Incomplete Re-endothelialization Research Articles

Features of re-endothelialization process during implantation of coronary stents (review of literature)

Method of endovascular treatment for coronary artery lesions has made its way from balloon angioplasty to implantation of drug-eluting stents. Today, drug-eluting stents are widely used for coronary artery revascularization and have significantly reduced the incidence of restenosis compared to bare metal stents. Delayed or incomplete re-endothelialization is a very specific problem for most drug-eluting stents, as after their implantation, there is a risk of developing late thrombosis associated not only with incomplete endothelialization of the stent struts or impaired endothelial function, but also with chronic inflammatory reaction of the vascular wall. The development of stents with ultrathin struts, the abluminal coating of the struts with polymer and drug contribute to the decrease of vascular wall trauma and of the inflammatory response and provide faster stent endothelialization. Keywords: coronary heart disease, coronary stents, re-endothelialization, microporous coating, polymer-free antiproliferative coating, Yukon stent

P3386Angioscopic comparison between polymer-free biolimus A9-coated stent and durable polymer drug-eluting stent 10 months after the implantation

Abstract Background Polymer-free biolimus A9-coated stent (DCS) has currently emerged as expected better arterial healing compared to durable polymer drug-eluting stent (DP-DES). However, superiority of DCS on arterial healing over DP-DES has not been well elucidated using intracoronary images. Methods This study examined 288 stents in 224 patients with de novo coronary artery lesions. We angioscopically compared 55 DCS from 35 patients with 233 DP-DES from 189 patients 10±2 months after the implantation. We assessed thrombus adhesion, which is a marker of incomplete re-endothelialization. Dominant neointimal coverage (NIC) grade, heterogeneity of NIC and maximum yellow color of plaque underneath the stent were also evaluated. Neointimal coverage was graded as follows: grade 0, stent struts exposed; grade 1, struts bulged into the lumen, although covered; grade 2, struts embedded by the neointima, but translucent; grade 3, struts fully embedded and invisible. NIC was judged as heterogeneous when differences in the NIC grade became apparent. Yellow plaque was graded as follows: grade 0, white; grade 1, light yellow; grade 2, yellow; grade 3, intensive yellow. Results Thrombus adhesion was similar between DCS and DP-DES (29% versus 23%, P=0.32). Dominant NIC was greater in DCS than in BP-DES (P<0.001), while NIC was more heterogeneous in DCS than in BP-DES (P=0.001, Figure). Maximum yellow color of stented segment was similar between DCS and DP-DES (P=0.09). Conclusion DCS provided similar thrombus adhesion to DP-DES, which suggested similar re-endothelialization 10 months after implantation. However, DCS showed thick and heterogeneous NIC compared to DP-DES. The specific feature of polymer-free and Biolimus A9 would cause the difference, and further investigation is necessary to evaluate the longer-term safety and efficacy. Acknowledgement/Funding None

Dimethylfumarate attenuates restenosis after acute vascular injury by cell-specific and Nrf2-dependent mechanisms

Excessive proliferation of vascular smooth muscle cells (VSMCs) and incomplete re-endothelialization is a major clinical problem limiting the long-term efficacy of percutaneous coronary angioplasty. We tested if dimethylfumarate (DMF), an anti-psoriasis drug, could inhibit abnormal vascular remodeling via NF−E2-related factor 2 (Nrf2)-NAD(P)H quinone oxidoreductase 1 (NQO1) activity. DMF significantly attenuated neointimal hyperplasia induced by balloon injury in rat carotid arteries via suppression of the G1 to S phase transition resulting from induction of p21 protein in VSMCs. Initially, DMF increased p21 protein stability through an enhancement in Nrf2 activity without an increase in p21 mRNA. Later on, DMF stimulated p21 mRNA expression through a process dependent on p53 activity. However, heme oxygenase-1 (HO-1) or NQO1 activity, well-known target genes induced by Nrf2, were dispensable for the DMF induction of p21 protein and the effect on the VSMC proliferation. Likewise, DMF protected endothelial cells from TNF-α-induced apoptosis and the dysfunction characterized by decreased eNOS expression. With knock-down of Nrf2 or NQO1, DMF failed to prevent TNF-α-induced cell apoptosis and decreased eNOS expression. Also, CD31 expression, an endothelial specific marker, was restored in vivo by DMF. In conclusion, DMF prevented abnormal proliferation in VSMCs by G1 cell cycle arrest via p21 upregulation driven by Nrf2 and p53 activity, and had a beneficial effect on TNF-α-induced apoptosis and dysfunction in endothelial cells through Nrf2–NQO1 activity suggesting that DMF might be a therapeutic drug for patients with vascular disease.

