility, elasticity, thermal conductivity, and transparency. To unleash PHA's full potential, bridging the gap between research and commercial viability is paramount. Successful PHA production scale-up hinges on establishing direct supply chains to specific application domains, including packaging, food and beverage materials, medical devices, and agriculture. This review underscores that PHA's future rests on ongoing exploration across these industries and more, paving the way for PHA to supplant conventional plastics and foster a circular economy.
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Affiliation(s)
- Helen Park
- School of Life Sciences, Tsinghua University, Beijing 100084, China; EPSRC/BBSRC Future Biomanufacturing Research Hub, BBSRC Synthetic Biology Research Centre, SYNBIOCHEM, Manchester Institute of Biotechnology and Department of Chemistry, School of Natural Sciences, The University of Manchester, Manchester M1 7DN, UK
| | - Hongtao He
- School of Life Sciences, Tsinghua University, Beijing 100084, China
| | - Xu Yan
- School of Life Sciences, Tsinghua University, Beijing 100084, China
| | - Xu Liu
- PhaBuilder Biotech Co. Ltd., Shunyi District, Zhaoquan Ying, Beijing 101309, China
| | - Nigel S Scrutton
- EPSRC/BBSRC Future Biomanufacturing Research Hub, BBSRC Synthetic Biology Research Centre, SYNBIOCHEM, Manchester Institute of Biotechnology and Department of Chemistry, School of Natural Sciences, The University of Manchester, Manchester M1 7DN, UK
| | - Guo-Qiang Chen
- School of Life Sciences, Tsinghua University, Beijing 100084, China; Center for Synthetic and Systems Biology, Tsinghua University, Beijing 100084, China; Tsinghua-Peking Center for Life Sciences, Beijing, China; MOE Key Lab of Industrial Biocatalysis, Dept Chemical Engineering, Tsinghua University, Beijing 100084, China.
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22
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Anjana, Rawat S, Goswami S. Synergistic approach for enhanced production of polyhydroxybutyrate by Bacillus pseudomycoides SAS-B1: Effective utilization of glycerol and acrylic acid through fed-batch fermentation and its environmental impact assessment. Int J Biol Macromol 2024; 258:128764. [PMID: 38103666 DOI: 10.1016/j.ijbiomac.2023.128764] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/01/2023] [Revised: 12/07/2023] [Accepted: 12/11/2023] [Indexed: 12/19/2023]
Abstract
The continual rise in the consumption of petroleum-based synthetic polymers raised a significant environmental concern. Bacillus pseudomycoides SAS-B1 is a gram-positive rod-shaped halophilic bacterium capable of accumulating Polyhydroxybutyrate (PHB)-an intracellular biodegradable polymer. In the present study, the optimal conditions for cell cultivation in the seed media were developed. The optimal factors included a preservation age of 14 to 21 days (with 105 to 106 cells/mL), inoculum size of 0.1 % (w/v), 1 % (w/v) glucose, and growth temperature of 30 °C. The cells were then cultivated in a two-stage fermentation process utilizing glycerol and Corn Steep Liquor (CSL) as carbon and nitrogen sources, respectively. PHB yield was effectively increased from 2.01 to 9.21 g/L through intermittent feeding of glycerol and CSL, along with acrylic acid. FTIR, TGA, DSC, and XRD characterization studies were employed to enumerate the recovered PHB and determine its physicochemical properties. Additionally, the study assessed the cradle-to-gate Life Cycle Assessment (LCA) of PHB production, considering net CO2 generation and covering all major environmental impact categories. The production of 1000 kg of PHB resulted in lower stratospheric ozone depletion and comparatively reduced carbon dioxide emissions (2022.7 kg CO2 eq.) and terrestrial ecotoxicity (9.54 kg 1,4-DCB eq.) than typical petrochemical polymers.
