Ultrastructural Investigations of Arterial Bypass Conduits after the Use of Different Harvesting Techniques Using an Electron Microscope.

OBJECTIVES: The aim of this study was to perform morphometric analysis of arterial conduits harvested by harmonic scalpel in coronary artery bypass grafting (CABG) patients. METHODS: From 100 CABG patients, 200 arterial conduits-100 radial arteries (RAs) and 100 left internal thoracic artery (LITAs)-were harvested. The patients had similar characteristics (mean age, sex ratio, comorbidities, etc.). We divided the patients into 2 groups according to harvesting technique. In group 1, a harmonic scalpel was used in 50 patients for harvesting arterial conduits (50 LITA and 50 RA). In group 2, conduits were harvested using low-voltage electrocautery. To prevent side effects of clipping, all conduits in both groups remained in perfused condition until anastomosis. A 10-mm length of conduit was cut for transmission electron microscopy investigation. We calculated duration of harvesting, blood flow changes, and histopathologic changes of the conduits according to a vessel scoring system. RESULTS: In the harmonic scalpel group, we detected pathologic findings-corruption of endothelial integrity, subendothelial damage, and endothelial pathology-in 5 specimens (10%) (3 LITA  [6%] and 2 RA [4%]). In group 2, pathologic findings were detected in 16 conduits (32%; 11 LITA, 22%, and 5 RA, 10%). Endothelial dissection, subendothelial disarrangement, cellular separation due to intercellular edema, and subadventitial hematoma were the main pathologic changes in the classic harvesting method. There was a significant difference between the groups (P = .001). Harvesting time of LITA was nearly similar in both groups: 26.9 ± 11.1 min (range 25-38) in group 1 and 21.3 ± 8.6 min (range 21-25) in group 2 (P = .049). RA harvesting time was significantly shorter with the harmonic scalpel technique (20.3 ± 3.9 versus 27.6 ± 5.4 min, P = .022). The blood flow of the conduits was similar, with no statistical difference for the 2 arterial conduits (LITA, P = .76; RA, P = .55). CONCLUSION: In the learning curve period, the use of a harmonic scalpel is time consuming and presents some difficulties during the harvesting of conduits. According to our study results, however, the harmonic scalpel technique may be useful because of decreased pathology, including spasm. In our opinion, graft occlusion or thrombus as a life-threatening condition and endothelial dysfunction may decrease with the use of this alternative harvesting technique.

Three-year efficacy and patency follow-up of decellularized human internal mammary artery as a novel vascular graft in animal models.

BACKGROUND: Various investigations have reported that the internal mammary artery (IMA) is an efficient and functional choice of conduit for vascular graft surgeries, especially for coronary artery bypass grafts; however, the quest to find an ideal vascular substitute remains. We hypothesized that acellular IMA could be an appropriate graft for small-diameter vascular bypasses that could be used in various surgeries including coronary artery bypass grafting. METHODS: We decellularized human IMAs and performed histologic evaluations and scanning electron microscopy to confirm the decellularization process and the preservation of the extracellular matrix. Subsequently, we grafted the scaffolds into the superficial femoral arteries of 8 New Zealand rabbits with an end-to-end anastomosis. Computed tomography angiograms were provided at 3, 12, and 36 months postoperatively. Subsequently, the animals were killed, and biopsies were taken for histologic and immunohistochemical assessments. RESULTS: Evaluation of the acellular tissue confirmed the efficacy of the decellularization protocol and the preservation of the extracellular matrix. All 8 animals survived the entire follow-up period. Doppler ultrasonography and computed tomography angiographies verified the conduit's patency. Histologic assessments depicted the recellularization of all 3 layers of the scaffold. Smooth muscle cells were detected in tunica media. Immunohistochemical assessments confirmed these findings. CONCLUSIONS: In conclusion, we demonstrated that acellular human IMA could be used as an efficient small-diameter vascular substitute with high patency. These findings could pave the path for future investigations on the clinical application of acellular IMA as a novel vascular graft for small-diameter bypass surgeries.

