Human arterial surface fluorescence: atherosclerotic plaque identification and effects of laser atheroma ablation

In vivo plaque recognition may be important for safe and precise intra-arterial atheroma ablation during laser coronary angioplasty. This study examined the feasibility and sensitivity of utilizing quantitative fluorescence spectroscopy and video-enhanced fluorescence imaging for plaque identification in atherosclerotic human necropsy arterial wall before and after laser atheroma ablation. With wide-band (450 to 490 nm) blue light excitation, the 540 nm fluorescence intensity ratio of normal to diseased sites (n = 13) was 2.09 +/- 0.82 (p less than 0.001) and video fluorescence imaging provided enhanced delineation of atheroma surface characteristics. Continuous argon and pulsed excimer (308 nm) laser ablation of atheroma decreased fluorescence intensity ratios by 42 and 20% (p less than 0.001), respectively (that is, from abnormal to nearly normal). Low power 325 nm laser-excited fluorescence spectroscopy from normal (n = 115) and abnormal (n = 146) necropsy sites revealed an average 45% decrease in atheroma fluorescence intensity (p less than 0.0001) and changes in fluorescence spectra appearance that corresponded to plaque morphologic subtypes. Studies using a dual laser system combining 325 nm laser-excited fluorescence plaque recognition and a 480 nm pulsed dye laser for tissue ablation with common optical fibers demonstrated normalization of both fluorescence intensity and spectra appearance after laser atheroma ablation. Thus, in vitro analysis of surface arterial fluorescence by quantitative spectroscopy and video fluorescence imaging reliably differentiate plaque from normal tissue and may provide the feedback signal needed to activate a laser source for selective plaque removal.     

Spatial fluorescence imaging of atherosclerotic plaque: contrast enhancement by 2 wavelength laser stimulation, digital image processing and dye marking]

A new video-enhanced fluorescence imaging technique has been used for the first time for in vitro differentiation of human atherosclerotic plaques vs normal arterial wall. Laser-induced superficial tissue fluorescence of specimens from human aorta was documented after alternating excitation with violet (405 nm +/- 5 nm) and blue (470 nm +/- 10 nm) krypton-ion laser light. Subsequent digital subtraction of the corresponding fluorescence images allowed to differentiate areas of atherosclerosis from normal intima. Fluorescence intensity was correlated with the morphological aspect of samples and histology of the plaque. Dihematoporphyrin-ether/ester (DHE) incubation enhanced fluorescence contrast of plaques in comparison to normal artery vessel wall. Depending on the concentration of the incubation solution (10, 20, and 40 micrograms DHE/ml NaCl solution), fluorescence increased. Fluorescence intensity was highest in fatty plaque areas, while calcific lesions showed no substantial DHE uptake.     

Characterization of human coronary artery atherosclerotic plaque fluorescence emission

Preliminary trials using fluorescence guidance of laser ablation in femoral arteries have been successful. There have, however, been few studies of the characteristics of fluorescence emissions from coronary arteries. A large series of fluorescence emission spectra from human coronary artery specimens was examined. Analysis included: fluorescence emission during excitation with ultraviolet and visible light; histologic correlations between plaque content and thickness, averaged spectra and fluorescence intensity ratios; and differences in specific plaque morphology with excitation of the same coronary specimens at 325 or 458 nm. Ratios of fluorescence emission intensity at selected wavelengths were calculated for both 325 and 458 nm excitation (13 wavelengths, 78 ratios for 325 nm; 11 wavelengths, 55 ratios for 458 nm). The following were found: atherosclerotic lesions in human coronary arteries were characterized by an increase in normalized fluorescence intensity at longer wavelengths when excited with either ultraviolet or visible light; calcific plaque content greater than 10% in lesions more than 1 mm thick was identified by increased normalized fluorescence intensity at 443 nm during excitation at 325 nm; and fatty plaque content correlated with fluorescence intensity ratios during 325 nm excitation, whereas fibrous and calcific content correlated well with fluorescence ratios during 458 nm excitation. It is concluded that characteristic fluorescence emission has the potential to correctly identify and characterize plaque morphology in human coronary arteries.     