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Affiliation(s)
- Anjana
- Division of Chemical Engineering, Centre of Innovative and Applied Bioprocessing, Knowledge City, Sector-81, Mohali, Punjab 140306, India; Department of Biotechnology, Regional Center for Biotechnology (RCB), Faridabad, Haryana 121001, India
| | - Shristhi Rawat
- Division of Chemical Engineering, Centre of Innovative and Applied Bioprocessing, Knowledge City, Sector-81, Mohali, Punjab 140306, India
| | - Saswata Goswami
- Division of Chemical Engineering, Centre of Innovative and Applied Bioprocessing, Knowledge City, Sector-81, Mohali, Punjab 140306, India; Department of Biotechnology, Regional Center for Biotechnology (RCB), Faridabad, Haryana 121001, India.
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23
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Durán-Lara EF, Rafael D, Andrade F, G OL, Vijayakumar S. Bacterial Polyhydroxyalkanoates-based Therapeutics-delivery Nano-systems. Curr Med Chem 2024; 31:5884-5897. [PMID: 37828676 DOI: 10.2174/0109298673268775231003111540] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/23/2023] [Revised: 08/05/2023] [Accepted: 09/01/2023] [Indexed: 10/14/2023]
Abstract
Microbial polyhydroxyalkanoates (PHAs) are bio-based aliphatic biopolyester produced by bacteria as an intracellular storage material of carbon and energy under stressed conditions. PHAs have been paid attention to due to their unique and impressive biological properties including high biodegradability, biocompatibility, low cytotoxicity, and different mechanical properties. Under this context, the development of drug-delivery nanosystems based on PHAs has been revealed to have numerous advantages compared with synthetic polymers that included biocompatibility, biodegradability, non-toxic, and low-cost production, among others. In this review article, we present the available state of the art of PHAs. Moreover, we discussed the potential benefits, weaknesses, and perspectives of PHAs to the develop drug delivery systems.
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Affiliation(s)
- Esteban F Durán-Lara
- Departamento de Microbiología, Facultad de Ciencias de la Salud, Universidad de Talca, Talca 3460000, Maule, Chile
- Bio & NanoMaterials Lab, Drug Delivery and Controlled Release, Universidad de Talca, Talca, 3460000, Maule, Chile
| | - Diana Rafael
- Drug Delivery and Targeting Group, Molecular Biology and Biochemistry Research Centre for Nanomedicine (CIBBIM-Nanomedicine), Vall d'Hebron Institut de Recerca, Universitat Autònoma de Barcelona, Barcelona, Spain
- Networking Research Centre for Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Instituto de Salud Carlos III, Madrid, Spain
| | - Fernanda Andrade
- Drug Delivery and Targeting Group, Molecular Biology and Biochemistry Research Centre for Nanomedicine (CIBBIM-Nanomedicine), Vall d'Hebron Institut de Recerca, Universitat Autònoma de Barcelona, Barcelona, Spain
- Networking Research Centre for Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Instituto de Salud Carlos III, Madrid, Spain
| | - Olga Lobos G
- Departamento de Microbiología, Facultad de Ciencias de la Salud, Universidad de Talca, Talca 3460000, Maule, Chile
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24
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Dubey AK, Mostafavi E. Biomaterials-mediated CRISPR/Cas9 delivery: recent challenges and opportunities in gene therapy. Front Chem 2023; 11:1259435. [PMID: 37841202 PMCID: PMC10568484 DOI: 10.3389/fchem.2023.1259435] [Citation(s) in RCA: 11] [Impact Index Per Article: 5.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/15/2023] [Accepted: 09/15/2023] [Indexed: 10/17/2023] Open
Abstract
The use of biomaterials in delivering CRISPR/Cas9 for gene therapy in infectious diseases holds tremendous potential. This innovative approach combines the advantages of CRISPR/Cas9 with the protective properties of biomaterials, enabling accurate and efficient gene editing while enhancing safety. Biomaterials play a vital role in shielding CRISPR/Cas9 components, such as lipid nanoparticles or viral vectors, from immunological processes and degradation, extending their effectiveness. By utilizing the flexibility of biomaterials, tailored systems can be designed to address specific genetic diseases, paving the way for personalized therapeutics. Furthermore, this delivery method offers promising avenues in combating viral illnesses by precisely modifying pathogen genomes, and reducing their pathogenicity. Biomaterials facilitate site-specific gene modifications, ensuring effective delivery to infected cells while minimizing off-target effects. However, challenges remain, including optimizing delivery efficiency, reducing off-target effects, ensuring long-term safety, and establishing scalable production techniques. Thorough research, pre-clinical investigations, and rigorous safety evaluations are imperative for successful translation from the laboratory to clinical applications. In this review, we discussed how CRISPR/Cas9 delivery using biomaterials revolutionizes gene therapy and infectious disease treatment, offering precise and safe editing capabilities with the potential to significantly improve human health and quality of life.
