组织工程支架的隔离层对新生软骨Ⅱ型胶原蛋白表达的影响

Effect of the insulation layer of tissue-engineering scaffold on collagen type Ⅱ expression in the neo-cartilage

目的研究隔离层在组织工程骨软骨复合支架中,对新生软骨Ⅱ型胶原蛋白表达的影响。方法将向软骨方向诱导后的兔骨髓间充质干细胞接种到骨软骨复合支架的软骨支架上,然后将具有隔离层和没有隔离层的骨软骨复合支架分别植入兔膝关节骨软骨全层缺损处。术后3月和6月分别取材,进行新生软骨Ⅱ型胶原蛋白蛋白的免疫组化染色、Western blot法检测Ⅱ型胶原蛋白的表达、实时荧光定量PCR(qRT-PCR)测定Ⅱ型胶原蛋白相关基因的表达。结果与无隔离层的对照组相比,具有隔离层的骨软骨复合支架所形成的新生软骨的Ⅱ型胶原蛋白和基因的表达均明显增强(P<0.05)。结论隔离层加入骨软骨复合支架后,能显著提高新生组织工程软骨Ⅱ型胶原蛋白表达的能力。

Objective To investigate the effect of the insulation layer in the tissue-engineering composite osteochondral scaffolds on collagen type ]] expression in the neo-cartilage. Methods Chondral phases of insulation layer-free or insulation layer-containing biphasic scaffolds were seeded with autogeneic chondrocyte-induced bone marrow-derived mesenchymal stem cells. Then, the biphasic scaffolds-cells constructs were implanted into osteochondral defects of rabbits＇ knees. The expression of type]] collagen in the tissue-engineering cartilages was evaluated by immunohistochemistry and Western blotting at 3 and 6 months after surgery, respectively. Moreover, the mRNA expression of collagen type I1 was also detected by qRT- PCR. Results The expression of the collagen type ]1 at both protein and mRNA levels in tissue-engineering neo-cartilages generated by the insulation layer-containing biphasic scaffold were significantly higher than those by the insulation layer-free biphasic scaffold in vivo （ P 〈 0.05 ）. Cenduslon After the insulation layer is added into the osteochondral composite scaffold, the collagen type ]] expression in the tissue-engineering neo-cartilage can be significantly enhanced.