Effects of hematoporphyrin derivative and photodynamic therapy on atherosclerotic rabbits

This study was performed to demonstrate selective uptake of hematoporphyrin derivative (HPD) within actively developing atheroma, to localize the site of uptake of HPD within the atheroma, and to determine the potential for photodynamic therapy (PDT) of atherosclerosis in the rabbit model. Fifteen rabbits were rendered atherosclerotic. Five rabbits received neither HPD nor PDT and 2 rabbits received HPD, 10 mg/kg intravenously, without subsequent irradiation. Eight other rabbits received 5 to 20 mg of HPD intravenously and subsequent intravascular 636-nm laser radiation to either the thoracic aorta or the aortic arch. A total of 32 to 288 J of laser energy was delivered through a 300-mu quartz fiber. All rabbits that received in vivo HPD had red fluorescence of their aortas when placed under ultraviolet light. The pattern of fluorescence corresponded precisely to the pattern of atheroma. In segments that received PDT, light microscopic examination revealed an accumulation of smooth muscle cells at the intimal surface. Fluorescence microscopy revealed a diminishing concentration gradient of HPD from intimal surface layers towards the media. Assessment of treated thoracic aortic segments revealed quantitative and qualitative differences compared with control segments. In the arch-treated segments, however, no changes were seen. It is concluded that HPD localizes within rabbit atheroma, can be detected by fluorescence and is deposited in a diminishing concentration gradient from lumen toward media. Irradiation with 636-nm light may induce qualitative and quantitative changes in atheroma.     

The interaction between excimer laser energy and vascular tissue

The effects of XeF1 excimer laser on isolated normal and atherosclerotic aorta were studied. Experiments were performed in flowing water at constant temperature, flow rate, water depth, pulse width (10 nsec), wavelength (351 nm), beam size (1 mm2) and focal length (50 cm). The number of pulses, the pulse energy, and the pulse frequency were varied, and the vascular tissue was studied histologically. The following observations were made: tissue ablation required a minimum threshold pulse energy and was nonlinearly proportional to the number of pulses and the pulse energy delivered; precise tissue ablation occurred at low pulse frequencies, but changes resembling a thermal process were seen as pulse frequency increased; calcified plaque was more photoresistant than atheroma or normal vessel; excimer laser energy was markedly attenuated by blood; and the time interval between pulses and high peak power are related to the precision of ablation by pulsed excimer laser. It is concluded that excimer laser can rapidly and precisely ablate vascular tissue by a photothermal process.     

Effect of laser irradiation on atherosclerosis vascular segments in relation to wave length and pulse width

The interaction of pulsed Nd:YAG laser radiation and atherosclerotic vessel wall is reported. The effects in the area of radiation were examined at an infrared, green, and ultraviolet wavelength at variable pulsewidths. The laser beam was focused directly on the plane surface of the tissue. The macroscopic and histologic examinations demonstrate that thermal damage of tissue is higher at an infrared wavelength than at a green wavelength and respectively at an ultraviolet wavelength. With increasing pulsewidth an increase of coagulation and necrosis of plaque material and adjacent vessel wall can be noted. The results show that for the precise ablation of plaque without adjacent vessel wall injury short pulsed ultraviolet radiation is preferable.     

XeCl excimer laser-induced fluorescence for selective ablation of atheromatous tissue.

In order to develop a reliable laser-induced fluorescence (LIF) guided laser angioplasty system, real time, pulse-by-pulse fluorescence spectra were recorded and the same fiberoptic was used for both detection of the fluorescence and for atheromatous tissue ablation. A 308 nm XeCl excimer laser served as the laser source for both the induction of fluorescence and the ablation. The fluorescence signal was induced at high laser energies during ablation without any serious change in the fluorescence pattern. A new characteristic fluorescence peak at 540 nm for atheromatous tissue was observed after treatment with chlortetracycline hydrochloride (CTC). This allowed the development of an algorithm and a subsequent index to discriminate the atheromatous tissue from the normal tissue. During atheromatous tissue ablation, this index changed as normal tissue was approached, thereby avoiding vessel perforation. Our results suggest that monitoring of this index through the catheter delivering the laser energy enhances selective ablation while simultaneously reducing the risk of vessel perforation.     