Ultrastructural and histomorphologic properties of the internal thoracic artery: implications for coronary revascularization.

Coronary artery disease represents a major health problem worldwide for which coronary artery bypass surgery remains a standard of care. Among the several grafts that are available, the internal thoracic artery (ITA) has long been considered the best as several advantages have been described compared with other vessels (e.g. saphenous vein or radial artery), namely, an absent to minor atherosclerotic development. In fact, several studies showed the presence of preatherosclerotic lesions, such as intimal and/or medial thickening, medial fibrosis, among others, in the presence of certain cardiovascular risk factors as well as established atherosclerotic lesions (i.e. type II or more lesions). This paper primarily aimed at reviewing the current knowledge on the histomorphological characteristics of ITA as well as the comparative histomorphology of ITA with other vessel grafts currently in use in coronary surgery. As some of the evidence is not clear or consensual, this paper also aimed at reviewing the main histopathological, histomorphometrical, and ultrastructural findings in ITAs from patients with known cardiovascular risk factors (e.g. aging, obesity, hypertension, diabetes, smoking, and others). As the presence of preatherosclerotic and/or atherosclerotic lesions may compromise the success of the myocardial revascularization and lead to graft failure, contributing toward the associated morbidity and/or mortality, it is essential to improve the scientific knowledge on the structural characterization of ITAs and its correlation with the cardiovascular risk profile.

Caveolin 2: a facultative marker of unfavourable prognosis in long-term patency rate of internal thoracic artery grafts used in coronary artery bypass grafting. Preliminary report.

OBJECTIVES: Intimal hyperplasia leading to graft failure in patients undergoing coronary artery bypass grafting (CABG) is related to vascular smooth muscle cells (SMCs) proliferation. SMCs respond to a variety of mediators, the most important of which is platelet-derived growth factor (PDGF). The platelet-derived growth factor-induced cellular response has been shown to be mediated by caveolins. The aim of this study was to analyze CAV1-3 expression in internal thoracic artery (ITA) grafts used in CABG and correlate their expression with graft occlusion. METHODS: Six hundred patients undergoing CABG with the use of ITA grafts between 2008 and 2014 were enrolled into this prospective study. CAV1-3 expression in the ITA grafts was analyzed prior to graft transplantation into the coronary circulation via immunohistochemistry. Estimated caveolins expression pattern was then correlated with the occurrence of ITA graft failure observed within 24-months of surgery. RESULTS: Thirty-four patients developed ITA graft failure (subgroup A) and 566 study participants presented no adverse events (subgroup B). CAV1 and CAV3 expression levels in SMCs of the tunica media of the ITA grafts did not differ between the study subgroups and were not associated with the risk of graft failure. CAV2 was expressed within SMCs of the ITA grafts in 94.1% of the patients from subgroup A and 2.5% from subgroup B, and its expression was associated with ITA graft occlusion observed within 24-months after CABG. CONCLUSIONS: CAV2 expression in SMCs of the tunica media in autologous ITA transplants might indicate the risk of graft failure.

Histologic comparison between the internal mammary artery and the deep inferior epigastric artery and clinical implications for microsurgical breast reconstruction.

BACKGROUND: The internal mammary artery (IMA) is one of the most popular recipients for microsurgical breast reconstruction. However, it is often separated into sleeve-like layers when it is handled. This study tried to explain this unique behaviour of the IMA through histologic observation. METHODS: Nine pairs of IMAs and DIEAs were harvested and subject for haematoxylin-eosin and Verhoeff's elastic staining. Thickness of the tunica media and the number of elastic lamellae were compared. Samples of the IMA, the DIEA, and the thoracodorsal artery from another patient were observed through the transmission electron microscope to further show the structural differences. RESULTS: The most notable difference was presence of multiple elastic lamellae in tunica media in the IMAs, which was barely present in the DIEAs. The mean number of elastic lamellae was 9.2 in the IMA group and 1.0 in the DIEA group (p < 10(-9)). A transmission electron microscope showed that the tunica media of the DIEA and the TDA was densely packed with smooth muscle cells, while the muscle cells distributed sparsely in the IMA. CONCLUSIONS: The IMA is an elastic artery which is characterised by multiple layers of elastic lamellae while relatively lacking in smooth muscle cells. The wall of the IMA is easily dissected between the tunica media and the adventitia, or at the outer 1/3 of the tunica media. The inner structure is easily torn if microsutures do not engage the tunica adventitia.