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Affiliation(s)
- Ankit Kumar Dubey
- Global Research and Publishing Foundation, New Delhi, India
- Institute of Scholars, Bengaluru, Karnataka, India
| | - Ebrahim Mostafavi
- Stanford Cardiovascular Institute, Stanford University School of Medicine, Stanford, CA, United States
- Department of Medicine, Stanford University School of Medicine, Stanford, CA, United States
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25
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Tyubaeva PM, Gasparyan KG, Fedotov AY, Lobzhanidze PV, Baranov OV, Egorov AA, Sirotinkin VP, Komlev VS, Olkhov AA. Development of Nonwoven Fibrous Materials Based on Poly-3-Hydroxybutyrate with a High Content of α-Tricalcium Phosphate. Polymers (Basel) 2023; 15:3167. [PMID: 37571064 PMCID: PMC10421182 DOI: 10.3390/polym15153167] [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: 06/30/2023] [Revised: 07/20/2023] [Accepted: 07/24/2023] [Indexed: 08/13/2023] Open
Abstract
α-tricalcium (α-TCP) phosphate is widely used as an osteoinductive biocompatible material, serving as an alternative to synthetic porous bone materials. The objective of this study is to obtain a highly filled fibrous nonwoven material composed of poly-3-hydroxybutyrate (PHB) and α-TCP and to investigate the morphology, structure, and properties of the composite obtained by the electrospinning method (ES). The addition of α-TCP had a significant effect on the supramolecular structure of the material, allowing it to control the crystallinity of the material, which was accompanied by changes in mechanical properties, FTIR spectra, and XRD curves. The obtained results open the way to the creation of new osteoconductive materials with a controlled release of the source of calcium into the living organism.
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Affiliation(s)
- Polina M. Tyubaeva
- Department of Physical Chemistry of Synthetic and Natural Polymer Compositions, Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 4 Kosygina Street, 119334 Moscow, Russia; (P.M.T.)
- Academic Department of Innovational Materials and Technologies Chemistry, Plekhanov Russian University of Economics, 36 Stremyanny Per., 117997 Moscow, Russia
| | - Kristina G. Gasparyan
- Department of Physical Chemistry of Synthetic and Natural Polymer Compositions, Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 4 Kosygina Street, 119334 Moscow, Russia; (P.M.T.)
- Academic Department of Innovational Materials and Technologies Chemistry, Plekhanov Russian University of Economics, 36 Stremyanny Per., 117997 Moscow, Russia
| | - Alexander Yu. Fedotov
- A.A. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Leninsky Prospect 49, 119334 Moscow, Russia (V.P.S.)
| | - Pavel V. Lobzhanidze
- A.A. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Leninsky Prospect 49, 119334 Moscow, Russia (V.P.S.)
| | - Oleg V. Baranov
- A.A. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Leninsky Prospect 49, 119334 Moscow, Russia (V.P.S.)
| | - Alexey A. Egorov
- A.A. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Leninsky Prospect 49, 119334 Moscow, Russia (V.P.S.)
| | - Vladimir P. Sirotinkin
- A.A. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Leninsky Prospect 49, 119334 Moscow, Russia (V.P.S.)
| | - Vladimir S. Komlev
- A.A. Baikov Institute of Metallurgy and Materials Science, Russian Academy of Sciences, Leninsky Prospect 49, 119334 Moscow, Russia (V.P.S.)
| | - Anatoly A. Olkhov
- Department of Physical Chemistry of Synthetic and Natural Polymer Compositions, Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 4 Kosygina Street, 119334 Moscow, Russia; (P.M.T.)
