During the last stages of wound healing, myofibroblasts differentiate mainly from fibroblasts. Myofibroblasts from normal skin wounds (Wmyo) can communicate with its surrounding using secreted factors. They also have the capacity to produce microvesicles (MVs), a type of extracellular vesicles, as mediators of intercellular communication. MVs cargo are potentially capable of regulating the behavior of targeted cells and tissues. The aim of this study is to evaluate the effect of Wmyo-derived MVs on dermal fibroblasts and to determine the responsible signaling molecule. Microvesicles were obtained from culture media of myofibroblasts and characterized using protein quantification, dynamic light scattering and transmission electron microscopy. Uptake of fluorescent MVs in fibroblasts was assessed by flow cytometry. Cytokines concentrations were quantified in MV samples by a multiplex ELISA. Different concentration of MVs or a selected cytokine were used as treatments over fibroblasts culture for 5 days. Following the treatments, parameters linked to the extracellular matrix were studied. Lastly, the selected cytokine was neutralized within MVs before evaluating collagen production. We showed that Wmyo derived-MVs were internalized by dermal fibroblasts. Cytokine array analysis revealed that a large amount of placental growth factor 1 (PLGF-1) (0.88 ± 0.63 pg/μg proteins in MVs) could be detected in MVs samples. Cutaneous fibroblasts treated with MVs or PLGF-1 showed significantly stimulated procollagen I level production (Fold change of 1.80 ± 0.18 and 2.07 ± 0.18, respectively). Finally, the neutralization of PLGF-1 in MVs significantly inhibited the production of procollagen I by fibroblasts. Our study shows that Wmyo derived-MVs are involved in intercellular communication by stimulating collagen production by fibroblasts during wound healing. This effect is possibly attained through PLGF-1 signalling. These findings represent a promising opportunity to gain insight into how MVs and Wmyo may mediate the healing of a skin wound.

在伤口愈合的最后阶段，成肌纤维细胞主要区别于成纤维细胞。正常皮肤伤口（Wmyo）产生的成肌纤维细胞可以利用分泌因子与其周围环境进行通讯。它们还具有产生微囊泡（MVs）（一种细胞外囊泡）的能力，作为细胞间通讯的介质。MVs货物有可能能够调节目标细胞和组织的行为。这项研究的目的是评估Wmyo衍生的MV对真皮成纤维细胞的作用，并确定负责任的信号分子。从成纤维细胞的培养基中获得微囊泡，并使用蛋白质定量，动态光散射和透射电子显微镜对其进行表征。通过流式细胞术评估成纤维细胞中荧光MV的摄取。通过多重ELISA对MV样品中的细胞因子浓度进行定量。在成纤维细胞培养物中使用不同浓度的MV或选定的细胞因子治疗5天。处理后，研究了与细胞外基质有关的参数。最后，在评估胶原蛋白产生之前，将选定的细胞因子在MV中中和。我们显示Wmyo衍生的MVs被真皮成纤维细胞内在化。细胞因子阵列分析显示，在MV样品中可以检测到大量的胎盘生长因子1（PLGF-1）（MV中为0.88±0.63 pg /μg蛋白）。MVs或PLGF-1处理的皮肤成纤维细胞显示出显着刺激的前胶原I水平产生（倍数变化分别为1.80±0.18和2.07±0.18）。最后，MVs中PLGF-1的中和作用显着抑制了成纤维细胞产生前胶原I。我们的研究表明Wmyo衍生的MVs通过刺激伤口愈合过程中成纤维细胞产生胶原蛋白而参与细胞间通讯。该作用可能是通过PLGF-1信号传导获得的。这些发现代表了一个有前途的机会，可以深入了解MV和Wmyo如何介导皮肤伤口的愈合。