论文部分内容阅读
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 repairing full-thickness skin defects of rats with human skin derived from syngeneic cells using inbred rat experimental model, and to lay the foundation for its clinical application. Methods Skin of SD rats was inoculated with Dispase-trypsin two-step digestion method. The skin of SD rats was taken and the acellular dermal matrix was prepared according to the hypertonic saline-SDS-trypsin chemical treatment. Followed by submerged culture and gas-liquid separation culture of the second generation of dermal cells and acellular dermal matrix composite cultured tissue engineering skin. Thirty-six inbred F344 rats of 4 to 5 weeks old were used to prepare full thickness skin defects symmetrically on both sides of the back of the rats. The defect area was 1.5 cm × 1.5 cm pre-determined. Thirty-two wounds were randomly divided into three groups (n = 24). The two sides were implanted with different repair materials: the experimental group was transplanted with tissue engineered skin and the negative control group was transplanted with allogeneic acellular dermal matrix. The positive control group was transplanted with full- Non-orthotopic transplantation). General observation after operation, skin survival rate, wound contraction rate measurement and histological observation, evaluation of the repair effect. Results In the experimental group, the wounds reached healing stage I 4 weeks after operation and were closely connected with the surrounding tissues. The appearance of the wounds was close to that of the normal surrounding skin. At 4 weeks after operation, the survival rate of the negative control group was 0. The experimental group was 62.5% (15/24), which was significantly different from that of the positive control group (91.7%, 22/24) (χ2 = 5.779, P = 0.016). At 4 weeks after operation, the wound shrinkage in the experimental group and the positive control group was significantly lower than that in the negative control group (P <0.05). There was no significant difference between the experimental group and the positive control group (P> 0.05). Histological observation showed that the experimental group had a slight inflammatory reaction 1 week after operation; the small blood vessels and fibroblasts in the dermis were hyperplasia and the epidermis gradually differentiated and matured 2 weeks after operation; along with new collagen deposition at 4 weeks after operation, The dermis appears collagen remodeling. CONCLUSION: Genomic cell-derived tissue engineered skin can effectively repair the full-thickness skin defect in rats and achieve the effect similar to autologous full-thickness skin grafts in preventing wound contractions and promoting wound healing.