Treatment Of Retinal Degeneration Research Articles

Statistical Evaluation of ERG Responses: A New Method to Validate Cycle-by-Cycle Recordings in Advanced Retinal Degenerations.

To describe and evaluate a novel method to determine the validity of measurements made using cycle-by-cycle (CxC) recording techniques in patients with advanced retinal degenerations (RD) having low-amplitude flicker electroretinogram (ERG) responses. The method extends the original CxC recording algorithm introduced by Sieving et al., retaining the original recording setup and the preliminary analysis of raw data. Novel features include extended use of spectrum analysis, reduction of errors due to known sources, and a comprehensive statistical assessment using three different tests. The method was applied to ERG recordings from seven patients with RD and two patients with CNGB3 achromatopsia. The method was implemented as a Windows application to processes raw data obtained from a commercial ERG system, and it features a computational toolkit for statistical assessment of ERG recordings with amplitudes as low as 1 µV, commonly found in advanced RD patients. When recorded using conditions specific for eliciting cone responses, none of the CNGB3 patients had a CxC validated response, indicating that no signal artifacts were present with our recording conditions. A comparison of the presented method with conventional 30 Hz ERG was performed. Bland-Altman plots indicated good agreement (mean difference, -0.045 µV; limits of agreement, 0.193 to -0.282 µV) between the resulting amplitudes. Within-session test-retest variability was 15%, comparing favorably to the variability of standard ERG amplitudes. This novel method extracts highly reliable clinical recordings of low-amplitude flicker ERGs and effectively detects artifactual responses. It has potential value both as a cone outcome variable and planning tool in clinical trials on natural history and treatment of advanced RDs.

Extracellular vesicles from organoid-derived human retinal progenitor cells prevent lipid overload-induced retinal pigment epithelium injury by regulating fatty acid metabolism.

Retinal degeneration (RD), a group of diseases leading to irreversible vision loss, is characterised by retinal pigment epithelium (RPE) or retinal neuron damage and loss. With fewer risks of immune rejection and tumorigenesis, stem cell-secreted extracellular vesicles (EVs) offer a new cell-free therapeutic paradigm for RD, which remains to be investigated. Human retinal organoid-derived retinal progenitor cells (hERO-RPCs) are an easily accessible and advanced cell source for RD treatment. However, hERO-RPCs-derived EVs require further characterisation. Here, we compared the characteristics of EVs from hERO-RPCs (hRPC-EVs) with those of human embryonic stem cell (hESC)-derived EVs (hESC-EVs) as controls. Based on in-depth proteomic analysis, we revealed remarkable differences between hRPC-EVs and hESC-EVs. A comparison between EVs and their respective cells of origin demonstrated that the protein loading of hRPC-EVs was more selective than that of hESC-EVs. In particular, hESC-EVs were enriched with proteins related to angiogenesis and cell cycle, whereas hRPC-EVs were enriched with proteins associated with immune modulation and retinal development. More importantly, compared with that of hESC-EVs, hRPC-EVs exhibited a lower correlation with cell proliferation and a unique capacity to regulate lipid metabolism. It was further confirmed that hRPC-EVs potentially eliminated lipid deposits, inhibited lipotoxicity and oxidative stress, and enhanced phagocytosis and survival of oleic acid-treated ARPE-19 cells. Mechanistically, hRPC-EVs are integrated into the mitochondrial network of oleic acid-treated ARPE-19 cells, and increased the level of mitochondrial fatty acid β-oxidation-related proteins. Thus, organoid-derived hRPC-EVs represent a promising source of cell-free therapy for RD, especially for blinding diseases related to abnormal lipid metabolism in RPE cells.

Nonviral base editing of KCNJ13 mutation preserves vision in a model of inherited retinal channelopathy.

