Translational Research for Orthopedic Bone Graft Development
Designing biomaterials for bone-substitute applications is still a challenge regarding the natural complex structure of hard tissues. Aiming at bone regeneration applications, scaffolds based on natural collagen and synthetic nanohydroxyapatite were developed, and they showed adequate mechanical and...
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| Format: | Article |
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MDPI AG
2021-07-01
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| Online Access: | https://www.mdpi.com/1996-1944/14/15/4130 |
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| author | Maria J. C. Vilela Bruno J. A. Colaço José Ventura Fernando J. M. Monteiro Christiane L. Salgado |
| author_facet | Maria J. C. Vilela Bruno J. A. Colaço José Ventura Fernando J. M. Monteiro Christiane L. Salgado |
| author_sort | Maria J. C. Vilela |
| collection | DOAJ |
| description | Designing biomaterials for bone-substitute applications is still a challenge regarding the natural complex structure of hard tissues. Aiming at bone regeneration applications, scaffolds based on natural collagen and synthetic nanohydroxyapatite were developed, and they showed adequate mechanical and biological properties. The objective of this work was to perform and evaluate a scaled-up production process of this porous biocomposite scaffold, which promotes bone regeneration and works as a barrier for both fibrosis and the proliferation of scar tissue. The material was produced using a prototype bioreactor at an industrial scale, instead of laboratory production at the bench, in order to produce an appropriate medical device for the orthopedic market. Prototypes were produced in porous membranes that were e-beam irradiated (the sterilization process) and then analysed by scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), dynamic mechanical analysis (DMA), cytotoxicity tests with mice fibroblasts (L929), human osteoblast-like cells (MG63) and human MSC osteogenic differentiation (HBMSC) with alkaline phosphatase (ALP) activity and qPCR for osteogenic gene expression. The prototypes were also implanted into critical-size bone defects (rabbits’ tibia) for 5 and 15 weeks, and after that were analysed by microCT and histology. The tests performed for the physical characterization of the materials showed the ability of the scaffolds to absorb and retain water-based solvents, as well as adequate mechanical resistance and viscoelastic properties. The cryogels had a heteroporous morphology with microporosity and macroporosity, which are essential conditions for the interaction between the cells and materials, and which consequently promote bone regeneration. Regarding the biological studies, all of the studied cryogels were non-cytotoxic by direct or indirect contact with cells. In fact, the scaffolds promoted the proliferation of the human MSCs, as well as the expression of the osteoblastic phenotype (osteogenic differentiation). The in vivo results showed bone tissue ingrowth and the materials’ degradation, filling the critical bone defect after 15 weeks. Before and after irradiation, the studied scaffolds showed similar properties when compared to the results published in the literature. In conclusion, the material production process upscaling was optimized and the obtained prototypes showed reproducible properties relative to the bench development, and should be able to be commercialized. Therefore, it was a successful effort to harness knowledge from the basic sciences to produce a new biomedical device and enhance human health and wellbeing. |
| first_indexed | 2024-03-10T09:12:38Z |
| format | Article |
| id | doaj.art-040ff1e8681d46edba59863dfea85973 |
| institution | Directory Open Access Journal |
| issn | 1996-1944 |
| language | English |
| last_indexed | 2024-03-10T09:12:38Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Materials |
| spelling | doaj.art-040ff1e8681d46edba59863dfea859732023-11-22T05:52:10ZengMDPI AGMaterials1996-19442021-07-011415413010.3390/ma14154130Translational Research for Orthopedic Bone Graft DevelopmentMaria J. C. Vilela0Bruno J. A. Colaço1José Ventura2Fernando J. M. Monteiro3Christiane L. Salgado4Instituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, 4200-135 Porto, PortugalDepartment of Animal Science, CECAV—Animal and Veterinary Research Centre UTAD, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, PortugalArtur Salgado S.A., 4250-288 Porto, PortugalInstituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, 4200-135 Porto, PortugalInstituto de Investigação e Inovação em Saúde (i3S), Universidade do Porto, 4200-135 Porto, PortugalDesigning biomaterials for bone-substitute applications is still a challenge regarding the natural complex structure of hard tissues. Aiming at bone regeneration applications, scaffolds based on natural collagen and synthetic nanohydroxyapatite were developed, and they showed adequate mechanical and biological properties. The objective of this work was to perform and evaluate a scaled-up production process of this porous biocomposite scaffold, which promotes bone regeneration and works as a barrier for both fibrosis and the proliferation of scar tissue. The material was produced using a prototype bioreactor at an industrial scale, instead of laboratory production at the bench, in order to produce an appropriate medical device for the orthopedic market. Prototypes were produced in porous membranes that were e-beam irradiated (the sterilization process) and then analysed by scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM), dynamic mechanical analysis (DMA), cytotoxicity tests with mice fibroblasts (L929), human osteoblast-like cells (MG63) and human MSC osteogenic differentiation (HBMSC) with alkaline phosphatase (ALP) activity and qPCR for osteogenic gene expression. The prototypes were also implanted into critical-size bone defects (rabbits’ tibia) for 5 and 15 weeks, and after that were analysed by microCT and histology. The tests performed for the physical characterization of the materials showed the ability of the scaffolds to absorb and retain water-based solvents, as well as adequate mechanical resistance and viscoelastic properties. The cryogels had a heteroporous morphology with microporosity and macroporosity, which are essential conditions for the interaction between the cells and materials, and which consequently promote bone regeneration. Regarding the biological studies, all of the studied cryogels were non-cytotoxic by direct or indirect contact with cells. In fact, the scaffolds promoted the proliferation of the human MSCs, as well as the expression of the osteoblastic phenotype (osteogenic differentiation). The in vivo results showed bone tissue ingrowth and the materials’ degradation, filling the critical bone defect after 15 weeks. Before and after irradiation, the studied scaffolds showed similar properties when compared to the results published in the literature. In conclusion, the material production process upscaling was optimized and the obtained prototypes showed reproducible properties relative to the bench development, and should be able to be commercialized. Therefore, it was a successful effort to harness knowledge from the basic sciences to produce a new biomedical device and enhance human health and wellbeing.https://www.mdpi.com/1996-1944/14/15/4130collagennanohydroxyapatitescaffoldbone regenerationbiomaterials |
| spellingShingle | Maria J. C. Vilela Bruno J. A. Colaço José Ventura Fernando J. M. Monteiro Christiane L. Salgado Translational Research for Orthopedic Bone Graft Development Materials collagen nanohydroxyapatite scaffold bone regeneration biomaterials |
| title | Translational Research for Orthopedic Bone Graft Development |
| title_full | Translational Research for Orthopedic Bone Graft Development |
| title_fullStr | Translational Research for Orthopedic Bone Graft Development |
| title_full_unstemmed | Translational Research for Orthopedic Bone Graft Development |
| title_short | Translational Research for Orthopedic Bone Graft Development |
| title_sort | translational research for orthopedic bone graft development |
| topic | collagen nanohydroxyapatite scaffold bone regeneration biomaterials |
| url | https://www.mdpi.com/1996-1944/14/15/4130 |
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