Electrospun Silk Fibroin Blended P(LLA-CL) Membrane As Potential Substrate For Epithelial Constructs Used In Ocular Surfac Bioengineerin

Dysfunction or loss of limbal epithelial stem cells produces varying degrees of limbal stem cell deficiency (LSCD), which may lead to decreased vision, ocular discomfort, pain, and an unstable ocular surface. Transplantation of ex-vivo expanded limbal epithelial cells (LECs) has been found as a promising procedure to treat corneas manifesting LSCD.This study sought to assess a electrospun silk fibroin (SF) and poly(L-lactic acid-co-e-caprolactone)(P(LLA-CL)) biomaterial as a substrate for corneal epithelial cell adhere, proliferation, and differentiation. To develop a synthetic stromal substrate for LECs expansion that can serve as a potential substrate. The main contents are as follows:Nanofibrous scaffolds of SF and poly(L-lacticacid-co-s-caprolactone)(P(LLA-CL)) blends were fabricated via electrospinning. The results showed that the The average nanofibrous diameter of SF/P(LLA-CL) in control group was the widest, and the ethanol treated fibre was the thinnest. Water contact angle measurements conformed that the greatest angle existed in the ethanol treated group, while others similar. The membranes in a wet state have significantly higher transparency when compared with dry PCL membrane. The transmittance of all the samples in a wet state was higher than a dry state using UV-Visible spectroscopy.Rabbit limbal epithelial cell on primary passage was planted on this membrane. The cells were showed to growth faster on the water vapor-treated SF/P(LLA-CL) nanofiber membranes via cell attachment analysis and MTT assay. The viability staining and HE staining suggested that the cells growed well on the membranes and form a comparable stratified graft with a2to3cell layering. IF results revealed that there was no change in the expression profile of LECs grown on nanofibers. SEM micrograph showed that the corneal epithelium displayed a polygonal pattern and was firmly anchored to the nanofiber surface. There were abundant microvilli distributed on the surface of the cells, which had wide connection with the adjacent cells.SF/P(LLA-CL) blended nanofibrous scaffolds were implanted in the comea of New Zealand rabbits and evaluated their tissue compatibility in vivo.The results showed by slit lamp observation and HE staining demonstrated that SF/P(LLA-CL) blended nanofibrous scaffolds had good tissue compatibility. This scaffold provides not only a milieu supporting LEC expansion, but also serves as a potential alternative substrate for ocular surface tissue engineering.