Research Institute of Clinical and Experimental Lymphology Branch of Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences, 2 Timakova Str., 630060 Novosibirsk, Russia.
Novosibirsk Branch of S. Fedorov Eye Microsurgery Federal State Institution, 10 Kolkhidskaya Str., 630096 Novosibirsk, Russia.
Int J Mol Sci. 2024 Jun 27;25(13):7053. doi: 10.3390/ijms25137053.
Cell-based therapies using corneal stromal stem cells (CSSC), corneal keratocytes, or a combination of both suppress corneal scarring. The number of quiescent keratocytes in the cornea is small; it is difficult to expand them in vitro in quantities suitable for transplantation. This study examined the therapeutic effect of corneal fibroblasts reversed into keratocytes (rCF) in a mouse model of mechanical corneal injury. The therapeutic effect of rCF was studied in vivo (slit lamp, optical coherence tomography) and ex vivo (transmission electron microscopy and immunofluorescence staining). Injection of rCF into the injured cornea was accompanied by recovery of corneal thickness, improvement of corneal transparency, reduction of type III collagen in the stroma, absence of myofibroblasts, and the improvement in the structural organization of collagen fibers. TEM results showed that 2 months after intrastromal injection of cells, there was a decrease in the fibril density and an increase in the fibril diameter and the average distance between collagen fibrils. The fibrils were well ordered and maintained the short-range order and the number of nearest-neighbor fibrils, although the averaged distance between them increased. Our results demonstrated that the cell therapy of rCF from ReLEx SMILe lenticules promotes the recovery of transparent corneal stroma after injury.
基于角膜基质干细胞 (CSSC)、角膜成纤维细胞或两者联合的细胞疗法可抑制角膜瘢痕形成。角膜中的静止性角膜成纤维细胞数量较少;很难在体外大量扩增它们,以达到适合移植的数量。本研究在机械性角膜损伤的小鼠模型中研究了角膜成纤维细胞逆转为角膜细胞(rCF)的治疗效果。在体内(裂隙灯、光学相干断层扫描)和体外(透射电子显微镜和免疫荧光染色)研究了 rCF 的治疗效果。将 rCF 注射到受伤的角膜中伴随着角膜厚度的恢复、角膜透明度的改善、基质中层 III 型胶原的减少、成肌纤维细胞的消失以及胶原纤维结构组织的改善。TEM 结果表明,细胞间质内注射后 2 个月,纤维密度降低,纤维直径和胶原纤维之间的平均距离增加。尽管它们之间的平均距离增加,但纤维排列整齐,保持了短程有序和最近邻纤维的数量。我们的研究结果表明,ReLEx SMILe 角膜透镜的 rCF 细胞疗法可促进损伤后透明角膜基质的恢复。