环氧氯丙烷对组织工程心脏瓣膜构建中MMP-9表达的影响

目的探讨环氧氯丙烷对组织工程心脏瓣膜构建中基质金属蛋白酶-9(Matrix metalloproteinase-9;MMP-9)表达的影响。方法采用去垢剂和胰蛋白酶消化制备脱细胞猪主动脉瓣膜支架,用3%环氧氯丙烷处理48h的去细胞支架材料作为实验组;用0.2%戊二醛处理48h的去细胞支架材料作为对照组。将培养的人骨髓基质干细胞(human bone marrow mesenchy-mal stem cells;hBMSCs)种植于脱细胞支架上构建组织工程心脏瓣膜(tissue engineered heart valves;TEHV),分别行石蜡包埋切片HE染色和扫描电镜观察TEHV的组织结构,免疫组化检测MMP-9表达的阳性率。结果hBMSCs在实验组脱细胞瓣膜表面生长良好,MMP-9的表达比对照组降低。结论环氧氯丙烷处理的脱细胞猪主动脉瓣膜支架,可以抑制hBMSCs的MMP-9表达,对防止组织工程心脏瓣膜钙化的形成有一定作用。

Effects of the Epoxy Chloropropane on the Expression of Matrix Metalloproteinase - 9 During Creating Tissue Engineered Heart Valves

Objective To investigate the effects of epoxychloropropane on the expression of matrix metalloproteinases- 9 (MMPs- 9) in creating tissue engineered heart valves (TEHV), on the tissue structures of TEHV, and to study the effects of epoxy chloropropane on the calcification of TEHV. Methods The porcine aortic valve leaflets were digested and decellularized by using detergent and trypsin. Those treated with 0.2% glutaraldehyde for 48 hours served as the control group; those treated with 3% epoxy chloropropane for 48 hours served as the experimental group. The cultured human bone marrow mesenchymal stem cells (hBMSCs) were seeded onto the decellularized scaffolds of TEHV. The histological studies were done with pathological sections and scanning electron microscopy. Immunohistochemistry was used to detect the expression of MMP-9.Results In the experimental group, the histology showed that the hBMSCs grew well into the pores and formed a confluent layer in decellularized scaffolds; the studies of immunohistochemistry indicated significantly attenuated expressions of MMPs,compared with the control(P<0.05). Conclusions The decellularized porcine aortic valves treated with 3% epoxy chloropropane may inhibit the expression of MMP-9; therefore epoxy chloropropane may prevent the calcification of tissue engineered heart valves.