title: In vitro generation of osteochondral differentiation of human marrow mesenchymal stem cells in novel collagen-hydroxyapatite layered scaffolds.
abstract: Integrated, layered osteochondral (OC) composite materials and/or engineered OC grafts are considered as promising strategies for the treatment of OC damage. A novel biomimetic collagen-hydroxyapatite (COL-HA) OC scaffold with different integrated layers has been generated by freeze-drying. The capacity of the upper COL layer and the lower COL/HA layer to promote the growth and differentiation of human mesenchymal stem cells (hMSCs) into chondrocytes and osteoblasts respectively was evaluated. Cell viability and proliferation on COL and COL/HA scaffolds were assessed by the MTT test. The chondrogenic differentiation of hMSCs on both scaffolds was evaluated by glucosaminoglycan (GAG) quantification, alcian blue staining, type II collagen immunocytochemistry assay and real-time polymerase chain reaction in chondrogenic medium for 21 days. Osteogenic differentiation was evaluated by alkaline phosphatase activity assay, type I collagen immunocytochemistry staining, alizarin S staining and mRNA expression of osteogenic gene for 14 days in osteogenic medium. The results indicated that hMSCs on both COL and COL/HA scaffolds were viable and able to proliferate over time. The COL layer was more efficient in inducing hMSC chondrogenic differentiation than the COL/HA layer, while the COL/HA layer possessed the superiority on promoting hMSC osteogenic induction over either COL layer or pure HA. In conclusion, the layered OC composite materials can effectively promote cartilage and bone tissue generation in vitro and are potentially usable for OC tissue engineering.
authors.affiliation: College of Materials Science and Engineering, South China University of Technology, Guangzhou, Guangdong 510640, People's Republic of China.
authors.forename: Jiaan
authors.initials: J
authors.lastname: Zhou
authors.forename: Caixia
authors.initials: C
authors.lastname: Xu
authors.forename: Gang
authors.initials: G
authors.lastname: Wu
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Cao
authors.forename: Liangming
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhichen
authors.initials: Z
authors.lastname: Zhai
authors.forename: Zhiwen
authors.initials: Z
authors.lastname: Zheng
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Chen
authors.forename: Yingjun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000328:Adult; D000943:Antigens, Differentiation / Q000096:biosynthesis; D001861:Bone Regeneration; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D020219:Chondrogenesis; D003094:Collagen / Q000737:chemistry*; D017886:Durapatite / Q000737:chemistry*; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D010012:Osteogenesis; D054457:Tissue Scaffolds
chemical_list: D000943:Antigens, Differentiation; D003094:Collagen; D017886:Durapatite