自动切割仪制备微粒皮修复大鼠全层皮肤缺损创面

Repairing full-thickness skin defects of rats with microskins prepared by automatic cutting instrument

目的 探索自主研发的自动切割仪切割微粒皮的效率与效果,并评估其修复大鼠全层皮肤缺损创面的疗效。方法 通过与手工剪切的微粒皮比较,评价自动切割仪制备微粒皮的效率及效果。采用H-E染色观察机械切割(实验组)和手工剪切(对照组)制备的微粒皮结构,CCK-8法检测单位质量的微粒皮组织活性。以1∶20的移植比例将实验组和对照组微粒皮移植到大鼠全层皮肤缺损创面模型,观察创面上皮化速度。采用Masson染色、Ⅳ型胶原免疫荧光染色、Ki-67免疫组织化学染色分别检测创面胶原纤维结构及排列、基底膜形成、创面细胞增殖情况。结果 切割2 cm×3 cm的皮片,使用自动切割仪所需时间为5 min,而使用组织剪手工剪切所需时间约为40 min。H-E染色结果显示实验组皮粒结构保留完好,而手工剪切的皮粒表皮层损伤严重、结构保留不完整。CCK-8法检测结果显示实验组微粒皮活性指数高于对照组[(0.067 981±0.004 467)mg-1vs(0.045 168±0.002 698)mg-1,P<0.01]。移植创面第24天,实验组创面上皮化率达到(78.6±2.3)%,高于对照组的(27.2±1.9)%(P<0.01)。Masson染色结果显示实验组创面肉芽组织厚度低于对照组,胶原排列疏松且规则。实验组创面基底膜形成良好,基底层细胞增殖明显,而对照组基底膜形成不良,基底层细胞增殖不明显。结论 应用自主研发的自动切割仪制备微粒皮的效率高,微粒皮结构保留完好、活性保留高,可以1∶20大比例快速修复全层皮肤缺损创面,展示了与传统手工剪切微粒皮不同的特性,具有良好的应用前景。

Objective To explore the efficiency and effect of self-developed automatic cutting instrument for cutting microskins,and evaluate its effect in repairing full-thickness skin defects of rats.Methods The efficiency and effect of the automatic cutting instrument in preparing microskins were evaluated by comparing with that cut by hand.The microskins prepared by mechanical cutting (experimental group) and manual cutting (control group) were observed by hematoxylineosin (H-E) staining,and the tissue activity of the unit mass microskins of the experimental group and the control group was detected by cell counting kit 8 (CCK-8) assay.The microskins of the experimental group and the control group were transplanted into the rat model of full-thickness skin defect at the ratio of 1∶20,and the rate of wound epithelialization was observed.Masson staining,collagenⅣimmuno?uorescence staining and Ki-67 immunohistochemical staining were used to detect the structure and arrangement of collagen?bers,the formation of basement membrane and the proliferation of wound cells.Results The time required for cutting 2 cm×3 cm skin by using the automatic cutting instrument was 5 min,and the manual cutting time with tissue scissors was about 40 min.The results of H-E staining showed that the microskins in the experimental group had intact structure,but those cut by hand was not,their epidermis was seriously damaged.CCK-8assay showed that the activity index of skin grafts in the experimental group was signi?cantly higher than that in the control group ([0.067 981±0.004 467]mg-1vs[0.045 168±0.002 698]mg-1,P<0.01).On the 24thday after transplantation,the rate of wound epithelialization in the experimental group was (78.6±2.3)%,which was signi?cantly higher than that in the control group ([27.2±1.9]%,P<0.01).Masson staining showed that the thickness of granulation tissue in the experimental group was significantly lower than that in the control group,and the arrangement of collagen was loose and regular.In the experimental group,the wound basement membrane formed well and the basal layer cells proliferated obviously,while in the control group,the basement membrane formed poorly and the basal layer cells did not proliferate obviously.Conclusion The self-developed automatic cutting instrument has the advantages of high efficiency of cutting microskins,intact structure and high tissue activity retention,and can quickly repair the animal model of full-thickness skin defect at1∶20 ampli?cation ratio.It has different characteristics from that of traditional manual cutting microskins,and has a good application prospect.