Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Int Immunopharmacol. 2022 Jan;102:108404. doi: 10.1016/j.intimp.2021.108404. Epub 2021 Dec 2.
Impaired chronic wound healing frequently occurs in diabetic patients. We hypothesized that menstrual blood-derived mesenchymal stem cells (MenSCs) in combination with bilayer scaffold consisted of human amniotic membrane (AM) and electrospun silk fibroin nanofibers could potentially promote wound healing in diabetic mice.
METHODS & METHODS: Two bilateral full-thickness wounds were created on dorsal skin of type-1 diabetic mice model and animals were equally divided in four groups including: no-treatment group (NT), amniotic membrane treated group (AM), bilayer scaffold treated group (bSC), and MenSCs-seeded bilayer scaffold treated group (bSC + MenSCs). Wound healing evaluations were performed at 3, 7, and 14 days after their treatment. The wound healing was analyzed by macroscopic and microscopic evaluations, and immunofluorescence staining of involucrin (IVL), type III collagen, CD31/ von Willebrand factor (vWF), and PGP9.5 were performed. Furthermore, number of neutrophils and macrophages and subpopulation of macrophages were assessed. In addition, the expression of Egr2, Mmp9, CXCL12, IDO1, Ptgs2 and VEGFA transcripts involved in wound repair were also analyzed.
After 14 days, the best epidermal and dermal regeneration belonged to the cases received bSC + MenSCs as wound dressing. Moreover, the wound healing was typically faster in this group compared to other groups. Immunofluorescence evaluation represented higher levels of CD31 and VWF, higher ratio of M2/M1 macrophages, greater expression of IVL, and higher levels of the PGP9.5 in the bSC + MenSCs group in comparison with other groups. Expression analysis of assessed genes also supported assumption of more regeneration and healing in the bSC + MenSCs group versus other groups.
These results indicate that enhanced immunomodulatory and reparative properties of MenSCs in conjunction with bilayer scaffold specified this cellular skin substitute for modulating wound chronicity and contribution to resolution of wound healing process in diabetic ulcer.
糖尿病患者常出现慢性伤口愈合受损。我们假设，月经血源性间充质干细胞（MenSCs）与由人羊膜（AM）和静电纺丝丝素纳米纤维组成的双层支架结合使用，可能有助于促进糖尿病小鼠的伤口愈合。
在 1 型糖尿病小鼠模型的背部皮肤上创建了两个双侧全层伤口，将动物等分为 4 组，包括：未治疗组（NT）、羊膜处理组（AM）、双层支架处理组（bSC）和 MenSCs 接种双层支架处理组（bSC+MenSCs）。在治疗后 3、7 和 14 天进行伤口愈合评估。通过宏观和微观评估以及钙调蛋白结合蛋白 9.5（PGP9.5）、IVL、III 型胶原蛋白、CD31/血管性血友病因子（vWF）的免疫荧光染色分析伤口愈合情况。此外，还评估了中性粒细胞和巨噬细胞的数量以及巨噬细胞亚群。此外，还分析了参与伤口修复的 Egr2、Mmp9、CXCL12、IDO1、Ptgs2 和 VEGFA 转录本的表达。
在 14 天内，作为伤口敷料，接受 bSC+MenSCs 的病例具有最佳的表皮和真皮再生。此外，与其他组相比，该组的伤口愈合通常更快。与其他组相比，免疫荧光评估显示 CD31 和 vWF 水平更高，M2/M1 巨噬细胞比例更高，IVL 表达更高，PGP9.5 水平更高。评估基因的表达分析也支持 bSC+MenSCs 组比其他组具有更强的再生和愈合能力的假设。
这些结果表明，MenSCs 的增强的免疫调节和修复特性与双层支架结合使用，使这种细胞皮肤替代物能够调节慢性伤口，并有助于糖尿病溃疡愈合过程的解决。
