title: In vitro generation of whole osteochondral constructs using rabbit bone marrow stromal cells, employing a two-chambered co-culture well design.
abstract: The regeneration of whole osteochondral constructs with a physiological structure has been a significant issue, both clinically and academically. In this study, we present a method using rabbit bone marrow stromal cells (BMSCs) cultured on a silk-RADA peptide scaffold in a specially designed two-chambered co-culture well for the generation of multilayered osteochondral constructs in vitro. This specially designed two-chambered well can simultaneously provide osteogenic and chondrogenic stimulation to cells located in different regions of the scaffold. We demonstrated that this co-culture approach could successfully provide specific chemical stimulation to BMSCs located on different layers within a single scaffold, resulting in the formation of multilayered osteochondral constructs containing cartilage-like and subchondral bone-like tissue, as well as the intermediate osteochondral interface. The cells in the intermediate region were found to be hypertrophic chondrocytes, embedded in a calcified extracellular matrix containing glycosaminoglycans and collagen types I, II and X. In conclusion, this study provides a single-step approach that highlights the feasibility of rabbit BMSCs as a single-cell source for multilayered osteochondral construct generation in vitro.
authors.affiliation: Department of Bioengineering, National University of Singapore.
authors.forename: Kelei
authors.initials: K
authors.lastname: Chen
authors.affiliation: Department of Bioengineering, National University of Singapore.
authors.forename: Kian Siang
authors.initials: KS
authors.lastname: Ng
authors.affiliation: Department of Bioengineering, National University of Singapore.
authors.forename: Sujata
authors.initials: S
authors.lastname: Ravi
authors.affiliation: Department of Bioengineering, National University of Singapore.|Department of Orthopaedic Surgery, National University of Singapore.
authors.forename: James C H
authors.initials: JC
authors.lastname: Goh
authors.affiliation: Department of Bioengineering, National University of Singapore.|Department of Mechanical Engineering, National University of Singapore.
authors.forename: Siew Lok
authors.initials: SL
authors.lastname: Toh
mesh_terms: D000818:Animals; D012831:Bombyx; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects; D002113:Calcification, Physiologic / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D020219:Chondrogenesis / Q000187:drug effects; D018920:Coculture Techniques / Q000379:methods*; D003094:Collagen / Q000378:metabolism; D019245:Compressive Strength / Q000187:drug effects; D004058:Diffusion; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D005786:Gene Expression Regulation / Q000187:drug effects; D006025:Glycosaminoglycans / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D010012:Osteogenesis; D010455:Peptides / Q000494:pharmacology*; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction; D047011:Silk / Q000494:pharmacology*; D054457:Tissue Scaffolds / Q000737:chemistry; D014057:Tomography, X-Ray Computed
chemical_list: D006025:Glycosaminoglycans; D010455:Peptides; D047011:Silk; D003094:Collagen
keywords: BMSCs; co-cultured well; osteochondral