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目的探讨利用深Ⅱ度烧伤皮肤(DBS)作为真皮替代物构建复合皮移植模型进行修复创面的可行性。方法 20只雄性Wistar大鼠随机分为深Ⅱ度烧伤真皮基质(DBDM)移植组(实验组)和无细胞真皮基质(ADM)移植组(对照组),每组10只。采用酶消化法,实验组取2只大鼠烫伤24 h后皮肤进行脱细胞等处理,制备DBDM;取新生24 h内乳鼠背部完整皮肤,修薄后与网状DBDM一次性重叠移植到大鼠背部约2 cm×2 cm全层皮肤缺损创面,14 d后观察复合皮生长情况。对照组采用Wistar大鼠正常皮肤制备的ADM替代DBDM,其他实验步骤同实验组。结果新鲜制备的DBDM呈浅黄白色,质地较柔软,有一定的韧性和伸展性,较ADM弱。镜下胶原纤维增粗和透明样变,排列紊乱、疏松,无细胞、血管和毛囊等成分存在。大鼠复合皮移植术后14 d,两组大鼠及移植物的成活率差异有统计学意义(P<0.05),双层复合皮与皮下组织紧密连接,深层DBDM或ADM呈粉红色,未见明显肿胀及炎性反应。与ADM相比,DBDM组移植物较早出现炎性细胞浸润和成纤维细胞、毛细血管长入;炎性细胞以中性粒细胞和巨噬细胞为主,新生胶原蛋白沉积较快,纤维排列规则、致密。结论 DBDM来源广泛、制备方法简单,移植物生物相容性好,无皮肤烧伤毒素样反应,可充当自体刃厚皮片的真皮替代物。本研究可为今后临床改造和利用烧伤皮肤提供新的思路。
Objective To explore the feasibility of constructing composite skin graft model by using deep second degree burn skin (DBS) as dermal substitute for repairing wounds. Methods Twenty male Wistar rats were randomly divided into deep second degree burn dermal matrix (DBDM) transplantation group (experimental group) and acellular dermal matrix (ADM) transplantation group (control group), 10 rats in each group. Using enzymatic digestion method, two rats in the experimental group scalded 24 hours after the skin was decellularized to prepare DBDM; take the intact skin of the back of neonatal rats within 24 hours of newborn, thinly overlap with reticulated DBDM after transplanting to large Full-thickness skin defect of about 2 cm × 2 cm on the back of the rats was observed. The growth of composite skin was observed after 14 days. In the control group, ADM prepared by normal skin of Wistar rats was used instead of DBDM. The other experimental steps were the same as the experimental group. Results Freshly prepared DBDM was light yellowish white, soft texture, a certain degree of toughness and extensibility, ADM weak. Microscopic collagen fibers thicker and transparent change, disordered, loose, no cells, blood vessels and hair follicles and other ingredients exist. There was significant difference in survival rate between the two groups of rats and grafts 14 days after composite skin transplantation (P <0.05), the double composite skin was closely connected with the subcutaneous tissue, the deep DBDM or ADM was pink, See obvious swelling and inflammatory reactions. Compared with ADM, the infiltration of inflammatory cells and fibroblasts and capillaries in DBDM group were earlier than those in ADM group. Inflammatory cells were mainly neutrophils and macrophages, with rapid deposition of nascent collagen, Rule, dense. Conclusion DBDM has a wide range of sources, simple preparation method, good biocompatibility of graft and no toxins-like reaction to skin burn, which can be used as dermal substitute of autologous blade edge skin graft. This study can provide new ideas for the clinical reform and utilization of burn skin.