Serial Optical Coherence Tomography Assessment of Xience V and Nobori Stents at 1-and 6-months Follow-up

Delayed neointimal healing and incomplete reendothelialization has gained attention as a cause of late stent thrombosis with first-generation drug-eluting stents (DES). Newer-generation DES, such as Xience V (Abbott Vascular, Santa Clara, CA) and Nobori stents (Terumo Corporation, Tokyo, Japan), are expected to provide minimal inflammatory reaction and earlier neointimal healing compared with first-generation DES. However, the process of neointimal healing after newer-generation DES implantation has not been fully evaluated.

Very late stent thrombosis

Drug-eluting stents (DES) are effective in reducing neointimal proliferation and in-stent restenosis. However, the procedure is complicated by early and late stent thrombosis that is related to incomplete healing, leading to stent malapposition and incomplete reendothelialization. Stent restenosis results in significant mortality and morbidity and portends a poor long-term outcome. In this article, we review recent findings regarding the prevalence of late and very late stent thrombosis (VLST) and the mechanisms that may be at play. Long-term follow-up from large registry studies and randomized controlled trials of DES implantation have recently created an awareness of the persistence of VLST (0.26%/year) for up to at least 5 years. Recent findings utilizing intravascular ultrasound and optical coherence tomography have also provided important insights into the mechanism of VLST and suggest that delayed healing and neoatherosclerosis are important. Finally, the development of novel stent scaffolds and antiplatelet agents holds much promise for reducing the risk of VLST. DES implantation continues to be complicated by the risk of VLST. Recent insights into the mechanisms of this significant clinical problem have important implications in preventing VLST.

Endothelium-Dependent Vasomotor Dysfunction in Pig Coronary Arteries With Paclitaxel-Eluting Stents Is Associated With Inflammation and Oxidative Stress

We sought to evaluate coronary epicardial and intramyocardial resistance, arterial vasomotor function, local inflammatory reaction, and superoxide anion (O(2)(.-)) production after overlapping paclitaxel-eluting stent (PES) implantation in a porcine model. PES implantation has been shown to elicit coronary vasomotor dysfunction. However, underlying mechanisms remain largely unknown. Nine pigs received overlapping PES and bare-metal stents (BMS) in the coronary arteries, and 3 sham animals were naïve. At 1 month, inflammatory response at the overlapped region was assessed by histopathology and scanning electron microscopy. Endothelial vasomotor function and O(2)(*-) at nonstented coronary reference segments were measured by angiography and organ chamber tensiometry, and lucigenin luminometry; vasomotor function of distal resistance arteries was measured by myography. Paclitaxel-eluting stents showed reduced late lumen loss, but inflammation and luminal inflammatory cell adherence were higher than for BMS (p < 0.001) at overlapped segments. Endothelium-dependent relaxation to substance P was significantly impaired in PES at nonstented coronary reference segments (>or=15 mm proximally and distally) and perfusion bed resistance arteries (p < 0.05). In contrast, endothelium-independent relaxation to nitroglycerin and sodium-nitroprusside was similar between groups. Local O(2)(*-) production at both proximal and distal nonstented coronary reference segments was elevated for PES when compared with O(2)(*-) production in BMS and naïve arteries (p < 0.001). Abnormal endothelium-dependent relaxation at both coronary conduit and resistance arteries was demonstrated after overlapping PES implantation. Profound localized inflammatory reaction, as well as enhanced local oxidative stress, may contribute to vasomotor dysfunction.