Effect of electrocautery on endothelial integrity of the internal thoracic artery: ultrastructural analysis with transmission electron microscopy.

The internal thoracic artery (ITA) is typically harvested from the chest wall by means of conventional electrocautery. We investigated the effects of electrocautery on endothelial-cell and vessel-wall morphology at the ultrastructural level during ITA harvesting. Internal thoracic artery specimens from 20 patients who underwent elective coronary artery bypass grafting were investigated in 2 groups. The ITA grafts were sharply dissected with use of a scalpel and clips in the control group (n=10) and were harvested by means of electrocautery in the study group (n=10). Each sample was evaluated for intimal, elastic-tissue, muscular-layer, and adventitial changes. Free flow was measured intraoperatively. Light microscopic examinations were performed after hematoxylin-eosin and Masson's trichrome staining. Transmission electron microscopy was used to evaluate ultrastructural changes in the endothelial cells and vessel walls of each ITA. In the sharp-dissection group, the endothelial surfaces were lined with normal amounts of original endothelium, endothelial cells were distinctly attached to the basal lamina, cytoplasmic organelles were evident, and intercellular junctional complexes were intact. Conversely, in the electrocautery group, the morphologic integrity of endothelial cells was distorted, with some cell separations and splits, contracted cells, numerous large cytoplasmic vacuoles, and no visible cytoplasmic organelles. The subendothelial layer exhibited disintegration. Free ITA flow was higher in the sharp-dissection group (P=0.04). The integrity of endothelial cells can be better preserved when the ITA is mobilized by means of sharp dissection, rather than solely by electrocautery; we recommend a combined approach.

Internal mammary artery atherosclerosis: an ultrastructural study of two cases.

Atherosclerosis of the internal mammary artery (IMA) is generally regarded as a rare (but existent) pathological entity with only a few cases reported in the most recent literature. The only study which to our knowledge has investigated the ultrastructural features of IMA atherosclerosis, demonstrate the presence of endothelial cells loss, defects of internal elastic lamina with no evidence of lipid accumulation. In the present study, we describe two cases of IMA atherosclerosis in which ultrastructural analysis revealed the presence of a typical atherosclerotic plaque morphology with infiltration of inflammatory cells, formation of intraplaque lipid pools, and accumulation of lipid-laden foam cells throughout the thickened intima, never described in this rare lesion before. Microscopically, the lesions were also characterized by intimal thickening, invagination of endothelial cells, migration of smooth muscle cells with splitting, fenestration and/or fragmentation of the elastic sheets. Our observations add new data to the scarce and contradictory literature and to this largely understudied vascular disorder.

Electron microscopic evaluation of internal thoracic artery endothelial morphology in diabetic coronary bypass patients.

