Alternating Current-Electric Field Inducing Chorio Allantoic Membrane (CAM) Angiogenesis through Exogenous Growth Factor Intervention
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Abstract
Angiogenesis is widely used in various therapies by promoting or inhibiting the formation of new blood vessels. The use of Alternating Current-Electric Fields (AC-EF) in Electro-Capacitive Cancer Therapy (ECCT) showed its potential as an anti-cancer device, and is characterized by its anti-proliferative and pro-apoptotic effects. However, the role of AC-EF in angiogenesis remains unclear. To investigate the effects of AC-EF on CAM angiogenesis, we used the ex ovo culture method of chorioallantoic membrane (CAM). A basic fibroblast growth factor (bFGF) dose of 30 ng/µL was administered as an exogenous growth factor. The ECCT device, generating AC-EF of 150 kHz and 18 Vpp, was exposed to the CAMs. Subsequently, the 24 CAMs of chick embryo were divided into four groups. Two groups were non-bFGF-induced CAM, while the other two were bFGF-induced CAM, and each group was exposed either with or without AC-EF. The vascularization was evaluated through macroscopic observation, while vascular endothelial growth factor A
(VEGFA) gene expression was measured using qPCR. The data were statistically analyzed using ANOVA with GraphPad Prism 9.5. The results showed that an AC-EF exposure had no effects on normal CAM angiogenesis (P>0.05). Moreover, VEGFA gene expression did not show significant upregulation (P>0.05) in the bFGF-induced CAM with or without AC-EF exposure. Interestingly, the number of new blood vessels was significantly higher (P<0.05) in the bFGF-induced with AC-EF exposure than in the non-bFGF-induced group. In conclusion, AC-EF of ECCT did not affect normal angiogenesis. AC-EF may trigger CAM angiogenesis with bFGF induction. This observation suggested that AC-EF of intermediate frequency could enhance angiogenesis by administration of external growth factors, offering a potential avenue for addressing obstructive vascular conditions.
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References
Carmeliet, P. and Jain, R.K., 2011. Molecular mechanisms and clinical applications of angiogenesis. Nature, 473(7347), 298-307, https://doi.org/10.1038/nature10144.
Mittal, V., 2018. Epithelial mesenchymal transition in tumor metastasis. Annual Review of Pathology, 13, 395-412, https://doi.org/10.1146/annurev-pathol-020117-043854.
Vogelstein, B. and Kinzler, K.W., 2004. Cancer genes and the pathways they control. Nature Medicine, 10(8), 789-799, https://doi.org/10.1038/nm1087.
Houtman, J.J., 2010. Acknowledgments, Life's Blood: Angiogenesis in Health and Disease. Breakthroughs in Bioscience, FASEB. [online] Available at: https://www.jhoutman.com/documents/Angiogenesis.pdf.
Pandya, N.M., Dhalla, N.S. and Santani, D.D., 2006. Angiogenesis-a new target for future therapy. Vascular Pharmacology, 44(5), 265-274, https://doi.org/10.1016/j.vph.2006.01.005.
Rajabi, M. and Mousa, S.A., 2017. The role of angiogenesis in cancer treatment. Biomedicines, 5(2), https://doi.org/10.3390/biomedicines5020034.
La Mendola, D., Trincavelli, M.L. and Martini, C., 2022. Angiogenesis in disease. International Journal of Molecular Sciences, 23(18), https://doi.org/10.3390/ijms231810962.
Carmeliet, P., 2000. Mechanisms of angiogenesis and arteriogenesis. Nature Medicine, 6(4), 389-395, https://doi.org/10.1038/74651.
Shibuya, M., 2011. Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) signaling in angiogenesis: a crucial target for anti- and pro-angiogenic therapies. Genes and Cancer, 2(12), 1097-1105, https://doi.org/10.1177/1947601911423031.
Alamsyah, F., Ajrina, I.N., Dewi, F.N.A., Iskandriati, D., Prabandari, S.A. and Taruno, W.P., 2015. Anti-proliferative effect of electric fields on breast tumor cells in vitro and in vivo. Indonesian Journal of Cancer Chemoprevention, 6(3), 71-77. http://dx.doi.org/10.14499/indonesianjcanchemoprev6iss3pp71-77.
Pratiwi, R., Antara, N.Y., Fadliansyah, L.G., Ardiansyah, S.A., Nurhidayat, L., Sholikhah, E.N., Sunarti, S., Widyarini, S., Fadhlurrahman, A.G., Fatmasari, H., Tunjung, W.A.S., Haryana, S.M., Alamsyah, F. and Taruno, W.P., 2019. CCL2 and IL18 expressions may associate with the anti-proliferative effect of noncontact electro capacitive cancer therapy in vivo. F1000Research, 8, https://doi.org/:10.12688/f1000research.20727.2.
Fabian, D., Eibl, M.D.P.G.P., Alnahhas, I., Sebastian, N., Giglio, P., Puduvalli, V., Gonzalez, J. and Palmer, J.D., 2019. Treatment of glioblastoma (GBM) with the addition of tumor-treating fields (TTF): a review. Cancers, 11(2), https://doi.org/10.3390/cancers11020174.
Kim, E.H., Song, H.S., Yoo, S.H. and Yoon, M., 2016. Tumor treating fields inhibit glioblastoma cell migration, invasion and angiogenesis. Oncotarget, 7(40), 65125-65136, https://doi.org/10.18632/oncotarget.11372.