Determinants of coronary artery compliance in subjects with and without angiographic coronary artery disease

OBJECTIVES: The goal of this study was to determine factors contributing to the biomechanical properties of coronary arteries in people with and without angiographic coronary artery disease (CAD). BACKGROUND: The stiffness of the aorta is known to increase with increasing age and in the presence of CAD. However, little is known about the mechanics of coronary arteries, which may have important clinical consequences. METHODS: Intravascular ultrasound was used to determine the mechanical properties of coronary arteries and plaque behavior in subjects with CAD (n = 38), those with chest pain but angiographically normal coronary arteries (N) (n = 9) and those early (<2 weeks) after cardiac transplant (T) (n = 14). RESULTS: Coronary arteries dilated during systole in all groups, but cross-sectional compliance and distensibility were lowest in the proximal left anterior descending artery (LAD) in the subjects with CAD compared with the N and T groups (compliance: 1.2 +/- 0.2 vs. 1.7 +/- 0.5 and 2.7 +/- 0.6 x 10(-2) mm(2) mm Hg(-1) [mean +/- SEM] respectively, p < 0.02 CAD vs. T; distensibility: 0.8 +/- 0.2 vs. 1.7 +/- 0.5 and 1.7 +/- 0.3 x 10(-3) mm Hg(-1), p < 0.05 CAD vs. T). There was extensive plaque in the CAD group, and plaque was also present in the N group, but minimal atheroma was present in the T group. Plaque cross-sectional area diminished significantly during systole in both the LAD and circumflex arteries. Absolute changes were: 0.50 +/- 0.30, 0.33 +/- 0.11 and 0.68 +/- 0.13 mm(2) in the proximal LAD, distal LAD and proximal circumflex arteries, respectively. In subjects with atheroma, there was a significant correlation between cross-sectional compliance and plaque compression at all sites, and plaque compression was a significant determinant of cross-sectional compliance at both proximal sites in multiple regression analyses with age, mean arterial pressure and extent of plaque as the other independent variables. CONCLUSIONS: A major determinant of the systolic increase in coronary luminal area in patients with atheroma is a reduction in plaque cross-sectional area during systole.     

激光诱导荧光光谱诊断动脉粥样斑块的实验研究

研究337nm 氮分子激光诱导荧光光谱(LIFS)方法诊断家兔动脉粥样硬化斑块及血液介质、四环素浸泡对荧光光谱的影响。结果显示用 LIFS 可区别正常和斑块动脉,区分的正确率为94%,斑块诊断敏感性92%,特异性100%,阳性预测值100%。氧合血红蛋白的荧光再吸收作用对LIFS 有明显影响,是 LIFS 中低谷产生的原因。血液介质使正常动脉和斑块动脉的 LIFS 两高峰的荧光强度比值变得没有差异(P>0.05)。正常动脉和斑块动脉经四环素溶液浸泡后均出现特征性的四环素荧光谱,未显示四环素对斑块的特异亲合性。     

Photoangioplasty with local motexafin lutetium delivery reduces macrophages in a rabbit post-balloon injury model

