Inhibition of tissue factor reduces thrombus formation and intimal hyperplasia after porcine coronary angioplasty

OBJECTIVES

We investigated the in vivo effects of tissue factor (TF) inhibition with recombinant tissue factor pathway inhibitor (rTFPI) on acute thrombus formation and intimal hyperplasia and the in vitro effects on smooth muscle cell migration and proliferation.

BACKGROUND

Inhibition of TF with TFPI has been shown to reduce intimal hyperplasia in experimental models. However, its effects after coronary angioplasty and the cellular mechanisms involved have not been investigated.

METHODS

Twenty-three swine underwent multivessel coronary angioplasty. Fifteen (n = 25 arteries) were euthanized at 72 h to assess thrombus formation and eight (n = 24 arteries) at 28 days to assess intimal hyperplasia. Animals in the 72-h time point received: 1) human rTFPI (0.5 mg bolus plus 25 μg/kg/min continuous infusion for 3 days) plus heparin (150 IU/kg intravenous bolus) plus acetyl salicylic acid (ASA) (325 mg/day); 2) rTFPI regimen plus ASA and 3) heparin (150 IU/kg intravenous bolus) plus ASA.

RESULTS

On histology the control group had evidence of mural thrombus (area 0.8 ± 0.4 mm²). Treatment with TFPI plus heparin abolished thrombus formation (mean area: 0.0 ± 0.0 mm², p < 0.05) but was associated with prolonged activated partial thromboplastin time and extravascular hemorrhage. Recombinant TFPI alone inhibited thrombosis without bleeding complications (mean area: 0.03 ± 0.02 mm², p < 0.05 vs. control). Animals in the 28-day time point received continuous intravenous infusion of rTFPI or control solution for 14 days. Tissue factor pathway inhibitor reduced neointimal formation with mean intimal area of 1.2 ± 0.3 mm² versus 3.2 ± 0.4 mm² in the control group; p < 0.01. Recombinant TFPI had no effect on human aortic smooth muscle cell growth but inhibited platelet-derived growth factor BB-induced migration.

CONCLUSIONS

Inhibition of TF with rTFPI can prevent acute thrombosis and intimal hyperplasia after injury. Tissue factor plasma inhibitor may prove useful as an adjunct to intracoronary interventions.     

Histopathologic analysis of in-stent neointimal regression in a porcine coronary model

BACKGROUND: Animal and clinical studies have demonstrated late regression of in-stent neointima. This study was performed to identify the temporal changes in the in-stent neointimal constituents responsible for late regression. METHODS: NIR stents were implanted in porcine coronary arteries (size of stent (in mm) to size of artery (in mm) approximately equal to 1.1) and harvested at 2 months and 6 months (n = 6 stents/group). Histopathologic analyses included morphometric analysis, smooth muscle cell density, and extracellular matrix contents. RESULTS: Compared with the findings at 2 months, at 6 months there was a significant reduction in area stenosed (from 21 +/- 3% to 14 +/- 1%, P < 0.05) and neointimal thickness (from 0.2 +/- 0.03 mm to 0.03 +/- 0.02 mm, P < 0.05), despite similar injury scores (0.05 +/- 0.06 at 2 months and 0.36 +/- 0.29 at 6 months). This regression was accompanied mainly by a reduction in proteoglycan (from 24 +/- 19% to 5 +/- 8%, P = 0.05), with no change in smooth muscle cell density (71 +/- 7 compared with 76 +/- 23/high power field) or collagen content (25 +/- 19% compared with 25 +/- 19%). CONCLUSIONS: The study confirmed the regression of in-stent neointima, which was mainly attributable to a reduction in proteoglycan content, resembling the natural healing response.     

