The effect of growth factors, cytokines, and extracellular matrix proteins on fibronectin production in human vascular smooth muscle cells.

PURPOSE: Although 60% to 80% of the mature intimal hyperplastic plaque is composed of extracellular matrix (ECM) proteins, little is known about the factors that stimulate smooth muscle cells (SMCs) to produce these proteins. A major component of the ECM protein is fibronectin. Thus we studied fibronectin production and its response to various growth factors, cytokines, and other ECM proteins that are released at the time of vascular injury. METHODS: Quiescent cultured human SMCs were stimulated for varying intervals with increasing concentrations of agonist. Fibronectin in the cell medium was assayed by immunoblotting with a fibronectin-specific antibody. RESULTS: After 72 hours of stimulation, transforming growth factor-beta (10 ng/mL) had the most profound effect on fibronectin production (9.6- +/- 2.1-fold; P <.05), followed by epidermal growth factor (100 ng/mL; 5.0- +/- 0.1-fold; P <.05, for both). Surprisingly, the platelet-derived growth factors (AA, AB, and BB) and fibroblast growth factor did not stimulate fibronectin production. Among the matrix proteins studied, only collagen type I (20 microg/mL) stimulated fibronectin production (1.9- +/- 0.1-fold; P <.05), whereas collagen type IV and laminin had no effect. The contractile protein angiotensin II (100 ng/mL) was a weak stimulant of fibronectin (1.6- +/- 0.2-fold; P <.05). Time course studies of fibronectin production up to 72 hours revealed kinetics that varied with each agonist. Transforming growth factor-beta stimulated significant early production of fibronectin, whereas fibronectin production in response to epidermal growth factor and collagen type I was initially modest but increased with time. The effect of angiotensin II did not become evident until 72 hours. CONCLUSION: Cytokines, growth factors, and matrix proteins have varying quantitative effects on ECM protein production by human vascular SMCs. Knowledge of the factors that influence ECM protein production may allow for the design of specific inhibitors that can prevent intimal hyperplasia.

The role of mitogen-activated protein kinase and protein kinase C in fibronectin production in human vascular smooth muscle cells.

BACKGROUND: After evaluating various growth factors, cytokines, and extracellular matrix (ECM) proteins, we found that the most potent agonists of smooth muscle cell (SMC) fibronectin (Fn) production were transforming growth factor-beta (TGF-beta) and epidermal growth factor (EGF). To determine the possible signaling pathways involved in the production of this matrix protein, we investigated the role of the intracellular proteins, protein kinase C (PKC) and mitogen-activated protein kinase (MAP-K), in TGF-beta- and EGF-induced human vascular SMC Fn production. MATERIALS AND METHODS: After stimulation of human SMCs with TGF-beta (10 ng/ml) and EGF (100 ng/ml), Fn in the cell medium was assayed by immunoblotting using a specific antibody. PKC was activated by brief stimulation of SMC with phorbol 12,13-dibutyrate (PDBu) and inhibited by downregulation with PDBu or the inhibitor, GF109203X. MAP-K was inhibited with PD098059. RESULTS: PKC activation increased basal and synergistically enhanced TGF-beta- and EGF-induced Fn production. However, inhibition of PKC by downregulation and GF109203X did not diminish Fn production by TGF-beta and EGF. Surprisingly, these two methods of inhibition slightly increased basal and agonist-induced Fn production. The MAP-K kinase inhibitor, PD098059, produced an almost complete inhibition of EGF and a partial inhibition of TGF-beta-induced Fn production. CONCLUSIONS: Activation of PKC stimulates Fn production; however, neither TGF-beta nor EGF produce Fn through a PKC-dependent pathway. EGF and TGF-beta both stimulate Fn production at least in part through the intracellular signaling protein MAP-K. Understanding the signaling pathways involved in extracellular matrix protein production will allow the design of specific inhibitors of intimal hyperplasia.

Endovascular femoropopliteal bypass: a cadaveric study.

OBJECTIVE: Patency rates of standard femoropopliteal bypass in infra-inguinal occlusive disease have yet to be matched by minimally invasive percutaneous procedures. We report a feasibility study of a less invasive endovascular femoropopliteal bypass technique. METHODS: (1) groin exposure of femoral artery, (2) guidewire passage and mechanical dilatation of superficial femoral artery (SFA), (3) expandable helical cutter endarterectomy of SFA, (4) transluminal placement of PTFE graft, (5) graft balloon dilatation to shape and set distal interface and (6) end-to-end anastomosis of proximal graft to femoral artery. Development and testing was undertaken in 48 limbs of 26 fresh human cadavers. Limbs with no demonstrable SFA disease were excluded. Seventeen limbs had mild, diffuse disease. Three limbs had a single, short, tight stenosis. Seventeen limbs had multiple, high grade stenotic lesions 12-40 cm long (mean 24 cm). Eleven limbs had occlusive lesions, 8-38 cm long (mean 24 cm). RESULTS: We successfully completed the procedure in 39 (81%) limbs. We failed to complete the procedure in nine limbs; four from failed guidewire passage, four from vessel avulsion, and one from graft deployment failure. Histology confirmed endarterectomy cleavage in the standard plane. Angiography and explants demonstrated a patent graft and popliteal artery, and smooth distal graft/arterial interface with no obvious defects in 24 (62%) cases. Defects included combinations of: contrast extravasation/reflux, graft malpositioned/incorrectly sized, distal graft fold, and distal intimal flap. CONCLUSION: Endovascular femoropopliteal bypass is feasible and warrants further studies for possible clinical application.

Endovascular femoropopliteal bypass: early human cadaver and animal studies.

We report herein a feasibility study of a minimally invasive endovascular femoropopliteal bypass procedure. The steps include the following: (1) a small groin incision to expose the femoral artery, (2) guidewire passage and mechanical dilatation of the diseased superficial femoral artery, (3) semiclosed endarterectomy of the superficial femoral artery using an expandable metal endarterectomy catheter that engages atheroma, (4) placement of a 6 mm thin-walled PTFE graft, (5) balloon dilatation of the graft to press the graft flat against the arterial wall, and (6) a standard end-to-end anastomosis of the proximal graft to the femoral artery. This technique was tested in 13 limbs from eight fresh (stored 1 to 5 days at 4 degrees C) human cadavers (seven females and one male). Five limbs had stenotic superficial femoral artery lesions, 1 to 15 cm (mean 7.6 cm). Four limbs had occlusive lesions, 9 to 38 cm long (mean 26.8 cm). Four limbs had no disease. We successfully completed the procedure in 10 of 13 limbs. Completion arteriography showed a widely patent graft and a popliteal artery with a smooth distal graft/arterial interface in 9 of 10 limbs; one had a distal graft fold due to a size mismatch. Histologic studies of the superficial femoral artery revealed intima, atheromatous plaque, and media. We failed to complete our procedure in three limbs: two because of inadequate instruments and one because of perforation of the artery. We also performed the same procedure unilaterally in six dogs, except that no endarterectomy was performed.(ABSTRACT TRUNCATED AT 250 WORDS)