Arteriosclerosis, Thrombosis, and Vascular Biology. 1998;18:379-388
© 1998 American Heart Association, Inc.
Original Contributions |
Calcification of Human Vascular Cells In Vitro Is Correlated With High Levels of Matrix Gla Protein and Low Levels of Osteopontin Expression
From the Department of Medicine, University of Cambridge, Addenbrooke's Hospital (D.P., C.M.S., P.L.W.), and the Department of Anatomy (J.N.S.), Cambridge, England.
Correspondence to D. Proudfoot, Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Level 5, Hills Rd, Cambridge CB2 2QQ, UK. E-mail dp{at}mole.bio.cam.ac.uk
Abstract—The cellular and molecular
events leading to calcification in atherosclerotic lesions are unknown.
We and others have shown that bone-associated proteins, particularly
matrix Gla protein (MGP) and osteopontin (OP), can be detected in
atherosclerotic lesions, thus suggesting an active calcification
process. In the present study, we aimed to determine whether human
vascular smooth muscle cells (VSMCs) could calcify in vitro and to
determine whether MGP and OP have a role in vascular calcification. We
established that human aortic VSMCs and placental microvascular
pericytes spontaneously form nodules in cell culture and induce
calcification, as detected by von Kossa's method, Alizarin red S
staining, and electron microscopy. The cells in calcifying nodules
differed from those in monolayer cultures by expressing higher levels
of the SMC markers 
-SM actin, SM22, and calponin. In addition,
Northern blot analysis revealed that in human VSMCs,
calcification was associated with increased levels of MGP mRNA. In
contrast, OP mRNA was barely detectable in calcified human VSMCs and
pericyte nodules, nor was OP protein detected, suggesting that OP was
not necessary for calcification to occur. These studies reveal that
human VSMCs are capable of inducing calcification and that MGP may have
a role in human vascular calcification.
Key Words: calcification • vascular smooth muscle • pericytes • osteopontin • matrix Gla protein
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