自动切割仪制备微粒皮修复大鼠全层皮肤缺损创面的实验研究

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,评估其修复大鼠全层皮肤缺损创面的效果。 方法 通过与手工剪切的微粒皮比较,评价自动切割仪的制备效果及效率。实验组(机械切割)和对照组(手工剪切)制备的微粒皮HE染色观察微粒皮结构,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。Masson染色、免疫荧光Ⅳ型胶原染色、Ki67免疫组化染色分别检测创面胶原纤维结构及排列、基底膜形成情况、创面细胞增殖情况。 结果 切割2 cm×3 cm的皮片,使用一代自动切割仪所需时间为10 min,二代机所需时间为5 min,而使用组织剪手工剪切所需时间约为40 min,使用自动切割仪可以显著减少切割时间,省时省工。CCK-8法检测发现自动切割组微粒皮活性指数明显高于手工切割组。移植创面第24天,实验组创面上皮化达到78.6%±2.3%,显著高于对照组27.2%±1.9%(P<0.001)。Masson染色发现自动机械切割组创面肉芽组织厚度明显低于手工微粒皮组,胶原排列疏松且规则。自动机械切割组创面基底膜形成良好,基底层细胞增殖明显,而手工切割组基底膜形成不良,基底层细胞增殖不明显。结论 自主研发的自动皮片切割仪制备微粒皮效率高、效果好,微粒皮活性保留高,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,具有很好的应用前景。

Experimental study on repairing full-thickness skin defects of rats with micro-skin prepared by automatic cutting instrument

Objective To explore the efficiency and effect of self-developed automatic cutting instrument for cutting micro-skin, and to evaluate its effect in repairing full-thickness skin defects of rats. Methods The effect and efficiency of preparing micro-skin by the automatic cutting instrument were evaluated by comparing with preparing the micro-skin cut by hand. The micro-skin prepared by the experimental group (mechanical cutting) and the control group (manual cutting) were observed by HE staining, and the tissue activity of the unit mass micro-skin of the experimental group and the control group was detected by CCK-8 reagent. The micro-skins 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 and recorded. Masson staining, Collagen IV immunofluorescence staining and Ki67 immunohistochemical staining were used to detect the structure and arrangement of collagen fibers, the formation of basement membrane and the proliferation of wound cells. Results The time required for cutting 2cm × 3cm skin by using the first-generation automatic cutter is 10min, the second-generation machine is 5min, and the manual cutting time with tissue scissors is about 40min. Using the automatic cutter can significantly reduce the cutting time and save labor. CCK-8 assay showed that the activity index of skin grafts in the automatic cutting group was significantly higher than that in the manual cutting group. On the 24th day, the wound epithelialization in the experimental group was 78.6% ±2.3%, which was significantly higher than that in the control group (27.2% ±1.9%) (P < 0.001). Masson staining showed that the thickness of granulation tissue in the automatic mechanical cutting group was significantly lower than that in the manual micro-skin group, and the arrangement of collagen was loose and regular. In the automatic mechanical cutting group, the wound basement membrane formed well and the basal layer cells proliferated obviously, while in the manual cutting group, the basement membrane formed poorly and the basal layer cells did not proliferate obviously. Conclusion The self-developed automatic skin cutting instrument has the advantages of high efficiency of cutting micro-skin, good effect and high tissue activity retention, and can quickly repair the animal model of full-thickness skin defect at 1:20 amplification ratio, which is different from that of traditional manual shearing micro-skin. The self-developed automatic skin cutting instrument shows a good application prospect.