BACKGROUND: Diabetes is a well- identified major risk factor for cardiovascular diseases. This study was performed to evaluate the effect of diabetes and impact of glycemic control on internal thoracic artery (ITA) morphology by electron microscopy. METHODS: Thirty patients scheduled for coronary artery bypass grafting were enrolled in this study. Samples of ITA were taken during the surgery for electron microscopic evaluation. Group I (n = 10) consisted of diabetics who have poor glycemic control (HbA1c > 7.5%), group II (n = 10) of well-regulated (HbA1c = 4.4% to 6.2%) diabetic patients, and group III (n = 10) of nondiabetic patients. Samples were prepared as ultrathin sections and an original semiquantitative method of scoring was applied to describe the morphologic changes of endothelium. Final scores were analyzed with analysis of variance and post hoc analysis. RESULTS: In group I large vacuoles, swollen mitochondria were seen in endothelial cells and subendothelial edema was prominent. Endothelia (2.5 ± 1.2), arterial wall (2.0 ± 0.0), and endothelial mitochondria (2.9 ± 1.3) scores of group I were significantly higher than the other 2 groups (p < 0.001). The samples of group II and group III did not show significant differences with each other. The correlation between HbA1c values and total endothelial scores statistically significant (r = 0.912; p < 0.001). CONCLUSIONS: There is a correlation between HbA1c values and morphologic changes of ITA graft. Uncontrolled diabetes is an important predictor of morphologic changes evidenced by the ultrastructural findings. These ultrastructural changes were not as prominent in the diabetes mellitus patients with well controlled metabolic statuses and patients without diabetes.

[Intraoperative assessment of composite bypass grafts by means of ultrasonic Doppler flowmetry].

The use of the both internal thoracic arteries while forming a composite bypass graft has been suggested as a method making it possible to carry out total myocardial revascularization. However, controversial still remain the problem of the distribution of blood in these bypass grafts, as well as that concerning T-grafts' capability of providing the sufficient blood inflow to the basin of the coronary arteries being bypassed. The method of ultrasonic Doppler flowmetry was used to perform the intraoperative assessment of 59 composite bypass grafts along such parameters as the pulsatility index (PI), mean volume blood flow velocity (Qmean), and percentage of diastolic volumetric filling (DF). The obtained findings were statistically processed making it possible to obtain the average parameters of blood flow for the composite bypass grafts and their branches. A dependence was established between blood flow distribution in the composite conduit and the degree of the lesion of the artery being bypassed.

Reduced negative surface charge on arterial endothelium explains accelerated atherosclerosis in type 2 diabetic patients.

Reduced endothelial surface charge markedly increases the rate of LDL uptake into blood vessels. Previous work in the streptozotocin diabetic rat reported reduced endothelial surface charge. We compared endothelial surface charge density in internal mammary artery rings from patients with type 2 diabetes (n = 12) and from non diabetic patients undergoing coronary artery bypass grafting, and observed a substantial (52%) reduction in the former. This was associated with higher plasma sialic acid levels suggesting loss of sialic acid residues from the glycocalyx as a possible mechanism.

Internal mammary perforator recipient vessels for breast reconstruction using free TRAM, DIEP, and SIEA flaps.

BACKGROUND: Use of internal mammary perforator recipient vessels may decrease recipient-site morbidity in free flap breast reconstruction. However, it is not known whether they are as reliable as the internal mammary vessels or whether their use would result in increased recipient-site complications. METHODS: A retrospective study of all 686 lower abdominal free flap breast reconstructions performed during a 4-year period at the authors' institution identified 38 cases in which internal mammary perforator vessels were used. A subset of 114 of the 686 cases was studied in which patients were treated by a single surgeon, internal mammary perforator vessel use was attempted, and vessel measurements were taken intraoperatively. RESULTS: Internal mammary perforator recipient vessels were used in 27 percent of cases attempted, 94 percent of which were located at the second or third intercostal space. Internal mammary perforator arteries were significantly smaller in diameter (1.9 mm) than internal mammary arteries (2.6 mm) (p < 0.0001). Despite this, the incidences of flap loss, fat necrosis, and mastectomy skin flap necrosis were not significantly different between the internal mammary and internal mammary perforator vessel groups. The internal mammary and internal mammary perforator veins were similar in diameter. CONCLUSIONS: To the authors' knowledge, this is the largest published series of internal mammary perforator vessels used as free flap recipient vessels. In selected cases, internal mammary perforator vessels can be used without increasing the risk of flap loss, fat necrosis, or mastectomy skin flap necrosis, and are equally reliable and require less invasive surgical dissection than internal mammary vessels. The use of internal mammary perforator vessels may safely minimize recipient-site morbidity.

