Abstract
The three-dimensional changes in the cytoskeleton and in cell proliferation of bovine pulmonary artery endothelial cells when exposed to sustained hydrostatic pressure were investigated in vitro using laser scanning confocal microscopy. Subconfluent endothelial cells on rigid substrates were exposed to 1.5, 5 and 10 cm H2O pressure under hydrostatic heads of culture medium for up to seven days. Confocal microscopic images were taken at distances of 0.4 μm through the thickness of the sample and visualised in multiplanar, stereopair and 90o rotation formats. The results of the study provide evidence of: increased proliferation after exposure to 10 cm H2O pressure for five and seven days; cell bilayering after exposure to 1.5 and 5 cm H2O pressure and trilayering after exposure to 10 cm H2O pressure for seven days; and F-actin filament reorganisation into centrally located, parallel, stress fibres in confluent cells, into peripheral bands in subconfluent, multilayered cells, and into multilayers in the plane perpendicular to the applied force.
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Acevedo, A.D., Bowser, S.S., Gerritsen, M.E. andBizios, R. (1993): ‘Morphological and proliferative responses of endothelial cells to hydrostatic pressure: Role of fibroblast growth factor’,J. Cell. Physiol.,157, pp. 603–614
Dewey, C.F. (1984): ‘Effects of fluid flow on living vascular cells’,J. Biomech. Engr.,106, pp. 31–35
Dewey, C.F., Bussolari, S.R., Gimbrone, M.A., andDavies, P.F. (1981): ‘The dynamic response of vascular endothelial cells to fluid shear stress,’J. Biomech. Engr.,103, pp. 177–185
Eskin, S.G., Ives, C.L., Frangos, J.A., andMcIntire, L.V. (1985): ‘Cultured endothelium: response to flow,’ASAIO,8, 109–112
Gabbiani, G., Badonnel, M.C. andRona G. (1975): ‘Cytoplasmic contractile apparatus in the aortic endothelial cells of hypertensive rats,’Lab. Invest.,32, pp. 227–234
Gotlieb, A.I., andWong, M.K.K. (1988): Current concepts of the role of the endothelial cytoskeleton in endothelial integrity, repair, and disfunction. inRyan, U.S. (Ed.): ‘Endothelial cells. Vol. II’ (CRC Press, Boca Raton), pp. 81–102
Guyton, A.C. (1991): ‘Pulmonary circulation; Pulmonary edema; Pleural fluid’in ‘Textbook of medical physiology’ (W.B. Saunders and Company, Philadelphia), pp. 414–421
Iba, T. andSumpio, B.E. (1991): ‘Morphological response of human endothelial cells subjected to cyclic strainin vitro,’Microvasc. Res.,42, 245–254
Kanda, K. andMatsuda, T. (1993): ‘Behavior of arterial wall cells cultured on periodically stretched substrates,’Cell Transplantation,2, 475–484
Kim, D.W., Gotlieb, A.I. andLangille, B.L. (1989): ‘In vivo modulation of endothelial F-actin microfilaments by experimental alterations in shear stress,’Arteriosclerosis,9, 439–445
Lin, W., Holmes, T., Ancin, H., Roysam, B., Szarowski, D.H., andTurner, J.N. (1993). ‘Three-dimensional light microscopy, optimized staining and automated image analysis of cell nuclei in thick tissue slices’, Proc 51st Ann Mtg Microsc Soc Am 268–269
Nerem, R.M., Levesque, M.J., andCornhill, J.F. (1981): ‘Vascular endothelial morphology as an indicator of the pattern of blood flow,’Trans. ASME,103, pp. 172–176
Pawley, J.B. (Ed.) (1995): ‘Handbook of biological confocal microscopy’ (Plenum Press, New York)
Robb, R.A., andBarillot, C. (1988): ‘Interactive 3-D image display and analysis,’Proc. SPIE Hybrid Image Signal Process 939, pp. 173–202
Robb, R.A., andBarillot, C. (1989): ‘Interactive display and analysis of 3-D medical images,’IEEE Trans. Med. Imaging 8, pp. 217–226
Schulte, E. (1990): ‘Standardization of the Feulgen reaction for absorption DNA image cytometry: A review,’Anal. Cellular Pathol. 3, pp. 167–182
Shotton, D.M. (1989): ‘Confocal scanning optical microscopy and its applications for biological specimens,’J. Cell Sci.,94 pp. 175–206
Tokunaga, O., Fan, J.L., andWatanabe, T. (1989): ‘Atherosclerosis and endothelium. II. Properties of aortic endothelial and smooth muscle cells cultured at various ambient pressures,’Acta Pathol. Jpn. 39, pp. 356–362
Turner, J.N., Szarowski, D.H., Ancin, H., Cohen, A., Lin, W., Roysam, B., andHolmes, T. (1994): ‘Three-dimensional imaging and image analysis of hippocampal neurons: confocal and digitally enhanced wide field microscopy”,Microsc. Res. Technol.,29, pp. 269–278
Wechezak, A.R., Wight, T.N., Viggers, R.F., andSauvage, L.R. (1989): ‘Endothelial adherence under shear stress is dependent upon microfilament reorganization,’J. Cell Physiol.,139, pp. 136–146
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Salwen, S.A., Szarowski, D.H., Turner, J.N. et al. Three-dimensional changes of the cytoskeleton of vascular endothelial cells exposed to sustained hydrostatic pressure. Med. Biol. Eng. Comput. 36, 520–527 (1998). https://doi.org/10.1007/BF02523225
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DOI: https://doi.org/10.1007/BF02523225