- Academic Department of Innovational Materials and Technologies Chemistry, Plekhanov Russian University of Economics, 36 Stremyanny Per., 117997 Moscow, Russia
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26
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Kalia VC, Patel SKS, Lee JK. Exploiting Polyhydroxyalkanoates for Biomedical Applications. Polymers (Basel) 2023; 15:polym15081937. [PMID: 37112084 PMCID: PMC10144186 DOI: 10.3390/polym15081937] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/21/2023] [Revised: 04/15/2023] [Accepted: 04/18/2023] [Indexed: 04/29/2023] Open
Abstract
Polyhydroxyalkanoates (PHA) are biodegradable plastic. Numerous bacteria produce PHAs under environmental stress conditions, such as excess carbon-rich organic matter and limitations of other nutritional elements such as potassium, magnesium, oxygen, phosphorus, and nitrogen. In addition to having physicochemical properties similar to fossil-fuel-based plastics, PHAs have unique features that make them ideal for medical devices, such as easy sterilization without damaging the material itself and easy dissolution following use. PHAs can replace traditional plastic materials used in the biomedical sector. PHAs can be used in a variety of biomedical applications, including medical devices, implants, drug delivery devices, wound dressings, artificial ligaments and tendons, and bone grafts. Unlike plastics, PHAs are not manufactured from petroleum products or fossil fuels and are, therefore, environment-friendly. In this review, a recent overview of applications of PHAs with special emphasis on biomedical sectors, including drug delivery, wound healing, tissue engineering, and biocontrols, are discussed.
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Affiliation(s)
- Vipin Chandra Kalia
- Department of Chemical Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
| | - Sanjay K S Patel
- Department of Chemical Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
| | - Jung-Kul Lee
- Department of Chemical Engineering, Konkuk University, 120 Neungdong-ro, Gwangjin-gu, Seoul 05029, Republic of Korea
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27
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Chen L, Wei L, Su X, Qin L, Xu Z, Huang X, Chen H, Hu N. Preparation and Characterization of Biomimetic Functional Scaffold with Gradient Structure for Osteochondral Defect Repair. Bioengineering (Basel) 2023; 10:bioengineering10020213. [PMID: 36829707 PMCID: PMC9952804 DOI: 10.3390/bioengineering10020213] [Citation(s) in RCA: 8] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/30/2022] [Revised: 01/20/2023] [Accepted: 02/03/2023] [Indexed: 02/08/2023] Open
Abstract
Osteochondral (OC) defects cannot adequately repair themselves due to their sophisticated layered structure and lack of blood supply in cartilage. Although therapeutic interventions are reaching an advanced stage, current clinical therapies to repair defects are in their infancy. Among the possible therapies, OC tissue engineering has shown considerable promise, and multiple approaches utilizing scaffolds, cells, and bioactive factors have been pursued. The most recent trend in OC tissue engineering has been to design gradient scaffolds using different materials and construction strategies (such as bi-layered, multi-layered, and continuous gradient structures) to mimic the physiological and mechanical properties of OC tissues while further enabling OC repair. This review focuses specifically on design and construction strategies for gradient scaffolds and their role in the successful engineering of OC tissues. The current dilemmas in the field of OC defect repair and the efforts of tissue engineering to address these challenges were reviewed. In addition, the advantages and limitations of the typical fabrication techniques for gradient scaffolds were discussed, with examples of recent studies summarizing the future prospects for integrated gradient scaffold construction. This updated and enlightening review could provide insights into our current understanding of gradient scaffolds in OC tissue engineering.
