血管细胞外基质和膀胱平滑肌细胞的生物相容性研究

Biocompatibility of vessel extracellular matrix with bladder smooth muscle cells of rabbits

目的探讨血管细胞外基质（VECM）和膀胱平滑肌细胞的细胞相容性和组织相容性，为临床应用提供依据。方法制备兔VECM，分离培养兔膀胱平滑肌细胞，并以1×10^6个细胞／ml种植于VECM上，相差显微镜和扫描电镜观察细胞的黏附及生长。制备VECM和乳胶的提取液，分别作为实验组和阳性对照组，以培养基为阴性对照组，以无细胞的单纯培养基作为空白对照组。取2～4代对数生长期的膀胱平滑肌细胞，四甲基偶氮唑盐（MTT）法测定VECM浸提液对培养平滑肌细胞的细胞毒性；将VECM植入异体兔皮下，观察其组织相容性。结果兔膀胱平滑肌细胞能在体外培养扩增；接种1h后已有部分细胞黏附于VECM上，随时间推移，细胞逐渐增多，并伸展成梭形；培养第1、3、5天，实验组细胞增殖率分别为95．61％、98．34％、102．91％；阴性对照组为100．00％；阳性对照组分别为35．14％、38．95％、32．66％；实验组细胞增殖率明显高于阳性对照组，差异有统计学意义（P〈0．01），与阴性对照组比较差异无统计学意义（P〉0．05）。动物皮下埋植实验无排斥反应发生。结论VECM具有良好的生物相容性，是一种较好的泌尿道修复支架材料。

Objective To evaluate the biocompatibility of vessel extracellular matrix （VECM） with bladder smooth muscle cells of rabbits, and to discuss the feasibility of vessel extracellular matrix as a matrix for urinary tract reconstruction. Methods Primary cultured bladder smooth muscle cells （RBSMCs） isolated from New Zealand rabbits were implanted on VECM （1 × 10^6 cells/ml）. The effect of VECM on meta- bolic activity, attachment, proliferation of RBSMCs were monitored in vitro by inverted light microscopy and scanning electron microscopy. The extracts of VECM and emulsion were prepared as experimental group and positive controls separately. The culture medium was used as negative control, and simple culture medium without cells was used as blank control. The cell viability was monitored by MTT method after 1-,3-,5-d seeding. The in vivo tissue response to VECM was investigated by implanting into the subcutaneous sites of the rabbits. Results VECM exhibited nontoxic and bioactive effect on RBSMCs. RBSMCs could be attached to and proliferated on VECM and remained their morphologies. The cell proliferation rates of experimental group were 95.61% 、98.34% 、 102.91% , respectively, after 1,3,5 d ; those of negative control group were 100.00% ,respectively;and those of positive control group were 35. 14% 、38.95% 、32.66% , respectively. There was significant difference in the rate between experimental group and positive control （P 〈 0.01 ） , and no significant difference in the rate between experimental group and negative control （ P 〉 0.05 ）. In vivo, VECM demonstrated favorable tissue compatibility without tissue necrosis and fibrosis. Conclusions VECM exhibits nontoxic and bioactive effects on primary cultured bladder smooth muscle cells. It is a suitable material for urinary tract reconstruction.