Ginger-Derived Nanoparticles Deliver a VEGFA-Targeting DNAzyme and Modulate Retinal VEGFA Expression and Inflammatory Markers in Experimental Diabetic Retinopathy

Tong Wang, Yupeng Zhang, Jian Cao, Jing Li, Yunxiang Chao, Wen Yang, Xiaoxiao Jin, Zhipeng YouDepartment of Ophthalmology, The Affiliated Eye Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi, 330006, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhipeng You, Department of Ophthalmology, The Affiliated Eye Hospital, Jiangxi Medical College, Nanchang University, 463 Bayi Road, Nanchang, Jiangxi, 330006, People’s Republic of China, Tel +86 13970032654, Email yzp74@sina.comPurpose: To address the limitations of current anti-vascular endothelial growth factor (anti-VEGF) therapies for diabetic retinopathy (DR), which do not effectively inhibit VEGFA synthesis at the genetic source and provide only marginal improvements in the chronic inflammatory microenvironment, this study employs ginger-derived nanoparticles (GDNPs) to create a deoxyribozyme (DNAzyme) delivery system (GDNPs@Dz) targeting VEGFA mRNA. It further investigates the combined effects of this system on gene silencing and anti-inflammatory activity in an experimental setting.Methods: GDNPs were isolated from fresh ginger using differential centrifugation followed by size-exclusion chromatography purification, and their physicochemical properties and biosafety were characterized. Phosphorothioate-modified 10– 23 DNAzyme was incorporated into GDNPs via ultrasonication. The uptake of GDNPs@Dz by human retinal microvascular endothelial cells was evaluated in vitro, alongside its inhibitory effects on cell migration and tube formation. In a streptozotocin-induced DR mouse model, the study assessed the intraocular retention of the AF647-labeled DNAzyme signal, VEGFA expression, and inflammatory markers (IL-1β, TNF-α) following a single intravitreal injection (2 μL/eye, 39 pmol DNAzyme + 4 μg GDNPs). Treatment doses in vitro were 500 nM free DNAzyme, 50 μg/mL empty GDNPs, or an equivalent combination. All in vitro experiments were performed in three independent biological replicates, with three technical replicates per condition, and in vivo analyses used n = 3 animals per group per time point, with VEGFA expression assessed at days 3, 7, 10, 14, and 21.Results: GDNPs@Dz was internalized by retinal microvascular endothelial cells and reduced high glucose-induced cell migration and tube formation in vitro. In the diabetic mouse model, GDNPs@Dz prolonged the ocular retention of the AF647-labeled DNAzyme signal compared with free DNAzyme and reduced retinal VEGFA mRNA for up to two weeks after a single injection. Representative immunofluorescence showed reduced IL-1β and TNF-α positive signals after empty GDNPs treatment. These findings indicate a combined effect of gene silencing and anti-inflammatory activity.Conclusion: This preclinical proof-of-concept study demonstrates that ginger-derived nanoparticles can deliver a VEGFA-targeting DNAzyme and modulate retinal VEGFA expression and inflammatory markers in an experimental diabetic setting. The system showed a favorable biosafety profile and combined gene-silencing and anti-inflammatory activity, providing a basis for further preclinical development.Keywords: ginger exosome-like nanoparticles, DNAzyme, diabetic retinopathy, VEGFA, anti-inflammatory