Local delivery of 17-beta-estradiol decreases neointimal hyperplasia after coronary angioplasty in a porcine model

BACKGROUND: Neointimal hyperplasia is an important mechanism of restenosis after percutaneous transluminal coronary angioplasty (PTCA). Systemically administered estrogen is known to inhibit neointimal formation after arterial injury. OBJECTIVES: We sought to assess the efficacy of locally delivered 17-beta-estradiol (BE) in inhibiting neointimal hyperplasia after PTCA. METHODS: Eighteen juvenile farm pigs were studied. Coronary angioplasty was performed in all three coronary arteries of each animal. After PTCA, each coronary artery in each pig was randomized to receive either local delivery of 600 microg BE, vehicle alone or PTCA only. Twelve animals were euthanized at 28 days for morphometric analysis, and four animals were euthanized at seven days for immunohistochemical analysis of vascular smooth muscle cell (SMC) proliferative activity. Two animals died a few days after PTCA and were excluded. RESULTS: On morphometric study, the arterial segments treated with BE demonstrated significantly less neointimal proliferation. Arteries treated with BE had reductions in several indexes of restenosis compared with arteries treated with vehicle alone or PTCA only: neointimal area (0.4+/-0.09 mm2 for BE vs. 1.14+/-0.33 mm2 for vehicle alone vs. 0.88+/-0.2 mm2 for PTCA only, p<0.05), percent neointima (12.16+/-2.57% vs. 25.46+/-4.73% vs. 23.02+/-3.97%, p<0.025), neointima/media area (0.59+/-0.14 vs. 1.75+/-0.41 vs. 1.67+/-0.43, p<0.01) and restenotic index (1.3+/-0.14 vs. 2.42+/-0.22 vs. 2.4+/-0.23, p<0.005). Immunohistochemistry showed decreased SMC proliferative activity in BE-treated arteries compared with the other two treatment groups (p<0.05). CONCLUSIONS: Local delivery of BE significantly decreases neointimal hyperplasia after PTCA in pigs, probably by the inhibition of SMC proliferation.     

Restenosis after balloon angioplasty. A practical proliferative model in porcine coronary arteries

A model of proliferative human restenosis was developed in domestic pigs by using deep injury to the coronary arterial media. Metal wire coils were delivered percutaneously to the coronary arteries of 11 pigs with an oversized, high-pressure (14 atm) balloon and were left in place for times ranging from 28 to 70 days. During placement, the balloon expanded the coils and delivered them securely within the arterial lumen. Light microscopic examination of the vessels confirmed fracture of the internal elastic lamina by the coil. An extensive proliferative response occurred in 10 of the 11 pigs and was associated with a luminal area narrowing of at least 50% in all but one pig. The histopathologic features of the proliferative response were identical to those observed in human cases of restenosis after angioplasty. Immunohistochemical studies confirmed the prominence of smooth muscle cells in the proliferative tissue. A similar response was obtained in two of five porcine coronary arteries in which balloon inflation only was performed, without coil implant. This model is practical and inexpensive and closely mimics the proliferative portion of human restenosis both grossly and microscopically. Thus, it may be useful for understanding human restenosis and for testing therapies aimed at preventing restenosis after balloon angioplasty or other coronary interventional procedures.     

Hypercholesterolemia impairs vascular remodelling after porcine coronary angioplasty

OBJECTIVE: To assess the effect of hypercholesterolemia on neointima formation and vascular remodelling after porcine coronary angioplasty. METHODS: Left anterior descending coronary angioplasty was carried out in five control and 16 age-matched hypercholesterolemic miniature pigs. Vascular remodelling was measured by intravascular ultrasound. Neointima size and composition were assessed by quantitative image analysis. Coronary smooth muscle cells (SMC) from control and diet pigs were collected 1 h after angioplasty for in vitro study of the effect of hypercholesterolemic serum on SMC migration and of macrophage-induced matrix degradation on SMC adhesion. RESULTS: Twenty-eight days after angioplasty, lumen increase was 0.08+/-1.7 mm(2) in diet and 2.7+/-2.7 mm(2) (P=0.016) in control pigs. Lumen increase correlated with vascular remodelling (IEL(post)/IEL(pre); R(2)=0.59; P<0.001) and with the circumferential gain relative to the neointima (R(2)=0.32; P<0.01) but not with neointimal area that was similar in control and diet pigs. Circumferential gain correlated with VSMC deposition at the site of the injury (R(2)=0.28; P<0.01) that correlated with organized collagen (R(2)=0.34; P<0.01). The VSMC and collagen content of neointima in diet pigs was lower whereas the macrophage content was higher. Hypercholesterolemic serum and oxidised LDL reduced migration of VSMC from diet pigs. Macrophage-induced degradation of VSMC extracellular matrix reduced VSMC adhesion (P=0.015). CONCLUSION: Hypercholesterolemia impairs vascular remodelling of balloon-treated coronary arteries. It decreases VSMC and collagen accumulation at the site of injury. Our in vitro data suggest that this decrease can be due to macrophage-induced matrix degradation and reduced VSMC adhesion and to impaired VSMC migration. Oxidised LDL mimics the inhibitory effect of hypercholesterolemic serum.     

