1 Surgical Research Unit, Centre for Eye Research Australia , Melbourne, Australia .
Tissue Eng Part A. 2014 Feb;20(3-4):646-55. doi: 10.1089/ten.TEA.2013.0089. Epub 2014 Jan 23.
Extensive damage to the limbal region of the cornea leads to a severe form of corneal blindness termed as limbal stem cell deficiency (LSCD). Whereas most cases of corneal opacity can be treated with full thickness corneal transplants, LSCD requires stem cell transplantation for successful ocular surface reconstruction. Current treatments for LSCD using limbal stem cell transplantation involve the use of murine NIH 3T3 cells and human amniotic membranes as culture substrates, which pose the threat of transmission of animal-derived pathogens and donor tissue-derived cryptic infections. In this study, we aimed to produce surface modified therapeutic contact lenses for the culture and delivery of corneal epithelial cells for the treatment of LSCD. This approach avoids the possibility of suture-related complications and is completely synthetic. We used plasma polymerization to deposit acid functional groups onto the lenses at various concentrations. Each surface was tested for its suitability to promote corneal epithelial cell adhesion, proliferation, retention of stem cells, and differentiation and found that acid-based chemistries promoted better cell adhesion and proliferation. We also found that the lenses coated with a higher percentage of acid functional groups resulted in a higher number of cells transferred onto the corneal wound bed in rabbit models of LSCD. Immunohistochemistry of the recipient cornea confirmed the presence of autologous, transplanted 5-bromo-2'-deoxyuridine (BrdU)-labeled cells. Hematoxylin staining has also revealed the presence of a stratified epithelium at 26 days post-transplantation. This study provides the first evidence for in vivo transfer and survival of cells transplanted from a contact lens to the wounded corneal surface. It also proposes the possibility of using plasma polymer-coated contact lenses with high acid functional groups as substrates for the culture and transfer of limbal cells in the treatment of LSCD.
角膜缘区域的广泛损伤会导致一种严重的角膜盲,称为角膜缘干细胞缺乏症(LSCD)。虽然大多数角膜混浊病例可以通过全层角膜移植治疗,但 LSCD 需要干细胞移植才能成功进行眼表面重建。目前使用角膜缘干细胞移植治疗 LSCD 的方法涉及使用鼠 NIH 3T3 细胞和人羊膜作为培养基质,这存在传播动物源性病原体和供体组织来源的隐匿性感染的威胁。在这项研究中,我们旨在生产表面改性治疗性隐形眼镜,用于培养和输送角膜上皮细胞,以治疗 LSCD。这种方法避免了与缝线相关的并发症的可能性,并且完全是合成的。我们使用等离子体聚合在镜片上沉积不同浓度的酸官能团。每个表面都经过测试,以确定其适合促进角膜上皮细胞黏附、增殖、保留干细胞以及分化的能力,结果发现基于酸的化学物质能更好地促进细胞黏附和增殖。我们还发现,涂覆更高百分比酸官能团的镜片在 LSCD 兔模型中导致更多数量的细胞转移到角膜伤口床上。受体角膜的免疫组织化学证实了存在自体、移植的 5-溴-2'-脱氧尿苷(BrdU)标记细胞。苏木精染色也显示出在移植后 26 天存在分层上皮。这项研究首次提供了从隐形眼镜移植到受伤角膜表面的细胞在体内转移和存活的证据。它还提出了使用具有高酸官能团的等离子体聚合物涂层隐形眼镜作为培养和转移角膜缘细胞的基质,用于治疗 LSCD 的可能性。