Effect of hyaluronic acid on the differentiation of chondrogenic cells cultured in type Ⅱ collagen

• Main Authors: Chi-Han Wang, 王紀涵
• Other Authors: Yng-Jiin Wang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/59514259055779063891

碩士 === 國立陽明大學 === 醫學工程研究所 === 97 === The repair of articular cartilage defect is probably the most challenging work of the orthopedic clinical surgery. Among the currently used materials for cartilage repair, hyaluronic acid (HA) is one of the best choices. However, the documented HA effects on the chondrogenesis are rather conflicting to each other. Therefore, the goal of this study was to clarify this controversy by examining the effect of different HA concentrations on the differentiation of articular cartilage cells, synovium MSC and periosteum MSC from SD rats. In order to simulate the environment of articular cartilage, the cells obtained from three different sources were cultured in 3-D type Ⅱ collagen matrix, and incubated in a medium containing TGFβ3 (tissue growth factor-β3), vitamin C (ascorbate-2- phosphate), dexamethasone and ITS Premix. The type Ⅱ collagen used in this study was treated with anion-exchange resin (DE52 column) to remove the residual GAG. There was about 4% w/w content of saccharides that were not GAG, therefore any HA-effect due to the GAG contamination in type Ⅱ collagen can be ruled out. The presence of lacunae was found in the matrix of all types of the cultured cells after 7-day induction. The intensity of alcian blue stain increased with culturing time without any observed effect by HA. The gene expressions of both aggrecan and type Ⅱ collagen increased with time, but there was apparently no positive effect of HA on chondrogenic differentiation. The expressions of aggrecan of MSCs from synovium and periosteum membranes were suppressed by HA at high concentration after 21-day induction. This was in consistent with the type Ⅹ collagen gene expression of synovium and periosteum MSCs cultured at high HA concentration after 21-day induction. Interestingly, the characteristics of bone formation were not found in the matrix of cartilage cells throughout the culture period.