Effects of exogenous glycosaminoglycans on human chondrocytes cultivated on type II collagen scaffolds
Effects of exogenous glycosaminoglycans on human chondrocytes cultivated on type II collagen scaffolds
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DOI：
10.1007/s10856-009-3889-8
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发表时间：
2010-02-01
影响因子：
3.7
通讯作者：
Chu, I-Ming
中科院分区：
文献类型：
--
作者：
Wu, Chun-Hsien;Ko, Chih-Sheng;Chu, I-Ming
软骨细胞外基质（ECM）主要由II型胶原（COL II）和含有糖胺聚糖（如透明质酸（HA）和硫酸软骨素（CS））的大蛋白聚糖（PG）网络组成。由于软骨几乎没有自我修复的倾向，因此损伤多年无法愈合，最终可能导致进一步的退化。在过去的几十年中，许多研究都在寻求刺激关节软骨修复或再生的技术。本研究评估了外源性糖胺聚糖（GAG），包括CS-A，CS-B，CS-C，硫酸乙酰肝素和HA，对人软骨细胞增殖和基质合成的影响，同时将细胞接种并生长在京尼平交联的II型胶原（COL II）支架上。DNA含量用Hoechst染料嵌入法测定，基质沉积用DMMB染料评价。通过RT-PCR，随后通过凝胶电泳评估II型胶原和聚集蛋白聚糖mRNA的表达。在28天的体外培养中，施用5 μ g/ml CS-A、50 μ g/ml CS-B、50 μ g/ml CS-C、5 μ g/ml HS和500 kDa HA导致PG的生物合成速率显著增加。CS-A、CS-C和HA均能上调聚集蛋白聚糖和II型胶原的基因表达。这些结果显示了GAG与用于组织工程和再生医学应用的体外/离体新软骨合成问题的相当大的相关性。
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 mu g/ml CS-A, 50 mu g/ml CS-B, 50 mu g/ml CS-C, 5 mu g/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.