Clinical genome editing is emerging for rare disease treatment, but one of the major limitations is the targeting of CRISPR editors' delivery. We delivered base editors to the retinal pigmented epithelium (RPE) in the mouse eye using silica nanocapsules (SNCs) as a treatment for retinal degeneration. Leber congenital amaurosis type 16 (LCA16) is a rare pediatric blindness caused by point mutations in the KCNJ13 gene, a loss of function inwardly rectifying potassium channel (Kir7.1) in the RPE. SNCs carrying adenine base editor 8e (ABE8e) mRNA and sgRNA precisely and efficiently corrected the KCNJ13W53X/W53X mutation. Editing in both patient fibroblasts (47%) and human induced pluripotent stem cell-derived RPE (LCA16-iPSC-RPE) (17%) showed minimal off-target editing. We detected functional Kir7.1 channels in the edited LCA16-iPSC-RPE. In the LCA16 mouse model (Kcnj13W53X/+ΔR), RPE cells targeted SNC delivery of ABE8e mRNA preserved normal vision, measured by full-field electroretinogram (ERG). Moreover, multifocal ERG confirmed the topographic measure of electrical activity primarily originating from the edited retinal area at the injection site. Preserved retina structure after treatment was established by optical coherence tomography (OCT). This preclinical validation of targeted ion channel functional rescue, a challenge for pharmacological and genomic interventions, reinforced the effectiveness of nonviral genome-editing therapy for rare inherited disorders.

Engineered Extracellular Vesicles for Delivery of an IL-1 Receptor Antagonist Promote Targeted Repair of Retinal Degeneration.

Retinal degeneration (RD) is an irreversible blinding disease that seriously affects patients' daily activities and mental health. Targeting hyperactivated microglia and regulating polarization are promising strategies for treating the disease. Mesenchymal stem cell (MSC) transplantation is proven to be an effective treatment due to its immunomodulatory and regenerative properties. However, the low efficiency of cell migration and integration of MSCs remains a major obstacle to clinical use. The goal of this study is to develop a nanodelivery system that targets hyperactivated microglia and inhibits their release of proinflammatory factors, to achieve durable neuroprotection. This approach is to engineer extracellular vesicles (EVs) isolated from MSC, modify them with a cyclic RGD (cRGD) peptide on their surface, and load them with an antagonist of the IL-1 receptor, anakinra. Comparing with non-engineered EVs, it is observed that engineered cRGD-EVs exhibit an increased targeting efficiency against hyperactivated microglia and strongly protected photoreceptors in experimental RD cells and animal models. This study provides a strategy to improve drug delivery to degenerated retinas and offers a promising approach to improve the treatment of RD through targeted modulation of the immune microenvironment via engineered cRGD-EVs.

MTOR Signalling Pathway: A Potential Therapeutic Target for Ocular Neurodegenerative Diseases.

Recent advances in the research of the mammalian target of the rapamycin (mTOR) signalling pathway demonstrated that mTOR is a robust therapeutic target for ocular degenerative diseases, including age-related macular degeneration (AMD), diabetic retinopathy (DR), and glaucoma. Although the exact mechanisms of individual ocular degenerative diseases are unclear, they share several common pathological processes, increased and prolonged oxidative stress in particular, which leads to functional and morphological impairment in photoreceptors, retinal ganglion cells (RGCs), or retinal pigment epithelium (RPE). mTOR not only modulates oxidative stress but is also affected by reactive oxygen species (ROS) activation. It is essential to understand the complicated relationship between the mTOR pathway and oxidative stress before its application in the treatment of retinal degeneration. Indeed, the substantial role of mTOR-mediated autophagy in the pathogenies of ocular degenerative diseases should be noted. In reviewing the latest studies, this article summarised the application of rapamycin, an mTOR signalling pathway inhibitor, in different retinal disease models, providing insight into the mechanism of rapamycin in the treatment of retinal neurodegeneration under oxidative stress. Besides basic research, this review also summarised and updated the results of the latest clinical trials of rapamycin in ocular neurodegenerative diseases. In combining the current basic and clinical research results, we provided a more complete picture of mTOR as a potential therapeutic target for ocular neurodegenerative diseases.

Blue Light Induces RPE Cell Necroptosis, Which Can Be Inhibited by Minocycline.

