Ⅰ、Ⅱ型胶原蛋白对人软骨细胞生物学特性的影响

Effect of type I or type Ⅱ collagen on biological characteristics of human chondrocytes

背景:实验证明胶原蛋白底物具有刺激成软骨的作用,但关于不同类型胶原蛋白刺激成软骨作用的能力仍存在争议。目的:观察Ⅰ、Ⅱ型胶原蛋白对体外培养人软骨细胞生物学特性的影响。方法:将P3代人软骨细胞分别加入普通培养板、Ⅰ型胶原蛋白包被培养板、Ⅱ型胶原蛋白包被培养板继续培养。培养10 d内,MTT法绘制细胞生长曲线;培养28 d后,采用ELISA法、聚合酶链反应、二甲基亚甲基蓝比色等方法检测3种培养板中软骨细胞分泌Ⅰ胶原蛋白、Ⅱ型胶原蛋白及糖胺多糖的量。结果与结论:Ⅱ型胶原蛋白包被培养板中软骨细胞数量最多,增殖速度为Ⅰ型胶原蛋白包被培养板的2倍、普通培养板的5倍。Ⅱ型胶原蛋白包被培养板中软骨细胞分泌Ⅰ型胶原蛋白最少,与普通培养板板检测结果差异有显著性意义(P<0.01),与Ⅰ型胶原蛋白包被培养板检测结果差异无显著性意义;Ⅱ型胶原蛋白包被培养板中软骨细胞分泌Ⅱ型胶原蛋白、糖胺多糖最多,与其他两种培养板检测结果差异有显著性意义(P<0.01)。表明胶原蛋白包被培养板培养软骨细胞优于普通培养板,其中Ⅱ型胶原蛋白包被培养板在培养软骨细胞时更能维持细胞形态,延长去分化现象出现的时间,更利于细胞再分化。

BACKGROUND： Experiments have shown that the collagen Substrate has the capability of stimulating cartilage generation, but the stimulating role of different types of collagen substrates remains controversial. OBJECTIVE： To investigate the effect of type I and type Ⅱ collagen on the biological characteristics of human chondrocytes cultured in vitro. METHODS： Human chondrocytes at passage were cultured onto the ordinary culture plates （ordinary plate）, type I collagen-coated culture plates （type I plate）, and type Ⅱ collagen-coated culture plates （type Ⅱ plate）. Cell growth curves were determined by MTT method after cells were cultured for 10 days. By ELISA, PCR, and 1,9-dimethyl methyleneblue technology, type I and type Ⅱ collagen and glycosaminoglycan contents were quantitatively detected in cartilage cells 28 days after culture. RESULTS AND CONCLUSION： The number of cartilage cells was the highest in type Ⅱ plate, which was twice of that in type I plate and five times of that in ordinary plate. Cartilage cells in type Ⅱ plate secreted the least amount of type I collagen, which showed significant differences compared with the ordinary plate （P 〈 0.01 ） and had no statistically significant difference with type I plate （P 〉 0.01 ）. Cartilage cells in type Ⅱ plate secreted the most amount of type Ⅱ collagen and glycosaminoglycan, showing significant differences compared with the other two plates （P 〈 0.01 ）. The cartilage cells cultured in collagen plates are better than that cultured in ordinary culture plate, type Ⅱ collagen culture plate is better than type I collagen culture plate in maintaining cell shape, extending the dedifferentiation pattern, and promoting cell differentiation.