Department of Orthopaedics, Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University, Taizhou 317000, Zhejiang, China.
Fengxian District Central Hospital Graduate Student Training Base, Jinzhou Medical University, Shanghai 201499, China.
J Wound Care. 2022 Mar 1;31(Sup3):S29-S38. doi: 10.12968/jowc.2022.31.Sup3.S29.
The purpose of this study was to explore the paracrine effects of adipose-derived stem cells (ASCs) on cutaneous wound healing in diabetic rats.
The ASCs were isolated and identified by immunofluorescent staining. The ASCs-conditioned medium (ASCs-CM) was harvested. Cell counting kit (CCK)-8 assay, scratch experiments, western blot and quantitative polymerase chain reaction (qPCR) were performed to observe the effects of ASCs-CM on fibroblasts. A full-thickness skin wound diabetic rat model was prepared, using 34 male, Sprague Dawley rats. ASCs-CM or negative-control medium (N-CM) was injected around the wound surface. The existing wound area was measured on days 4, 8, 12 and 16 after the postoperative day, and the wound tissues were collected for immunohistochemical staining and qPCR quantitative study.
In this experiment, the isolated cells were characterised as ASCs. The results of CCK-8 assay, cell scratch test, western blot and qPCR showed ASCs-CM could significantly promote the proliferation, migration and differentiation of fibroblasts. Simultaneously, the healing rate of full-thickness skin wounds in diabetic rats was significantly higher in the ASCs-CM group than the N-CM group on days 4, 8, 12 and 16. Immunohistochemical staining and qPCR results showed that the expression of vascular endothelial growth factor (VEGF, days 4 and 8), α-smooth muscle actin (SMA) (days 4 and 16), transforming growth factor (TGF)-β1 (days 4, 8 and 12) were higher in the ASCs-CM group than that of the N-CM group (p<0.05).
This experiment demonstrated that ASCs-CM may accelerate wound healing in diabetic rats by promoting the secretion of TGF-β1, VEGF and the proliferation, migration and differentiation of fibroblasts.
本研究旨在探讨脂肪干细胞（ASCs）对糖尿病大鼠皮肤伤口愈合的旁分泌作用。
通过免疫荧光染色分离和鉴定 ASCs。收集 ASCs 条件培养基（ASCs-CM）。通过细胞计数试剂盒（CCK-8）检测、划痕实验、Western blot 和定量聚合酶链反应（qPCR）观察 ASCs-CM 对成纤维细胞的影响。制备全层皮肤创伤糖尿病大鼠模型，使用 34 只雄性 Sprague Dawley 大鼠。在术后第 4、8、12 和 16 天，在伤口表面周围注射 ASCs-CM 或阴性对照培养基（N-CM）。测量术后第 4、8、12 和 16 天的现有伤口面积，并收集伤口组织进行免疫组化染色和 qPCR 定量研究。
在本实验中，分离的细胞被鉴定为 ASCs。CCK-8 检测、细胞划痕实验、Western blot 和 qPCR 的结果表明，ASCs-CM 可显著促进成纤维细胞的增殖、迁移和分化。同时，在 ASCs-CM 组，糖尿病大鼠全层皮肤伤口的愈合率在术后第 4、8、12 和 16 天均明显高于 N-CM 组。免疫组化染色和 qPCR 结果显示，ASCs-CM 组血管内皮生长因子（VEGF，第 4 和 8 天）、α-平滑肌肌动蛋白（SMA，第 4 和 16 天）和转化生长因子（TGF）-β1（第 4、8 和 12 天）的表达均高于 N-CM 组（p<0.05）。
本实验表明，ASCs-CM 通过促进 TGF-β1、VEGF 的分泌以及成纤维细胞的增殖、迁移和分化，可能加速糖尿病大鼠的伤口愈合。
