title: A novel collagen scaffold supports human osteogenesis--applications for bone tissue engineering.
abstract: Collagen glycosaminoglycan (CG) scaffolds have been clinically approved as an application for skin regeneration. The goal of this study has been to examine whether a CG scaffold is a suitable biomaterial for generating human bone tissue. Specifically, we have asked the following questions: (1) can the scaffold support human osteoblast growth and differentiation and (2) how might recombinant human transforming growth factor-beta (TGF-beta(1)) enhance long-term in vitro bone formation? We show human osteoblast attachment, infiltration and uniform distribution throughout the construct, reaching the centre within 14 days of seeding. We have identified the fully differentiated osteoblast phenotype categorised by the temporal expression of alkaline phosphatase, collagen type 1, osteonectin, bone sialo protein, biglycan and osteocalcin. Mineralised bone formation has been identified at 35 days post-seeding by using von Kossa and Alizarin S Red staining. Both gene expression and mineral staining suggest the benefit of introducing an initial high treatment of TGF-beta(1) (10 ng/ml) followed by a low continuous treatment (0.2 ng/ml) to enhance human osteogenesis on the scaffold. Osteogenesis coincides with a reduction in scaffold size and shape (up to 70% that of original). A notable finding is core degradation at the centre of the tissue-engineered construct after 49 days of culture. This is not observed at earlier time points. Therefore, a maximum of 35 days in culture is appropriate for in vitro studies of these scaffolds. We conclude that the CG scaffold shows excellent potential as a biomaterial for human bone tissue engineering.
authors.affiliation: Division of Biology, Royal College of Surgeons in Ireland, 123 St. Stephen's Green, Dublin 2, Ireland. mkeogh@rcsi.ie
authors.forename: Michael B
authors.initials: MB
authors.lastname: Keogh
authors.forename: Fergal J
authors.initials: FJ
authors.lastname: O' Brien
authors.forename: Jacqueline S
authors.initials: JS
authors.lastname: Daly
mesh_terms: D001672:Biocompatible Materials / Q000627:therapeutic use*; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D001847:Bone Diseases / Q000628:therapy; D001861:Bone Regeneration / Q000187:drug effects / Q000502:physiology*; D002113:Calcification, Physiologic / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003094:Collagen / Q000627:therapeutic use*; D004305:Dose-Response Relationship, Drug; D006025:Glycosaminoglycans / Q000627:therapeutic use; D006801:Humans; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D010012:Osteogenesis / Q000187:drug effects / Q000502:physiology*; D010641:Phenotype; D013194:Staining and Labeling; D013997:Time Factors; D046509:Tissue Culture Techniques; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology / Q000627:therapeutic use
chemical_list: D001672:Biocompatible Materials; D015415:Biomarkers; D006025:Glycosaminoglycans; D053773:Transforming Growth Factor beta1; D003094:Collagen