Abstract
Biocompatible polymer scaffolds are promising as potential carriers for the delivery of retinal progenitor cells (RPCs) in cell replacement therapy for the repair of damaged or diseased retinas. The primary goal of the present study was to investigate the effects of blended electrospun nanofibrous membranes of silk fibroin (SF) and poly(L-lactic acid-co-ε-caprolactone) (PLCL), a novel scaffold, on the biological behaviour of RPCs in vitro. To assess the cell-scaffold interaction, RPCs were cultured on SF/PLCL scaffolds for indicated durations. Our data revealed that all the SF/PLCL scaffolds were thoroughly cytocompatible, and the SF:PLCL (1:1) scaffolds yielded the best RPC growth. The in vitro proliferation assays showed that RPCs proliferated more quickly on the SF:PLCL (1:1) than on the other scaffolds and the control. Quantitative polymerase chain reaction (qPCR) and immunocytochemistry analyses demonstrated that RPCs grown on the SF:PLCL (1:1) scaffolds preferentially differentiated toward retinal neurons, including, most interestingly, photoreceptors. In summary, we demonstrated that the SF:PLCL (1:1) scaffolds can not only markedly promote RPC proliferation with cytocompatibility for RPC growth but also robustly enhance RPCs’ differentiation toward specific retinal neurons of interest in vitro, suggesting that SF:PLCL (1:1) scaffolds may have potential applications in retinal cell replacement therapy in the future.
Highlights
Bipolar cells[14,15,16]
scanning electron microscope (SEM) images depicting the micromorphology of the electrospun nanofibrous scaffolds are shown in Fig. 1B–F, and the average fibre diameter of the scaffolds gradually decreased from 432.7 nm to 137.8 nm with increasing Silk fibroin (SF) content (Fig. 1G–K), demonstrating that all five types of scaffolds were constructed of randomly displayed fibres and completely interconnected pore structures
The equilibrium swelling ratio (ESR) results are shown in Supplementary Fig. S1, and the results showed that the swelling ratio of the scaffolds decreased with an increase in the poly(L-lactic acid-co-ε -caprolactone) (PLCL) content
Summary
Bipolar cells[14,15,16]. The second problem for the future clinical application of RPCs is how to effectively deliver RPCs to the retina and ensure their ability to integrate into the retina and differentiate into retinal neuronal cells. Electrospun PLCL nanofibres have been demonstrated to support the growth and proliferation of many cell types while showing inadequate cell affinity due to the absence of recognition sites for cell adhesion. We investigated the effects of electrospun interconnected and porous nanofibrous scaffolds composed of SF and PLCL on retinal progenitor growth. Our data demonstrate that different concentrations of SF/PLCL nanofibrous scaffolds performed well but showed different bioactivities for RPC growth. The data obtained with the SF:PLCL (1:1) scaffolds demonstrate that these can enhance RPC proliferation and promote RPC differentiation toward retinal neurons, such as rhodopsin-positive photoreceptor cells, indicating that electrospun SF:PLCL (1:1) scaffolds may be useful in retinal cell replacement therapies
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