ABSTRACT
AIMS: Inflammatory mediators, including blood cells and their products, contribute critically to atherogenesis, but the igniting triggers of inflammation remain elusive. Atherosclerosis develops at sites of flow perturbation, where the enhanced haemodynamic stress could initiate the atherogenic inflammatory process due to the occurrence of mechanic injury. We investigated the role of haemodynamic stress-induced breaches, allowing the entry of blood cells in the arterial intima, in triggering inflammation-driven atherogenesis. METHODS AND RESULTS: Human coronary samples isolated from explanted hearts, (n = 47) displayed signs of blood entry (detected by the presence of iron, ferritin, and glycophorin A) in the subintimal space (54%) as assessed by histology, immunofluorescence, high resolution episcopic microscopy, and scanning electron microscopy. Computational flow dynamic analysis showed that intimal haemorrhagic events occurred at sites of flow disturbance. Experimental carotid arteries from Apoe deficient mice showed discrete endothelial breaches and intimal haemorrhagic events specifically occurring at the site of flow perturbation, within 3 days after the exacerbation of the local haemodynamic stress. Endothelial tearing was associated with increased VCAM-1 expression and, within 7 days, substantial Ly6G+ leucocytes accumulated at the sites of erythrocyte-derived iron and lipids droplets accumulation, pathological intimal thickening and positive oil red O staining. The formation of fatty streaks at the sites of intimal breaches was prevented by the depletion of Ly6G+ leucocytes, suggesting that the local injury driven by haemodynamic stress-induced breaches triggers atherogenic inflammation. CONCLUSION: Haemodynamic-driven breaches of the arterial intima drive atherogenic inflammation by triggering the recruitment of leucocyte at sites of disturbed arterial flow.
Subject(s)
Atherosclerosis/metabolism , Hemodynamics/physiology , Inflammation/pathology , Tunica Intima/pathology , Animals , Antigens, Ly/metabolism , Apolipoproteins E/deficiency , Blood Flow Velocity , Carotid Arteries/metabolism , Carotid Arteries/pathology , Coronary Vessels/metabolism , Coronary Vessels/pathology , Coronary Vessels/ultrastructure , Endothelial Cells/metabolism , Endothelial Cells/pathology , Humans , Leukocytes/pathology , Male , Mice , Mice, Inbred C57BL , Stress, Mechanical , Tunica Intima/injuries , Vascular Cell Adhesion Molecule-1/metabolismABSTRACT
The fish heart ventricle has varied morphology and may have a specific morpho-functional design in species adapted to extreme environmental conditions. In general, the Amazonian ichthyofauna undergoes constant variations in water temperature, pH and oxygen saturation, which makes these species useful for investigations of cardiac morphology. Arapaima gigas, a member of the ancient teleost group Osteoglossomorpha, is one of the largest freshwater fish in the world. This species has a specific heart metabolism that uses fat as the main fuel when O2 supplies are abundant but also can change to glycogen fermentation when O2 content is limiting. However, no information is available regarding its heart morphology. Here, we describe the heart of A. gigas, with emphasis on the ventricular anatomy and myoarchitecture. Specimens of A. gigas weighing between 0.3 and 4040 g were grouped into three developmental stages. The hearts were collected and the anatomy analyzed with a stereomicroscope, ultrastructure with a scanning electron microscope, and histology using toluidine blue, Masson's trichrome and Sirius red stains. The ventricle undergoes morphological changes throughout its development, from the initial saccular shape with a fully trabeculated myocardium and coronary vessel restricted to the subepicardium (Type I) (group 1) to a pyramidal shape with mixed myocardium and coronary vessels that penetrate only to the level of the compact layer (Type II) (groups 2 and 3). The trabeculated myocardium has a distinct net-like organization in all the specimens, differing from that described for other teleosts. This arrangement delimits lacunae with a similar shape and distribution, which seems to allow a more uniform blood distribution through this myocardial layer.
