Yildiz Technical University, Department of Bioengineering, Istanbul, Turkey; Center for Regenerative Medicine and Stem Cell Research & Manufacturing (LivMedCell), Istanbul, Turkey.
Yildiz Technical University, Department of Bioengineering, Istanbul, Turkey.
J Pharm Sci. 2022 Dec;111(12):3232-3242. doi: 10.1016/j.xphs.2022.08.016. Epub 2022 Aug 20.
The aim of the present study was to explore the antileishmanial performance and wound healing effect of exosomes isolated from Wharton Jelly derived mesenchymal stem cells (WJ-MSCs) in combination with aloe-emodin. MSCs obtained from Wharton Jelly were characterized by flow cytometry. Exosomes were isolated from cultivated stem cells by ultacentrifugation method. Scanning Electron Microscopy (SEM), Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA) and flow cytometry were used for characterization of obtained exosomes. The cytotoxicities of characterized exosomes and aloe-emodin at different concentrations were investigated on L929 and J774 cell lines. Non-toxic concentrations of each agent were combined and their inhibitory efficacies on L.major promastigotes and amastigotes were investigated by different techniques such as MTT, parasite count and measurements of infection index. Finally, wound healing activities of combinations were examined on in vitro artifical wound model and compared with the use of exosomes alone. According to outcome of flow cytometic analysis, vesicles isolated from WJ-MSCs highly expressed the markers such as CD63 special for exosome profile. SEM and NTA results demonstrated that derived exosomes possessed dimensions between 150 to 200 nanometers and elicited the cup-shape specific to exosomes. Combinations including non-toxic dosages of exosomes and aloe-emodin demonstrated superior antileishmanial effectivenesses both on promastigotes and amastigotes in contrast to use of exosome alone since they lead to inhibition of promastigotes and amastigotes for 4 and 10-folds in comparison to control, respectively. Additionally, combinations elicited more rapidly and effective in vitro wound-healing performance in contrast to use of exosome alone. At the end of 24 h incubation application of combinations gave rise to wound closure at a rate of 72 %, while in the control group 52 % of wound area has not been healed, yet. These results reflect that mentioned combination has great potential to be used in treatment of cutaneus leishmaniasis (CL) since they have magnificient capacity to inhibit Leishmania parasites while enhancing wound healing.
本研究旨在探讨与大黄素联合使用从牙髓间充质干细胞(WJ-MSCs)分离的外泌体的抗利什曼原虫性能和伤口愈合效果。通过流式细胞术对从牙髓中获得的 MSC 进行表征。通过超速离心法从培养的干细胞中分离出外泌体。扫描电子显微镜(SEM)、动态光散射(DLS)、纳米颗粒跟踪分析(NTA)和流式细胞术用于表征获得的外泌体。在 L929 和 J774 细胞系上研究了表征的外泌体和大黄素在不同浓度下的细胞毒性。将每种药物的无毒浓度组合,并通过 MTT、寄生虫计数和感染指数测量等不同技术研究其对 L. major 前鞭毛体和无鞭毛体的抑制作用。最后,在体外人工伤口模型上检查组合的伤口愈合活性,并将其与单独使用外泌体进行比较。根据流式细胞术分析的结果,从 WJ-MSCs 分离的囊泡高度表达 CD63 等特殊的外泌体标志物。SEM 和 NTA 结果表明,衍生的外泌体具有 150 至 200 纳米之间的尺寸,并呈现出特有的杯状结构。与单独使用外泌体相比,包括外泌体和大黄素的无毒剂量组合对前鞭毛体和无鞭毛体均表现出更高的抗利什曼原虫作用,因为它们分别导致前鞭毛体和无鞭毛体的抑制率达到对照的 4 倍和 10 倍。此外,与单独使用外泌体相比,组合在体外更迅速有效地发挥伤口愈合作用。在 24 小时孵育结束时,组合的应用导致伤口闭合率达到 72%,而在对照组中,52%的伤口面积尚未愈合。这些结果表明,所述组合具有很大的潜力用于治疗皮肤利什曼病(CL),因为它们具有抑制利什曼原虫的巨大能力,同时增强伤口愈合。