OBJECTIVE: Motexafin lutetium (Lu-Tex, Antrin Injection) is a photosensitizer that selectively accumulates in atheromatous plaque where it can be activated by far-red light. The localization and retention of intra-arterially administered Lu-Tex and its efficacy following activation by endovascularly delivered light (photoangioplasty) was evaluated. METHODS: Bilateral iliac artery lesions were induced in 17 rabbits by balloon denudation, followed by a high cholesterol diet. Lu-Tex distribution within the atheroma was examined (n=8) following local injection. Fluorescence spectral imaging and chemical extraction techniques were used to measure Lu-Tex levels within the atheroma and adjacent normal tissue. Photoactivation was performed 15 min following Lu-Tex administration (180 J/cm fiber at 200 mW/cm fiber). Two weeks post photoangioplasty, vessels were harvested and hematoxylin and eosin (H&E) and RAM11 (macrophages) staining was performed. RESULTS: Local delivery of Lu-Tex achieved immediate high concentrations within plaque (mean 40x control iliac atheroma). Mean percent plaque area in the treated segments was significantly lower than in the non-treated contralateral lesions (73 vs. 82%, P<0.01). No medial damage was observed. Quantitative analysis using RAM11 positive cells revealed significant reduction of macrophages in treated lesions in both the intima (5 vs. 22%, P<0.01) and in media (8 vs. 23%, P<0.01) compared to untreated contralateral segments. CONCLUSIONS: Local delivery provides high levels of Lu-Tex selectively within atheroma. Photoactivation results in a significant decrease in macrophage and a small decrease in atheroma burden without damage to the normal vessel wall.     

Rate of progression of coronary atherosclerotic plaque in women.

OBJECTIVES: The purpose of this study was to determine the relationship between gender and the extent of coronary atherosclerosis assessed by intravascular ultrasound (IVUS) and its rate of progression in subjects treated with established medical therapies. BACKGROUND: It is uncertain whether the pathophysiology of coronary artery disease (CAD) differs between genders. METHODS: A systematic analysis was performed of 978 subjects who participated in serial studies of atheroma progression. Genders were compared with regard to the extent of coronary atheroma at baseline and subsequent change in response to use of established medical therapies. RESULTS: Women were more likely to have a history of hypertension and higher levels of body mass index, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, C-reactive protein, and systolic and diastolic blood pressure. Despite this, women had less plaque in terms of percent atheroma volume (PAV) (33.9 +/- 10.2% vs. 37.8 +/- 10.3%, p < 0.001) and total atheroma volume (TAV) (148.7 +/- 66.6 mm3 vs. 194.7 +/- 84.3 mm3, p < 0.001). With medical therapy, the rate of change of PAV (0.7 +/- 0.6% vs. 0.7 +/- 0.5%, p = 0.92) and TAV (-2.3 +/- 3.2 mm3 vs. -1.9 +/- 2.9 mm3, p = 0.84) did not differ between genders. In the setting of intensive risk factor modification, there was no significant difference between genders with regard to the rates of plaque progression or regression. CONCLUSIONS: Despite the presence of more risk factors, the extent of atheroma in women with angiographic CAD is less than in men in subjects participating in clinical trials that employed serial assessments with IVUS. The finding that the rate of plaque progression or regression does not differ between genders in the setting of intensive risk factor modification supports the use of established medical therapies in women with CAD.     

Measurement of atherosclerotic plaque volume in hyperlipidemic rabbit aorta by intravascular ultrasound

OBJECTIVES: Atherosclerotic changes in the rabbit have been evaluated by various methods. Although most previous studies have analyzed atherosclerotic plaque in the femoral, carotid and iliac arteries of rabbits by intravascular ultrasound (IVUS) because of easier access, we established a method for the precise measurement of plaque volume as well as plaque area in the thoracic descending aorta in the Watanabe heritable hyperlipidemic (WHHL) rabbit, which has severe atherosclerosis. METHODS: WHHL and Japanese White (JW)rabbits were used. An IVUS catheter was inserted into the right femoral artery and advanced to the left subclavian artery, which was used as an anatomical landmark. After IVUS image acquisition, the catheter was removed. Vessel volume, lumen volume and plaque volume were analyzed. RESULTS: Atheroma of the aorta was easily detected in WHHL rabbits by IVUS examination, whereas atherosclerosis was not observed in JW rabbits. The atheroma showed a low-echoic lesion compared to the adventitia, with morphological characteristics similar to human lipid-rich, soft atheromatous plaques. In 15-month-old WHHL rabbits, the vessel volume, lumen volume and plaque volume in the thoracic descending aorta were 815 +/- 109, 559 +/- 107 and 256 +/- 10 mm3/ 3 cm, respectively. CONCLUSIONS: We established a method for the precise quantitation of plaque volume by IVUS technology in WHHL rabbits aorta for the first time. This method is useful for evaluating several locally or generally delivered therapeutic agents in a hyperlipidemic animal model.     