C6-Ceramide-Coated Catheters Promote Re-Endothelialization of Stretch- Injured Arteries

OBJECTIVE: Drug eluting stents have recently been associated with the increased risk of adverse thrombogenic events and/or late luminal loss, which is highly associated with incomplete re-endothelialization. The increased risks behoove the design of alternative delivery modalities and/or drugs that do not compromise the re-endotheliaization process. The objective of the present study is to elucidate the biological mechanism(s) by which non-stent-based delivery modalities for the anti-proliferative lipid metabolite, C(6)-ceramide, could lead to a reduction in arterial injury after angioplasty. RESULTS: Immunohistochemical studies in rabbit and porcine models suggest that C(6)-ceramide-coated balloon catheters limit arterial stenosis without inhibiting endothelial wound healing responses. Specifically, C(6)-ceramide-coated balloon catheters reduce internal elastica injury with a corresponding reduction in medial fracture length in a 28-day porcine coronary artery stretch model. In addition, C(6)-ceramide decreases the formation of the fibrin matrix to possibly augment the subsequent wound healing response. We hypothesized that differential metabolism of exogenous ceramide by coronary endothelial and smooth muscle cells could explain the apparent discrepancy between the anti-proliferative actions of ceramide and the pro-wound healing responses of ceramide. Human coronary artery endothelial cells (HCAEC), in contrast to human coronary artery smooth muscle cells (HCASMC), preferentially express ceramide kinase and form ceramide-1-phosphate, which promotes endothelial cell survival. CONCLUSION: Differential metabolism of ceramide between HCASMC and HCAEC offers a mechanism by which ceramide preferentially limits smooth muscle cell growth, in the presence of active wound healing. The combinatorial ability of ceramide to limit vascular smooth muscle proliferation and promote re-endothelialization, offers the potential for C(6)-ceramide-coated catheters to serve as adjuncts to stent-based modalities or as a stand-alone treatment.

Late Coronary Stent Thrombosis

Introduced >20 years ago,1 coronary artery stents have improved the safety and particularly the efficacy of percutaneous coronary interventions (PCIs).2 Abrupt vessel closure, complicating 6% to 8% of balloon angioplasty procedures, was associated with a 5% mortality, 40% rate of myocardial infarction (MI), and 40% rate of emergency coronary artery bypass grafting.3 Stents significantly reduced these adverse events (Figure 1).4,5 The reduction of restenosis afforded by bare metal stents (BMS) was modest (30% to 40%). Repeat revascularization still occurred in 15% to 20% of cases.6 Drug-eluting stents (DES) with antiproliferative drugs attached via polymers on the stent surface to minimize smooth muscle proliferation have reduced restenosis and rates of target lesion revascularization by 50% to 70% compared with BMS across nearly all lesion and patient subsets.7 Initially8 and again more recently,9–16 safety concerns were raised about DES, particularly about late stent thrombosis (ST). Figure 1. Outcome of 112 patients with abrupt vessel closure in the prestent balloon angioplasty experience of a registry (1985 to 1986; total, 1801 patients)3 and 339 patients with abrupt vessel closure during balloon angioplasty treated with BMS (Palmaz-Schatz) (total, 4596 patients).5 CABG indicates coronary artery bypass grafting. ST was a bane of stent implantation from the beginning. The initial experience with Wallstents in the late 1980s was overshadowed by ST rates approaching 24%.17 Subsequent series with Palmaz-Schatz and Gianturco-Roubin stents (still predominantly bailout stenting) observed ST in 6% to 12% of cases.4,18 The postprocedural antithrombotic regimen at the time consisted of aspirin, often in conjunction with oral anticoagulation. Dual antiplatelet therapy of aspirin and the thienopyridine ticlopidine in conjunction with a shift from bailout to elective stenting resulted in a significant reduction of ST to <2%.5,19 Earlier oral antiplatelet drug loading and …

High-dose external beam irradiation inhibits neointima formation in stented pig coronary arteries

