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[目的]观察不同代次正常软骨细胞和关节炎软骨细胞对琼脂糖-BMSCs向软骨细胞分化的促进作用。[方法]分离新西兰兔BMSCs,正常软骨细胞。制作兔关节炎模型,提取兔关节炎软骨细胞。将BMSCs和低熔点琼脂糖复合成凝胶块,放在自制的六孔板网格架上,构建兔软骨细胞-BMCSs共培养系统。在3、7、14 d取各组琼脂糖-BMSCs凝胶块进行实时定量PCR、GAG含量检测。[结果]兔关节炎模型制作成功,关节面色泽较灰暗,关节软骨粗糙。Normal P0-BMSCs组的II型胶原基因表达明显增强,Normal P3-BMSCs组Ⅰ、Ⅱ型胶原及蛋白聚糖基因表达均未见明显增强,OA P0-BMSCs组蛋白聚糖基因表达明显增强,OA P3-BMSCs组I型胶原基因表达水平在3个时间点均低于对照组。Normal P0-BMSCs组的GAG含量为5.7±0.49μg/mg(湿重),较对照组有明显上升。OA P0-BMSCs组GAG含量与对照组相比差异无统计学意义(P>0.05),其余三组与对照组相比差异均具有统计学意义(P<0.05)。[结论]兔正常P0软骨细胞与兔关节炎P0软骨细胞所分泌的形态发生素能够有效促进BMSCs向软骨细胞分化,而兔正常P3软骨细胞的促分化作用微弱,兔关节炎P3软骨细胞不能促进BMSCs向软骨细胞分化。
[Objective] To observe the effect of different generations of normal chondrocytes and arthritis chondrocytes on the differentiation of agarose-BMSCs into chondrocytes. [Method] New Zealand rabbit BMSCs and normal chondrocytes were isolated. Rabbit arthritis model was made to extract rabbit arthritis chondrocytes. The BMSCs and low-melting point agarose complexed into gel blocks, placed on a homemade six-hole grid frame, the construction of rabbit chondrocytes-BMCSs co-culture system. At 3, 7, and 14 days, the agarose-BMSCs were collected for real-time quantitative PCR and GAG assay. [Result] Rabbit arthritis model was successfully made, the color of articular surface was gloomy and the articular cartilage was rough. The expression of type II collagen gene in normal P0-BMSCs group was significantly increased. The expression of type I, type II collagen and proteoglycan gene in Normal P3-BMSCs group was not significantly increased, and the expression of histone deacetylase gene in OA P0-BMSCs group was significantly increased. OA The expression of type I collagen gene in P3-BMSCs group was lower than that in control group at three time points. The GAG content of Normal P0-BMSCs group was 5.7 ± 0.49μg / mg (wet weight), which was significantly higher than that of the control group. There was no significant difference in the GAG content between the OA P0-BMSCs group and the control group (P> 0.05). The differences between the other three groups and the control group were statistically significant (P <0.05). [Conclusion] Morphogenetic hormone secreted by P0 chondrocytes in rabbit normal P0 chondrocytes and rabbit arthritis osteochondrosis can effectively promote the differentiation of BMSCs to chondrocytes. However, the differentiation of rabbit normal P3 chondrocytes is weak, and the P3 chondrocytes of rabbit arthritis can not be promoted BMSCs differentiate into chondrocytes.