脂肪间充质干细胞聚合物-细胞外基质复合物补片修复兔外耳道全层皮肤损伤的实验研究

Application of adipose-derived stem cell aggregate-extracellular matrix composite patch to repair full-thickness skin trauma in external ear canal of New Zealand rabbits

目的探讨脂肪间充质干细胞聚合物(adipose-derived stem cell aggregates,ADSC aggregates)-细胞外基质(extracellular matrix,ECM)复合物补片(ADSC aggregates-ECM)在兔外耳道全层皮肤损伤修复中的应用。方法将新西兰大白兔随机分为3组,空白组、对照组、实验组,每组10只。在大白兔的耳腹侧耳根处构建外耳道内壁全层皮肤缺损模型。对照组、实验组采用内植法将ECM支架、ADSC aggregates-ECM支架铺于明胶海绵与创面边缘之间,缝合伤口前确保支架完全覆盖创面。空白组不做处理。术后14 d,比较各组动物创面愈合率、创面组织细菌菌落阳性比例;免疫组化检测各组创面组织中细胞核增殖抗原(proliferating cell nuclear antigen,Ki67)的表达;免疫印迹实验检测各组创面组织中白细胞介素1β(IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达。结果与空白组相比,实验组、对照组动物的创面愈合率、Ki67阳性细胞比例明显升高,细菌菌落阳性比例、创面组织中IL-1β、TNF-α的表达明显下降,差异均有统计意义(P<0.05);与对照组相比,实验组动物的创面愈合率明显升高、Ki67阳性细胞比例明显升高,细菌菌落阳性比例、创面组织中IL-1β、TNF-α的表达明显下降,差异均有统计意义(P<0.05)。结论ADSC aggregates-ECM补片能明显促进大白兔外耳道内壁全层皮肤缺损的修复。

Objective To explore the application of an adipose-derived stem cell(ADSC)aggregate-extracellular matrix(ECM)composite patch in full-thickness skin repair of the external auditory meatus of rabbits.Methods New Zealand white rabbits were randomly divided into 3 groups of 10 rabbits each:a blank group,a control group,and an experimental group.A full-thickness skin defect of the inner wall of the external auditory meatus was created at the ventral root of the ear.In the control group and experimental group,the ECM scaffold and the ADSC aggregate-ECM scaffold were laid between the gelatin sponge and the edge of the wound surface using the in-plant method to ensure that the scaffold completely covered the wound surface before the wound was sutured.The blank group underwent no treatment.After postoperative 14 days,the wound healing rate and the positive ratio of bacterial colonies in the wound tissues were compared among the groups.Immunohistochemical examination was used to detect the expression of proliferating cell nuclear antigen(Ki-67)in the wound tissues of each group,and Western blot was used to detect the expression of the inflammatory factors interleukin 1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the wound tissues of each group.Results The wound healing rate and the proportion of Ki-67–positive cells were significantly higher in the experimental group and control group than those in the blank group,whereas the positive ratio of bacterial colonies and the expression of IL-1βand TNF-αin the wound tissue were significantly lower.Compared with the control group,the wound healing rate and proportion of Ki-67-positive cells were significantly higher whereas the positive ratio of bacterial colonies and the expression of IL-1βand TNF-αin the wound tissue were significantly lower in the experimental group.These differences were also statistically significant(P<0.05).Conclusions The ADSC aggregate-ECM composite patch can effectively promote the repair of full-thickness skin defects of the inner wall of the external auditory meatus of rabbits.