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Jong Hwa Jun, Rebecca Kim; An In Vitro Pseudozonule Model of Posterior Capsular Opacification: Effects of Anti-Vascular Endothelial Growth Factor-A Humanized Monoclonal Antibody (Bevacizumab®) on Epithelial-to-Mesenchymal Transition in Lens Epithelium. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2954.
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© ARVO (1962-2015); The Authors (2016-present)
This report presents a novel capsular bag model of an in vitro organ culture for investigating effects of anti-vascular endothelial growth factor-A (VEGF-A) humanized monoclonal antibody (Bevacizumab®) on epithelial-to-mesenchymal transition (EMT) in lens epithelial cells (LEC).
After dissecting the zonule of the lens, whole lens explants were embedded in 2% PBS-based low melting point (LMP) agarose gel in 20 C57BL/6 mice eyes. Sham cataract extraction was performed in support of pseudozonule that was provided by LMP agarose gel. Remained capsular bag-LEC complex (cbLEC) in organ culture model mimicked in vivo configuration of cbLECs. A graded concentration of anti-VEGF-A monoclonal antibody was added in culture medium, cbLECs of capsular bag were cultured in each concentration group. The migration of cbLECs was quantified and 5-bromo-2’-deoxyuridine (BrdU) labeling was evaluated in each group. The expression of EMT markers, the α-smooth muscle actin and collagen type I, and the loss of epithelial markers, pancytokeratin, cytokeratin 8/18 were analyzed by whole mount immunohistochemistry, and western blot analyses.
The cbLECs migration was decreased over 500ng/ml of anti-VEGF-A concentration group. Proliferation rate, production of EMT markers, and loss of epithelial markers were decreased in capsular bag model that were investigated by BrdU labeling, immunohistochemistry and western blot analyses. Bevacizumab effectively inhibited migration, proliferation, and transdifferentiation of cbLECs.
The In vitro pseudozonule model effectively mimicked physiological configuration of lens epithelial cell-capsular bag complex for the evaluation of the effects of anti-VEGF monoclonal antibody on EMT. VEGF-A seems to be related to signaling cascade of migration, proliferation, and transdifferentiation of LECs and anti-VEGF monoclonal antibody effectively decreases the posterior capsular opacification in vitro.
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