Di Girolamo Nick, Chui Jeanie, Wakefield Denis, Coroneo Minas T
Inflammatory Diseases Research Unit, Department of Pathology, School of Medical Sciences, University of New South Wales, Sydney, NSW 2052, Australia.
Br J Ophthalmol. 2007 Apr;91(4):459-64. doi: 10.1136/bjo.2006.103895. Epub 2006 Sep 20.
This study was initiated after observation of some intriguing epithelial growth properties of contact lenses used as a bandage for patients after pterygium surgery.
To determine the efficacy of culturing human ocular surface epithelial cells on therapeutic contact lenses in autologous serum with a view of using this system to transfer epithelial cells to patients with persistent corneal or limbal defects.
Excess graft tissue resected from patients undergoing pterygium surgery (n = 3) consisting of limbal epithelium was placed on siloxane-hydrogel contact lenses (lotrafilcon A and balafilcon A). Limbal explants were cultured in media with 10% autologous serum. Morphology, proliferative capacity and cytokeratin profile were determined by phase contrast, light and electron microscopy, and immunohistochemical analysis.
Lotrafilcon A contact lenses sustained proliferation and migration from limbal tissue. Cells became confluent after 10-14 days and consisted of 2-3 layers with a corneal phenotype (CK3(+)/CK12(+)/CK19(-)) and a propensity to proliferate (p63(+)). Electron microscopy showed microvilli on the apical surface with adhesive projections, indicating that these cells were stable and likely to survive for a long term. Growth was not observed from limbal explants cultured on balafilcon A contact lenses.
A method for culturing human ocular surface epithelium on contact lenses that may facilitate expansion and transfer of autologous limbal epithelial cells while avoiding the risks associated with transplanting allogeneic tissue has been developed. This technique may be potentially useful for the treatment of patients with limbal stem cell deficiency.
本研究始于观察到用于翼状胬肉手术后患者包扎的隐形眼镜具有一些有趣的上皮生长特性之后。
确定在自体血清中于治疗性隐形眼镜上培养人眼表上皮细胞的效果,以期利用该系统将上皮细胞转移至患有持续性角膜或角膜缘缺损的患者。
将从接受翼状胬肉手术的患者(n = 3)切除的多余移植组织(由角膜缘上皮组成)置于硅氧烷水凝胶隐形眼镜(lotrafilcon A和balafilcon A)上。角膜缘外植体在含10%自体血清的培养基中培养。通过相差显微镜、光学显微镜和电子显微镜以及免疫组织化学分析来确定形态、增殖能力和细胞角蛋白谱。
Lotrafilcon A隐形眼镜能维持角膜缘组织的增殖和迁移。细胞在10 - 14天后汇合,由2 - 3层具有角膜表型(CK3(+)/CK12(+)/CK19(-))且有增殖倾向(p63(+))的细胞组成。电子显微镜显示顶端表面有微绒毛和黏附突起,表明这些细胞稳定且可能长期存活。在balafilcon A隐形眼镜上培养的角膜缘外植体未观察到生长。
已开发出一种在隐形眼镜上培养人眼表上皮的方法,该方法可能有助于自体角膜缘上皮细胞的扩增和转移,同时避免与移植异体组织相关的风险。这项技术可能对治疗角膜缘干细胞缺乏症患者有潜在用途。