Baradaran-Rafii Alireza, Biazar Esmaeil, Heidari-keshel Saeed
a Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences , Tehran , Iran .
b Department of Biomaterials Engineering , Islamic Azad University , Tonekabon Branch , Iran , and.
Curr Eye Res. 2016;41(3):326-33. doi: 10.3109/02713683.2015.1019004. Epub 2015 Apr 21.
The aim of this study was to develop nanofibrous polycaprolactone (PCL) substrate for limbal stem cell (LSC) expansion that can serve as a potential alternative substrate to replace human amniotic membrane (AM).
The human limbus stem cell was used to evaluate the biocompatibility of substrates (nanofibrous scaffold and, human AM) based on their phenotypic profile, viability, proliferation and attachment ability.
Biocompatibility results indicated that the all substrates were highly biocompatible, as LSCs could favorably attach and proliferate on the nanofibrous surface. Microscopic figures showed that the human LSCs were firmly anchored to the substrates and were able to retain a normal corneal stem cell phenotype. Microscopic analyses illustrated that cells infiltrated the nanofibers and successfully formed a three-dimensional corneal epithelium, which was viable for two weeks. Immunocytochemistry (ICC) and real time-PCR results revealed no change in the expression profile of LECs grown on nanofibrous substrate when compared to those grown on human AM.
In addition, electrospun nanofibrous PCL substrate provides not only a milieu supporting LSCs expansion, but also serve as a useful alternative carrier for ocular surface tissue engineering and could be used as an alternative substrate to AM.
本研究的目的是开发用于角膜缘干细胞(LSC)扩增的纳米纤维聚己内酯(PCL)基质,其可作为替代人羊膜(AM)的潜在替代基质。
用人角膜缘干细胞基于其表型特征、活力、增殖和附着能力来评估基质(纳米纤维支架和人羊膜)的生物相容性。
生物相容性结果表明,所有基质都具有高度生物相容性,因为角膜缘干细胞能够在纳米纤维表面良好地附着和增殖。显微镜图像显示,人角膜缘干细胞牢固地锚定在基质上,并能够保持正常的角膜干细胞表型。显微镜分析表明,细胞浸润纳米纤维并成功形成三维角膜上皮,其在两周内保持活力。免疫细胞化学(ICC)和实时PCR结果显示,与在人羊膜上生长的角膜缘上皮细胞相比,在纳米纤维基质上生长的角膜缘上皮细胞的表达谱没有变化。
此外,电纺纳米纤维PCL基质不仅提供了支持角膜缘干细胞扩增的环境,而且还可作为眼表组织工程的有用替代载体,并可作为羊膜的替代基质。