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Affiliation(s)
| | | | | | | | | | - Xiao Huang
- Correspondence: (X.H.); (H.C.); (N.H.); Tel.: +86-023-89011202 (X.H. & H.C. & N.H.)
| | - Hong Chen
- Correspondence: (X.H.); (H.C.); (N.H.); Tel.: +86-023-89011202 (X.H. & H.C. & N.H.)
| | - Ning Hu
- Correspondence: (X.H.); (H.C.); (N.H.); Tel.: +86-023-89011202 (X.H. & H.C. & N.H.)
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28
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Máčalová D, Janalíková M, Sedlaříková J, Rektoříková I, Koutný M, Pleva P. Genotypic and Phenotypic Detection of Polyhydroxyalkanoate Production in Bacterial Isolates from Food. Int J Mol Sci 2023; 24:ijms24021250. [PMID: 36674766 PMCID: PMC9864133 DOI: 10.3390/ijms24021250] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2022] [Revised: 12/22/2022] [Accepted: 01/05/2023] [Indexed: 01/11/2023] Open
Abstract
Polyhydroxyalkanoates (PHAs) are widely used in medical and potentially in other applications due to their biocompatibility and biodegradability. Understanding PHA biosynthetic pathways may lead to the detection of appropriate conditions (substrates) for producing a particular PHA type by a specific microbial strain. The aim of this study was to establish a method enabling potentially interesting PHA bacterial producers to be found. In the study, all four classes of PHA synthases and other genes involved in PHA formation (fabG, phaA, phaB, phaG, and phaJ) were detected by PCR in 64 bacterial collection strains and food isolates. Acinetobacter, Bacillus, Cupriavidus, Escherichia, Klebsiella, Lelliottia, Lysinibacillus, Mammaliicoccus, Oceanobacillus, Pantoea, Peribacillus, Priestia, Pseudomonas, Rahnella, Staphylococcus, and Stenotrophomonas genera were found among these strains. Fructose, glucose, sunflower oil, and propionic acid were utilized as carbon sources and PHA production was detected by Sudan black staining, Nile blue staining, and FTIR methods. The class I synthase and phaA genes were the most frequently found, indicating the strains' ability to synthesize PHA from carbohydrates. Among the tested bacterial strains, the Pseudomonas genus was identified as able to utilize all tested carbon sources. The Pseudomonas extremorientalis strain was determined as a prospect for biotechnology applications.
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Affiliation(s)
- Daniela Máčalová
- Department of Environmental Protection Engineering, Faculty of Technology, Tomas Bata University in Zlin, 275 Vavreckova, 76001 Zlin, Czech Republic
| | - Magda Janalíková
- Department of Environmental Protection Engineering, Faculty of Technology, Tomas Bata University in Zlin, 275 Vavreckova, 76001 Zlin, Czech Republic
| | - Jana Sedlaříková
- Department of Fat, Surfactant and Cosmetics Technology, Faculty of Technology, Tomas Bata University in Zlin, 275 Vavreckova, 76001 Zlin, Czech Republic
| | - Iveta Rektoříková
- Department of Environmental Protection Engineering, Faculty of Technology, Tomas Bata University in Zlin, 275 Vavreckova, 76001 Zlin, Czech Republic
| | - Marek Koutný
- Department of Environmental Protection Engineering, Faculty of Technology, Tomas Bata University in Zlin, 275 Vavreckova, 76001 Zlin, Czech Republic
| | - Pavel Pleva
- Department of Environmental Protection Engineering, Faculty of Technology, Tomas Bata University in Zlin, 275 Vavreckova, 76001 Zlin, Czech Republic
- Correspondence:
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29
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Ene N, Savoiu VG, Spiridon M, Paraschiv CI, Vamanu E. The General Composition of Polyhydroxyalkanoates and Factors that Influence their Production and Biosynthesis. Curr Pharm Des 2023; 29:3089-3102. [PMID: 38099526 DOI: 10.2174/0113816128263175231102061920] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2023] [Accepted: 09/26/2023] [Indexed: 01/05/2024]
Abstract
