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VEGF-A165 Potently Induces Human Blood–Nerve Barrier Endothelial Cell Proliferation, Angiogenesis, and Wound Healing In Vitro

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Abstract

Several mitogens such as vascular endothelial growth factor (VEGF) have been implicated in mammalian vascular proliferation and repair. However, the molecular mediators of human blood-nerve barrier (BNB) development and specialization are unknown. Primary human endoneurial endothelial cells (pHEndECs) were expanded in vitro and specific mitogen receptors detected by western blot. pHEndECs were cultured with basal medium containing different mitogen concentrations with or without heparin. Non-radioactive cell proliferation, Matrigel™-induced angiogenesis and sterile micropipette injury wound healing assays were performed. Proliferation rates, number and total length of induced microvessels, and rate of endothelial cell monolayer wound healing were determined and compared to basal conditions. VEGF-A165 in the presence of heparin, was the most potent inducer of pHEndEC proliferation, angiogenesis, and wound healing in vitro. 1.31 nM VEGF-A165 induced ~110 % increase in cell proliferation relative to basal conditions (∼51 % without heparin). 2.62 pM VEGF-A165 induced a three-fold increase in mean number of microvessels and 3.9-fold increase in total capillary length/field relative to basal conditions. In addition, 0.26 nM VEGF-A165 induced ∼1.3-fold increased average rate of endothelial wound healing 4–18 h after endothelial monolayer injury, mediated by increased cell migration. VEGF-A165 was the only mitogen capable of complete wound closure, occurring within 30 h following injury via increased cell proliferation. This study demonstrates that VEGF-A165, in the presence of heparin, is a potent inducer of pHEndEC proliferation, angiogenesis, and wound healing in vitro. VEGF-A165 may be an important mitogen necessary for human BNB development and recovery in response to peripheral nerve injury.

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Acknowledgments

Special thanks to Dr. Monique Stins for providing THBMECs. Aspects of this study were presented in part in abstract form at the 2011 American Academy of Neurology meeting, Honolulu, Hawaii, USA and the 2011 Peripheral Nerve Society meeting, Potomac, Maryland, USA. This study was supported by a Baylor College of Medicine New Investigator Start-Up Award (2007–2011). The Neuromuscular Immunopathology Research Laboratory is currently supported by the National Institutes of Health Grants R21 NS073702, R21 NS078226, and R01 NS075212, and a subaward P30 AI27767 to E.E.U. The funding sources had no involvement in the conduct of the research, manuscript preparation, data collection/analyses or decision to submit this work for publication. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

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  1. Nejla Yosef

    Present address: Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX, USA

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  1. Neuromuscular Immunopathology Research Laboratory, Department of Neurology, Baylor College of Medicine, One Baylor Plaza, Mailstop NB302, Houston, TX, 77030-3411, USA

    Chetan Lakshmana Reddy, Nejla Yosef & Eroboghene E. Ubogu

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Reddy, C.L., Yosef, N. & Ubogu, E.E. VEGF-A165 Potently Induces Human Blood–Nerve Barrier Endothelial Cell Proliferation, Angiogenesis, and Wound Healing In Vitro. Cell Mol Neurobiol 33, 789–801 (2013). https://doi.org/10.1007/s10571-013-9946-3

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