复合雪旺细胞的猪小肠粘膜下层对大鼠海绵体神经损伤勃起功能恢复的实验研究

Compound graft of porcine small intestinal submucosa with Schwann cells to reconstruct injured cavernous nerves and restore erectile function

目的:研究雪旺细胞(SCs)与猪小肠粘膜下层(SIS)构成的人工神经移植替代损伤的大鼠双侧海绵体神经(CN)恢复大鼠的勃起功能。方法:体外培养SCs并复合于SIS,成功制备成SIS与SCs的复合体,选取健康SD大鼠33只,随机分为3组:假手术组(n=11)、CN切断组(n=11)、SCs+SIS组(n=11)。各组大鼠于术后3个月进行阿朴吗啡试验;然后取阴茎海绵体中后部海绵体组织,进行nNOS免疫组织化学染色。结果:①联合使用机械剥离、混合酶消化、差速贴壁法、Ara-C短期使用等方法可获得较高纯度的SCs,满足神经组织工程的需要;②经过机械刮除、化学方法消毒制备的SIS作为一种天然支架材料,与SCs具有良好的细胞生物相容性,适合SCs在其表面粘附、生长、增殖和分化;③术后3个月行阿朴吗啡试验:无论是勃起率还是勃起次数,SCs+SIS组均显著高于CN切断组(P<0.01),但低于假手术组(P<0.01);④nNOS神经纤维数目:SCs+SIS组与CN切断组差异有统计学意义(P<0.01),但二者均低于假手术组(P<0.01)。结论:SCs+SIS作为神经移植物可以修复缺损的CN,并在一定程度上恢复阴茎勃起功能。

Objective：To investigate the restoration of rat penile erection by reconstructing injured cavernous nerves（CN） with a compound graft prepared from porcine small intestinal submucosa（SIS） and Schwann cells（SC）.Methods：SCs were cultured in vitro and a compound graft was prepared from the SCs and SIS.Thirty-three healthy SD rats were randomly divided into three groups of equal number,sham-operation,CN ablation,and SIS ＋ SC graft.Three months after the operation,all the rats underwent the apomorphine test,followed by immunohistochemical staining of the tissues from the middle part of the corpus cavernosum penis.Results：Combined use of mechanical stripping,mixed-enzyme digestion,different-speed adhesion,short-term Ara-C and some other methods yielded SCs of a purity high enough for nerve tissue engineering.The SIS prepared by mechanical and chemical methods exhibited a good biocompatibility with SCs,which could adhere,grow,propagate and differentiate on its surface.The apomorphine test showed that both the rate and frequency of penile erection were significantly higher in the SIS ＋ SC graft than in the CN ablation group（P〈0.01）,but lower than in the sham operation group（P〈0.01）.The number of nNOS positive nerve fibers in the SIS ＋ SC graft group was significantly different from that of the CN ablation（P〈0.01）,but both were smaller than that of the sham-operation group（P〈0.01）.Conclusion：The compound of SIS with SCs,as a nerve graft,can be used to reconstruct injured cavernous nerves,and to some extent,restore penile erectile function.