Purpose: To evaluate high-dose external beam irradiation (EBRT) in a pig coronary stent preparation because low and intermediate-dose EBRT failed to show inhibition of neointima formation in stented animal models.Methods and Materials: Thirty-five stents were implanted in the coronary arteries of 17 pigs. Seven pigs were exposed to a single dose of 21 Gy EBRT immediately after stenting. Ten stented, nonirradiated pigs served as controls. After 4 weeks, the study arteries and myocardium were examined by light and scanning electron microscopy.Results: Compared with controls, 21 Gy EBRT resulted in a larger lumen area (7.57 ± 1.67 mm2 vs. 4.00 ± 1.63 mm2, p <0.001), a smaller neointima area (0.47 ± 0.43 mm2 vs. 3.36 ± 2.26 mm2, p <0.001) and a smaller maximal intimal thickness (0.16 ± 0.09 mm vs. 0.68 ± 0.31 mm, p <0.001). Unresorbed intramural hemorrhages and adherent mural thrombi were present in the irradiated vessels, which also showed incomplete re-endothelialization. The irradiated hearts demonstrated diffuse interstitial and perivascular inflammation and fibrosis.Conclusions: EBRT at 21 Gy to the entire heart significantly inhibited neointima formation in stented pig coronary arteries but also resulted in incomplete re-endothelialization, myocardial inflammation, and fibrosis. Improvements in localization and delivery techniques are required to allow clinical implementation of this technique.

Delayed re-endothelialization and T-cell infiltration following intracoronary radiation therapy in the porcine model

Purpose: To evaluate the late induction of apoptosis following intracoronary radiation (IR) and the effects of IR on inflammatory cells. Methods and Materials: Porcine coronaries were injured by balloon overstretch followed by either 0 or 15 Gy of 192Ir prescribed to 2 mm from the center of the source. Swine were euthanized at 3, 7, and 14 days posttreatment, and arteries were stained for markers of smooth muscle cells (SMCs α-actin), T cells (CD3), macrophages, endothelial cells, and apoptotic nuclei (terminal uridine nick end labeling, TUNEL). Intimal area (IA) and IA corrected for medial fracture length (IA/FL) were quantified by digital image analysis, which was also used to quantify the distribution of immunostain-positive cells in the adventitia, media, and neointima, respectively. Results: IA/FL was significantly reduced following treatment with 15 Gy, in association with decreased SMC density. Following injury and IR, TUNEL- and CD3-positive cell density increased significantly, and density of macrophages was increased in the adventitia and neointima. Staining for endothelial cells revealed a delay of re-endothelialization after radiation treatment. Conclusion: Increased T-cell infiltration at the medial tear following IR, perhaps due to incomplete re-endothelialization, may indicate incomplete healing. The elevated apoptosis of these infiltrating T cells may indicate a mechanism for the resolution of inflammation.

Inhibition of Endothelial Cell Migration by Cigarette Smoke Condensate

Cigarette smoking is among the leading risk factors in the etiology of atherosclerotic vascular disease. The mechanism, however, that links cigarette smoking to an increased incidence of atherosclerosis is poorly understood. Endothelial cell (EC) integrity is critical in preventing vascular lesion formation, and after a loss of EC integrity reendothelialization must be rapid and complete. We therefore investigated whether cigarette smoke affected the ECs ability to migrate or altered the intracellular signals generated during migration. The DMSO-soluble fraction of cigarette smoke condensate (CSC), derived from the standard research cigarette, was tested on cultured ECs (HUVEC) derived from human umbilical vein. The addition of CSC caused a dose-dependent decrease in the ability of EC to migrate as measured over a 24-h time period. Nicotine and cadmium sulfate, two constituents of cigarette smoke, individually or in combination, had no effect on migration. Examination of the tyrosine phosphorylation state of various intracellular proteins by Western blot analysis showed that CSC caused the hyperphosphorylation of a 130-kDa protein. In addition, other intracellular proteins showed changes in their phosphorylation states after CSC addition. These results support the hypothesis that CSC is detrimental to normal EC function in maintaining vascular integrity and suggest that smokers are more likely to develop complications of vascular disease due to delayed or incomplete reendothelialization as a consequence of decreased EC migration.