Purpose:Damage to and death of the retinal pigment epithelium (RPE) are closely related to retinal degeneration. Blue light is a high-energy light that causes RPE damage and triggers inflammatory responses. This study investigates whether blue light induces RPE necroptosis, explores pharmacologic therapy and specific mechanisms, and provides hints for research on retinal degeneration.MethodsThe human RPE cell line ARPE-19 was cultured and subjected to blue light insult in vitro. Annexin V/PI was used to evaluate RPE survival. Minocycline was applied to inhibit the death of RPE. Proteomic measurement was used to analyze protein expression. Inhibitors of necroptosis and apoptosis were applied to assess the death mode. Immunofluorescence of protein markers was detected to analyze the mechanism of cell death. Subcellular structural changes were detected by transmission electron microscopy. Reactive oxygen species (ROS) was tested by DCFH-DA. Mitochondrial membrane potential (Δψm) was detected by JC-1. BALB/c mice received bule light exposure, and RPE flatmounts were stained for verification in vivo.ResultsBlue light illumination induced RPE death, and minocycline significantly diminished RPE death. Proteomic measurement showed that minocycline effectively mitigated protein hydrolysis and protein synthesis disorders. Necroptosis inhibitors (Nec-1s, GSK-872) increased the survival of RPE cells, but apoptosis inhibitors (Z-VAD-FMK) did not. After blue light illumination, high-mobility group box-1 (HMGB1) was released from the nucleus, receptor-interacting protein kinase 3 (RIPK3) aggregated, and mixed-lineage kinase domain-like protein (MLKL) increased in the RPE. The application of minocycline alleviated the above phenomena. After blue light illumination, RPE cells exhibited necrotic characteristics accompanied by destruction of cell membranes and vacuole formation, but nuclear membranes remained intact. Minocycline improved the morphology of RPE. Blue light increased ROS and decreased Δψm of RPE, minocycline did not reduce ROS but kept Δψm stable. In vivo, HMGB1 release and RIPK3 aggregation appeared in the RPE of BALB/c mice after blue light illumination, and minocycline alleviated this effect.ConclusionsBlue light exposure causes RPE necroptosis. Minocycline reduces the death of RPE by keeping Δψm stable, inhibiting necroptosis, and preventing HMGB1 release. These results provide new ideas for the pathogenesis and treatment of retinal degeneration.

Bone marrow-derived mononuclear stem cells in the treatment of retinal degenerations.

Retinal degenerative diseases affecting the outer retina in its many forms (inherited, acquired or induced) are characterized by photoreceptor loss, and represent currently a leading cause of irreversible vision loss in the world. At present, there are very few treatments capable of preventing, recovering or reversing photoreceptor degeneration or the secondary retinal remodeling, which follows photoreceptor loss and can also cause the death of other retinal cells. Thus, these diseases are nowadays one of the greatest challenges in the field of ophthalmological research. Bone marrow derived-mononuclear stem cell transplantation has shown promising results for the treatment of photoreceptor degenerations. These cells may have the potential to slow down photoreceptor loss, and therefore should be applied in the early stages of photoreceptor degenerations. Furthermore, because of their possible paracrine effects, they may have a wide range of clinical applications, since they can potentially impact on several retinal cell types at once and photoreceptor degenerations can involve different cells and/or begin in one cell type and then affect adjacent cells. The intraocular injection of bone marrow derived-mononuclear stem cells also enhances the outcomes of other treatments aimed to protect photoreceptors. Therefore, it is likely that future investigations may combine bone marrow derived-mononuclear stem cell therapy with other systemic or intraocular treatments to obtain greater therapeutic effects in degenerative retinal diseases.

A column-based deep learning method for the detection and quantification of atrophy associated with AMD in OCT scans.

The objective quantification of retinal atrophy associated with age-related macular degeneration (AMD) is required for clinical diagnosis, follow-up, treatment efficacy evaluation, and clinical research. Spectral Domain Optical Coherence Tomography (OCT) has become an essential imaging technology to evaluate the macula. This paper describes a novel automatic method for the identification and quantification of atrophy associated with AMD in OCT scans and its visualization in the corresponding infrared imaging (IR) image. The method is based on the classification of light scattering patterns in vertical pixel-wide columns (A-scans) in OCT slices (B-scans) in which atrophy appears with a custom column-based convolutional neural network (CNN). The network classifies individual columns with 3D column patches formed by adjacent neighboring columns from the volumetric OCT scan. Subsequent atrophy columns form atrophy segments which are then projected onto the IR image and are used to identify and segment atrophy lesions in the IR image and to measure their areas and distances from the fovea. Experimental results on 106 clinical OCT scans (5,207 slices) in which cRORA atrophy (the end point of advanced dry AMD) was identified in 2,952 atrophy segments and 1,046 atrophy lesions yield a mean F1 score of 0.78 (std 0.06) and an AUC of 0.937, both close to the observer variability. Automated computer-based detection and quantification of atrophy associated with AMD using a column-based CNN classification in OCT scans can be performed at expert level and may be a useful clinical decision support and research tool for the diagnosis, follow-up and treatment of retinal degenerations and dystrophies.

