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While cell transplantation therapies show great promise as treatments for retinal degeneration, the challenge of low cell survival upon transplantation motivates exploration of materials that may serve as cell delivery vehicles and promote survival and differentiation. In this study, we explored the native matrix that surrounds the outer segments of photoreceptors and promotes their homeostasis, interphotoreceptor matrix (IPM), as a substrate for human retinal progenitor cells (hRPCs). Bovine IPM was characterized to determine its structure and biochemical composition, and processed to develop substrates for cells. Cell viability, morphology, proliferation and expression of photoreceptors marker genes were studied on IPM-based substrates in vitro. We explored different preparations of IPM as a scaffold. Lectin staining revealed that a distinct honeycomb structure of native IPM is lost during centrifugation to prepare a more concentrated suspension of matrix. Biochemical analysis of bovine IPM indicated presence of glycosaminoglycans and proteins. IPM mediated hRPC attachment and spreading with no signs of cytotoxicity. Cells proliferated more on native IPM substrates compared to IPM that was centrifuged to create a concentrated suspension. Cells cultured on IPM substrates expressed markers of photoreceptors: rhodopsin, NRL and ROM1. Together this data supports further exploration of IPM as a tool for retinal tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 891-899, 2018. [ABSTRACT FROM AUTHOR]

Purpose: Degenerative diseases of the retina, such as retinitis pigmentosa and age-related macular degeneration, are characterized by the irreversible loss of photoreceptors. Several growth factors, including glial cell derived neurotrophic factor (GDNF), have been shown to rescue retinal neurons. An alternative strategy to direct GDNF administration is its induction in host retina by small molecules. Here we studied the ability of a novel small molecule GSK812 to induce GDNF in vitro/in vivo and rescue photoreceptors.Methods: GDNF induction in vitro was assessed in human ARPE-19, human retinal progenitor cells (RPCs) and mouse pluripotent cell-derived eyecups. For time course pharmacokinetic and GDNF induction studies in C57Bl/6 mice, GSK812 sustained release formulation was injected intravitreally. The same delivery approach was used in the rhodopsin knockout mice and Royal College of Surgeon (RCS) rats to assess long-term GDNF induction and photoreceptor rescue.Results: The suspension provided sustained GSK812 delivery with 28 μg of drug remaining in the eye 2 weeks after a single injection. GSK812 suspension injection in C57Bl/6 mice resulted in significant upregulation of GDNF mRNA (>1.8-fold) and protein levels (>2.8-fold). Importantly, GSK812 treatment resulted in outer nuclear layer preservation in rho-/- mice with a 2-fold difference in photoreceptor number. In the RCS rat, the GSK812 injection provided long-term rescue of photoreceptors and outer segments, accompanied by function preservation as well.Conclusions: GSK812 is a potent neuroprotectant that can induce GDNF in normal and diseased retina. This induction results in photoreceptor rescue in 2 models of retinal degeneration. [ABSTRACT FROM AUTHOR]

Many advances have been made in an attempt to treat retinal degenerative diseases, such as age-related macular degeneration and retinitis pigmentosa. The irreversible loss of photoreceptors is common to both, and currently no restorative clinical treatment exists. It has been shown that retinal progenitor and photoreceptor precursor cell transplantation can rescue the retinal structure and function. Importantly, retinal progenitor cells can be collected from the developing neural retina with further expansion and additional modification in vitro, and the delivery into the degenerative host can be performed as a single-cell suspension injection or as a complex graft transplantation. Previously, we have described several polymer scaffolds for culture and transplantation of retinal progenitor cells of both mouse and human origin. This tissue engineering strategy increases donor cell survival and integration. We have also shown that biodegradable poly(ɛ-caprolactone) induces mature photoreceptor differentiation from human retinal progenitor cells. However, poor adhesive properties limit its use, and therefore it requires additional surface modification. The aim of this work was to study vitronectin-mimicking oligopeptides (Synthemax II-SC) poly(ɛ-caprolactone) films and their effects on human retinal progenitor cell adhesion, proliferation, and differentiation. Here, we show that the incorporation of vitronectin-mimicking oligopeptide into poly(ɛ-caprolactone) leads to dose-dependent increases in cell adhesion; the optimum dose identified as 30 µg/ml. Inhibition of human retinal progenitor cells proliferation was seen on poly(ɛ-caprolactone) and was maintained with the hybrid scaffold. This has been shown to be beneficial for driving cell differentiation. Additionally, we observed equal expression of Nrl, rhodopsin, recoverin, and rod outer membrane 1 after differentiation on the hybrid scaffold as compared to the standard fibronectin coating of poly(ɛ-caprolactone). After transplantation into rd1 retina degenerative mice, human retinal progenitor cells were able to migrate to the outer nuclear layer and survive for three weeks. We conclude that Synthemax II-SC can be incorporated into poly(ɛ-caprolactone) to create a hybrid chemically defined scaffold for clinical application. [ABSTRACT FROM AUTHOR]

Background: Human retinal progenitor cells (hRPCs) are expandable in vitro and represent a possible therapy for retinal degenerative diseases. Results: In a rat model of retinal degeneration, transplantation of hRPCs preserved photoreceptors and visual function. Conclusion: Subretinal injection of hRPCs rescues photoreceptors without causing adverse effects. Significance: This study provides proof of concept for hRPC transplantation and paves the way for further studies and human trials. [ABSTRACT FROM AUTHOR]

Tissue engineering has been widely applied in different areas of regenerative medicine, including retinal regeneration. Typically, artificial biopolymers require additional surface modification (e.g. with arginine-glycine-aspartate-containing peptides or adsorption of protein, such as fibronectin), before cell seeding. Here, we describe an alternative approach for scaffold design: the manufacture of hybrid interphotoreceptor matrix-poly (ε-caprolactone) scaffolds, in which the insoluble extracellular matrix of the retina is incorporated into a biodegradable polymer well suited for transplantation. The incorporation of interphotoreceptor matrix did not change the topography of polycaprolactone film, although it led to a slight increase in hydrophilic properties (water contact angle measurements). This hybrid scaffold provided sufficient stimuli for human retinal progenitor cell adhesion and inhibited proliferation, leading to differentiation toward photoreceptor cells (expression of Crx, Nrl, rhodopsin, ROM1). This scaffold may be used for transplantation of retinal progenitor cells and their progeny to treat retinal degenerative disorders. [ABSTRACT FROM AUTHOR]