Relationship between Pericytes and Endothelial Cells in Retinal Neovascularization: A Histological and Immunofluorescent Study of Retinal Angiogenesis
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To evaluate the relationship between pericytes and endothelial cells in retinal neovascularizationthrough histological and immunofluorescent studies. Methods: C57BL/6J mice were exposed to hyperoxia from postnatal day (P) 7 to P12 and were returned toroom air at P12 to induce a model of oxygen-induced retinopathy (OIR). The cross sections of enucleatedeyes were processed with hematoxylin and eosin. Immunofluorescent staining of pericytes, endothelial cells,and N-cadherin was performed. Microfluidic devices were fabricated out of polydimethylsiloxane using softlithography and replica molding. Human retinal microvascular endothelial cells, human brain microvascularendothelial cells, human umbilical vein endothelial cells and human placenta pericyte were mixed and co-cultured. Results: Unlike the three-layered vascular plexus found in retinal angiogenesis of a normal mouse, angiogenesisin the OIR model is identified by the neovascular tuft extending into the vitreous. Neovascular tufts and thethree-layered vascular plexus were both covered with pericytes in the OIR model. In this pathologic vascularization,N-cadherin, known to be crucial intercellular adhesion molecule, was also present. Further evaluationusing the microfluidic in vitro model, successfully developed a microvascular network of endothelial cells coveredwith pericytes, mimicking normal retinal angiogenesis within 6 days. Conclusions: Pericytes covering endothelial cells were observed not only in vasculature of normal retina butalso pathologic neovascularization of OIR mouse at P17. Factors involved in the endothelial cell-pericyte interactioncan be evaluated as an attractive novel treatment target. These future studies can be performed usingmicrofluidic systems, which can shorten the study time and provide three-dimensional structural evaluation.
Abstract
To evaluate the relationship between pericytes and endothelial cells in retinal neovascularizationthrough histological and immunofluorescent studies. Methods: C57BL/6J mice were exposed to hyperoxia from postnatal day (P) 7 to P12 and were returned toroom air at P12 to induce a model of oxygen-induced retinopathy (OIR). The cross sections of enucleatedeyes were processed with hematoxylin and eosin. Immunofluorescent staining of pericytes, endothelial cells,and N-cadherin was performed. Microfluidic devices were fabricated out of polydimethylsiloxane using softlithography and replica molding. Human retinal microvascular endothelial cells, human brain microvascularendothelial cells, human umbilical vein endothelial cells and human placenta pericyte were mixed and co-cultured. Results: Unlike the three-layered vascular plexus found in retinal angiogenesis of a normal mouse, angiogenesisin the OIR model is identified by the neovascular tuft extending into the vitreous. Neovascular tufts and thethree-layered vascular plexus were both covered with pericytes in the OIR model. In this pathologic vascularization,N-cadherin, known to be crucial intercellular adhesion molecule, was also present. Further evaluationusing the microfluidic in vitro model, successfully developed a microvascular network of endothelial cells coveredwith pericytes, mimicking normal retinal angiogenesis within 6 days. Conclusions: Pericytes covering endothelial cells were observed not only in vasculature of normal retina butalso pathologic neovascularization of OIR mouse at P17. Factors involved in the endothelial cell-pericyte interactioncan be evaluated as an attractive novel treatment target. These future studies can be performed usingmicrofluidic systems, which can shorten the study time and provide three-dimensional structural evaluation.
참고문헌 (38)
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