Polyhydroxyalkanoates (PHAs) have been a current research topic for many years. PHAs are biopolymers produced by bacteria under unfavorable growth conditions. They are biomaterials that exhibit a variety of properties, including biocompatibility, biodegradability, and high mechanical strength, making them suitable for future applications. This review aimed to provide general information on PHAs, such as their structure, classification, and parameters that affect the production process. In addition, the most commonly used bacterial strains that produce PHAs are highlighted, and details are provided on the type of carbon source used and how to optimize the parameters for bioprocesses. PHAs present a challenge to researchers because a variety of parameters affect biosynthesis, including the variety of carbon sources, bacterial strains, and culture media. Nevertheless, PHAs represent an opportunity to replace plastics, because they can be produced quickly and at a relatively low cost. With growing environmental concerns and declining oil reserves, polyhydroxyalkanoates are a potential replacement for nonbiodegradable polymers. Therefore, the study of PHA production remains a hot topic, as many substrates can be used as carbon sources. Both researchers and industry are interested in facilitating the production, commercialization, and application of PHAs as potential replacements for nonbiodegradable polymers. The fact that they are biocompatible, environmentally biodegradable, and adaptable makes PHAs one of the most important materials available in the market. They are preferred in various industries, such as agriculture (for bioremediation of oil-polluted sites, minimizing the toxicity of pollutants, and environmental impact) or medicine (as medical devices). The various bioprocess technologies mentioned earlier will be further investigated, such as the carbon source (to obtain a biopolymer with the lowest possible cost, such as glucose, various fatty acids, and especially renewable sources), pretreatment of the substrate (to increase the availability of the carbon source), and supplementation of the growth environment with different substances and minerals). Consequently, the study of PHA production remains a current topic because many substrates can be used as carbon sources. Obtaining PHA from renewable substrates (waste oil, coffee grounds, plant husks, etc.) contributes significantly to reducing PHA costs. Therefore, in this review, pure bacterial cultures (Bacillus megaterium, Ralstonia eutropha, Cupriavidus necator, and Pseudomonas putida) have been investigated for their potential to utilize by-products as cheap feedstocks. The advantage of these bioprocesses is that a significant amount of PHA can be obtained using renewable carbon sources. The main disadvantage is that the chemical structure of the obtained biopolymer cannot be determined in advance, as is the case with bioprocesses using a conventional carbon source. Polyhydroxyalkanoates are materials that can be used in many fields, such as the medical field (skin grafts, implantable medical devices, scaffolds, drug-controlled release devices), agriculture (for polluted water cleaning), cosmetics and food (biodegradable packaging, gentle biosurfactants with suitable skin for cosmetics), and industry (production of biodegradable biopolymers that replace conventional plastic). Nonetheless, PHA biopolymers continue to be researched and improved and play an important role in various industrial sectors. The properties of this material allow its use as a biodegradable material in the cosmetics industry (for packaging), in the production of biodegradable plastics, or in biomedical engineering, as various prostheses or implantable scaffolds.
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Affiliation(s)
- Nicoleta Ene
- Department of Industrial Biotechnology, Faculty of Biotechnology, University of Agronomical Sciences and Veterinary Medicine, Bucharest, Romania
- Department of Pharmacology, National Institute for Chemical Pharmaceutical Research and Development- ICCF, Vitan Avenue 112, Bucharest 031299, Romania
| | - Valeria Gabriela Savoiu
- Department of Biotechnology, National Institute For Chemical Pharmaceutical Research and Development, Bucharest 031299, Romania
| | - Maria Spiridon
- Department of Biotechnology, National Institute For Chemical Pharmaceutical Research and Development, Bucharest 031299, Romania
| | - Catalina Ileana Paraschiv
- Department of Chemistry, National Institute for Chemical Pharmaceutical Research and Development, Bucharest 031299, Romania
| | - Emanuel Vamanu
- Department of Industrial Biotechnology, Faculty of Biotechnology, University of Agronomical Sciences and Veterinary Medicine, Bucharest, Romania
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30
