Mirzadegan Ebrahim, Golshahi Hannaneh, Saffarian Zahra, Edalatkhah Haleh, Darzi Maryam, Khorasani Somayeh, Saliminejad Kioomars, Kazemnejad Somaieh
Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Avicenna J Med Biotechnol. 2023 Jul-Sep;15(3):139-156. doi: 10.18502/ajmb.v15i3.12923.
In this study we differentially showed the effects of cell-seeded bilayer scaffold wound dressing in accelerating healing process in diabetic ulcers that still remains as a major clinical challenge. The aim of the study was to compare immunomodulatory and angiogenic activity, and regenerative effect differences between Menstrual blood-derived Stem Cells (MenSCs) and foreskin-derived keratinocytes/fibroblasts.
The streptozotocin-induced diabetic mice model was developed in male C57/BL6 mice. A bilayer scaffold was fabricated by electrospining silk fibroin nano-fibers on human Amniotic Membrane (AM). Dermal fibroblasts and keratinocyte isolated from neonatal foreskin and MenSCs were isolated from the menstrual blood of healthy women. The diabetic mice were randomly divided into three groups including no treatment group, fibroblast/keratinocyte-seeded bilayer scaffold group (bSC+FK), and MenSCs-seeded bilayer scaffold group. The healing of full-thickness excisional wounds evaluations in the diabetic mice model in each group were evaluated at 3, 7, and 14 days after treatment.
The gross and histological data sets significantly showed wound healing promotion re-epithelialization and wound contraction along with enhanced regeneration in MenSCs-seeded bilayer scaffold group with the most similarity to adjacent intact tissue. Immunofluorescence staining of mouse skin depicted a descending trend of type III collagen along with the higher expression of involucrin as keratinocyte marker in the MenSCs-seeded bilayer nanofibrous scaffold group in comparison with other treatment groups from day 7 to day 14. Moreover, higher levels of CD31 and von Willebrand factor (VWF), and also a higher ratio of M2/M1 macrophages in association with higher levels of the neural marker were observed in the bSC+MenSCs group in comparison with bSC+FK and no treatment groups.
Healing symptoms in wounds dressed with keratinocyte/fibroblast-seeded bilayer scaffold was significantly lower than MenSCs-seeded bilayer scaffold done on impaired diabetic wound chronicity.
在本研究中，我们差异化地展示了接种细胞的双层支架伤口敷料在加速糖尿病溃疡愈合过程中的作用，糖尿病溃疡仍是一项重大的临床挑战。本研究的目的是比较月经血来源的干细胞（MenSCs）与包皮来源的角质形成细胞/成纤维细胞之间的免疫调节和血管生成活性以及再生效果差异。
在雄性C57/BL6小鼠中建立链脲佐菌素诱导的糖尿病小鼠模型。通过将丝素蛋白纳米纤维静电纺丝在人羊膜（AM）上制备双层支架。从新生儿包皮中分离出真皮成纤维细胞和角质形成细胞，从健康女性的月经血中分离出MenSCs。将糖尿病小鼠随机分为三组，包括未治疗组、接种成纤维细胞/角质形成细胞的双层支架组（bSC+FK）和接种MenSCs的双层支架组。在治疗后3、7和14天对每组糖尿病小鼠模型中的全层切除伤口愈合情况进行评估。
大体和组织学数据集显著显示，接种MenSCs的双层支架组促进了伤口愈合、上皮再形成和伤口收缩，同时再生增强，与相邻完整组织最为相似。与其他治疗组相比，从第7天到第14天，接种MenSCs的双层纳米纤维支架组小鼠皮肤的免疫荧光染色显示III型胶原蛋白呈下降趋势，同时作为角质形成细胞标志物的内披蛋白表达较高。此外，与bSC+FK组和未治疗组相比，bSC+MenSCs组中CD31和血管性血友病因子（VWF）水平更高，并且M2/M1巨噬细胞比例更高，同时神经标志物水平也更高。
用接种角质形成细胞/成纤维细胞的双层支架处理的伤口愈合症状明显低于对受损糖尿病伤口慢性期使用接种MenSCs的双层支架处理的情况。