Prevention of distal embolization and no-reflow in patients with acute myocardial infarction and total occlusion in the infarct-related vessel: a subgroup analysis of the cohort of acute revascularization in myocardial infarction with excimer laser-CARMEL multicenter study.

To overcome the adverse complications of percutaneous coronary interventions in thrombus laden lesions (i.e., distal embolization, platelet activation, no-reflow phenomenon), mechanical removal of the thrombus or distal embolization protection devices are frequently required. Pulsed-wave ultraviolet excimer laser light at 308 nm can vaporize thrombus, suppress platelet aggregation, and, unlike other thrombectomy devices, ablate the underlying plaque. The following multicenter registry was instituted to evaluate the safety and efficacy of laser ablation in patients presenting with acute myocardial infarction (AMI) complicated by persistent thrombotic occlusions. Patients with AMI and complete thrombotic occlusion of the infarct-related vessel were included in eight participating centers. Patients with further compromising conditions (i.e., cardiogenic shock, thrombolysis failures) were also included. Primary endpoint was procedural respective laser success; secondary combined endpoints were TIMI flow and % stenosis by quantitative coronary analysis and visual assessment at 1-month follow-up. Eighty-four percent of all patients enrolled (n = 56) had a very large thrombus burden (TIMI thrombus scale > or = 3), and 49% were compromised by complex clinical presentation, i.e., cardiogenic shock (21%), degenerated saphenous vein grafts (26%), or thrombolysis failures (5%). Laser success was achieved in 89%, angiographic success in 93%, and the overall procedural success rate was 86%. The angiographic prelaser total occlusion was reduced angiographically to 58% +/- 25% after laser treatment and to 4% +/- 13% final residual stenosis after adjunctive balloon angioplasty and/or stent placement. TIMI flow increased significantly from grade 0 to 2.7 +/- 0.5 following laser ablation (P < 0.001) and 3.0 +/- 0.2 upon completion of the angioplasty procedure (P > 0.001 vs. baseline). Distal embolizations occurred in 4%, no-reflow was observed in 2%, and perforations in 0.6% of cases. Laser-associated major dissections occurred in 4% of cases, and total MACE was 13%. The safety and efficacy of excimer laser for thrombus dissolution in a cohort of high-risk patients presenting with AMI and total thrombotic occlusion in the infarct-related vessel are encouraging and should lead to further investigation.     

Fluorescence-guided laser-assisted balloon angioplasty in patients with femoropopliteal occlusions

In 12 patients (aged 64 +/- 10 years) with femoropopliteal occlusions (1-27 cm; average, 8.4 cm length) that could not be recanalized by standard guidewire-balloon angioplasty techniques, percutaneous laser-assisted balloon angioplasty was performed by use of a new fluorescence-guided dual-laser system. Plaque detection by 325-nm laser-excited fluorescence spectroscopy provided real-time feedback control to a 480-nm pulsed dye laser (2-microseconds pulses) for atheroma ablation. By means of a common 200-microns optical fiber, after diagnostic fluorescence sensing, computer algorithms directed a fire or no-fire signal (5 Hz) to the treatment laser for selective plaque removal. Laser recanalization (15-50 mJ/pulse) was successful in 10 of 12 patients; this procedure was followed by definitive balloon angioplasty in seven of 12 patients with increased ankle/arm indexes (from 0.60 +/- 0.12 at baseline to 0.84 +/- 0.11 after treatment, p = 0.0043). In laser and balloon angioplasty failures, all femoropopliteal occlusions were heavily calcified, and there were two mechanical guidewire perforations without clinical sequelae. Ablation of calcified lesions required higher pulse energies and greater total energy per centimeter of recanalized tissue (1,837 +/- 1,251 mJ/cm vs. 90 +/- 39 mJ/cm, p = 0.0036). Fluorescence spectroscopy (n = 219 sites) was helpful in flush occlusions and correctly identified plaque, underlying media, and thrombus by changes in fluorescence intensity, shape, and peak position. Thus, when fluorescence-guided laser angioplasty was used in a subgroup of patients refractory to standard angioplasty techniques, primary recanalization and subsequent balloon angioplasty of femoropopliteal occlusions was successful in 83% and 58% of the patients, respectively. Importantly, treatment of heavily calcified lesions accounted for all of the failures and will require modified delivery systems to create larger primary channels and to increase catheter-tip control, which should improve clinical results in the future.     