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Bhatia SK, Gurav R, Kim B, Kim S, Cho DH, Jung H, Kim YG, Kim JS, Yang YH. Coproduction of exopolysaccharide and polyhydroxyalkanoates from Sphingobium yanoikuyae BBL01 using biochar pretreated plant biomass hydrolysate. BIORESOURCE TECHNOLOGY 2022; 361:127753. [PMID: 35944863 DOI: 10.1016/j.biortech.2022.127753] [Citation(s) in RCA: 11] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/07/2022] [Revised: 08/02/2022] [Accepted: 08/03/2022] [Indexed: 06/15/2023]
Abstract
Sphingobium yanoikuyae BBL01 can produce exopolysaccharides (EPS) and polyhydroxyalkanoates (PHAs). The effect of side products (furfural, hydroxymethylfurfural (HMF), vanillin, and acetate) produced during pretreatment of biomass was evaluated on S. yanoikuyae BBL01. It was observed that a certain concentration range (0.01-0.03 %) of these compounds can improve growth, EPS production, and polyhydroxybutyrate (PHB) accumulation. The addition of HMF increases glucose and xylose utilization while other side products have a negative effect. The C/N of 5 favors EPS production (3.24 ± 0.05 g/L), while a higher C/N ratio of 30 promotes PHB accumulation (38.7 ± 0.08 % w/w), when commercial sugar is used as a carbon source. Pine biomass-derived biochar was able to remove 40 ± 2.1 % of total phenolic. Various biomass hydrolysates were evaluated and the use of detoxified pine biomass hydrolysate (DPH) as a carbon source resulted in the higher coproduction of EPS (2.83 ± 0.03 g/L) and PHB (40.8 ± 2.4 % w/w).
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Affiliation(s)
- Shashi Kant Bhatia
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea; Institute for Ubiquitous Information Technology and Applications, Seoul 05029, Republic of Korea
| | - Ranjit Gurav
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea
| | - Byungchan Kim
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea
| | - Suhyun Kim
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea
| | - Do-Hyun Cho
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea
| | - Heeju Jung
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea
| | - Yun-Gon Kim
- Department of Chemical Engineering, Soongsil University, Seoul 06978, Republic of Korea
| | - Jae-Seok Kim
- Department of Laboratory Medicine, Kangdong Sacred Heart Hospital, Hallym University College of Medicine, Seoul 05355, Republic of Korea
| | - Yung-Hun Yang
- Department of Biological Engineering, College of Engineering, Konkuk University, Seoul 05029, Republic of Korea; Institute for Ubiquitous Information Technology and Applications, Seoul 05029, Republic of Korea.
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Biosynthesis of P(3HB-co-3HHx) Copolymers by a Newly Engineered Strain of Cupriavidus necator PHB−4/pBBR_CnPro-phaCRp for Skin Tissue Engineering Application. Polymers (Basel) 2022; 14:polym14194074. [PMID: 36236022 PMCID: PMC9570888 DOI: 10.3390/polym14194074] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/31/2022] [Revised: 09/14/2022] [Accepted: 09/18/2022] [Indexed: 11/17/2022] Open
Abstract
Polyhydroxyalkanoates (PHAs) are biodegradable polymers synthesized by certain bacteria and archaea with functions comparable to conventional plastics. Previously, our research group reported a newly PHA-producing bacterial strain, Rhodococcus pyridinivorans BSRT1-1, from the soil in Thailand. However, this strain’s PHA synthase (phaCRp) gene has not yet been characterized. Thus, this study aims to synthesize PHA using a newly engineered bacterial strain, Cupriavidus necator PHB−4/pBBR_CnPro-phaCRp, which harbors the phaCRp from strain BSRT1-1, and characterize the properties of PHA for skin tissue engineering application. To the best of our knowledge, this is the first study on the characterization of the PhaC from R. pyridinivorans species. The results demonstrated that the expression of the phaCRp in C. necator PHB−4 had developed in PHA production up to 3.1 ± 0.3 g/L when using 10 g/L of crude palm kernel oil (CPKO) as a sole carbon source. Interestingly, the engineered strain produced a 3-hydroxybutyrate (3HB) with 2 mol% of 3-hydroxyhexanoate (3HHx) monomer without adding precursor substrates. In addition, the 70 L stirrer bioreactor improved P(3HB-co-2 mol% 3HHx) yield 1.4-fold over the flask scale without altering monomer composition. Furthermore, the characterization of copolymer properties showed that this copolymer is promising for skin tissue engineering applications.