Intracoronary irradiation markedly reduces restenosis after balloon angioplasty in a porcine model

Objectives. This study examined the effects of intracoronary Irradiation on neointimal proliferation after overstretch balloon angioplasty in a normolipemic swine model of restenosis.Background. Restenosis after percutaneous transluminal coronary angioplasty represents, in part, a proliferative response of vascular smooth muscle at the site of injury. We have previously shown that ionizing radiation, delivered by means of an intracoronary source, causes focal medial fibrosis. We therefore hypothesized that ntracoronary irradiation delivered at the time of balloon angtoplasty might impair the restenosis process.Methods. Nineteen juvenile swine underwent coronary angiography; a segment of the coronary artery was chosen as a target for balloon injury. In 10 swine, a ribbon of iridiun-192 was positioned at the target segment, and 2,000 cGy was delivered at the vessel wall. Subsequently, overdilation balloon angioplasty was perfromed at the irradiated segment. In nine control swine, overdilation balloon angioplasty was performed without previous irradiation. Eighteen animals survived and were killed at 30 days. Histopathologic analysis was performed by a pathologist in blinded manner. The area of maximal lumen compromise within the target segment was analyzed by computer-assisted planimetry.Results. In the control group, mean (± SD) neointimal area was 0.84 ± 0.60 mm²compared with that in the irradiated group, 0.24 ± 0.13 mm²(p = 0.01). In the control group, mean percent area stenosis was 47.6 ± 20.7%, whereas that in the irradiated group was 17.6 ± 10.5% (p = 0.001). This represents a 71.4% reduction in neointimal area and a 63.0% reduction in percent area stenosis in the irradiated group. Adjacent coronary segments and surrounding myocardium were unaffected.Conclusions. Intracoronary irradiation (2,000 cGy) delivered to a target porcine coronary segment before balloon overdilation markedly reduces neointima formation at 30 days and thus significantly impairs the restenosis process.     

Triage of patients for short term observation after elective coronary angioplasty

OBJECTIVE—To evaluate triage of patients for short term observation after elective percutaneous transluminal coronary angioplasty (PTCA), as appropriate selection of patients for short term observation after angioplasty may facilitate early discharge.METHODS—1015 consecutive patients scheduled for elective PTCA were prospectively included for short term observation. Patients with unstable angina Braunwald class III were excluded. There were no angiographic exclusion criteria. Patients were discharged from the interventional centre when considered stable during 4 hours of observation after PTCA. It was left to the operator''s discretion whether to prolong the observation period. Procedural complications were defined as death, coronary bypass surgery, early repeat PTCA, and myocardial infarction.OUTCOME MEASURES—The need for prolonged observation (> 4 hours) and the occurrence of complications. Predictors for prolonged observation and the occurrence of complications after the 4 hours observation were assessed by univariate and multivariate analysis.RESULTS—Two patients died, including one of six patients who underwent emergency bypass surgery. In all, 922 patients (90.8%) were triaged to short term observation and had an uncomplicated three day follow up. Observation was prolonged in 87 patients (8.6%), and 40 patients had a complicated course. Independent predictors of procedural complications were acute closure (odds ratio (OR) 9.7; 95% confidence interval 4.4 to 21.4), side branch occlusion (OR 8.9; 3.4 to 23.7), no angiographic success (OR 5.1; 2.4 to 11.0), female sex (OR 3.1, 1.7 to 5.7), any unplanned stent (OR 2.8, 1.4 to 5.9), and ostial lesion (OR 2.2, 1.0 to 4.7).CONCLUSIONS—A 4 hour observation period is safe after elective coronary angioplasty. As procedural variables are the strongest predictors of postprocedural complications, the immediate procedural results allow effective triage of patients for short term or prolonged observation in order to anticipate complications.     