Subject(s)
Coronary Vessels/cytology , Fishes/anatomy & histology , Heart Ventricles/cytology , Heart/anatomy & histology , Myocardium/ultrastructure , Animals , Coronary Vessels/anatomy & histology , Coronary Vessels/ultrastructure , Heart Ventricles/anatomy & histology , Histological Techniques , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Myocardium/cytology , Staining and Labeling/methodsABSTRACT
INTRODUCTION: Coronary microcirculation disturbance plays an important role in chronic heart failure (CHF). High thoracic sympathetic block (HTSB) is effective to treat CHF, but its impact on coronary microcirculation is unclear. METHODS: Forty male Wistar rats were subcutaneously injected with isoproterenol (340â¯mg/kg) for 2â¯days. Eight weeks later, 24 surviving rats were randomized to the CHF and HTSB groups and 10 rats were used as the control group. 50⯵l of saline and ropivacaine (0.2%) were epidurally infused everyday in the CHF and HTSB group respectively. Four weeks later, echocardiography and pathological and ultrastructural examination, capillary histochemical staining and vascular endothelial growth factor (VEGF) immunohistochemical staining in left ventricular (LV) subendocardial myocardium were performed. RESULTS: Compared with the control group, LV dilation and dysfunction, myocardial focal necrosis, capillary spasm appeared in the CHF group. HTSB ameliorated LV dilation and dysfunction, myocardial necrosis and capillary spasm. Rats in the CHF group had less myocardial capillary density and more VEGF expression than in the control group (1591⯱â¯99 vs. 1972⯱â¯118/mm2, 0.62⯱â¯0.13 vs. 0.33⯱â¯0.10 optic density, all pâ¯<â¯0.05). Myocardial capillary density (1782⯱â¯96/mm2) was more and VEGF expression (0.47⯱â¯0.12 optic density) was less in the HTSB group than in the CHF group (all pâ¯<â¯0.05). CONCLUSION: HTSB improves coronary microcirculation disturbance in CHF, which may contribute to reversing myocardial remodeling and dysfunction. HTSB stimulates myocardial capillary growth independent of VEGF.
Subject(s)
Anesthetics, Local/administration & dosage , Capillaries/physiopathology , Coronary Circulation , Coronary Vessels/physiopathology , Heart Failure/therapy , Microcirculation , Nerve Block/methods , Ropivacaine/administration & dosage , Thoracic Nerves , Animals , Capillaries/diagnostic imaging , Capillaries/metabolism , Capillaries/ultrastructure , Chronic Disease , Coronary Vessels/diagnostic imaging , Coronary Vessels/metabolism , Coronary Vessels/ultrastructure , Disease Models, Animal , Heart Failure/diagnostic imaging , Heart Failure/metabolism , Heart Failure/physiopathology , Male , Myocardium/metabolism , Myocardium/pathology , Neovascularization, Physiologic , Rats, Wistar , Vascular Endothelial Growth Factor A/metabolism , Ventricular Function, Left , Ventricular RemodelingABSTRACT
We have developed a new method for obtaining information on whole tissues by light microscopy (LM) and ultrastructural features by transmission electron microscopy (TEM). This method uses serial sections of a stented artery embedded in resin. Stents were implanted in porcine coronary arteries in this study. The heart was perfusion fixed in a 2% paraformaldehyde and 1.25% glutaraldehyde mixed solution. The stented artery was then removed, fixed in 1% osmium, embedded in Quetol 651 resin, and sectioned serially. For LM, the black color of osmium was removed from the section by immersion in periodic acid and hydrogen peroxide after deplasticization. These sections were stained with hematoxylin and eosin and Elastica-Masson trichrome stain. For TEM, thin sections were re-embedded in Quetol 812 resin by the resupinate method and cut into ultrathin sections. A clear, fine structure was obtained, and organelles, microvilli, and cell junctions in the endothelium were easily observed. The combined observation of adjacent specimens by LM and TEM enabled us to relate histopathological changes in the millimeter scale to those in the nanometer scale.
Subject(s)
Coronary Vessels/ultrastructure , Epoxy Resins/chemistry , Histological Techniques/instrumentation , Histological Techniques/methods , Methacrylates/chemistry , Stents/adverse effects , Animals , Coronary Vessels/pathology , Microscopy , Microscopy, Electron, Transmission , Microtomy/methods , SwineABSTRACT
BACKGROUND: Although drug-eluting stents have dramatically reduced the rates of restenosis and target lesion revascularization, they are associated with stent thrombosis (ST), a catastrophic event likely due to delayed endothelialization and chronic inflammation caused by the polymer and the metal scaffolds. To increase the safety and efficacy of stents, polymer-free dual drug-eluting stents (DDES) have been developed. METHODS: A total 160 stents (Bare-metal stents (BMS), polymer-free probucol stents (PrES), sirolimus-eluting stents (SES) and DDES) were randomly implanted in the coronary arteries of 80 pigs. 14, 28, 90 and 191 days after implantation, QCA and OCT evaluations were performed in 20 pigs respectively, and the arteries were harvested for scanning electron microscope (SEM), histomorphology, histopathology analyses and for the relative expression of CD31, CD34 and CD133 on mRNA and protein levels. RESULTS: At the 14-day time point, there were significant differences in the strut rate coverage (p = 0.011), with greater coverage in the PrES than in the SES group (53.2%vs. 20.3%, p = 0.002). As well, there were no significant differences in the expression of CD31, CD34 and CD133 between groups in mRNA and protein level. CONCLUSIONS: DDES were as safe as BMS and SES, but they did not further improve the endothelialization of the stented coronary arteries in the porcine model.
