Institute of Oral Biology, National Yang-Ming University, Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan, ROC.
Biomaterials. 2012 Nov;33(32):8003-16. doi: 10.1016/j.biomaterials.2012.07.029. Epub 2012 Jul 31.
Induced pluripotent stem cells (iPSCs) have promising potential in regenerative medicine, but whether iPSCs can promote corneal reconstruction remains undetermined. In this study, we successfully reprogrammed human corneal keratocytes into iPSCs. To prevent feeder cell contamination, these iPSCs were cultured onto a serum- and feeder-free system in which they remained stable through 30 passages and showed ESC-like pluripotent property. To investigate the availability of iPSCs as bioengineered substitutes in corneal repair, we developed a thermo-gelling injectable amphiphatic carboxymethyl-hexanoyl chitosan (CHC) nanoscale hydrogel and found that such gel increased the viability and CD44+proportion of iPSCs, and maintained their stem-cell like gene expression, in the presence of culture media. Combined treatment of iPSC with CHC hydrogel (iPSC/CHC hydrogel) facilitated wound healing in surgical abrasion-injured corneas. In severe corneal damage induced by alkaline, iPSC/CHC hydrogel enhanced corneal reconstruction by downregulating oxidative stress and recruiting endogenous epithelial cells to restore corneal epithelial thickness. Therefore, we demonstrated that these human keratocyte-reprogrammed iPSCs, when combined with CHC hydrogel, can be used as a rapid delivery system to efficiently enhance corneal wound healing. In addition, iPSCs reprogrammed from corneal surgical residues may serve as an alternative cell source for personalized therapies for human corneal damage.
诱导多能干细胞（iPSCs）在再生医学中有很大的潜力，但 iPSCs 是否能促进角膜重建尚未确定。在这项研究中，我们成功地将人角膜成纤维细胞重编程为 iPSCs。为了防止饲养细胞污染，我们将这些 iPSCs 培养在无血清和无饲养细胞的系统中，在 30 代传代过程中保持稳定，并表现出 ESC 样多能性。为了研究 iPSCs 作为角膜修复的生物工程替代物的可用性，我们开发了一种热凝胶化可注射两亲性羧甲基-己酰壳聚糖（CHC）纳米水凝胶，发现这种凝胶在存在培养基的情况下增加了 iPSCs 的活力和 CD44+比例，并维持了它们的干细胞样基因表达。iPSC 与 CHC 水凝胶（iPSC/CHC 水凝胶）联合治疗促进了手术磨损性角膜损伤的愈合。在碱性诱导的严重角膜损伤中，iPSC/CHC 水凝胶通过下调氧化应激和招募内源性上皮细胞来恢复角膜上皮厚度，促进了角膜重建。因此，我们证明了这些人角膜成纤维细胞重编程的 iPSCs，与 CHC 水凝胶结合使用时，可以作为一种快速传递系统，有效地促进角膜伤口愈合。此外，从角膜手术残余物中重编程的 iPSCs 可以作为治疗人角膜损伤的个性化治疗的替代细胞来源。
