三种不同材料修复大鼠口腔黏膜缺损创面的对比实验研究

Comparative Experimental Study on Three Different Materials for Repairing Oral Mucosa Defects in Rats

目的:探究同种(人皮)脱细胞基质、异种(牛皮)脱细胞基质、纳米纤维丝素蛋白三种材料修复大鼠口腔黏膜缺损创面的效果.方法:选择80只3月龄雄性Wister清洁级大鼠,随机分为A(纳米纤维丝素组,n=20)、B(牛皮脱细胞基质组,n=20)、C(人皮脱细胞基质组,n=20)、D组(空白对照组,n=20),各组大鼠均切除直径10mm的全层口腔颊黏膜,A、B、C组大鼠分别取同一直径、经辐照灭菌的纳米纤维丝素蛋白、异种(牛皮)脱细胞基质、同种(人皮)脱细胞基质覆盖于创面,D组大鼠用凡士林纱布覆盖,比较各组大鼠术后1、2、3、4周时创面面积,采用HE染色及免疫组化分析大鼠各时间点成纤维细胞、新生血管内皮、炎性细胞数量及内皮细胞生长情况.结果:①各组大鼠术后创面无感染,术后1周A、B、C组受植区可见透明膜状物,D组创缘稍红肿,术后1、2、3周时D组大鼠创面直径显著大于A、B、C组(P<0.05);A、B、C组大鼠各时间点创面大小组间比较差异无统计学意义(P>0.05).②HE染色显示,A、B、C组大鼠不同时间点成纤维细胞数、炎性细胞数均显著低于D组(P<0.05),A、B、C组大鼠各时间点组间比较差异无统计学意义(P>0.05).③免疫组化示,A、B、C组术后1、2、3、4周时大鼠CK阳性细胞表达数显著高于D组(P<0.05);术后1、2、3周时CD34阳性细胞表达数显著高于D组(P<0.05),A、B、C组各时间点组间比较差异无统计学意义(P>0.05).结论:纳米纤维丝素蛋白、自体、异体细胞基质对大鼠口腔黏膜损伤具有相似的修复效果,有较高的临床应用价值.

Objective:To investigate the effects of three kinds of materials,namely,homologous(human skin)acellular matrix,heterogeneous(cowhide)acellular matrix,and nanocellulose protein in the repair of oral mucosa defects in rats.Methods:803-month-old male Wister clean rats were randomly divided into three groups:group A(nanocellulose silk fibroin group,n=20),group B(cowhide acellular matrix group,n=20),group C(human skin acellular matrix group,n=20),and group D(blank control group,n=20).Full-thickness buccal mucosa with a diameter of 10 mm was excised from rats in each group.Wounds of rats in groups A,B,and C were respectively covered with silk fibroin,heterogeneous(cowhide)acellular matrix,and homologous(human skin)acellular matrix which had the same diameter and were treated with radicidation.The wounds of rats in group D were covered with vaseline gauze.The wound areas were compared among groups at 1 week,2 weeks,3 weeks and 4 weeks after surgery.The numbers of fibroblasts,neovascular endothelial cells and inflammatory cells,and growth of endothelial cells in rats at different time points were analyzed through HE staining and immunohistochemistry.Results:①There was no wound infection in each group.Transparent membranes could be observed in the grafted area of group A,B,and C at 1 week after surgery.The wound margin of group D was slightly red and swollen.The wound diameter of group D was significantly larger than that of group A,B,and C at 1 week,2 weeks,and 3 weeks after surgery(P<0.05),without significant difference in wound size among group A,B,and C at each time point(P>0.05).②HE staining showed that the numbers of fibroblasts and inflammatory cells in group A,B,and C at different time points were significantly smaller than those in group D(P<0.05),without significant difference among group A,B,and C at each time point(P>0.05).③Immunohistochemistry showed that the expression of CK-positive cells in group A,B,and C was significantly higher than that in group D at 1 week,2 weeks,3 weeks and 4 weeks after surgery(P<0.05).The number of CD34-positive cells was significantly larger than that in group D at 1 week,2 weeks,and 3 weeks after surgery(P<0.05),without significant difference among group A,B,and C at each time point(P>0.05).Conclusion:The effects of nanocellulose protein,autologous and allogeneic cell matrices are similar in repair of oral mucosa damage in rats.