Subject(s)
Cardiovascular Agents/administration & dosage , Coronary Vessels , Drug-Eluting Stents , Metals , Percutaneous Coronary Intervention/instrumentation , Probucol/administration & dosage , Sirolimus/administration & dosage , Stents , AC133 Antigen/genetics , AC133 Antigen/metabolism , Animals , Antigens, CD34/genetics , Antigens, CD34/metabolism , Coronary Angiography , Coronary Thrombosis/diagnostic imaging , Coronary Thrombosis/etiology , Coronary Thrombosis/metabolism , Coronary Thrombosis/pathology , Coronary Vessels/diagnostic imaging , Coronary Vessels/metabolism , Coronary Vessels/ultrastructure , Female , Male , Microscopy, Electron, Scanning , Models, Animal , Percutaneous Coronary Intervention/adverse effects , Platelet Endothelial Cell Adhesion Molecule-1/genetics , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Prosthesis Design , RNA, Messenger/genetics , RNA, Messenger/metabolism , Swine , Swine, Miniature , Time Factors , Tomography, Optical CoherenceABSTRACT
Static cold storage is accompanied with a partial safe ischemic interval for donor hearts. In this current study, a machine perfusion system was built to provide a better preservation for the donor heart and assessment for myocardial function. Chinese mini-swine (weight 30-35 kg, n = 16) were randomly divided into HTK, Celsior, and Heartbeat groups. All donor hearts were respectively preserved for 8 hours under static cold storage or machine perfusion. The perfusion solution is aimed to maintain its homeostasis based on monitoring the Heartbeat group. The ultrastructure of myocardium suggests better myocardial protection in the Heartbeat group compared with HTK or Celsior-preserved hearts. The myocardial and coronary artery structural and functional integrity was evaluated by immunofluorescence and Western blots in the Heartbeat. In the Heartbeat group, donor hearts maintained a high adenosine triphosphate level. Bcl-2 and Beclin-1 protein demonstrates high expression in the Celsior group. The Heartbeat system can be used to preserve donor hearts, and it could guarantee the myocardial and endothelial function of hearts during machine perfusion. Translating Heartbeat into clinical practice, it is such as to impact on donor heart preservation for cardiac transplantation.
Subject(s)
Extracorporeal Membrane Oxygenation/instrumentation , Heart , Hemofiltration/instrumentation , Organ Preservation/instrumentation , Perfusion/instrumentation , Animals , Cold Ischemia , Cold Temperature , Coronary Vessels/drug effects , Coronary Vessels/ultrastructure , Cytoprotection , Disaccharides/pharmacology , Electrolytes/pharmacology , Energy Metabolism , Equipment Design , Extracorporeal Membrane Oxygenation/adverse effects , Extracorporeal Membrane Oxygenation/methods , Glucose/pharmacology , Glutamates/pharmacology , Glutathione/pharmacology , Heart/drug effects , Heart/physiopathology , Hemofiltration/adverse effects , Hemofiltration/methods , Histidine/pharmacology , Mannitol/pharmacology , Models, Animal , Myocardial Contraction , Myocardium/metabolism , Myocardium/ultrastructure , Organ Preservation/adverse effects , Organ Preservation/methods , Organ Preservation Solutions/pharmacology , Perfusion/adverse effects , Perfusion/methods , Potassium Chloride/pharmacology , Procaine/pharmacology , Swine , Swine, Miniature , Time FactorsABSTRACT
The sustained or controlled release of nitric oxide (NO) can be the most promising approach for the suppression or prevention of restenosis and thrombosis caused by stent implantation. The aim of this study is to investigate the feasibility in the potential use of layer-by-layer (LBL) coating with a NO donor-containing liposomes to control the release rate of NO from a metallic stent. Microscopic observation and surface characterizations of LBL-modified stents demonstrate successful LBL coating with liposomes on a stent. Release profiles of NO show that the release rate is sustained up to 5 d. In vitro cell study demonstrates that NO release significantly enhances endothelial cell proliferation, whereas it markedly inhibits smooth muscle cell proliferation. Finally, in vivo study conducted with a porcine coronary injury model proves the therapeutic efficacy of the NO-releasing stents coated by liposomal LBL technique, supported by improved results in luminal healing, inflammation, and neointimal thickening except thrombo-resistant effect. As a result, all these results demonstrate that highly optimized release rate and therapeutic dose of NO can be achieved by LBL coating and liposomal encapsulation, followed by significantly efficacious outcome in vivo.