An Alternative to Synthetic Aortic Grafts Using Jejunum

Synthetic materials currently used for arterial grafts can provoke serious complications such as infection, rejection, and incomplete reendothelialization. Porcine small intestinal submucosa (SIS) allografts (homografts) may not share these disadvantages yet still may provide an effective graft alternative. This study was designed to provide additional information concerning the performance of this material when used as an allo-patch graft. A porcine model was selected to enable hemodynamic comparisons to the adult human physiology. Slaughterhouse small intestine was used to prepare the SIS graft material. A well-vascularized section ofjejunum was resected, rinsed, and maintained in 0.9% NaCl solution containing 10% gentamicin. The graft was prepared by removing the serosal layer, inverting the material, and discarding the mucosal surface in a similar manner. The remaining submucosa and stratum compactum constituted the graft material and was subdivided into sizes from 1.5 to 3.0 cm2 tacked to foil for manageability, and was returned to the gentamicin solution. The surgery was initiated with a femoral arterial line in the anesthetized swine, followed by isolation and preparation of the infrarenal aorta for grafting. Cross-clamps were applied and a 1.0 x 1.5-cm section of aortic wall was resected. Two sides of allograft were freed from the foil and sewn into the aorta. Then the remaining sides were removed from the foil and final graft placement was completed. The graft was harvested at postoperative day 28 and presented for microscopic examination. Results confirmed that reendothelialization with neovascularization had occurred during this interval.

The effect of endovascular irradiation on platelet recruitment at sites of balloon angioplasty in pig coronary arteries.

Endovascular irradiation (EI) inhibits balloon-induced neointima formation in animals and is now in clinical trials for restenosis prevention. However, little is known of the effect of EI on vessel thrombogenicity due to delayed arterial healing. We investigated EI effects on platelet recruitment in pig coronary arteries. EI was performed using (90)Sr/Y at 0 Gray (Gy), 15Gy, or 30Gy at 2 mm after balloon overstretch injury. At 1 day, 1 week, and 1 month, platelet recruitment and thrombus formation were assessed using autologous (111)In-oxine-platelet labeling and light and scanning electron microscopy. In balloon-injured nonirradiated vessels, there was complete reendothelialization at 1 month, and platelet recruitment was similar to normal uninjured arteries. In irradiated vessels, scanning electron microscopy showed incomplete reendothelialization at 1 month, and these areas demonstrated attachment of activated platelets. Light microscopy of irradiated coronaries showed adherent partially organized thrombi and incomplete resolution of intramural hemorrhages. There was a significant increase in platelet recruitment at 1 month in arteries receiving EI at 15Gy (5.1+/-2. 8x10(6), P=0.02) or 30Gy (12.5+/-9.9x10(6), P=0.005) compared with nonirradiated controls (2.7+/-1.5x10(6)); 30Gy was also higher than 15Gy (P=0.05). Platelet recruitment was also increased for 30Gy compared with control at 1 day. Endovascular irradiation at 15Gy or 30Gy after balloon angioplasty results in incomplete endothelial recovery, impaired resolution of intramural hemorrhage, and a dose-dependent increase in platelet recruitment at 1 month.

Catheter-induced intimal injury during routine coronary catheterization in dogs.

The location and progression of changes in arterial permeability and structure were studied in nine dogs over a 13-week period following left coronary catheterization with standard coronary catheters. Changes in arterial permeability were analyzed by quantitating Evans blue dye (EBD) uptake over the aortic luminal surface (blue areas). Structural changes were assessed by light and electron microscopy. In the catheterized animals, compared to uncatheterized controls, we observed a significant increase in aortic luminal EBD uptake that was maximal 4 hours after the procedure but still present up to 13 weeks later. Microscopic analysis of "blue areas" shortly after the procedure revealed widespread endothelial denudation, platelet and leukocyte adherence, with occasional intimal avulsions, disruption of the internal elastica, and thrombi. The EBD uptake patterns in association with the electron microscopic findings in these animals suggested that 67-89% of the aortic endothelium was removed by the catheter during the procedure. The proliferative response that occurred following this catheter-induced injury produced fibrocellular intimal thickening in the aortas and left main coronary arteries of animals studied 10 to 92 days after the procedure. In the aortic root, such thickening was associated with incomplete re-endothelialization, thrombogenicity, and grossly abnormal permeability patterns. We conclude that significant catheter-induced intimal injury can occur during coronary angiography. In the canine model, such injury is associated with widespread fibrocellular intimal thickening and abnormal permeability that persists for at least 13 weeks after the procedure.