Bruch's-Mimetic Nanofibrous Membranes Functionalized with the Integrin-Binding Peptides as a Promising Approach for Human Retinal Pigment Epithelium Cell Transplantation.

Shaocheng Wang,Bo Xue,Siyong Lin,Yuntao Hu,Yueyan Guan,Chenyu Wang,Nana Yan,Xuejun Wen

doi:10.3390/molecules27041429

Abstract

Background: This study aimed to develop an ultrathin nanofibrous membrane able to, firstly, mimic the natural fibrous architecture of human Bruch’s membrane (BM) and, secondly, promote survival of retinal pigment epithelial (RPE) cells after surface functionalization of fibrous membranes. Methods: Integrin-binding peptides (IBPs) that specifically interact with appropriate adhesion receptors on RPEs were immobilized on Bruch’s-mimetic membranes to promote coverage of RPEs. Surface morphologies, Fourier-transform infrared spectroscopy spectra, contact angle analysis, Alamar Blue assay, live/dead assay, immunofluorescence staining, and scanning electron microscopy were used to evaluate the outcome. Results: Results showed that coated membranes maintained the original morphology of nanofibers. After coating with IBPs, the water contact angle of the membrane surfaces varied from 92.38 ± 0.67 degrees to 20.16 ± 0.81 degrees. RPE cells seeded on IBP-coated membranes showed the highest viability at all time points (Day 1, p < 0.05; Day 3, p < 0.01; Days 7 and 14, p < 0.001). The proliferation rate of RPE cells on uncoated poly(ε-caprolactone) (PCL) membranes was significantly lower than that of IBP-coated membranes (p < 0.001). SEM images showed a well-organized hexa/polygonal monolayer of RPE cells on IBP-coated membranes. RPE cells proliferated rapidly, contacted, and became confluent. RPE cells formed a tight adhesion with nanofibers under high-magnification SEM. Our findings confirmed that the IBP-coated PCL membrane improved the attachment, proliferation, and viability of RPE cells. In addition, in this study, we used serum-free culture for RPE cells and short IBPs without immunogenicity to prevent graft rejection and immunogenicity during transplantation. Conclusions: These results indicated that the biomimic BM-IBP-RPE nanofibrous graft might be a new, practicable approach to increase the success rate of RPE cell transplantation.

Highlights

Degenerative retinal diseases such as age-related macular degeneration (AMD) or retinitis pigmentosa (RP) are the leading cause of irreversible vision loss in people over 50 [1,2,3,4]
We investigated the physical, chemical, and cytocompatibility properties of biomimic Bruch’s membrane (BM)-Integrin-binding peptides (IBPs)-retinal pigment epithelial (RPE) nanofibrous membranes
scanning electron microscopy (SEM) images of the nanofibrous membranes are shown in Figure 1, including images of electrospun PCL membranes without coating (Figure 1A–C) and IBP-coated membranes (Figure 1D–F)

Summary

Introduction

Degenerative retinal diseases such as age-related macular degeneration (AMD) or retinitis pigmentosa (RP) are the leading cause of irreversible vision loss in people over 50 [1,2,3,4]. Retinal pigment epithelium (RPE) cell dysfunction or loss is one of the main pathological changes leading to such a wide range of degenerative retinal diseases. RPE cells seeded on IBP-coated membranes showed the highest viability at all time points (Day 1, p < 0.05; Day 3, p < 0.01; Days 7 and 14, p < 0.001). SEM images showed a well-organized hexa/polygonal monolayer of RPE cells on IBP-coated membranes. Our findings confirmed that the IBP-coated PCL membrane improved the attachment, proliferation, and viability of RPE cells. Conclusions: These results indicated that the biomimic BM-IBP-RPE nanofibrous graft might be a new, practicable approach to increase the success rate of RPE cell transplantation

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