title: Effects of exogenous glycosaminoglycans on human chondrocytes cultivated on type II collagen scaffolds.
abstract: Cartilage extracellular matrix (ECM) is composed primarily of type II collagen (COL II) and large, networks of proteoglycans (PGs) that contain glycosaminoglycans such as hyaluronic acid (HA) and chondroitin sulfate (CS). Since cartilage shows little tendency for self-repair, injuries are kept unhealed for years and can eventually lead to further degeneration. During the past decades, many investigations have pursued techniques to stimulate articular cartilage repair or regeneration. The current study assessed the effects of exogenous glycosaminoglycans (GAGs) including CS-A, CS-B, CS-C, heparan sulfate and HA, administration on human chondrocytes in terms of proliferation and matrix synthesis, while the cells were seeded and grown on the genipin-crosslinked collagen type II (COL II) scaffold. DNA content was measured by Hoechst dye intercalation, matrix deposition was evaluated by DMMB dye. Expression of collagen II and aggrecan mRNAs was assessed by RT-PCR, followed by gel electrophoresis. In a 28-day in vitro culture, administration of 5 microg/ml CS-A, 50 microg/ml CS-B, 50 microg/ml CS-C, 5 microg/ml HS, and 500 kDa HA led to significant increase in biosynthesis rate of PGs. Gene expression of aggrecan and collagen II were upregulated by CS-A, CS-C and HA. These results showed considerable relevance of GAGs to the issue of in vitro/ex vivo neo-cartilage synthesis for tissue engineering and regenerative medical applications.
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, 101, Section 2, Kuang-Fu Road, Hsinchu, 30013, Taiwan, ROC.
authors.forename: Chun-Hsien
authors.initials: CH
authors.lastname: Wu
authors.forename: Chih-Sheng
authors.initials: CS
authors.lastname: Ko
authors.forename: Jia-Wen
authors.initials: JW
authors.lastname: Huang
authors.forename: Hsin-Jie
authors.initials: HJ
authors.lastname: Huang
authors.forename: I-Ming
authors.initials: IM
authors.lastname: Chu
mesh_terms: D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D024043:Collagen Type II / Q000737:chemistry* / Q000493:pharmacokinetics*; D006025:Glycosaminoglycans / Q000008:administration & dosage*; D006801:Humans; D008422:Materials Testing; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D001672:Biocompatible Materials; D024043:Collagen Type II; D006025:Glycosaminoglycans