A quantitative in vitro model of smooth muscle cell migration through the arterial wall using the human amniotic membrane.
OBJECTIVE
The development of intimal hyperplasia involves smooth muscle cell (SMC) migration into the intima and proliferation. Matrix metalloproteinases and their tissue inhibitors play important roles in this process. In this study, we describe a novel in vitro model for studying SMC migration through the vessel wall.


RESULTS
Human aortic SMCs (hASMCs) labeled with 125I-iododeoxyuridine or unlabeled were grown on the stromal aspect of the human amniotic membrane. Mechanical damage to endothelial cells grown on the basement membrane and addition of growth factors or platelets were characterized for their effect on SMC migration into the stroma both by histological methods and by measuring the radioactivity associated with the membrane after removal of noninvasive SMCs. To assess the reliability of the model, the cells were infected with a recombinant adenovirus encoding the tissue inhibitor of metalloproteinase-1 (TIMP-1). Addition of a platelet-derived growth factor gradient stimulated hASMC infiltration into the stroma. This effect was abolished with TIMP-1-transduced hASMC, confirming that TIMP-1 overexpression blocks SMC invasion of the stroma.


CONCLUSIONS
This in vitro model of SMC migration in the vessel wall provides an inexpensive, quantitative, and reliable tool to study the molecular and cellular mechanisms of intimal hyperplasia.
Kallenbach
Klaus
K
Seymour Cohn Cardiovascular Surgery Research Laboratory, New York University School of Medicine, New York, NY 10016, USA.
Fernandez
Harold A
HA
Seghezzi
Graziano
G
Baumann
F Gregory
FG
Patel
Sundeep
S
Grossi
Eugene A
EA
Galloway
Aubrey C
AC
Mignatti
Paolo
P
D000256:Adenoviridae / Q000235:genetics; D000650:Amnion; D001011:Aorta / Q000166:cytology; D001485:Basement Membrane; D002465:Cell Movement; D002478:Cells, Cultured / Q000166:cytology; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000166:cytology; D004730:Endothelium, Vascular / Q000166:cytology; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D020782:Matrix Metalloproteinases / Q000502:physiology; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D011993:Recombinant Fusion Proteins / Q000502:physiology; D013314:Stress, Mechanical; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D014161:Transduction, Genetic
D011993:Recombinant Fusion Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D020782:Matrix Metalloproteinases