Synaptic repair and vision restoration in advanced degenerating eyes by transplantation of retinal progenitor cells.

SummaryStem cell transplantation shows enormous potential for treatment of incurable retinal degeneration (RD). To determine if and how grafts connect with the neural circuits of the advanced degenerative retina (ADR) and improve vision, we perform calcium imaging of GCaMP5-positive grafts in retinal slices. The organoid-derived C-Kit+/SSEA1− (C-Kit+) retinal progenitor cells (RPCs) become synaptically organized and build spontaneously active synaptic networks in three major layers of ADR. Light stimulation of the host photoreceptors elicits distinct neuronal responses throughout the graft RPCs. The graft RPCs and their differentiated offspring cells in inner nuclear layer synchronize their activities with the host cells and exhibit presynaptic calcium flux patterns that resemble intact retinal neurons. Once graft-to-host network is established, progressive vision loss is stabilized while control eyes continually lose vision. Therefore, transplantation of organoid-derived C-Kit+ RPCs can form functional synaptic networks within ADR and it holds promising avenue for advanced RD treatment.

Targeting molecular pathways for the treatment of inherited retinal degeneration.

Inherited retinal degeneration is a major cause of incurable blindness characterized by loss of retinal photoreceptor cells. Inherited retinal degeneration is characterized by high genetic and phenotypic heterogeneity with several genes mutated in patients affected by these genetic diseases. The high genetic heterogeneity of these diseases hampers the development of effective therapeutic interventions for the cure of a large cohort of patients. Common cell demise mechanisms can be envisioned as targets to treat patients regardless the specific mutation. One of these targets is the increase of intracellular calcium ions, that has been detected in several murine models of inherited retinal degeneration. Recently, neurotrophic factors that favor the efflux of calcium ions to concentrations below toxic levels have been identified as promising molecules that should be evaluated as new treatments for retinal degeneration. Here, we discuss therapeutic options for inherited retinal degeneration and we will focus on neuroprotective approaches, such as the neuroprotective activity of the Pigment epithelium-derived factor. The characterization of specific targets for neuroprotection opens new perspectives together with many questions that require deep analyses to take advantage of this knowledge and develop new therapeutic approaches. We believe that minimizing cell demise by neuroprotection may represent a promising treatment strategy for retinal degeneration.

Exosomes derived from neural progenitor cells preserve photoreceptors during retinal degeneration by inactivating microglia

ABSTRACT Retinal degeneration (RD) is one of the most common causes of visual impairment and blindness and is characterized by progressive degeneration of photoreceptors. Transplantation of neural stem/progenitor cells (NPCs) is a promising treatment for RD, although the mechanisms underlying the efficacy remain unclear. Accumulated evidence supports the notion that paracrine effects of transplanted stem cells is likely the major approach to rescuing early degeneration, rather than cell replacement. NPC-derived exosomes (NPC-exos), a type of extracellular vesicles (EVs) released from NPCs, are thought to carry functional molecules to recipient cells and play therapeutic roles. In present study, we found that grafted human NPCs (hNPCs) secreted EVs and exosomes in the subretinal space (SRS) of RCS rats, an RD model. And direct administration of mouse neural progenitor cell-derived exosomes (mNPC-exos) delayed photoreceptor degeneration, preserved visual function, prevented thinning of the outer nuclear layer (ONL), and decreased apoptosis of photoreceptors in RCS rats. Mechanistically, mNPC-exos were specifically internalized by retinal microglia and suppressed their activation in vitro and in vivo. RNA sequencing and miRNA profiling revealed a set of 17 miRNAs contained in mNPC-exos that markedly inhibited inflammatory signal pathways by targeting TNF-α, IL-1β, and COX-2 in activated microglia. The exosomes derived from hNPC (hNPC-exos) contained similar miRNAs to mNPC-exos that inhibited microglial activation. We demonstrated that NPC-exos markedly suppressed microglial activation to protect photoreceptors from apoptosis, suggesting that NPC-exos and their contents may be the mechanism of stem cell therapy for treating RD.