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Demcisakova Z, Luptakova L, Tirpakova Z, Kvasilova A, Medvecky L, De Spiegelaere W, Petrovova E. Evaluation of Angiogenesis in an Acellular Porous Biomaterial Based on Polyhydroxybutyrate and Chitosan Using the Chicken Ex Ovo Chorioallantoic Membrane Model. Cancers (Basel) 2022; 14:cancers14174194. [PMID: 36077732 PMCID: PMC9454696 DOI: 10.3390/cancers14174194] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2022] [Revised: 08/25/2022] [Accepted: 08/26/2022] [Indexed: 12/04/2022] Open
Abstract
Simple Summary The chorioallantoic membrane (CAM) is an avian extraembryonic membrane widely used as an experimental assay to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the CAM has become popular in scientific studies focused on the use of its potential for the study of biocompatibility of materials for regenerative strategies and tissue engineering applications. Great research efforts are being made to develop innovative biomaterials able to treat hard tissue defects, including diseases such as a bone cancer. In this article, we describe an approach to detect the formation of blood vessels inside the porous acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold using the CAM assay as an in vivo alternative animal model, including macroscopic, histological, immunohistochemical, and molecular evaluation of the biocompatibility. Abstract The chorioallantoic membrane (CAM) is a highly vascularized avian extraembryonic membrane widely used as an in vivo model to study angiogenesis and its inhibition in response to tissues, cells, or soluble factors. In recent years, the use of CAM has become an integral part of the biocompatibility testing process for developing biomaterials intended for regenerative strategies and tissue engineering applications. In this study, we used the chicken ex ovo CAM assay to investigate the angiogenic potential of innovative acellular biopolymer polyhydroxybutyrate/chitosan (PHB/CHIT) scaffold, which is intended for the treatment of hard tissue defects, depending on treatment with pro- and anti-angiogenic substances. On embryonic day (ED) 7, the experimental biomaterials were placed on the CAM alone or soaked in vascular endothelial growth factor (VEGF-A), saline solution (PHY), or tyrosine kinase inhibitor (SU5402). After 72 h, the formation of vessels was analyzed in the surrounding area of the scaffold and inside the pores of the implants, using markers of embryonic endothelium (WGA, SNA), myofibroblasts (α-SMA), and macrophages (KUL-01). The morphological and histochemical analysis showed strong angiogenic potential of untreated scaffolds without additional effect of the angiogenic factor, VEGF-A. The lowest angiogenic potential was observed in scaffolds soaked with SU5402. Gene expression of pro-angiogenic growth factors, i.e., VEGF-A, ANG-2, and VE-CAD, was upregulated in untreated scaffolds after 72 h, indicating a pro-angiogenic environment. We concluded that the PHB/CHIT has a strong endogenous angiogenic potential and could be promising biomaterial for the treatment of hard tissue defects.
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Affiliation(s)
- Zuzana Demcisakova
- Department of Morphological Disciplines, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia
- Correspondence: (Z.D.); (E.P.)
| | - Lenka Luptakova
- Department of Biology and Physiology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia
| | - Zuzana Tirpakova
- Department of Biology and Physiology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia
| | - Alena Kvasilova
- Institute of Anatomy, Charles University, U Nemocnice 3, 12800 Prague, Czech Republic
| | - Lubomir Medvecky
- Department of Morphological Disciplines, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia
- Institute of Materials Research, The Slovak Academy of Sciences, Watsonova 1935/47, 04001 Kosice, Slovakia
| | - Ward De Spiegelaere
- Laboratory of Veterinary Morphology, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium
| | - Eva Petrovova
- Department of Morphological Disciplines, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 04181 Kosice, Slovakia
- Correspondence: (Z.D.); (E.P.)
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