Inflammation and Infection Research Centre, School of Medical Sciences, University of New South Wales, Sydney, Australia.
Invest Ophthalmol Vis Sci. 2011 Oct 17;52(11):8138-47. doi: 10.1167/iovs.11-8140.
The authors recently developed a therapeutic technique for patients with limbal stem cell deficiency by harvesting ocular surface stem cells (SCs), expanding them on therapeutic contact lenses (CLs), and applying them to diseased corneas. The present study determined the proteins that bind to CLs and whether such factors, along with transplanted cells, are critical determinants for corneal rehabilitation using this method.
Therapeutic CLs were exposed to human serum, and adherent proteins were analyzed by proteomics. The distribution of vitronectin (VN) on the ocular surface was determined with specific antibodies. Cadaveric human corneas were chemically wounded, and cell transfer by CLs was assessed in organ culture.
VN was identified as a serum factor that binds and desorbs from CLs. VN localized to the limbal and basement membranes (BM) of other SC-harboring organs. Clonogenic assays demonstrated higher colony-forming efficiency on VN compared with uncoated surfaces. Cell transfer from CLs was achieved through in vitro models and was abrogated by RGD peptides and inhibitory antibodies to VN and its receptor.
Identification of VN within the limbal BM, its effect on limbal SC activity, and the discovery of this factor on serum-exposed CL polymers implies a role in supporting progenitor cells and facilitating corneal regeneration.
作者最近开发了一种治疗眼表干细胞缺乏症患者的技术,方法是从眼表面采集干细胞(SCs),在治疗性隐形眼镜(CLs)上扩增,然后将其应用于患病的角膜。本研究旨在确定与 CL 结合的蛋白,以及这些因素与移植细胞一起,是否是该方法用于角膜康复的关键决定因素。
将治疗性 CL 暴露于人血清中,并通过蛋白质组学分析黏附蛋白。用特异性抗体测定 VN 在眼表面的分布。用人尸体角膜进行化学性创伤,在器官培养中评估 CL 介导的细胞转移。
VN 被鉴定为一种与 CL 结合并从 CL 上解吸的血清因子。VN 定位于角膜缘和其他含有 SC 的器官的基膜(BM)。克隆形成试验表明,VN 表面的集落形成效率高于未涂层表面。通过体外模型实现了从 CL 转移的细胞,并且 RGD 肽以及针对 VN 和其受体的抑制性抗体可阻断这种转移。
在角膜缘 BM 内鉴定出 VN,其对角膜缘 SC 活性的影响,以及在暴露于血清的 CL 聚合物上发现该因子,提示其在支持祖细胞和促进角膜再生方面具有重要作用。