Subject(s)
Coated Materials, Biocompatible/chemistry , Coronary Vessels/metabolism , Liposomes/chemistry , Nitric Oxide/metabolism , Stents , Adsorption , Animals , Coronary Vessels/pathology , Coronary Vessels/ultrastructure , Fibrinogen/metabolism , Human Umbilical Vein Endothelial Cells/metabolism , Humans , Male , Nitroso Compounds/chemistry , Quartz Crystal Microbalance Techniques , Sus scrofaABSTRACT
Candida glabrata (CG) is an opportunistic fungal pathogen that initiates infection by binding to host cells via specific lectin-like adhesin proteins. We have previously shown the importance of lectin-oligosaccharide binding in cardiac responses to flow and agonists. Because of the lectinic-oligosaccharide nature of CG binding, we tested the ability of CG to alter the agonist- and flow-induced changes in cardiac function in isolated perfused guinea pig hearts. Both transmission and scanning electron microscopy showed strong attachment of CG to the coronary endothelium, even after extensive washing. CG shifted the coronary flow vs. auricular-ventricular (AV) delay relationship upward, indicating that greater flow was required to achieve the same AV delay. This effect was completely reversed with mannose, partially reversed with galactose and N-acetylgalactosamine, but hyaluronan had no effect. Western blot analysis was used to determine binding of CG to isolated coronary endothelial luminal membrane (CELM) receptors, and the results indicate that flow-sensitive CELM receptors, ANG II type I, α-adrenergic 1A receptor, endothelin-2, and VCAM-1 bind to CG. In addition, CG inhibited agonist-induced effects of bradykinin, angiotensin, and phenylephrine on AV delay, coronary perfusion pressure, and left ventricular pressure. Mannose reversed the inhibitory effects of CG on the agonist responses. These results suggest that CG directly binds to flow-sensitive CELM receptors via lectinic-oligosaccharide interactions with mannose and disrupts the lectin-oligosaccharide binding necessary for flow-induced cardiac responses.
Subject(s)
Candida glabrata/metabolism , Candidiasis/metabolism , Cell Membrane/metabolism , Coronary Circulation , Coronary Vessels/metabolism , Endothelial Cells/metabolism , Receptors, G-Protein-Coupled/metabolism , Ventricular Function, Left , Angiotensin II/pharmacology , Animals , Bradykinin/pharmacology , Candida glabrata/genetics , Candida glabrata/ultrastructure , Candidiasis/genetics , Candidiasis/microbiology , Candidiasis/physiopathology , Cell Membrane/microbiology , Cell Membrane/ultrastructure , Coronary Circulation/drug effects , Coronary Vessels/microbiology , Coronary Vessels/physiopathology , Coronary Vessels/ultrastructure , Endothelial Cells/microbiology , Endothelial Cells/ultrastructure , Glycosylation , Guinea Pigs , Host-Pathogen Interactions , Isolated Heart Preparation , Mannose/metabolism , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Mutation , Myocardial Contraction , Phenylephrine/pharmacology , Receptor, Angiotensin, Type 1/metabolism , Receptors, Adrenergic, alpha-1/metabolism , Receptors, Endothelin/metabolism , Receptors, G-Protein-Coupled/agonists , Vascular Cell Adhesion Molecule-1/metabolism , Ventricular Function, Left/drug effects , Ventricular PressureABSTRACT
OBJECTIVE: Despite advances in stent technology for vascular interventions, in-stent restenosis (ISR) because of myointimal hyperplasia remains a major complication. APPROACH AND RESULTS: We investigated the regulatory role of microRNAs in myointimal hyperplasia/ISR, using a humanized animal model in which balloon-injured human internal mammary arteries with or without stenting were transplanted into Rowett nude rats, followed by microRNA profiling. miR-21 was the only significantly upregulated candidate. In addition, miR-21 expression was increased in human tissue samples from patients with ISR compared with coronary artery disease specimen. We systemically repressed miR-21 via intravenous fluorescein-tagged-locked nucleic acid-anti-miR-21 (anti-21) in our humanized myointimal hyperplasia model. As expected, suppression of vascular miR-21 correlated dose dependently with reduced luminal obliteration. Furthermore, anti-21 did not impede reendothelialization. However, systemic anti-miR-21 had substantial off-target effects, lowering miR-21 expression in liver, heart, lung, and kidney with concomitant increase in serum creatinine levels. We therefore assessed the feasibility of local miR-21 suppression using anti-21-coated stents. Compared with bare-metal stents, anti-21-coated stents effectively reduced ISR, whereas no significant off-target effects could be observed. CONCLUSION: This study demonstrates the efficacy of an anti-miR-coated stent for the reduction of ISR.