Pulsed laser and thermal ablation of atherosclerotic plaque: morphometrically defined surface thrombogenicity in studies using an annular perfusion chamber.

Although clinical trials using laser and thermal angioplasty devices have been underway, the effects of pulsed laser and thermal ablation of atherosclerotic plaque on surface thrombogenicity are poorly understood. This study examined the changes in platelet adherence and thrombus formation on freshly harvested atherosclerotic aorta segments from Watanabe-heritable hyperlipidemic rabbits after ablation by two pulsed laser sources (308-nm xenon chloride excimer and 2,940-nm erbium:yttrium-aluminum-garnet [YAG] lasers) and a prototype catalytic hot-tip catheter. Specimens were placed in a modified Baumgartner annular chamber and perfused with citrated whole human blood, followed by quantitative morphometric analysis to determine the percent surface coverage by adherent platelets and thrombi in the treated and contiguous control areas. Pulsed excimer laser ablation of plaque did not change platelet adherence or thrombus formation in the treated versus control zones. However, photothermal plaque ablation with a pulsed erbium:YAG laser resulted in a 67% reduction in platelet adherence, compared with levels in control areas (from 16.7 +/- 2.2% to 5.5 +/- 1.8%; p less than 0.005). Similarly, after plaque ablation using a catalytic thermal angioplasty device, there was a 74% reduction in platelet adherence (from 29.2 +/- 5.1% to 7.7 +/- 1.6%; p less than 0.005) and a virtual absence of platelet thrombi (from 8.6 +/- 2.3% to 0.03 +/- 0.03%; p less than 0.005). This reduced surface thrombogenicity after plaque ablation with either an erbium:YAG laser or a catalytic hot-tip catheter suggests that thermal modifications in the arterial surface ultrastructure or thermal denaturation of surface proteins, or both, may be responsible for reduced platelet adherence. These in vitro findings indicate that controlled thermal plaque ablation by catheter-based techniques may elicit endovascular responses that can reduce early thrombus formation during angioplasty procedures.     

In vitro photodynamic diagnosis of atherosclerotic wall changes with the use of mono-l-aspartyl chlorin e6. A preliminary report

BACKGROUND: Although several methods for atherosclerosis detection are available, none of them seems to be accurate enough to identify the vulnerable atheromatous plaque. Photodynamic diagnosis (PDD) and therapy (PDT) - a new method evaluated for neoplasm treatment, is a modern approach for detecting and treating atherosclerosis. AIM: To asses in vitro the capability of PDD with the use of chlorin e6 to detect atherosclerotic plaque and the usefulness of this method as a feedback system for photoangioplasty treatment. METHODS: 30 specimens of human aorta and 15 specimens of human coronary arteries were examined. The samples were soaked with chlorin e6 and then washed out. The luminescence spectra were then collected. All samples were examined with light microscopy. RESULTS: Tissue fluorescence is seen as green light. We noted a very strong red fluorescence of chlorin e6 originating from lipid-rich plaque. We established a quantitative factor (R) which is the ratio of chlorin e6 red intensity in its 660 nm maximum to the area of green luminescence centred at 515 nm. The highest value of R was reached at the atheromatous samples, followed by calcified and normal ones R(2)=3.51+/-0.62, R(3)=1.63+/-0.31, and R(1)=1.51+/-0.15, respectively. A statistically significant difference was noted between groups two and one, and between groups two and three (R(2)=3.51+/-0.62 vs. R(3)=1.63+/-0.31, p<0.05; and R(2)=3.51+/-0.62 vs. R(1)=1.51+/-0.15, p<0.05, respectively). CONCLUSIONS: This in vitro study confirms that photosensitiser chlorin e6 accumulates within atheromatous plaque. It may be a specific tool for atheromatous and normal or calcified segments discrimination. The advantage of the above method is the possibility of a real-time imaging followed by targeted therapy of various forms and stages of atherosclerosis.     