Direct stenting limits sirolimus-eluting stent edge neointimal thickening.

OBJECTIVE: The use of sirolimus eluting stent (SES) has strongly limited the incidence of in-stent restenosis that still remains a problem at the stent edge. The aim of this study was to analyze the neointimal thickening after implantation of SES and to assess the influence of the stent implantation procedure on the neointimal thickening in the in-stent segment and at the edge of the stent in an ex-vivo model of stented human artery. METHODS: Both balloon expandable SES and the corresponding bare metal stent (BMS) were used in a model of human mammary artery culture. Stents were implanted either directly or after predilatation (10 atm, 60 seconds) and analysis of arterial segments were performed at 28 days poststenting. Cell proliferation and neointimal thickening were assessed by immunohistochemistry, western blotting, and histomorphometry, both in the in-stent segment and at the edge of the stent. Neointimal thickening was expressed as the ratio ([neointimal area/neointimal area + media area]). RESULTS: The in-stent neointimal thickening was dramatically inhibited in the SES group compared with the BMS group whatever the stenting technique was (predilatation: 0.22 +/- 0.05 vs 0.30 +/- 0.10; P < .04; direct stenting 0.16 +/- 0.04 vs 0.30 +/- 0.13; P <.01). This effect of SES was associated with a smallest expression of the small G protein RhoA and an increase of p27kip expression. In the BMS group, predilatation and direct stenting gave similar in-stent neointimal thickening. In contrast, in the SES group, in-stent neointimal thickening was significantly reduced when direct stenting was performed (0.16 +/- 0.04 [direct stenting] vs 0.22 +/- 0.05 [predilatation], P < .03). At the stent edge, a similar neointimal thickening was observed with both type of stent when predilatation was performed on the entire segment of the artery. Direct stenting significantly reduced the neointimal thickness at the stent edge when SES where used (0.06 +/- 0.01 [direct stenting] vs 0.19 +/- 0.06 [predilatation]; P < .001) but not in the BMS group. CONCLUSION: These results confirm the efficiency of sirolimus released form SES to inhibit RhoA expression and to increase p27kip level in the arterial wall and show the benefit of direct stenting to limit the edge effect with SES.

Telomere dynamics in arteries and mononuclear cells of diabetic patients: effect of diabetes and of glycemic control.

Telomeres serve as a mitotic clock and biological marker of senescence. Diabetes mellitus (DM) is associated with damage to target organs and premature aging. We assessed the effect of glycemic control on telomere dynamics in arterial cells of 58 patients undergoing coronary artery bypass and in mononuclear blood cells of other diabetic (32 type I and 47 type II) patients comparing well controlled to uncontrolled patients. All were compared to age-dependent curve of healthy controls. Telomeres were significantly shorter in the arteries of diabetic versus non-diabetic patients (p=0.049) and in mononuclear cells of both type I and type II diabetes. In all study groups good glycemic control attenuated shortening of the telomeres. In arterial cells good glycemic control attenuated, but not abolished, the telomere shortening. In type II DM the mononuclear telomere attrition was completely prevented by adequate glycemic control. Telomere shortening in mononuclear cells of type I diabetic patients was attenuated but not prevented by good glycemic control. Results of this study suggest that diabetes is associated with premature cellular senescence which can be prevented by good glycemic control in type II DM and reduced in type I DM.

Comparison of the pretreatment effects of mixed vasodilators (3-D solution) on radial and internal thoracic arteries by using a 3-dimensional anaglyph electron microscope technique.

