摘要
目的利用近交系大鼠实验模型评价同基因细胞源组织工程皮肤修复大鼠全层皮肤缺损的效果,为其临床应用奠定基础。方法取近交系F344大鼠乳鼠皮肤,采用Dispase-胰蛋白酶两步消化法分离培养表皮细胞;取SD大鼠皮肤,按照高渗盐-SDS-胰蛋白酶化学处理法制备脱细胞真皮基质;依次利用浸没式培养和气液分离培养法将第2代表皮细胞与脱细胞真皮基质复合体外培养构建组织工程皮肤。取4～5周龄近交系F344大鼠36只,于大鼠背部两侧对称制备全层皮肤缺损,缺损面积为预实验确定的1.5 cm×1.5 cm。将72个创面随机分为3组(n=24),两侧移植不同修复材料:实验组移植组织工程皮肤、阴性对照组移植同种异体脱细胞真皮基质、阳性对照组移植自体全层皮肤(非原位移植)。术后行大体观察、皮片成活率、创面收缩率测量及组织学观察,评价修复效果。结果实验组术后4周创面可达到Ⅰ期愈合,与周围组织紧密连接,外观接近周围正常皮肤。术后4周,阴性对照组皮片成活率为0;实验组为62.5%(15/24),与阳性对照组91.7%(22/24)比较差异有统计学意义(χ2=5.779,P=0.016)。术后4周,实验组和阳性对照组创面收缩率显著低于阴性对照组(P<0.05),实验组与阳性对照组间差异无统计学意义(P>0.05)。组织学观察示实验组术后1周有轻微炎性反应;术后2周真皮层小血管和成纤维细胞大量增生,表皮层逐渐分化成熟;术后4周时伴随新生胶原纤维沉积,移植物真皮层出现胶原改建。结论同基因细胞源组织工程皮肤能够有效修复大鼠全层皮肤缺损,在防止创面收缩和促进创面愈合等方面均可达到类似于自体全层皮肤移植的效果。
Objective To evaluate the effect of tissue engineered skin with isogeneic cells on repairing skin defects in inbred rat model so as to provide relevant evidences for the clinical application.Methods The skins of newborn inbred F344 rats were harvested and treated with Dispase trypsin to isolate the epidermal cells.The skins of adult Sprague Dawley rats were obtained and treated with hypertonic sodium-SDS-trypsin to prepare the acellular dermal matrix.The tissue engineered skin was reconstructed by submerging culturing and air-liquid interface culturing in vitro.The full-thickness skin defects of 1.5cm×1.5cm in size were prepared along the dorsal both sides of 36 adult inbred F344 rats,and 72 defects were repaired with tissue engineered skin in experimental group（n=24）,with allogeneic acellular dermal matrix in negative control group（n=24）,and with autologous full-thickness skin in positive control group（n=24）.Finally the gross observation,the survival rate,wound contraction rate,and histological observation were used to evaluate the effect.Results The wound healed by first intension at 4 weeks postoperatively in the experimental group;the grafts connected with the adjacent tissue tightly and had normal appearance.At 4 weeks after operation,the survival rate of the graft was 0 in the negative control group;the survival rates were 62.5%（15/24） in the experimental group and 91.7%（22/24） in the positive control group,showing significant difference between 2 groups（χ2=5.779,P=0.016）.The wound contraction rates of the experimental group and positive control group were significantly lower than that of the negative control group（P 0.05）,but no significant difference was found between the experimental group and positive control group（P0.05）.Histological observation showed that slight inflammation reaction appeared at 1 week postoperatively in the experimental group;the regeneration of the blood vessel and the proliferation of the fibroblasts in dermis and the gradual maturation of epidermis were observed at 2 weeks,and new collagen deposition and collagen remodeling in the dermis of the graft were found at 4 weeks postoperatively.Conclusion The tissue engineered skin is able to repair full-thickness skin defect of rats effectively,it has similar effect to the autologous full-thickness skin in preventing the wound contraction and promoting the wound healing,which provides experimental evidences for the clinical application.
出处
《中国修复重建外科杂志》
CAS
CSCD
北大核心
2012年第11期1375-1380,共6页
Chinese Journal of Reparative and Reconstructive Surgery
基金
山西省青年科技研究基金资助项目(2012021033-1)~~
