Altug Burcugul, Soykan Merve Nur, Eyubova Sevinc, Eker Sariboyaci Ayla, Dogan Cezmi, Ozalp Onur, Atalay Eray
Cellular Therapy and Stem Cell Production Application, Research Centre (ESTEM), Eskisehir Osmangazi University, Eskisehir, Türkiye.
Department of Stem Cell, Institute of Health Sciences, Eskisehir Osmangazi University, Eskisehir, Türkiye.
Biofactors. 2024 Nov-Dec;50(6):1287-1297. doi: 10.1002/biof.2085. Epub 2024 May 28.
Inflammatory mediators that infiltrate the corneal stroma after corneal infections, trauma or refractive surgery can trigger the transformation of corneal keratocytes into myofibroblasts, resulting in highly irregular collagen deposition and subsequently corneal scarring. Mesenchymal stem cells (MSCs) can be used as therapeutic agents to regenerate corneal and conjunctival tissue damage, regulate inflammation, and reduce the development of limbal stem cell failure. The use of MSC-derived exosomes as a cell-free therapeutic vector is a novel therapeutic approach. This study aimed to assess the effect of exosomes obtained from melatonin (Mel)-treated human limbal mesenchymal stem cells (hLMSCs) on naïve hLMSCs and to determine their influence on the antifibrotic and pro-regenerative pathways involved in corneal scarring. hLMSCs were treated with varying concentrations of Mel, followed by isolation and characterization of the procured exosomes (Mel-prExos). These exosomes were added to the cell culture media of naïve hLMSCs to examine their antifibrotic and pro-regenerative effects. The expression of miR-155, miR-29, TGFβ1, TGFβ3, PPARγ, and α-SMA miRNAs and genes were compared between Mel-treated hLMSCs and Mel-prExo-treated hLMSCs by using real-time PCR. We found that at 1 μM Mel and in the presence of Mel-prExos, TGFβ1 was expressed 0.001-fold, while TGFβ3 was expressed 0.6-fold. miR-29 expression was increased 38-fold in the control-Exo group compared to that in the control group. Changes in TGFβ1/β3 and α-SMA expression are associated with miR-29 and miR-155. This approach could prove beneficial for ocular surface tissue engineering applications.
角膜感染、创伤或屈光手术后浸润角膜基质的炎症介质可触发角膜基质细胞向肌成纤维细胞转化，导致高度不规则的胶原沉积，进而形成角膜瘢痕。间充质干细胞（MSCs）可作为治疗剂用于再生角膜和结膜组织损伤、调节炎症以及减少角膜缘干细胞衰竭的发生。使用MSC来源的外泌体作为无细胞治疗载体是一种新型治疗方法。本研究旨在评估从褪黑素（Mel）处理的人角膜缘间充质干细胞（hLMSCs）获得的外泌体对未处理的hLMSCs的影响，并确定它们对角膜瘢痕形成中抗纤维化和促再生途径的影响。用不同浓度的Mel处理hLMSCs，随后分离并鉴定获得的外泌体（Mel-prExos）。将这些外泌体添加到未处理的hLMSCs的细胞培养基中，以检查它们的抗纤维化和促再生作用。通过实时PCR比较Mel处理的hLMSCs和Mel-prExo处理的hLMSCs之间miR-155、miR-29、TGFβ1、TGFβ3、PPARγ和α-SMA的miRNA和基因表达。我们发现，在1μM Mel及Mel-prExos存在的情况下，TGFβ1表达为0.001倍，而TGFβ3表达为0.6倍。与对照组相比，对照组外泌体组中miR-29表达增加了38倍。TGFβ1/β3和α-SMA表达的变化与miR-29和miR-155有关。这种方法可能对眼表组织工程应用有益。
