Gore Manish, Tiwari Ankit, Jahagirdar Devashree, Narayanasamy Angayarkanni, Jain Ratnesh, Dandekar Prajakta
Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai, 400 019, India.
Department of Biochemistry and Cell Biology, Vision Research Foundation, Sankara Nethralaya, Chennai, 600 006, India.
Curr Res Pharmacol Drug Discov. 2022 May 21;3:100111. doi: 10.1016/j.crphar.2022.100111. eCollection 2022.
Diabetic retinopathy (DR) is a primary microvascular complication of and a vision-threatening condition. Vascular endothelial growth factor (VEGF) induces neovascularization and causes metabolic damage to the retinal and choroidal vasculature in diabetic patients. Existing drug screening models and treatment strategies for DR need to be refined through the establishment of relevant pre-clinical models, which may enable development of effective and safe therapies. The present study discusses the development of an three-dimensional (3D) spheroid model, using RF/6A choroid-retinal vascular endothelial cells, to closely mimic the disease condition. Compact, reproducibly-sized, viable and proliferating RF/6A spheroids were fabricated, as confirmed by microscopy, live/dead assay, cell proliferation assay and histological staining. angiogenesis was studied by evaluating individual effects of VEGF and an anti-VEGF monoclonal antibody, Bevacizumab, and their combination on cellular proliferation and 3D endothelial sprout formation. VEGF stimulated angiogenic sprouting while Bevacizumab demonstrated a dose-dependent anti-angiogenic effect, as determined from the cellular proliferation observed and extent and length of sprouting. These investigations validated the potential of RF/6A spheroids in providing an alternative-to-animal, pathophysiologically-relevant model to facilitate pre-clinical and biomedical research related to DR.
糖尿病视网膜病变（DR）是糖尿病的一种主要微血管并发症，也是一种威胁视力的疾病。血管内皮生长因子（VEGF）可诱导新生血管形成，并对糖尿病患者的视网膜和脉络膜血管造成代谢损伤。现有的DR药物筛选模型和治疗策略需要通过建立相关临床前模型来完善，这可能有助于开发有效且安全的治疗方法。本研究讨论了一种使用RF/6A脉络膜视网膜血管内皮细胞的三维（3D）球体模型的构建，以紧密模拟疾病状况。通过显微镜检查、活/死检测、细胞增殖检测和组织学染色证实，制备出了紧凑、大小可重复、存活且增殖的RF/6A球体。通过评估VEGF和抗VEGF单克隆抗体贝伐单抗及其组合对细胞增殖和3D内皮芽形成的个体影响，研究了血管生成情况。从观察到的细胞增殖以及芽形成的程度和长度来看，VEGF刺激血管生成芽，而贝伐单抗表现出剂量依赖性抗血管生成作用。这些研究验证了RF/6A球体在提供一种替代动物的、病理生理相关模型以促进与DR相关的临床前和生物医学研究方面的潜力。