Wu, L., Yao, C. Xiong, Z., Zhang, R., Wang, Z., Wu, Y., Qin, Q. and Hua, Y., 2016. The effects of a picosecond pulsed electric field on angiogenesis in the cervical cancer xenograft models. Gynecologic Oncology, 141(1), 175-181, https://doi.org/10.1016/j.ygyno.2016.02.001.
Wu, L., Wu, Y., Xiong, Z., Yao, C, Zeng, M., Zhang, R. and Hua, Y., 2018. Effects and possible mechanism of a picosecond pulsed electric field on angiogenesis in cervical cancer in vitro. Oncology Letters, 17(2), 1517-1522, https://doi.org/10.3892/ol.2018.9782.
Nowak-Sliwinska, P., Segura, T. and Iruela-Arispe, M.L., 2014. The chicken chorioallantoic membrane model in biology, medicine and bioengineering. Angiogenesis, 17(4), 779-804, https://doi.org/10.1007/s10456-014-9440-7.
Ribatti, D,. 2010. The chick embryo chorioallantoic membrane as an in vivo assay to study antiangiogenesis. Pharmaceuticals, 3(3), 482-513, https://doi.org/10.3390/ph3030482.
Ribatti, D., 2016. The chick embryo chorioallantoic membrane (CAM). A multifaceted experimental model. Mechanisms of development, 141, 70-77, https://doi.org/10.1016/j.mod.2016.05.003.
Merckx, G., Tay, H., Monaco, M.L., van Zandvoort, M., De Spiegelaere, W., Lambrichts, I. and Bronckaers, A., 2020. Chorioallantoic membrane assay as model for angiogenesis in tissue engineering: focus on stem cells. Tissue Engineering. Part B: Reviews, 26(6), 519-539, https://doi.org/10.1089/ten.teb.2020.0048.
Dohle, D.S., Pasa, S.D., Gustmann, S., Laub, M., Wissler, J.H., Jennissen, H.P. and Dünker, N., 2010. Chick ex ovo culture and ex ovo CAM assay: how it really works. Journal of Visualized Experiments, 33, https://doi.org/10.3791/1620.
Federer, W.T., 1963. Experimental Design : Theory and Application. New York: Macmillan.
Oktavia, S., Wijayanti, N. and Retnoaji, B., 2017. Anti-angiogenic effect of Artocarpus heterophyllus seed methanolic extract in ex ovo chicken chorioallantoic membrane. Asian Pacific Journal of Tropical Biomedicine, 7(3), 240-244, https://doi.org/10.1016/j.apjtb.2016.12.024.
Neshat, S.B., Tehranipour, M. and Balanejad, S.Z., 2017. The effect of aqueous phase and hydroalcoholic extract of Stachys lavandulifolia on VEGF gene expression changes and angiogenesis of chick embryo chorioallantoic membrane. Journal of Kermanshah University of Medical Sciences, 20(4), 117-123, https://doi.org/10.22110/jkums.v20i4.3198.
Livak, K.J. and Schmittgen, T.D., 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4), 402-408, https://doi.org/10.1006/meth.2001.1262.
Cross, M.J. and Claesson-Welsh, L., 2001. FGF and VEGF function in angiogenesis: Signalling pathways, biological responses and therapeutic inhibition. Trends in Pharmacological Sciences, 22(4), 201-207, https://doi.org/10.1016/s0165-6147(00)01676-x.
Kobori, T., Hamasaki, S., Kitaura, A., Yamazaki, Y., Nishinaka, T., Niwa, A., Nakao, S., Wake, H., Mori, S., Yoshino, T., Nishibori, M. and Takahashi, H., 2018. Interleukin-18 amplifies macrophage polarization and morphological alteration, leading to excessive angiogenesis. Frontiers in Immunology, 9, https://doi.org/10.3389/fimmu.2018.00334.
Guo, S., Colbert, L.S., Fuller, M., Zhang, Y. and Gonzalez-Perez, R.R., 2010. Vascular endothelial growth factor receptor-2 in breast cancer. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer, 1806(1), 108-121, https://doi.org/10.1016/j.bbcan.2010.04.004.
Eguchi, R., Kawabe, J.I. and Wakabayashi, I., 2022. VEGF-independent angiogenic factors: beyond VEGF/VEGFR2 signaling. Journal of Vascular Research, 59(2), 78-89, https://doi.org/10.1159/000521584.
Bai, H., Forrester, J.V. and Zhao, M., 2011. DC electric stimulation upregulates angiogenic factors in endothelial cells through activation of VEGF receptors. Cytokine, 55(1), 110-115, https://doi.org/10.1016/j.cyto.2011.03.003.
Zhao, Z., Qin, L., Reid, B., Pu, J., Hara, T. and Zhao, M., 2012. Directing migration of endothelial progenitor cells with applied DC electric fields. Stem Cell Research, 8(1), 38-48, https://doi.org/10.1016/j.scr.2011.08.001.
Wei, X., Guan, L., Fan, P., Liu, X., Liu, R., Liu, Y. and Bai, H., 2020. Direct current electric field stimulates nitric oxide production and promotes NO-dependent angiogenesis: involvement of the PI3K/Akt signaling pathway. Journal of Vascular Research, 57(4), 195-205, https://doi.org/10.1159/000506517.
Chen, Y., Ye, L., Guan, L., Fan, P., Liu, R., Liu, H., Chen, J., Zhu, Y., Wei, X., Liu, Y. and Bai, H., 2018. Physiological electric field works via the VEGF receptor to stimulate neovessel formation of vascular endothelial cells in a 3D environment. Biology Open, 7(9), https://doi.org/10.1242/bio.035204.