OBJECTIVE: Currently, internal thoracic arteries (ITA) and radial arteries (RA) are the first choice of conduits for coronary artery bypass grafts (CABG). Because the perioperative vasospasm continues to be a major problem, a number of pharmacologic agents such as papaverine, calcium receptor blockers, nitroglycerine, and phenoxybenzamine have been suggested as topical antispasmodics that may be used in the pre- and postoperative periods. In the present study, we investigated the quantitative efficacies of the mixed solution, which included verapamil, nitroglycerin, and papaverine, on RA and ITA using a scanning electron microscope with a 3-dimensional anaglyph technique. METHODS: Diameter changes of RA and ITA in response to clinically important vasodilators were measured on 40 RA and 40 ITA rings from patients who had been subjected to coronary artery bypass procedure after 20 minutes of ex vivo incubation with verapamil (45 microg/L), nitroglycerin (45 microg/L), papaverine solution (266 micromol/L or 0.1 micro/mL), and 30 mL autologous heparinized whole blood (individual patient's blood obtained before cardiopulmonary bypass contained 100 IU of heparin per kg of patient weight). The pretreatment action was assessed by measuring the response to vasodilators. RESULTS: In all cases, we did not observe graft vasospasm in any of the conduits during the intraoperative period between postanastomosis and sternal closure. In the postoperative period, we did not record any evidence of ischemic change in patients' electrocardiographic and myocardial enzyme analyses. None of the cases required inotropic support after the operation. The diameters of the pretreated RA and ITA were: minimum, 2.1 mm; maximum, 4.0 mm; and mean value, 2.80 +/- 0.46 mm. The diameters of the pretreated ITA were: minimum, 1.2 mm; maximum, 2.5 mm; and mean value, 1.76 +/- 0.35 mm. Incubated arterial segment diameters for the RA were: minimum, 2.8 mm; maximum, 5.2 mm; and mean value, 3.95 +/- 0.65 mm. These values for the ITA were: minimum, 1.5 mm; maximum, 3.9 mm; and mean value, 2.37 +/- 0.50 mm. These findings were statistically significant for both arterial grafts (P <.05). CONCLUSIONS: According to our study findings, the mixed solution demonstrates a broad range of efficacy. We conclude that the described vasodilator solution with heparinized autologous blood seems to be very effective and may be used as a pretreatment agent in CABG conduits. Although papaverine has the shortest duration of action, its efficiency is increased by verapamil and nitroglycerin, in our opinion. To the best of our knowledge, high-quality imaging of CABG conduits with the 3-D anaglyph technique using a scanning electron microscope was a first in the literature. This technical approach may be used for confirming the ultrastructural anatomy and the quantitative vasodilator effects of arterial conduits. We believe that valuable anatomo-pathologic details of the CABG conduit can be obtained by this technique.

[Comparative study on the ultrastructures of radial and internal mammary arteries used for coronary artery bypass grafting].

OBJECTIVE: The radial artery differs from internal mammary artery in its vascular biology and long-term patency after coronary artery bypass grafting (CABG). This study was designed to investigate their ultrastructural differences that may have implications in arterial remodeling and graft failure. METHODS: Thirty-four radial artery and 11 internal mammary artery samples were obtained from patients underwent CABG, and subjected to routine electron microscopic examination. A semi-quantitative method was used to evaluate secretary endothelial cells, endothelial denudation, synthetic smooth muscle cells (SMCs), matrix accumulation, lipid deposition and medial submicroscopic calcification. RESULTS: Compared with internal mammary arteries, the radial arteries had more secretory endothelial cells (47.1%, 16/34 vs 27.2%, 3/ 11) and synthetic type SMCs in a background (14.4% vs 0.9%) that had more intimal lipid deposition and matrix accumulation (14.7%, 5/34 vs 9.1%, 1/11). Matrix vesicles and calcifications were frequently present in the media of both types of arteries. The calcifications, however, could not be visualized by routine histological stains, and therefore, named as submicroscopic calcification in this study. Fewer endothelial denudations were observed in the radial arteries, but no differences in medial lipid deposition and submicroscopic calcification were observed between these two types of arteries. The ultrastructural features and the arrangement of medial SMCs in radial arteries were similar to those of internal mammary arteries. CONCLUSIONS: Radial arteries have a higher SMC proliferative potential and more actively secretory status of endothelial cells, which may enhance the remodeling process and correlate with a decreased long-term patency. Better preservation of endothelial cells in radial arteries could be attributed to the "no touch" technique utilized in surgical harvesting. The significance of submicroscopic medial calcification during graft remodeling requires further investigations.

