title: In vitro ligament-bone interface regeneration using a trilineage coculture system on a hybrid silk scaffold.
abstract: The ligament-bone interface is a complex structure that comprises ligament, fibrocartilage, and bone. We hypothesize that mesenchymal stem cells cocultured in between ligament and bone cells, on a hybrid silk scaffold with sections suitable for each cell type, would differentiate into fibrocartilage. The section of scaffold for osteoblast seeding was coated with hydroxyapatite. A trilineage coculture system (osteoblasts-BMSCs-fibroblasts) on a hybrid silk scaffold was established. RT-PCR results and immunohistochemistry results demonstrated that BMSCs cocultured between fibroblasts and osteoblasts had differentiated into the fibrocartilaginous lineage. The morphological change was also observed by SEM observation. A gradual transition from the uncalcified to the calcified region was formed in the cocultured BMSCs from the region that directly interacted with fibroblasts to the region that directly interacted with osteoblasts. The role of transforming growth factor β3 (TGF-β3) in this trilineage coculture model was also investigated by supplementing the coculture system with 10 ng/mL TGF-β3. The TGF-treated group showed similar results of fibrocartilaginous differentiation of BMSCs with coculture group without TGF-β3 supplement. However, no calcium deposition was found in the cocultured BMSCs in the TGF-treated group. This may indicate TGF-β3 delayed the mineralization process of chondrocytes.
authors.affiliation: Department of Bioengineering, National University of Singapore, Singapore 117576.
authors.forename: Pengfei
authors.initials: P
authors.lastname: He
authors.forename: Kian Siang
authors.initials: KS
authors.lastname: Ng
authors.forename: Siew Lok
authors.initials: SL
authors.lastname: Toh
authors.forename: James Cho Hong
authors.initials: JC
authors.lastname: Goh
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D001861:Bone Regeneration / Q000187:drug effects; D001842:Bone and Bones / Q000166:cytology; D002454:Cell Differentiation / Q000187:drug effects; D020099:Coated Materials, Biocompatible / Q000737:chemistry* / Q000494:pharmacology; D018920:Coculture Techniques; D017886:Durapatite; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051445:Fibrocartilage / Q000166:cytology / Q000187:drug effects / Q000254:growth & development; D008022:Ligaments / Q000166:cytology / Q000187:drug effects / Q000254:growth & development; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D061251:Primary Cell Culture; D011817:Rabbits; D047011:Silk / Q000737:chemistry*; D023822:Tissue Engineering; D054457:Tissue Scaffolds; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D020099:Coated Materials, Biocompatible; D047011:Silk; D053782:Transforming Growth Factor beta3; D017886:Durapatite