骨髓间充质干细胞与自体腹膜联合修复盆腔自主神经损伤的实验研究

目的观察骨髓间充质干细胞(BMSC)与自体腹膜桥接管联合移植修复盆腔自主神经损伤的效果.方法建立比格犬盆腔自主神经损伤模型,实验组以自体腹膜管填充胶原蛋白海绵+BMSC桥接于缺损神经两端;对照组改BMSC为生理盐水.正常对照组为自体神经移植.术后12周取材,标本切片行HE染色和神经纤维(NF)免疫组化染色.应用图像分析系统对选定数据进行测量.透射电镜观察实验组再生神经纤维超微结构.结果实验组与对照组比较再生神经纤维总数[(1742±185)根比(1131±262)根,P<0.01]、密度[(168±14)根/104μm2比(124±17)根/104μm2,P<0.01]、直径[3.83±0.22)μm比(3.28±0.41)μm,P<0.05]、面积百分比(0.32±0.07比0.21±0.08,P<0.05)差异均有统计学意义,与正常对照组比较差异均无统计学意义(P>0.05).实验组再生神经纤维结构清晰,形态接近正常.结论 BMSC与自体腹膜桥接管联合移植修复盆腔自主神经缺损再生神经纤维生长好,方法可行,与自体神经移植修复效果相当.

Experimental study on repairing pelvic autonomic nerve injury by bone marrow mesenchymal stem cells and autologous peritoneum

Objective To evaluate the effect of using the bridge consisting of bone marrow mesenchymal stem cells(BMSCs) and autologous peritoneum to repair the injured pelvic autonomic nerve. Methods Injury models were made on beagle dogs and divided into three groups: collagen protein sponge and BMSCs were inseted in peritoneal canal to bridge the gap in experimental group, BMSCs was replaced by saline in control group, and autologous nerve was cutted previously and inversed 180° to bridge the gap in normal control group. Specimens were obtained after 12 weeks, and observed by HE and NF immunohistochemical staining. The chosen features was measured using image analytic system. Uhramierostructure in experimental group was observed by transmission electron microscope. Results The sum of regenerative nerve fiber (1742±185 vs 1131±262, P<0.01 ), density [ (168±14) (104μm2)-1 vs (124±17) (104μm2)-1, P<0.01 ], diameter [ (3.83±0.22) μm vs (3.28±0.41) μm, P<0.05] and area percent (0.32±0.07 vs 0.21±0.08, P<0.05 ) were significantly different between experimental group and control group respectively, and there were no significant differences between experimental group and normal control group ( P>0.05 ). The morphous of regenerative nerve fiber was near normal and all constitution was clear in experimental group. Conclusion The regenerative nerve is grown well. This bridge consisting of BMSCs and autologous peritoneum to repair the injured pelvic autonomic nerve. This method is feasible,and the result is the same as using autologous nerve to transplant.