Implantation in coronary circulation induces morphofunctional transformation of radial grafts from muscular to elastomuscular.

BACKGROUND: The purpose of this research was to investigate the in vivo morphofunctional changes induced in the radial artery (RA) by its use as coronary artery bypass conduit by comparing the morphological features and vasoreactivity of the native RA versus the coronary RA graft in the same patient. METHODS AND RESULTS: Ten years after surgery, 10 patients were submitted to intravascular ultrasound examination of the RA graft of the controlateral (in situ) RA and of the internal thoracic artery (ITA) graft and to vasoactive challenges with acetylcholine and serotonin. Quantitative angiographic assessment showed that the mean diameter of the RA coronary grafts was significantly larger than that of the in situ RA and of the ITA (2.89+/-0.40 mm RA grafts, 2.14+/-0.52 mm in situ RA, 2.25+/-0.53 mm ITA grafts; P<0.001). The in situ RA demonstrated a typical muscular architecture, whereas RA coronary grafts showed a clear reduction of the thickness of the medial layer and had a less well-defined muscular component of the media with interposition of elastic tissue. Serotonin endovascular infusion elicited a strong spastic reaction in in situ RAs; the same challenge induced only moderate constriction in RA and ITA coronary grafts. CONCLUSIONS: Implantation in the coronary circulation leads to major anatomic and vasoreactive modifications of the RAs that tend to lose the morphofunctional features of a muscular conduit and assume those of an elastomuscular artery, such as the ITA.

Molecular imaging of atherosclerotic plaques using a human antibody against the extra-domain B of fibronectin.

Current imaging modalities of human atherosclerosis, such as angiography, ultrasound, and computed tomography, visualize plaque morphology. However, methods that provide insight into plaque biology using molecular tools are still insufficient. The extra-domain B (ED-B) is inserted into the fibronectin molecule by alternative splicing during angiogenesis and tissue remodeling but is virtually undetectable in normal adult tissues. Angiogenesis and tissue repair are also hallmarks of advanced plaques. For imaging atherosclerotic plaques, the human antibody L19 (specific against ED-B) and a negative control antibody were labeled with radioiodine or infrared fluorophores and injected intravenously into atherosclerotic apolipoprotein E-null (ApoE-/-) or normal wild-type mice. Aortas isolated 4 hours, 24 hours, and 3 days after injection exhibited a selective and stable uptake of L19 when using radiographic or fluorescent imaging. L19 binding was confined to the plaques as assessed by fat staining. Comparisons between fat staining and autoradiographies 24 hours after 125I-labeled L19 revealed a significant correlation (r=0.89; P<0.0001). Minimal antibody uptake was observed in normal vessels from wild-type mice receiving the L19 antibody and in atherosclerotic vessels from ApoE-/- mice receiving the negative control antibody. Immunohistochemical studies revealed increased expression of ED-B not only in murine but also in human plaques, in which it was found predominantly around vasa vasorum and plaque matrix. In summary, we demonstrate selective targeting of atheromas in mice using the human antibody to the ED-B domain of fibronectin. Thus, our findings may set the stage for antibody-based molecular imaging of atherosclerotic plaques in the intact organism.

Endothelial integrity of ultrasonically skeletonized internal thoracic artery: morphological analysis with scanning electron microscopy.