Dissolution of a huge spontaneous coronary artery thrombus with a new antiplatelet agent and coronary angioplasty

A huge thrombus was observed in the proximal portion of the right coronary artery (RCA) in a 75-year-old patient. He presented with symptoms of cardiogenic shock due to acute myocardial infarction. Glycoprotein IIb/IIIa inhibitor (Tirofiban) was administered intra-coronary first, coronary angioplasty was performed later and, dramatic thrombus dissolution was observed on control coronary angiography.     

Neointimal response following rotational atherectomy compared to balloon angioplasty in a porcine model of coronary in-stent restenosis

In-stent restenosis remains a clinical therapeutic challenge. Rotational atherectomy (RA) is an attractive treatment option as it may cause less vascular injury than balloon angioplasty (BA) and, therefore, limit further neointimal response. In an animal model of coronary in-stent restenosis, thermal injury and stenting created neointima (old NI). The treatment of in-stent restenosis with either BA (n = 9) or RA (n = 11) also generated neointima (new NI). The average areas (mm²) of old NI in the BA and RA groups were similar (3.77 ± 0.40 vs. 3.67 ± 0.53; P = 0.32). However, new NI formed after treatment of in-stent restenosis was significantly less in the RA as compared to the BA group (0.33 ± 0.12 vs. 0.73 ± 36, P < 0.01). In this porcine coronary artery model of in-stent restenosis, treatment with rotational atherectomy resulted in significantly less recurrent neointimal hyperplasia than balloon angioplasty. This animal study, thus, provides a rationale for the clinical use of rotablation in the treatment of in-stent restenosis. Cathet. Cardiovasc. Diagn. 45:332–336, 1998. © 1998 Wiley-Liss, Inc.     

Long-term endothelial dysfunction is more pronounced after stenting than after balloon angioplasty in porcine coronary arteries

Objectives. To compare percutaneous transluminal coronary angioplasty (PTCA) and stent implantation with respect to the long-term changes they induce in the newly formed endothelium in porcine coronary arteries by studying both morphological and functional parameters of the endothelium at 2 weeks and 3 months after intervention.Background. Problems affecting PTCA or stent implantation have been overcome to a large extent by means of better techniques and the availability of new drugs. Late problems, however, still exist in that restenosis affects a large number of patients. With an increasing number of patients being treated with stents, the problem of in-stent restenosis is of even greater concern, as this seems difficult to treat. A functional endothelial lining is thought to be important in controlling the growth of the underlying vascular tissue. We hypothesized that the enhanced neointimal hyperplasia observed after stenting is associated with a more pronounced and prolonged endothelial dysfunction.Methods. Arteries were analyzed using a dye-exclusion test and planimetry of permeable areas. Thereafter, the arteries were processed for light and scanning electron microscopy for assessment of morphology and proliferative response.Results. Leakage of the endothelium for molecules such as Evans blue-albumin as well as prolonged endothelial proliferation is observed as late as 3 months after the intervention, and is more pronounced after stenting. Permeability is associated with distinct morphologic characteristics: endothelial retraction, the expression of surface folds, and the adhesion of leukocytes.Conclusions. Stenting especially decreases long-term vascular integrity with respect to permeability and endothelial proliferation, and is associated with distinct morphologic characteristics.     

Embolization complicating coronary angioplasty in the presence of an intracoronary thrombus

Two cases of coronary occlusion and subsequent embolization during percutaneous coronary angioplasty (PTCA) are described. Prior to PTCA, angiographic evidence of intracoronary thrombus was present. Abrupt reclosure after dilation was treated by successful redilation. However, coronary embolization of thrombus debris occurred downstream in one patient and into an adjacent coronary branch in the second patient.     

