Abstract
Our aim was to further characterize the interstitial cell phenotypes of normal porcine and human semilunar valves, information necessary for the design of bioengineered valves and for the understanding of valve disease processes such as aortic valve sclerosis. Existence of fibroblasts, myofibroblasts, and smooth muscle-like cells within semilunar heart valves has been established. However, the nature of the smooth muscle cell population has been controversial. We used immunochemical and western blotting methods to determine the status of smoothelin and smooth muscle α-actin in the valve. Our examination of valve interstitial cells confirmed the presence of terminally differentiated, contractile smooth muscle cells in situ. They were arranged in small bundles of 5–35 cells within the ventricularis or as individual cells scattered throughout the valvular layers in vivo, and were present in cells explanted from the valves in vitro. Colocalization of these proteins in semilunar heart valves was achieved with double-labeling experiments. Protein extraction, followed by coimmunoprecipitation, electrophoresis, and western blotting confirmed the immunochemical analysis and suggested that smooth muscle α-actin and smoothelin interact, as has been previously postulated. The presence of contractile smooth muscle within the valve may be an important factor in understanding valve pathology and in the design of tissue engineering efforts.
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Acknowledgements
M. Cimini would like to acknowledge the scholarship support from the Heart and Stroke Foundation of Canada. The authors would like to thank A.M. Ferreira and J. Ronald for critical reading of the manuscript and technical assistance and advice, L. Aldington for cell culture assistance, J. Dunmore-Buyze and Dr. T. Tweedie for sample collection, and N. Bechard for her imaging expertise. The authors would also like to thank the Ralf Bros and Mt. Brydges abattoirs for porcine samples. This study was supported by the Heart and Stroke Foundation of Ontario grant number T3573/4707 and Canadian Institutes for Health Research grant number MOP57839.
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Cimini, M., Rogers, K.A. & Boughner, D.R. Smoothelin-positive cells in human and porcine semilunar valves. Histochem Cell Biol 120, 307–317 (2003). https://doi.org/10.1007/s00418-003-0570-z
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DOI: https://doi.org/10.1007/s00418-003-0570-z