Pourjabbar Bahareh, Shams Forough, Heidari Keshel Saeed, Biazar Esmaeil
Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Regen Med. 2024 Nov;19(11):549-560. doi: 10.1080/17460751.2024.2427513. Epub 2024 Nov 18.
To address the limitations of Cultivated Limbal Epithelial Transplantation (CLET) and the use of amniotic membrane (AM) in treating Limbal Stem Cell Deficiency (LSCD), we aimed to develop a Collagen/Silk Fibroin (Co/SF) scaffold enriched with Platelet-Rich Growth Factor (PRGF) to support the proliferation, maintenance, and differentiation of Wharton's jelly-derived mesenchymal stem cells (WJMSCs) into corneal epithelial cells (CECs).
Scaffolds loaded with PRGF were evaluated through release studies, cytotoxicity assays, and cell differentiation. The proliferation and differentiation of WJMSCs and Limbal Epithelial Stem Cells (LESCs) were investigated using MTT assays, real-time PCR and immunostaining.
The PRGF-loaded Co/SF scaffold significantly promoted the proliferation of both WJMSCs and LESCs in a concentration-dependent manner. Real-time PCR and immune staining revealed a significant increase in the expression of P63, ABCG2, and cytokeratin 3/12 markers in WJMSCs, a significant decrease in the expression of P63 and ABCG2, and a significant increase in the expression of cytokeratin 3/12 markers indicating successful differentiation into CECs.
The WJMSC cultured on PRGF-enriched Co/SF scaffold demonstrates potential as a viable alternative to conventional CLET, offering a promising strategy for corneal tissue regeneration.
为解决培养的角膜缘上皮移植（CLET）及羊膜（AM）在治疗角膜缘干细胞缺乏症（LSCD）方面的局限性，我们旨在开发一种富含富血小板生长因子（PRGF）的胶原/丝素蛋白（Co/SF）支架，以支持脐带来源的间充质干细胞（WJMSCs）增殖、维持并分化为角膜上皮细胞（CECs）。
通过释放研究、细胞毒性测定和细胞分化对负载PRGF的支架进行评估。使用MTT测定、实时聚合酶链反应（PCR）和免疫染色研究WJMSCs和角膜缘上皮干细胞（LESCs）的增殖与分化。
负载PRGF的Co/SF支架以浓度依赖的方式显著促进了WJMSCs和LESCs的增殖。实时PCR和免疫染色显示，WJMSCs中P63、ABCG2和细胞角蛋白3/12标志物的表达显著增加，P63和ABCG2的表达显著降低，细胞角蛋白3/12标志物的表达显著增加，表明成功分化为CECs。
在富含PRGF的Co/SF支架上培养的WJMSC显示出作为传统CLET可行替代方案的潜力，为角膜组织再生提供了一种有前景的策略。