Targeting cGMP‐signalling to protect photoreceptors in hereditary retinal degeneration

AbstractcGMP‐signalling is fundamental to the phototransduction cascade in retinal photoreceptors. Paradoxically, high levels of cGMP lead to photoreceptor loss in hereditary retinal degeneration (RD) caused by a variety of different gene mutations. This makes cGMP‐signalling a prime target for the development of new treatment approaches. The presentation will give details about the development of compounds and drug delivery technology designed for a targeted interference with cGMP‐signalling in photoreceptors. As an example, in vivo treatment data on a liposomal formulation of the inhibitory cGMP analogue DF003/CN03 will be presented. Overall, cGMP analogues may constitute a new class of compounds for RD treatment and, when combined with suitable drug delivery vehicles, they provide a clear perspective for clinical translation, potentially bringing the very first pharmacological treatment to patients suffering from RD.

GSK-3 Inhibitors: From theBrain to theRetina and Back Again.

Enzyme glycogen synthase kinase-3 (GSK-3) is a candidate pharmacological target for the treatment of neurodegenerative diseases of the brain. Given the many molecular, cellular, and functional features shared by the brain and the retina in both physiological and pathological processes, drugs originally designed to treat neurodegenerative diseases of the brain could be useful candidates for the treatment of retinal degenerative pathologies. Moreover, the accessibility of the eye to noninvasive, quantitative diagnostic techniques allows for easier evaluation of the efficacy of candidate therapies in clinical trials. In this chapter, we discuss the potential of GSK-3 inhibitors in the treatment of retinal degeneration.

Mussel-inspired injectable hydrogel and its counterpart for actuating proliferation and neuronal differentiation of retinal progenitor cells

Biomaterials-mediated retinal progenitor cell (RPC)-based transplantation therapy has shown substantial potential for retinal degeneration (RD), but it is limited by the poor RPC survival, proliferation and differentiation. Herein, the gelatin-hyaluronic acid (Gel-HA)-based hydrogels formed via moderate Michael-type addition reaction with or without the introduction of mussel-inspired polydopamine (PDA), i.e. Gel-HA-PDA and its counterpart Gel-HA hydrogels are developed, and their effects on the biological behaviour of RPCs, including adhesion, survival, proliferation, differentiation, delivery and migration are investigated. The hybrid hydrogels can adopt the intricate structure of the retina with suitable mechanical strength, degradation rate and biological activity to support cellular adhesion, survival and delivery. Meanwhile, Gel-HA hydrogel can remarkably promote RPC proliferation with much larger cell clusters, while Gel-HA-PDA hydrogel significantly enhances RPC adhesion and migration, and directs RPCs to preferentially differentiate toward retinal neurons such as photoreceptors (the most crucial cell-type for RD treatment), which is mainly induced by the activation of integrin α5β1-phosphatidylinositol-3-kinase (PI3K) pathway. This study demonstrates that Gel-HA hydrogel possesses great potential for RPC proliferation, while mussel-inspired PDA-modified Gel-HA hydrogel with superior biocompatibility can significantly promote RPC neuronal differentiation, providing new insights for developing biomedical materials applied for RPC-based transplantation therapy.

Advances of stem cell transplantation in the treatment of retinal degeneration

Retinal degeneration mainly include age-related macular degeneration, retinitispigmentosa and Stargardt’s disease. Although its expression is slightly different, its pathogenesis is photoreceptor cells and/or retinal pigment epithelial (RPE) cell damage or degeneration. Because of the 1ack of self-repairing and renewal of retinal photoreceptor cells and RPE cells, cell replacement therapy is one of the most effective methods for treating such diseases.The stem cells currently used for the treatment of retinal degeneration include embryonicstem cells (ESC) and various adult stem cells, such as retinal stem cells (RSC), induced pluripotent stem cells (iPSC). and mesenchymal stem cells (MSC). Understanding the currentbasic and clinical application progress of ESC, iPSC, RSC, MSC can provide a new idea for the treatment of retinal degeneration. Key words: Retinal diseases/therapy; Embryonic stem cells; Adult stem cells; Review

Intravitreal Injection of Therapeutic Protein for Treatment of Retinal Degeneration

Intravitreal Injection of Therapeutic Protein for Treatment of Retinal Degeneration

Transplantation of rat embryonic stem cell-derived retinal cells restores visual function in the Royal College of Surgeons rats.

