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Platelet-derived growth factor and extracellular matrix proteins provide a synergistic stimulus for human vascular
P R Nelson1, S Yamamura, K C Kent
1Department of Surgery (Division of Vascular Surgery), Beth Israel Hospital, Harvard Medical School, Boston, MA, USA.
Purpose:
Smooth muscle cell (SMC) migration contributes significantly to the hyperplastic response that follows arterial injury. In vitro studies have shown that a number of growth factors and extracellular matrix (ECM) proteins individually stimulate vascular SMC migration. However, after arterial injury, SMCs exist in a complex environment in which they are exposed to many of these proteins simultaneously. The response of SMCs to multiple simultaneous stimuli may differ significantly from their response to any single individual stimulus. In this study, we evaluated the chemotactic response of human vascular SMCs to various combinations of growth factors and ECM proteins.
Methods:
Human saphenous vein SMCs were used for all experiments. Using a 4-hour modified Boyden-chamber assay, we evaluated the effect on SMC chemotaxis of combinations of one of three growth factors (platelet-derived growth factor [PDGF]-AB, basic fibroblast growth factor [bFGF], or epidermal growth factor [EGF]), and one of four ECM proteins (fibronectin, laminin, or collagen type I or IV). A standard fluorimetric assay was used to assess changes in intracellular calcium ([Ca2+]i) in response to the various combinations of growth factors and ECM proteins.
Results:
A simple additive effect was seen between ECM proteins and bFGF or EGF. However, when SMCs were simultaneously exposed to PDGF and ECM proteins, we observed a synergistic increase in chemotaxis. This synergy was evident for all concentrations of collagen type I and IV but only with higher concentrations of fibronectin and laminin. We evaluated whether intracellular calcium may be the signaling pathway through which this synergistic effect is mediated. Although ECM proteins alone did not stimulate a rise in [Ca2+]i, ECM proteins enhanced the early peak in [Ca2+]i induced by PDGF.
Conclusion:
These data show that PDGF acts synergistically with the ECM proteins to promote SMC migration; this effect appears to be specific for PDGF and was not observed with other growth factors. The mechanism responsible for this phenomenon may be a synergistic increase in [Ca2+]i in SMCs simultaneously exposed to both proteins.