title: In vitro generation of a multilayered osteochondral construct with an osteochondral interface using rabbit bone marrow stromal cells and a silk peptide-based scaffold.
abstract: Tissue engineering of a biological osteochondral multilayered construct with a cartilage-interface subchondral bone layer is a key challenge. This study presented a rabbit bone marrow stromal cell (BMSC)/silk fibroin scaffold-based co-culture approach to generate tissue-engineered osteochondral grafts with an interface. BMSC-seeded scaffolds were first cultured separately in osteogenic and chondrogenic stimulation media. The two differentiated pieces were then combined using an RADA self-assembling peptide and subsequently co-cultured. Gene expression, histological and biochemical analyses were used to evaluate the multilayered structure of the osteochondral graft. A complete osteochondral construct with a cartilage-subchondral bone interface was regenerated and BMSCs were used as the only cell source for the osteochondral construct and interface regeneration. Furthermore, in the intermediate region of co-cultured samples, hypertrophic chondrogenic gene markers type X collagen and MMP-13 were found on both chondrogenic and osteogenic section edges after co-culture. However, significant differences gene expression profile were found in distinct zones of the construct during co-culture and the section in the intermediate region had significantly higher hypertrophic chondrocyte gene expression. Biochemical analyses and histology results further supported this observation. This study showed that specific stimulation from osteogenic and chondrogenic BMSCs affected each other in this co-culture system and induced the formation of an osteochondral interface. Moreover, this system provided a possible approach for generating multilayered osteochondral constructs.
authors: National University of Singapore, Department of Bioengineering, Singapore, Singapore.
authors: Kelei
authors: K
authors: Chen
authors: National University of Singapore, Department of Orthopaedic Surgery, Singapore, Singapore.
authors: Pujiang
authors: P
authors: Shi
authors: National University of Singapore, Department of Bioengineering, Singapore, Singapore.
authors: Thomas Kok Hiong
authors: TK
authors: Teh
authors: National University of Singapore, Department of Bioengineering, Singapore, Singapore.|National University of Singapore, Department of Mechanical Engineering, Singapore, Singapore.
authors: Siew Lok
authors: SL
authors: Toh
authors: National University of Singapore, Department of Bioengineering, Singapore, Singapore.|National University of Singapore, Department of Orthopaedic Surgery, Singapore, Singapore.
authors: James Ch
authors: JCh
authors: Goh
mesh_terms: D000818:Animals; D012831:Bombyx; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D020219:Chondrogenesis / Q000187:drug effects; D018920:Coculture Techniques; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D005786:Gene Expression Regulation / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D010012:Osteogenesis / Q000187:drug effects; D010455:Peptides / Q000494:pharmacology*; D011817:Rabbits; D047011:Silk / Q000494:pharmacology*; D054457:Tissue Scaffolds / Q000737:chemistry*
chemical_list: D006025:Glycosaminoglycans; D010455:Peptides; D047011:Silk
keywords: BMSCs; co-culture; multilayer; osteochondral