Fei Feng1, Yan Cheng2, Qing-Huai Liu3. 1. Department of Ophthalmology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China ; Department of Ophthalmology, Taixing People's Hospital, Taixing 225400, Jiangsu Province, China. 2. Department of Ophthalmology, Taixing People's Hospital, Taixing 225400, Jiangsu Province, China. 3. Department of Ophthalmology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China.
Abstract
AIM: To evaluate the effect of different bevacizumab concentrations on retinal neovascularization in a retinopathy of prematurity (ROP) mouse model. METHODS: A total of 60 of C57BL/6 J mice were exposed to 75%±2% oxygen from postnatal d7 to postnatal d12. Fifteen nonexposed mice served as negative controls (group A). On d12, 30 mice (group C) were injected with 2.5 µg intravitreal bevacizumab (IVB), 30 mice (group D) were injected with 1.25 µg IVB in one eye. The contralateral eyes were injected with balanced salt solution (BSS) (control group=group B). The adenosine diphosphatase (ADPase) histochemical technique was used for retinal flat mount to assess the oxygen-induced changes of retinal vessels. Neovascularization was quantified by counting the endothelial cell proliferation on the vitreal side of the inner limiting membrane of the retina. Histological changes were examined by light microscopy. The mRNA levels of vascular endothelial growth factor (VEGF) were quantified by Real-time PCR. Western-blotting analysis was performed to examine the expression of P-VEGFR. RESULTS: Comparing with the control group B, regular distributions and reduced tortuosity of vessels were observed in our retinal flat mounts in groups C and D. The endothelial cell count per histological section was lower in groups C (P<0.0001) and D (P<0.0001) compared with the control group B. Histological evaluation showed no retinal toxicity in any group. In all oxygen treated groups VEGF mRNA expression was significantly increased as compared to age-matched controls. No significant change in VEGF mRNA expression could be achieved in either of the treatments or the oxygen controls. The results of the Western blot were consistent with that of the Real-time PCR analysis. CONCLUSION: An intravitreal injection of Bevacizumab is able to reduce angioproliferative retinopathy in a mouse model for oxygen-induced retinopathy.
AIM: To evaluate the effect of different bevacizumab concentrations on retinal neovascularization in a retinopathy of prematurity (ROP) mouse model. METHODS: A total of 60 of C57BL/6 J mice were exposed to 75%±2% oxygen from postnatal d7 to postnatal d12. Fifteen nonexposed mice served as negative controls (group A). On d12, 30 mice (group C) were injected with 2.5 µg intravitreal bevacizumab (IVB), 30 mice (group D) were injected with 1.25 µg IVB in one eye. The contralateral eyes were injected with balanced salt solution (BSS) (control group=group B). The adenosine diphosphatase (ADPase) histochemical technique was used for retinal flat mount to assess the oxygen-induced changes of retinal vessels. Neovascularization was quantified by counting the endothelial cell proliferation on the vitreal side of the inner limiting membrane of the retina. Histological changes were examined by light microscopy. The mRNA levels of vascular endothelial growth factor (VEGF) were quantified by Real-time PCR. Western-blotting analysis was performed to examine the expression of P-VEGFR. RESULTS: Comparing with the control group B, regular distributions and reduced tortuosity of vessels were observed in our retinal flat mounts in groups C and D. The endothelial cell count per histological section was lower in groups C (P<0.0001) and D (P<0.0001) compared with the control group B. Histological evaluation showed no retinal toxicity in any group. In all oxygen treated groups VEGF mRNA expression was significantly increased as compared to age-matched controls. No significant change in VEGF mRNA expression could be achieved in either of the treatments or the oxygen controls. The results of the Western blot were consistent with that of the Real-time PCR analysis. CONCLUSION: An intravitreal injection of Bevacizumab is able to reduce angioproliferative retinopathy in a mouse model for oxygen-induced retinopathy.
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