Subject(s)
Antibodies, Antinuclear/pharmacology , Coated Materials, Biocompatible , Coronary Restenosis/prevention & control , Gene Expression Regulation , Graft Occlusion, Vascular/prevention & control , MicroRNAs/genetics , Animals , Cell Proliferation/drug effects , Coronary Restenosis/genetics , Coronary Restenosis/metabolism , Coronary Vessels/drug effects , Coronary Vessels/metabolism , Coronary Vessels/ultrastructure , Disease Models, Animal , Drug-Eluting Stents , Female , Graft Occlusion, Vascular/genetics , Graft Occlusion, Vascular/metabolism , Humans , Male , MicroRNAs/biosynthesis , MicroRNAs/immunology , Microscopy, Electron, Scanning , Middle Aged , Muscle, Smooth, Vascular/drug effects , Muscle, Smooth, Vascular/ultrastructure , Neointima/metabolism , Neointima/pathology , Prosthesis Design , Rats , Rats, NudeABSTRACT
Consumption of fatty acids from fishes is widely regarded as beneficial for preventing cardiovascular disorders. Nevertheless, salmonids themselves are victims of vascular diseases. As the pathogenesis and nature of these changes are elusive, they are here addressed using novel morphological and transcriptional approaches. Coronary arteries of wild Atlantic salmon Salmo salar L., (n = 12) were investigated using histological and immunohistochemical techniques, and RT-qPCR was employed to investigate expression of stretch-induced genes. In an experimental trial, fish were fed diets with different fatty acids composition, and histological features of the coronary arteries (n = 36) were investigated. In addition, the heart fatty acid profile (n = 60) was analysed. There were no differences in morphological or immunological features between wild fish and groups of experimental fish. Arteriosclerotic lesions consisted of smooth muscle cells in dissimilar differential stages embedded in considerable amounts of extracellular matrix in a similar fashion to what is seen in early stages of human atherosclerosis. No fat accumulations were observed, and very few inflammatory cells were present. In affected arteries, there was an induction of stretch-related genes, pointing to a stress-related response. We suggest that salmon may have a natural resistance to developing atherosclerosis, which corresponds well with their high investment in lipid metabolism.
Subject(s)
Coronary Artery Disease/veterinary , Coronary Vessels/pathology , Dietary Fats/administration & dosage , Fatty Acids/administration & dosage , Fish Diseases/pathology , Salmo salar , Animals , Coronary Artery Disease/etiology , Coronary Artery Disease/pathology , Coronary Vessels/ultrastructure , Diet/veterinary , Dietary Fats/analysis , Extracellular Matrix/pathology , Fatty Acids/analysis , Fatty Acids/chemistry , Fish Diseases/etiology , Fish Diseases/genetics , Gene Expression Regulation, Bacterial , Immunohistochemistry/veterinary , Microscopy, Electron, Scanning/veterinary , Microscopy, Electron, Transmission/veterinary , Myocardium/chemistry , Myocardium/pathology , Myocardium/ultrastructure , Neointima/pathology , Up-RegulationABSTRACT
Peripheral vascular resistance is increased in essential hypertension. This involves structural changes of resistance arteries and stiffening of the arterial wall, including remodeling of the extracellular matrix. We hypothesized that biopsies of the human parietal pericardium, obtained during coronary artery bypass grafting or cardiac valve replacement surgeries, can serve as a source of resistance arteries for structural research in cardiovascular disease patients. We applied two-photon excitation fluorescence microscopy to study the parietal pericardium and isolated pericardial resistance arteries with a focus on the collagen and elastin components of the extracellular matrix. Initial findings in pig tissue were confirmed in patient biopsies. The microarchitecture of the internal elastic lamina in both the pig and patient pericardial resistance arteries (studied at a transmural pressure of 100 mm Hg) is fiber like, and no prominent external elastic lamina could be observed. This microarchitecture is very different from that in rat mesenteric arteries frequently used for resistance artery research. In conclusion, we add three-dimensional information on the structure of the extracellular matrix in resistance arteries from cardiovascular disease patients and propose further use of patient pericardial resistance arteries for studies of the human microvasculature.