Influence of contrast media on thrombus formation during coronary angioplasty

The influence of contrast media on thrombus formation during percutaneous transluminal coronary angioplasty was assessed in 124 consecutive patients undergoing coronary angioplasty and receiving either ionic (n = 57) (Group I) or nonionic (n = 67) (Group II) contrast medium. The presence of thrombus was assessed by qualitative analysis of angiograms in identical pre- and postangioplasty projections by four observers who had no knowledge of other data. Quantitation of stenosis severity before and after angioplasty and qualitative analysis of lesion eccentricity and complexity and of the presence of dissection were also performed. Although the baseline clinical characteristics of the two groups (including presenting syndromes and procedural and angiographic variables) did not differ, more patients in Group II than Group I developed new thrombus during coronary angioplasty (18% vs. 4%, p less than 0.02). In particular, patients with a presenting syndrome of recent myocardial infarction or rest angina, or both, and patients with an eccentric coronary plaque were more likely to develop new thrombus if they received nonionic than if they received ionic contrast medium (p less than 0.05). Patients with new thrombus formation and patients with thrombus present both before and after angioplasty had a high incidence of acute procedural complications (36% and 23%, respectively). Patients in Groups I and II had a similar incidence of ischemic events during follow-up.     

Aspiration of coronary thrombus during angioplasty for postmyocardial infarction ischemia

We report the use of direct coronary aspiration to remove obstructing thrombus from a proximal right coronary artery after failure of balloon angioplasty to resolve the thrombus responsible for postmyocardial infarction ischemia. © 1995 Wiley-Liss, Inc.     

Rho-kinase inhibitor suppressed restenosis in porcine coronary balloon angioplasty

BACKGROUND: Constrictive remodeling is thought to be more important than neointimal formation in coronary restenosis after balloon angioplasty. The inhibition of Rho-kinase prevents neointimal proliferation, but now this inhibition that affects constrictive remodeling remains unknown. To explore this issue further, we investigated whether a specific Rho-kinase inhibitor, Y-27632, could suppress restenosis after coronary balloon angioplasty in a porcine model. METHODS: Balloon angioplasty with local administration of Y-27632 (Y group) or vehicle (C group) was performed at 2 and 3 weeks after overstretch injury in a porcine coronary artery. Quantitative coronary angiography (QCA) and quantitative coronary ultrasound (QCU) were performed to assess the coronary lesion segment. A morphometrical analysis was performed in a histological study. Proliferative cells and p27(Kip1)-positive cells were evaluated in the arterial wall using immunohistochemistry. RESULTS: QCA and QCU demonstrated that the minimal lumen diameter and minimal lumen area were greater, and % stenosis was less in the Y group than in the C group. The QCU analysis also revealed a significant inhibition in the increase of the intimal area and a prevention of constrictive remodeling by Y-27632. In the histological study, the intimal, adventitial and collagen areas were significantly smaller in the Y group than in the C group. The Y group also exhibited significantly less proliferative activity and a significantly higher percentage of cells expressing p27(Kip1) in the arterial wall. CONCLUSION: Local delivery of Y-27632 suppressed constrictive remodeling as well as neointimal formation after coronary balloon angioplasty in pigs.     

Role of intracoronary thrombus in acute complications during percutaneous transluminal coronary angioplasty

Coronary angiograms from 2,372 consecutive patients who underwent percutaneous transluminal coronary angioplasty (PTCA) were retrospectively reviewed for the presence of intracoronary thrombus (ICT) before dilatation. Patients with evolving acute myocardial infarction and those receiving thrombolytic therapy were excluded from analysis. Coronary artery thrombus was present in 126 patients (6%) (group 1). When compared to 2,246 patients (group 2) without ICT, group 1 had a higher incidence of unstable angina, 74% vs. 66% (less than 0.06), previous myocardial infarction, 59% vs. 37% (P less than .0001), and history of a recent myocardial infarction, 28% vs. 9% (P less than .0001). Patients with predilatation intracoronary thrombus had a higher risk for acute occlusion, 6% vs. 2% (P less than .002); however, the incidence of emergency coronary bypass surgery and myocardial infarction was similar in both groups. Therefore, the presence of predilatation intracoronary thrombus heralds an increased risk of acute occlusion, but not myocardial infarction or emergency coronary artery bypass surgery.