OBJECTIVE: The skeletonized internal thoracic artery (ITA) has several advantages over a pedicled one in coronary artery bypass grafting (CABG). An ultrasonic scalpel makes ITA skeletonization easy and speedy, however, the ultrasonic energy that is transmitted to the artery itself can occasionally injure the endothelium. Therefore, the endothelial integrity of the ultrasonically skeletonized ITA is a major concern related to this technique. The purpose of this study is to assess the endothelial integrity of the ultrasonically skeletonized ITA. METHODS: We skeletonized the left ITA with an ultrasonic scalpel in nine patients who underwent CABG, and thereafter the terminal portion of this artery was subjected to a morphological study. The endothelial integrity of this artery was morphologically assessed using scanning electron microscopy, and the results were compared to that of the left ITA skeletonized with fine scissors. RESULTS: All ITA specimens showed a completely confluent endothelium, and no endothelial injury was observed by the scanning electron microscopic study. CONCLUSIONS: The skeletonization of the ITA with an ultrasonic scalpel had no deleterious effect on the endothelium. This morphological study confirmed the safety and the reliability of this technique, and we therefore recommend its clinical use in the skeletonization of the ITA for CABG.

Reduced high shear platelet adhesion to the vascular media: defective von Willebrand factor binding to the interstitial collagen.

Fibrillar collagen serves as a thrombogenic surface for platelet adhesion mediated by von Willebrand factor (vWf) at high shear. Although abundant throughout the arterial wall, vWf-dependent platelet deposition to artery cross-sections from perfused citrated blood is localized to the adventitia of the vessel wall. Here we describe a similarly skewed distribution of vWf-binding sites in artery cross-sections. Binding of vWf-coated fluorescent beads, as well as detection of plasma vWf bound to artery cross-section at 3350 s(-1) shear rate with indirect particle-immunofluorescence or immunoelectron microscopy demonstrate vWf binding sites in the adventitia, but not in the media. A monoclonal anti-vWf antibody that interferes with vWf-binding to collagen in a microplate ELISA inhibits vWf-binding to both the adventitia and sections of collagen fibrils. Our data suggest that the media, despite its fibrillar collagen content, evidenced by electron microscopy, is defective for vWf-binding, which may explain its thromboresistant nature at high shear rates.

Histologic and physiologic evaluation of skeletonized internal thoracic artery harvesting with an ultrasonic scalpel.

OBJECTIVES: The safety and reliability of a method of skeletonized internal thoracic artery harvesting with an ultrasonic scalpel (Harmonic Scalpel; Ethicon Endo-Surgery, CVG, Cincinnati, Ohio) were evaluated. METHODS: The mural branches of the internal thoracic artery were cut by means of 3 methods, differentiated by distance from the site of application of the Harmonic Scalpel blade to the internal thoracic artery. A total of 15 branches were cut from the internal thoracic artery at (0 mm) the origin (group I) or at 1 mm (group II) or 2 mm (group III) distal to the origin. Tissue preparations were examined for successful vessel closure and severity of tissue damage. The length of stump (L) and the length of tissue damage from the stump (D) were determined by a computer image analysis system, and pressure testing was performed to evaluate the physical strength of vessel closure. RESULTS: In group I, 8 of the 15 branches exhibited discontinuity of the vascular wall structure, probably because of insufficient sealing of the divided section, and 12 of the 15 branches exhibited tissue denaturation on the internal thoracic artery wall adjacent to areas of origin, which was probably caused by the heat transferred from the branches during the process of coagulation. In groups II and III, continuity of wall structure of stumps suggestive of stable closure of branches was confirmed. The lengths of tissue damage from the stump (D) were 0.96, 0.58, and 0.63 mm in groups I, II, and III, respectively, and the lengths of intact area (L - D) in the corresponding groups were -0.78, 0.61, and 1.51 mm. The negative figure in group I indicates the presence of tissue damage in the internal thoracic artery itself. By contrast, in groups II and III the internal thoracic arteries were intact, with a safety margin of greater than 0.5 mm. On physiologic evaluation of vessel closure, 2 of the 24 (8.3%) branches burst under a pressure lower than 350 mm Hg because of insufficient vessel coagulation, but the remaining 22 branches (91.7%) remained intact under pressures up to 350 mm Hg. CONCLUSION: The internal thoracic artery skeletonization method with an ultrasonic scalpel (Harmonic Scalpel: output level 2) appears to be a safe and reliable method of skeletonized internal thoracic artery harvesting when branches are sectioned at least 1 mm distal to their origin at a sufficiently slow speed.