Induced pluripotent stem cells-derived microvesicles accelerate deep second-degree burn wound healing in mice through miR-16-5p-mediated promotion of keratinocytes migration
Induced pluripotent stem cells-derived microvesicles accelerate deep second-degree burn wound healing in mice through miR-16-5p-mediated promotion of keratinocytes migration
复制标题
诱导多能干细胞来源的微泡通过 miR-16-5p 介导的角质形成细胞迁移加速小鼠深二度烧伤伤口愈合
DOI：
10.7150/thno.46639
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Zhang, Lin
中科院分区：
文献类型：
--
作者：
Yan, Yuan;Wu, Ruijun;Zhang, Lin
背景：诱导多能干细胞（iPSC）已成为皮肤伤口的一种有前途的治疗模式。细胞外囊泡现在被认为是有益的干细胞旁分泌效应的关键介质。在这项研究中，我们研究了iPSCs-derived microvesicles（iPSCs-MVs）对深Ⅱ度烧伤创面愈合的影响，并探讨了其潜在的机制。方法：从诱导多能干细胞的条件培养液中分离纯化多能干细胞，并通过电镜和粒径分布进行鉴定。在深二度烧伤模型中，在伤口部位周围皮下注射iPSCs-MV，并通过测量伤口闭合面积、组织学检查和免疫组织化学染色来评估功效。在体外，CCK-8、EdU染色和划痕试验用于评估iPSCs-MV对角质形成细胞增殖和迁移的影响。接下来，我们通过高通量microRNA测序探索了潜在的机制。评估miR-16- 5 p在由iPSCs-MV诱导的角质形成细胞功能的调节中的作用。此外，还检测了介导miR-16- 5 p在角质形成细胞中生物学效应的靶基因。最后，我们检查了局部miR-16- 5 p治疗对小鼠深二度烧伤伤口愈合的影响。结果如下：将iPSC-MV局部移植到烧伤创面床中导致加速的伤口闭合，包括增加的上皮再形成。在体外，iPSCs-MVs可以促进角质形成细胞的迁移。我们还发现miR-16- 5 p是iPSCs-MV通过靶向桥粒芯糖蛋白3（Dsg 3）激活p38/MARK通路而诱导的角质形成细胞体外迁移促进中的关键因子。最后，我们证实了局部miR-16- 5 p治疗可以促进烧伤伤口愈合期间的上皮再形成。结论：因此，我们的研究结果表明，iPSCs-MVs衍生的miR-16- 5 p可能是深二度烧伤创面愈合的新治疗方法。
Background: Induced pluripotent stem cells (iPSCs) have emerged as a promising treatment paradigm for skin wounds. Extracellular vesicles are now recognized as key mediators of beneficial stem cells paracrine effects. In this study, we investigated the effect of iPSCs-derived microvesicles (iPSCs-MVs) on deep second-degree burn wound healing and explored the underlying mechanism. Methods: iPSCs-MVs were isolated and purified from conditioned medium of iPSCs and confirmed by electron micrograph and size distribution. In deep second-degree burn model, iPSCs-MVs were injected subcutaneously around wound sites and the efficacy was assessed by measuring wound closure areas, histological examination and immunohistochemistry staining. In vitro, CCK-8, EdU staining and scratch assays were used to assess the effects of iPSCs-MVs on proliferation and migration of keratinocytes. Next, we explored the underlying mechanisms by high-throughput microRNA sequencing. The roles of the miR-16-5p in regulation of keratinocytes function induced by iPSCs-MVs were assessed. Moreover, the target gene which mediated the biological effects of miR-16-5p in keratinocytes was also been detected. Finally, we examined the effect of local miR-16-5p treatment on deep second degree-burns wound healing in mice. Results: The local transplantation of iPSCs-MVs into the burn wound bed resulted in accelerated wound closure including the increased re-epithelialization. In vitro, iPSCs-MVs could promote the migration of keratinocytes. We also found that miR-16-5p is a critical factor in iPSCs-MVs-induced promotion of keratinocytes migration in vitro through activating p38/MARK pathway by targeting Desmoglein 3 (Dsg3). Finally, we confirmed that local miR-16-5p treatment could boost re-epithelialization during burn wound healing. Conclusion: Therefore, our results indicate that iPSCs-MVs-derived miR-16-5p may be a novel therapeutic approach for deep second-degree burn wound healing.