Medical Center of Burn Plastic and Wound Repair, The First Affiliated Hospital of Nanchang University, 17 Yongwai Street, Nanchang, 330006, Jiangxi, People's Republic of China.
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, People's Republic of China.
Tissue Eng Regen Med. 2023 Apr;20(2):295-308. doi: 10.1007/s13770-022-00513-w. Epub 2023 Jan 25.
Diabetic wound healing remains a major challenge due to the impaired functionality of angiogenesis by persistent hyperglycemia. Mesenchymal stem cell exosomes are appropriate candidates for regulating the formation of angiogenesis in tissue repair and regeneration. Here, we explored the effects of exosomes derived from human amniotic mesenchymal stem cell (hAMSC-Exos) on the biological activities of human umbilical vein endothelial cells (HUVECs) treated with high glucose and on diabetic wound healing and investigate lncRNAs related to angiogenesis in hAMSC-Exos.
hAMSCs and hAMSC-Exos were isolated and identified by flow cytometry or western blot. A series of functional assays such as cell counting kit-8, scratching, transwell and tube formation assays were performed to evaluate the potential effect of hAMSC-Exos on high glucose-treated HUVECs. The effect of hAMSC-Exos on diabetic wound healing were tested by measuring wound closure rates and immunohistochemical staining of CD31. Subsequently, the lncRNAs profiles in hAMSC-Exos and hAMSCs were examined to screen the lncRNAs related to angiogenesis.
The isolated hAMSC-Exos had a size range of 30-150 nm and were positive for CD9, CD63 and CD81. The hAMSC-Exos facilitate the functional properties of high glucose-treated HUVECs including the proliferation, migration and the angiogenic activities as well as wound closure and angiogenesis in diabetic wound. hAMSC-Exos were enriched lncRNAs that related to angiogenesis, including PANTR1, H19, OIP5-AS1 and NR2F1-AS1.
Our findings demonstrated hAMSC-Exos facilitate diabetic wound healing by angiogenesis and contain several exosomal lncRNAs related to angiogenesis, which may represent a promising strategy for diabetic wound healing.
由于持续的高血糖导致血管生成功能受损，糖尿病创面愈合仍然是一个主要挑战。间充质干细胞外泌体是调节组织修复和再生中血管生成的合适候选物。在这里，我们探讨了人羊膜间充质干细胞（hAMSC-Exos）衍生的外泌体对高糖处理的人脐静脉内皮细胞（HUVEC）的生物学活性的影响，以及对糖尿病创面愈合的影响，并研究了 hAMSC-Exos 中与血管生成相关的 lncRNAs。
通过流式细胞术或 Western blot 分离和鉴定 hAMSCs 和 hAMSC-Exos。通过细胞计数试剂盒-8 检测、划痕、Transwell 和管形成检测等一系列功能检测，评估 hAMSC-Exos 对高糖处理的 HUVEC 的潜在影响。通过测量伤口闭合率和 CD31 的免疫组织化学染色来检测 hAMSC-Exos 对糖尿病创面愈合的影响。随后，检查 hAMSC-Exos 和 hAMSCs 中的 lncRNA 谱，以筛选与血管生成相关的 lncRNA。
分离的 hAMSC-Exos 的大小范围为 30-150nm，并且对 CD9、CD63 和 CD81 呈阳性。hAMSC-Exos 促进了高糖处理的 HUVEC 的功能特性，包括增殖、迁移和血管生成活性以及糖尿病创面的伤口闭合和血管生成。hAMSC-Exos 富含与血管生成相关的 lncRNAs，包括 PANTR1、H19、OIP5-AS1 和 NR2F1-AS1。
我们的研究结果表明，hAMSC-Exos 通过血管生成促进糖尿病创面愈合，并含有几种与血管生成相关的外泌体 lncRNA，这可能为糖尿病创面愈合提供一种有前途的策略。
