Effects Of SF/PLCL Nanofibrous Scaffolds And BTC On The Proliferation And Differentiation Of Retina Progenitor Cells

Objectives In this study,we characterized the role of Silk fibroin(SF)/ poly(L-lactic acid-co-ε-caprolactone)(PLCL)nanofibrous scaffold and betacellulin(BTC)on RPC proliferation and differentiation in vitro,which may provide a new insight for the treatment of degenerative retinal diseases.Methods 1.Testing different proportions of SF/PLCL nanofiber membrane for electron microscopy morphology,fiber diameter distribution,porosity,swelling rate,mechanical properties,hydrophilic Angles.Electron microscope and inverted microscope were used to detect the morphology of the cells on the membranes.Analyses of gene and protein expression level of retinal progenitor-related markers and retinal development-related makers were performed using q PCR and immunocytochemistry.2.Detection of the effect of BTC on RPC proliferation by CCK-8 analysis,immunocytochemistry and Western blot.q PCR and western blot analyses were performed to investigate the potential fates of the BTC-treated RPCs.Results 1.Our data demonstrate that the SF:PLCL(1:1)nanofibrous scaffolds present cytocompatibility for RPC growth.Moreover,the SF:PLCL(1:1)scaffolds can markedly promote RPC proliferation and can robustly accelerate the differentiation of RPCs into β3-tubulin and rhodopsin positive retinal neuronal cells.2.BTC plays important roles in the proliferation and differentiation of RPCs.Our results showed that BTC can significantly promote the proliferation of RPCs more efficiently than EGF.BTC stimulated RPC proliferation more powerfully through the EGFR/Erb B2/Erb B4-Akt/Erk pathway.Meanwhile,under differentiated conditions,the BTC-pretreated RPCs were preferentially differentiated into retinal neurons,including photoreceptors,compared to the EGF-pretreated RPCs.Conclusion 1.The SF:PLCL(1:1)nanofibrous scaffolds exhibited cytocompatibility for RPC growth and these scaffolds can not only markedly promote RPC expansion but also robustly enhance RPC differentiation toward retinal neurons,which suggests that SF: PLCL(1:1)may have potential application in the cell replacement therapy of retina degenerative diseases.2.BTC can not only markedly promote RPC expansion through the EGFR/Erb B2/Erb B4-Akt/Erk pathway but also enhance RPC differentiation into retinal neuronal cells,including photoreceptors,the most interesting class of retinal neuronal cells for retinal cell replacement therapy.The present study of the effects of BTC on RPCs may provide us with some new insights into the controlled proliferation and differentiation of RPCs in vitro and provide new insights for the cell replacement therapy for retinal degenerative diseases.