Subject(s)
Cardiovascular Diseases/pathology , Elastic Tissue/ultrastructure , Elastin/analysis , Pericardium , Sus scrofa/anatomy & histology , Aged , Animals , Cardiovascular Diseases/metabolism , Coronary Vessels/ultrastructure , Extracellular Matrix/chemistry , Extracellular Matrix/ultrastructure , Female , Humans , Male , Mesenteric Arteries/ultrastructure , Microscopy, Fluorescence, Multiphoton , Middle Aged , Rats , Species Specificity , Swine , Vascular ResistanceABSTRACT
BACKGROUND: Optical coherence tomography (OCT) is a new intracoronary imaging modality that has excellent resolution and image quality and has been used to image neointimal coverage after stent implantation. OCT has been compared to histologic, intravascular ultrasound, and scanning electron microscopy (SEM) studies. However, OCT has not been compared with SEM for imaging stent coverage over side branches. OBJECTIVE: The aim of this study was to compare OCT with SEM in imaging neointimal coverage over stent struts bridging coronary side-branch ostia. METHODS: Using a balloon-overstretch in-stent restenosis model, we deployed 38 everolimus-eluting stents across coronary bifurcations in nine pigs. We performed OCT immediately after stenting and 4 weeks later; SEM was performed after euthanizing the pigs. OCT images of each stent were compared to the corresponding SEM image. RESULTS: We analyzed OCT frames (n=111) for strut-level neointimal coverage and compared them to corresponding SEM images. The concordance correlation coefficient was 0.809 (95%CI; 0.734-0.864) and 0.951 (95%CI; 0.930-0.966) for covered and uncovered struts, respectively. CONCLUSIONS: In a non-atherosclerotic pig model, we showed strong agreement between OCT and SEM in imaging coverage of stent struts bridging side-branch ostia.
Subject(s)
Angioplasty, Balloon, Coronary/instrumentation , Coronary Restenosis/pathology , Coronary Vessels/pathology , Microscopy, Electron, Scanning , Stents , Tomography, Optical Coherence , Angioplasty, Balloon, Coronary/adverse effects , Animals , Coronary Restenosis/etiology , Coronary Vessels/ultrastructure , Disease Models, Animal , Neointima , Predictive Value of Tests , Reproducibility of Results , Sus scrofaABSTRACT
BACKGROUND: Coronary adventitia harbors a wide variety of components, such as inflammatory cells and vasa vasorum (VV). Adventitial VV initiates the development of coronary artery diseases as an outside-in supply route of inflammation. We have recently demonstrated that drug-eluting stent implantation causes the enhancement of VV formation, with extending to the stent edges in the porcine coronary arteries, and also that optical frequency domain imaging (OFDI) is capable of visualizing VV in humans in vivo. However, it remains to be fully validated whether OFDI enables the precise measurement of VV formation in pigs and humans. METHODS AND RESULTS: In the pig protocol, a total of 6 bare-metal stents and 12 drug-eluting stents were implanted into the coronary arteries, and at 1 month, the stented coronary arteries were imaged by OFDI ex vivo. OFDI data including the measurement of VV area at the stent edge portions were compared with histological data. There was a significant positive correlation between VV area on OFDI and that on histology (R=0.91, P<0.01). In the human protocol, OFDI enabled the measurement of the VV area at the stent edges after coronary stent implantation in vivo. CONCLUSIONS: These results provide the first direct evidence that OFDI enables the precise measurement of the VV area in coronary arteries after stent implantation in pigs and humans.
