Shear stress modulates the proliferation rate, protein synthesis, and mitogenic activity of arterial smooth muscle cells.
BACKGROUND
The aim of this study was to determine the correlation between hemodynamic forces and proliferation of smooth muscle cells (SMC).


METHODS
Bovine arterial SMC were seeded in a fibronectin-coated polystyrene cylinder at 5 x 10(5) cells/tube and allowed to reach confluence and to adhere for 48 hours. The experimental groups were subjected to a laminar flow of 150 ml/min (9 dyne/cm2), 100 ml/min (6 dyne/cm2), and 50 ml/min (3 dyne/cm2) for 24 hours. The control group was subjected to similar incubation conditions without flow. The cells in the experiments remained attached and viable. All experiments were performed in triplicate or more.


RESULTS
Shear stress significantly reduced (p < 0.001) the 24-hour incorporation of tritiated thymidine and cell proliferation. This effect was proportional to the level of shear stress and was still evident 24 hours after flow cessation. Results of flow cytometry confirmed a lower percentage of SMC in S phase with increasing shear stress. Synthesis of cell-associated proteins was increased twofold (p < 0.01) in SMC subjected to laminar flow. SMC subjected to shear stress released a higher quantity of mitogens, including a platelet-derived growth factor (PDGF)-like substance as detected by immunologic testing. Fifty percent volume per volume conditioned serum-free medium from SMC subjected to shear stress increased threefold the tritiated thymidine uptake in PDGF receptor-bearing Swiss 3T3 cells as compared with conditioned serum-free medium from control SMC not subjected to shear stress and twelvefold as compared with standard control. The release of mitogens was proportional to the level of shear stress and was still evident 24 hours after flow cessation. The mitogenic activity was partially reduced (30%, p < 0.01) by an excess of monospecific anti-PDGF antibody.


CONCLUSIONS
We conclude that (1) increasing shear stress inhibits SMC proliferation and stimulates the synthesis of cell-associated proteins and the release of mitogens and (2) decreasing shear stress facilitates proliferation of SMC. Thus, in situations of arterial flow separation, the increased release of mitogens from SMC subjected to high shear stress and the increased proliferation rate and susceptibility to mitogens of SMC subjected to very low shear stress may generate a critical condition that predisposes to the development of atherosclerosis with early plaque formation in regions of low-flow shear stress.
I Istituto Clinica Chirurgica, University of Rome La Sapienza, School of Medicine, Italy.
A V
AV
Sterpetti
A
A
Cucina
L S
LS
D'Angelo
B
B
Cardillo
A
A
Cavallaro
D000818:Animals; D001013:Aorta, Thoracic; D002417:Cattle; D002455:Cell Division; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D005434:Flow Cytometry; D008934:Mitogens / Q000032:analysis* / Q000096:biosynthesis; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000032:analysis / Q000276:immunology; D014176:Protein Biosynthesis; D013314:Stress, Mechanical
D017077:Culture Media, Conditioned; D008934:Mitogens; D010982:Platelet-Derived Growth Factor