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Updated: Apr 5, 2026
Limbal Approach-Subretinal Injection of Viral Vectors for Gene Therapy in Mice Retinal Pigment Epithelium
Published on: August 7, 2015
[Targeting VEGF siRNA transfection by new polymeric liposomes to inhibit retinal neovascularization]
Yan Gao1, Xinling Liu, Chunhui Li
1Department of Ophthalmology, Shanxi Eye Hospital, Taiyuan 030002, China.
Objective:
To formulate and evaluate polymeric liposomes (PL) nanoparticles as a novel non-viral gene delivery. To explore its applicability and feasibility as a non-viral vector for gene transportation.
Method:
Experimental study. To construct the orongoxygen induced retinopathy (OIR) mouse (C57BL/6J) model on the basis of improved Smith's methods. Western blot was used to measure VEGF protein expression in retinal tissue at P17 and P22. HE staining and fluorescein-dextran angiography of retinal vascular were performed to observe the morphologic alterations of retinal neovascularization. Frozen-section was used to show the membrane translocation of PL.
Results:
In fluorescence angiograms, irregular neovascularization and fluoresce leakage were observed in OIR model. The results of Western blot showed that VEGF protein in retinal tissues were significantly different among groups (F = 158.207, P = 0.000) at P17 and P22 (F = 25.695, P = 0.000). The protein level was lower in both PL (0.70 ± 0.03) and Lipo group (0.66 ± 0.04) at P17 (P = 0.092), and the lower level was presented at P22 in PL group (0.50 ± 0.03) than in Lipo group (0.53 ± 0.05) (P < 0.05). HE staining were performed to observe the significantly improvements of retinal neovascularization in PL (28.0 ± 3.44) and Lipo group (24.50 ± 3.06) at P17. Moreover, inhibitory effects maintained at P22 in PL group (11.70 ± 3.09) as HE staining showed. Fluorescein angiography of retinal vascular showed retinal non-perfusion and neovascularization areas were smaller in both PL and Lipo group at P17. Frozen-section examination showed the property of membrane translocation. GFP expression could be seen in vitreous cavity just at first day post-intravitreal administration in PL and Lipo group, which could reach their peaks in external retina nearby RPE layer at P17, remaining at P22 in PL group.
Conclusion:
The PL performed excellent ability of membrane translocation and it was a kind of slow steady released gene vector.