Subject(s)
Adventitia/blood supply , Coronary Stenosis/surgery , Coronary Vessels/physiopathology , Everolimus/therapeutic use , Neovascularization, Physiologic , Prosthesis Implantation , Sirolimus/analogs & derivatives , Stents , Tomography, Optical Coherence/methods , Vasa Vasorum/physiopathology , Adventitia/ultrastructure , Aged , Aged, 80 and over , Animals , Aspirin/therapeutic use , Clopidogrel , Coronary Stenosis/physiopathology , Coronary Vessels/drug effects , Coronary Vessels/ultrastructure , Disease Progression , Drug-Eluting Stents , Everolimus/administration & dosage , Everolimus/pharmacology , Female , Humans , Interferometry/methods , Male , Middle Aged , Neointima/pathology , Neovascularization, Physiologic/drug effects , Platelet Aggregation Inhibitors/therapeutic use , Postoperative Period , Sirolimus/administration & dosage , Sirolimus/pharmacology , Swine , Swine, Miniature , Ticlopidine/analogs & derivatives , Ticlopidine/therapeutic use , Vasa Vasorum/drug effects , Vasculitis/complications , Vasculitis/pathology , Vasculitis/physiopathologyABSTRACT
It has been considered that concentrations of certain amino acids in the egg are not sufficient to fully support embryonic development of modern broilers. In this study we evaluated embryo growth and development with particular emphasis on one of the major components of connective tissue, collagen. Experiments were performed on Ross 308 chicken embryos from 160 fertilised eggs. Experimental solutions of silver nanoparticles (Ag), hydroxyproline solution (Hyp) and a complex of silver nanoparticles with hydroxyproline (AgHyp) were injected into albumen, and embryos were incubated until day 20. An assessment of the mass of embryo and selected organs was carried out followed by measurements of the expression of the key signalling factors' fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor-A (VEGF-A). Finally, an evaluation of collagen microstructure using scanning electron microscopy was performed. Our results clearly indicate that Hyp, Ag and AgHyp administered in ovo to chicken embryos did not harm embryos. Comparing to the control group, Hyp, Ag and the AgHyp complex significantly upregulated expression of the FGF-2 at the mRNA and protein levels. Moreover, Hyp, Ag and, in particular, the complex of AgHyp significantly increased blood vessel size, cartilage collagen fibre lattice size and bundle thickness. The general conclusion from this study is that AgHyp treatment may help to build a stronger and longer lasting form of collagen fibres.
Subject(s)
Chick Embryo/drug effects , Hydroxyproline/pharmacology , Metal Nanoparticles/chemistry , Osteochondrodysplasias/metabolism , Silver/pharmacology , Animals , Chick Embryo/blood supply , Chick Embryo/metabolism , Coronary Vessels/embryology , Coronary Vessels/ultrastructure , Gene Expression Regulation, Developmental/drug effects , Hydroxyproline/administration & dosage , Microscopy, Electron, Scanning , Silver/chemistryABSTRACT
Current trends are toward actively developing approaches of tissue engineering, aimed at creating vascular grafts of small diameter. This is due to the existing in cardiovascular surgery demand for prostheses to be used in coronary artery bypass grafting. The present work was undertaken in order to assess possibilities of using smalldiameter vascular grafts made of biodegradable polymer polycaprolactone by means of electrospinning. The authors studied physico-mechanical properties and structure of polycaprolactone grafts, as well as their thromboresistance and patency after implantation into the vascular bed of rats. The obtained results demonstrated optimal physicomechanical properties of the vascular grafts, their biocompatibility, endothelialisation of the internal surface, and infiltration of the graft's wall by cells with the formation of new tissue, accompanied and followed by the development of an extensive intimal layer in the zones of the anastomoses. Hence, the study showed possibilities of using polycaprolactone grafts as vascular prostheses, however requiring their further modification which would promote and contribute to a decrease in hyperplasia of connective tissue in the graft's lumen.
Subject(s)
Blood Vessel Prosthesis , Coronary Artery Bypass/instrumentation , Coronary Artery Disease/surgery , Coronary Vessels/surgery , Polyesters , Tissue Engineering/methods , Animals , Cattle , Coronary Artery Disease/pathology , Coronary Restenosis/prevention & control , Coronary Vessels/ultrastructure , Disease Models, Animal , Follow-Up Studies , Male , Microscopy, Electron , Prosthesis Design , Rats , Rats, WistarABSTRACT
Here, we here present scanning electron microscope data for the existent telocytes (TCs) on the endothelial surface of the wall of pig coronary arteries, internal thoracic arteries and carotid arteries. These cells have a small (8.39 ± 1.97 µm/4.95 ± 0.91 µm) cell body of different shapes (from round to triangular, depending on the number of cellular prolongations) with very long (of about 30 µm) and thin cellular processes called telopodes (Tps), which have uneven calibre. The number of Tps ranges between 2 and 6. Tps typically present the alternation of podoms and podomers, and also have a dichotomic branching pattern. These data could influence the current attempts for elucidating the role(s) of TCs.