To evaluate the feasibility of transplantation of embryonic stem cell (ESC)-derived retinal cells in the treatment of retinal degeneration. Rat ESCs were isolated and induced into retinal progenitor cells (RPCs) in vitro, which were subsequently induced into retinal pigment epithelium cells (RPEs) and photoreceptors (PRCs). All cells were identified by Western blot detection of their specific markers. RPEs and PRCs were, respectively, injected into the retina of Royal College of Surgeons (RCSs) rats. Control group was injected with PBS. Post-transplantation visual function was determined by electroretinography (ERG). The histology of the whole eye was compared by H&E staining. RPEs and PRCs were successfully derived from rat ESCs through the two-step differentiation as indicated by the presence of ESC- (Oct-3/4, Nanog, TRA-1-60 and TRA-1-81), RPC- (Rx, Mitf, Pax6 and Chx10), RPE- (RPE65 and keratin) and PRC-specific markers (blue opsin, red/green opsin, recoverin and rhodopsin) in Western blot. The amplitude of ERG a- and b-wave in RPE- and PRC-transplanted groups at week 2 and 10 after transplantation was markedly higher compared with PBS controls. Retinal injury and vascular stress response was not detected in any of the RCS rats after transplantation. The developed stepwise protocol can derive retinal cells from ESCs. Transplantation of these retinal cells can restore visual function of RCS rats. Our study provides evidence for potential clinical application of ESC-based cell therapy for retinal degeneration.

Human Mesenchymal Stem Cells Expressing Erythropoietin Enhance Survivability of Retinal Neurons Against Oxidative Stress: An In Vitro Study.

Retinal degeneration is a prominent feature in ocular disorders. In exploring possible treatments, Mesenchymal Stem Cells (MSCs) have been recognized to yield therapeutic role for retinal degenerative diseases. Studies have also displayed that erythropoietin (EPO) administration into degenerative retina models confers significant neuroprotective actions in limiting pathological cell death. In this study, we aimed to use MSCs to deliver EPO and to evaluate the ability of EPO to rescue retinal neurons from dying upon reactive oxidative stress induction. We derived human MSCs from Wharton’s jelly (hWJMSCs) of the umbilical cord and cells were transduced with lentivirus particles encoding EPO and a reporter gene of green fluorescent protein (GFP). The supernatants of both transduced and non-transduced cells were collected and used as a pre-conditioning medium for Y79 retinoblastoma cells (retinal neuron cell line) following exposure to glutamate induction. Retinal cells exposed to glutamate showed reduced mitochondrial depolarization and enhanced improvement in cell viability when incubated with pre-conditioned media of transduced cells. Our results established a proof-of-concept that MSCs could be used as a candidate for the delivery of EPO therapeutic gene in the treatment of retinal degenerations.

Research on natural antioxidants in the treatment of retinal degeneration

Retinal degeneration (RD) is a common eye disease which may result in blindness. At present, there is no effective clinical treatment for RD. It is known that apoptosis causes the occurrence and development of retinal degeneration, therefore antiapoptotic/antioxidative treatment has always been the focus in the treatment of RD. There are many kinds of antioxidant drugs, among which natural antioxidants are featured with reliable sources, high safety and less side effects. Hence researchers across the world made tremendous efforts in finding the safe and effective natural antioxidants. This paper focuses on the research progress of natural antioxidants in the treatment of retinal degeneration, in an effort to provide some valuable reference for clinical research. (Chin J Ophthalmol, 2018, 54: 312-315).

Interphotoreceptor matrix based biomaterial: Impact on human retinal progenitor cell attachment and differentiation.

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.