Effect of stent implantation on upstream coronary artery compliance--a cause of late plaque rupture?

Stent implantation in the rabbit aorta has been shown to increase vessel wall compliance at the inflow to the stent, but it is uncertain whether similar effects might be seen in the coronary arteries of humans and whether this would have any significant clinical consequences. First, we measured vessel compliance (systolic lumen area--diastolic lumen area/pulse pressure) before, immediately after, and at the 6-month follow-up visit at a site 5 mm upstream of the proximal edge of an implanted coronary stent in patients undergoing coronary intervention using motorized pull-back intravascular ultrasound recordings. Compliance in the upstream segment increased significantly immediately after stenting (before 7.13 +/- 1.49 vs after 10.73 +/- 1.36 mm2/mm Hg, p = 0.03), an effect that was unchanged at 6 months of follow-up (11.84 +/- 2.11 mm2/mm Hg, p = 0.08 vs before stenting). Second, we examined the site of plaque rupture in all patients presenting with an acute coronary syndrome in whom the culprit lesion was in a vessel that had had a stent implanted >12 months previously (n = 31). Plaque rupture was statistically more likely at the inflow to the stent (n = 22) than at other sites within the culprit vessel (n = 9, p <0.01). We conclude that stenting causes an increase in vessel compliance immediately proximal to the stent, and that when a vessel has been previously stented, plaque rupture is most likely to occur at the stent inflow site.     

Vascular injury and time course of smooth muscle cell proliferation after experimental holmium laser angioplasty.

BACKGROUND. In vitro experiments have shown that holmium laser energy can effectively ablate even calcified plaque in human arterial vessels. Because high-energy densities from holmium lasers can easily be transmitted through quartz fibers, this solid-state laser has been suggested as an alternative intraluminal treatment of atherosclerotic plaque. METHODS AND RESULTS. To develop an intimal plaque, 35 New Zealand White rabbits underwent electrical stimulation of their right carotid artery for 28 days. Subsequently, in 25 rabbits, holmium laser angioplasty (wavelength, 2.12 microns; pulse duration, 150 microseconds; energy density, 350 mJ/mm2) was performed. To study the morphological results, the vessels were excised after 7, 14, 28, and 42 days. Cross sections were analyzed in regard to laser-specific injury. Staining of alpha-actin was used to identify smooth muscle cells (SMCs). After bromodeoxyuridine labeling, the extent of proliferation (number of cells undergoing DNA synthesis) was determined by using a monoclonal antibody. Holmium laser ablation resulted in an initial decrease of the numbers of intimal cell layers in the early group (7 days after treatment: 5 +/- 1 cell layers with 76 +/- 39 microns; control: 13 +/- 3 cell layers with 144 +/- 44 microns). Quantification of SMCs undergoing DNA synthesis in the intima (control: 51 +/- 19 cells/mm2) showed a significant increase of labeled cells after 7 (216 +/- 74 cells/mm2, p = 0.003) and 14 days (281 +/- 139 cells/mm2, p = 0.011). Integrity of the internal elastic lamina was disrupted in all animals after intervention. Seven and 14 days after treatment, a considerable reduction of medial cell nuclei was found in 10 of 12 animals. SMC proliferation in the medial layer was increased within the first 2 weeks after laser ablation (168 +/- 113 cells/mm2; control: 8 +/- 4 cells/mm2; p = 0.023). Six weeks after holmium laser angioplasty, SMC proliferation had returned to control levels in the intima and remained increased in the medial layer. This proliferative response resulted in a significant increase of intimal thickening within 6 weeks after laser ablation (30 +/- 6 cell layers, 375 +/- 97 microns resp.; p = 0.001 each). CONCLUSIONS. Holmium laser treatment leads to considerable vessel wall injury and results in SMC proliferation in the intimal and medial layer with a maximum of proliferative activity within the first 2 weeks. Subsequently, this results in considerable intimal and medial hyperplasia within 6 weeks after treatment.     