Subject(s)
Carotid Arteries/ultrastructure , Coronary Vessels/ultrastructure , Endothelium, Vascular/ultrastructure , Microscopy, Electron, Scanning/methods , Thoracic Arteries/ultrastructure , Animals , SwineABSTRACT
The epicardium and coronary vessels originate from progenitor cells in the proepicardium. Here we show that Tbx18, a T-box family member highly expressed in the proepicardium, controls critical early steps in coronary development. In Tbx18(-/-) mouse embryos, both the epicardium and coronary vessels exhibit structural and functional defects. At E12.5, the Tbx18-deficient epicardium contains protrusions and cyst-like structures overlying a disorganized coronary vascular plexus that contains ectopic structures resembling blood islands. At E13.5, the left and right coronary stems form correctly in mutant hearts. However, analysis of PECAM-1 whole mount immunostaining, distribution of SM22α(lacZ/+) activity, and analysis of coronary vascular casts suggest that defective vascular plexus remodeling produces a compromised arterial network at birth consisting of fewer distributing conduit arteries with smaller lumens and a reduced capacity to conduct blood flow. Gene expression profiles of Tbx18(-/-) hearts at E12.5 reveal altered expression of 79 genes that are associated with development of the vascular system including sonic hedgehog signaling components patched and smoothened, VEGF-A, angiopoietin-1, endoglin, and Wnt factors compared to wild type hearts. Thus, formation of coronary vasculature is responsive to Tbx18-dependent gene targets in the epicardium, and a poorly structured network of coronary conduit vessels is formed in Tbx18 null hearts due to defects in epicardial cell signaling and fate during heart development. Lastly, we demonstrate that Tbx18 possesses a SRF/CArG box dependent repressor activity capable of inhibiting progenitor cell differentiation into smooth muscle cells, suggesting a potential function of Tbx18 in maintaining the progenitor status of epicardial-derived cells.
Subject(s)
Coronary Vessels/embryology , Coronary Vessels/metabolism , Pericardium/embryology , Pericardium/metabolism , T-Box Domain Proteins/metabolism , Animals , Animals, Newborn , Cell Differentiation , Coronary Circulation , Coronary Vessels/pathology , Coronary Vessels/ultrastructure , Embryo, Mammalian/metabolism , Embryo, Mammalian/pathology , Gene Expression Profiling , Gene Expression Regulation, Developmental , Mice , Myocytes, Smooth Muscle/metabolism , Pericardium/pathology , Pericardium/ultrastructure , Repressor Proteins/metabolism , Serum Response Factor/chemistry , T-Box Domain Proteins/deficiency , T-Box Domain Proteins/genetics , Transcription, Genetic , beta-Galactosidase/metabolismABSTRACT
The accumulation of calcified material in cardiovascular tissue is thought to involve cytochemical, extracellular matrix and systemic signals; however, its precise composition and nanoscale architecture remain largely unexplored. Using nano-analytical electron microscopy techniques, we examined valves, aortae and coronary arteries from patients with and without calcific cardiovascular disease and detected spherical calcium phosphate particles, regardless of the presence of calcific lesions. We also examined lesions after sectioning with a focused ion beam and found that the spherical particles are composed of highly crystalline hydroxyapatite that crystallographically and structurally differs from bone mineral. Taken together, these data suggest that mineralized spherical particles may play a fundamental role in calcific lesion formation. Their ubiquitous presence in varied cardiovascular tissues and from patients with a spectrum of diseases further suggests that lesion formation may follow a common process. Indeed, applying materials science techniques to ectopic and orthotopic calcification has great potential to lend critical insights into pathophysiological processes underlying calcific cardiovascular disease.
Subject(s)
Calcinosis/pathology , Cardiomyopathies/pathology , Microscopy, Electron/methods , Aorta/pathology , Aorta/ultrastructure , Calcification, Physiologic , Calcium Phosphates/analysis , Coronary Vessels/pathology , Coronary Vessels/ultrastructure , Durapatite/analysis , Heart Valve Diseases/pathology , