异体阔筋膜细胞外基质作为肾修补材料的生物相容性研究

目的 研究异体阔筋膜细胞外基质（ECM）的生物相容性,探讨开发一种新的肾损伤修补材料。方法将犬的阔筋膜用脱细胞液及DNA酶、RNA酶等处理,制成异体阔筋膜ECM。制备阔筋膜ECM浸提液,对昆明种小白鼠行急性全身毒性试验;用四甲基偶氮唑盐（MTT）比色法对小鼠成纤维细胞进行细胞毒性检测。将犬左肾制成肾损伤模型,用异体阔筋膜ECM进行修补,分别于术后不同时段于修补局部取材行光镜及扫描电镜检查。结果阔筋膜ECM浸提液注射后4h,小鼠无异常反应,第1～3天体重均呈增长趋势,未发现死亡和中毒反应,与阴性对照生理盐水组无明显差别。MTT法显示细胞相对增殖率（RGR）为112％（第2天）、96％（第4天）及97％（第7天）,毒性评定相应为0、1、1级,材料无明显细胞毒性。术后各时段ECM材料修补局部均无明显的炎性细胞浸润,与周围组织无明显粘连。结论 异体阔筋膜ECM具有良好的生物相容性,有望成为理想的肾损伤修复材料。

Biocompatibility of extracellular matrix of homologous fascia lata as material for renal trauma repair

OBJECTIVE: To study the biocompatibility of extracellular matrix (ECM) of homologous fascia lata, so as to identify whether this material can be used in repair of renal trauma. METHODS: The fascia latae of 2 dogs were stripped and underwent treatment of decellularizing liquid, DNAse, RNAse, etc. so as to produce ECM. Twenty Kunming mice were randomly divided into 2 equal groups to be injected with the extractive liquid of the ECM or normal saline. 4, 24, 48, and 72 hours later the mice were observed so as to examine the toxicity of the ECM. Mouse fibroblasts of the line L929 were cultured. ECM was added into the suspension of the cells, and 2, 4, and 7 days later the relative growth rate (RGR) of the cells was observed by MTT method. The left kidneys of 10 dogs were cut by knife so as to establish kidney injury models, and the wounds were covered by prepared ECM pieces immediately. 1 and 2 weeks, and 1, 2, and 4 months after the operation respectively the dogs were killed and light microscopy and electron microscopy were conducted to observe the condition of the wounds. RESULTS: No abnormal situation was found in the mice after the extractive liquids of the ECM was injected into their bodies at any time point. MTT method showed that no obvious cytotoxicity was seen in the L292 cells, the RGR levels 2, 4, and 7 days were 112%, 96%, and 97% respectively, and the cytotoxic grade were 0, 1, and 1 respectively. No obvious infiltration of inflammatory cells into the local site of renal repair could be seen, only mild adhesion to the surrounding tissues was found at different time-points after operation. CONCLUSION: With good biocompatibility, ECM of homologous fascia lata may be used as an ideal tissue-engineering material for renal repair.