Biochemical basis for the difference between normal and atherosclerotic arterial fluorescence

The observation that laser-induced fluorescence (LIF) spectra of atherosclerotic and normal artery are different has been proposed as the basis for guiding a "smart" laser angioplasty system. The purpose of this study was to investigate the causes of this difference in LIF. Helium-cadmium laser-induced (325 nm) fluorescence was recorded from pure samples of known constituents of normal and atherosclerotic artery including collagen, elastin, calcium, cholesterol, and glycosaminoglycans. Similarities between the LIF spectra of atherosclerotic plaque and collagen and normal aorta and elastin were noted. LIF spectroscopy was then performed on specimens of atherosclerotic aortic plaque (n = 9) and normal aorta (n = 13) and on their extracted lipid, collagen, and elastin. Lipid extraction did not significantly alter atherosclerotic plaque or normal aortic LIF, suggesting a minor contribution of lipid to arterial LIF. The LIF spectra of normal aorta wall was similar to the spectra of the extracted elastin, whereas the LIF spectra of atherosclerotic aortic plaque was similar to the spectra of the extracted collagen. These observations are consistent with the reported relative collagen-to-elastin content ratio of 0.5 for normal arterial wall and 7.3 for atherosclerotic plaque. A classification algorithm was developed to discriminate normal and atherosclerotic aortic spectra based on an elastin and collagen spectral decomposition. A discriminant score was formed by the difference of elastin and collagen (E-C) coefficients and used to classify 182 aortic fluorescence spectra. The mean E-C value was +0.83 +/- 0.04 for normal and -0.48 +/- 0.07 for atherosclerotic aorta (p less than 0.001). Classification accuracy was 92%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of 17-beta estradiol on pre-existing atherosclerotic lesions: role of the endothelium.

The atheroprotective effects of estrogen during the process of atherogenesis is well documented, whereas limited information is available about the effect of estrogen on pre-existing atherosclerotic lesions. After bilateral ovariectomy, 24 New Zealand White rabbits were randomized into three groups of eight animals each and subsequently fed a 0.5% cholesterol diet. In group I, the vessels were excised at day 84, whereas in group II, the cholesterol diet was continued for a total of 168 days. In group III, the animals were first fed with a cholesterol diet for 84 days; in the second phase of the experiment, the cholesterol diet was continued for a further 84 days with a combined estrogen treatment (1 mg estradiol valerate per kg body weight per week intramuscularly). At the end of the experiment, the proximal aortic arch, right carotid artery, thoracical aorta and abdominal aorta of each animal were excised and prepared for histological and immunohistological examination. By day 168, morphometrical analysis displayed a significantly lower plaque development under estrogen therapy in the carotid artery (0.08+/-0.18 mm(2) vs. 0.60+/-0.39 mm(2)), the thoracic aorta (0.56+/-0.94 mm(2) vs. 3.63+/-2.06 mm(2)), and in the abdominal aorta (0.55+/-0.70 mm(2) vs. 1.71+/-1.05 mm(2)) in comparison with the corresponding 168 day control group. However, estrogen treatment has failed to reduce further atherosclerotic plaque development in the aortic arch (9.42+/-1.79 mm(2) vs. 11. 64+/-3.29 mm(2)). Immunohistological detection of the 'anti-human factor VIII related antigen', i.e. the 'von Willebrand factor' (vWF), showed a significantly lower number of luminal cells positive for vWF in the aortic arch in the 84-day cholesterol group, compared with the corresponding controls of normocholesterolemic rabbits (65. 9+/-12.4% vs. 83.1+/-6.2%; P<0.05). Estradiol was able to inhibit the further progression of atherosclerosis when moderate vessel wall alterations were present, whereas pre-existing severe atherosclerosis was associated with a failure of the anti-atherosclerotic estrogen action. As suggested by the in situ detection of vWF as a morphological marker for endothelial cells, an intact endothelial layer might play an important role in mediating the beneficial effect of estrogen in the process of atherosclerosis.