Enhanced differentiation of mesenchymal stem cells co-cultured with ligament fibroblasts on gelatin/silk fibroin hybrid scaffold.
The differentiation of mesenchymal stem cells (MSCs) towards fibroblasts is a crucial issue in ligament tissue engineering. This study aims to investigate the feasibility of using co-culture system to induce the differentiation of MSCs for constructing the tissue-engineered ligament in vitro. A kind of silk cable-reinforced gelatin/silk fibroin hybrid scaffold was used to provide three-dimensional (3-D) culture environments for MSCs. The 3-D co-culture system was set up by culturing MSCs/scaffold and ligament fibroblasts in the transwell insert and lower chamber, respectively. The regulatory effects of fibroblasts on MSCs were determined. After 2 weeks of co-culture the MSCs showed faster proliferation and higher DNA content compared with MSCs non-co-cultured. The MSCs were distributed uniformly throughout the scaffold and showed good viability. The collagen production also increased significantly with culture time. The MSCs in co-culture system were proved to differentiate into ligament fibroblasts by expressing ligament extra-cellular matrix (ECM)-specific genes including collagen I, collagen III, and tenascin-C in mRNA and protein level. The immunohistochemistry staining also confirmed the synthesis of key ligament ECM components. This study reveals that specific regulatory signals released from fibroblasts in 3-D co-culture system can enhance the differentiation of MSCs for ligament tissue engineering.
Fan
Hongbin
H
Department of Orthopaedic Surgery, National University of Singapore, 10 Kent Ridge Crescent, Singapore, Singapore.
Liu
Haifeng
H
Toh
Siew Lok
SL
Goh
James C H
JC
D000818:Animals; D002450:Cell Communication; D002454:Cell Differentiation; D049109:Cell Proliferation; D002470:Cell Survival; D018920:Coculture Techniques; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000032:analysis; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005349:Fibroins / Q000737:chemistry; D005780:Gelatin / Q000737:chemistry; D015870:Gene Expression; D007150:Immunohistochemistry; D008022:Ligaments / Q000166:cytology*; D059630:Mesenchymal Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D019063:Tenascin / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds
D024042:Collagen Type I; D024061:Collagen Type III; D012333:RNA, Messenger; D019063:Tenascin; D005780:Gelatin; D003094:Collagen; D004247:DNA; D005349:Fibroins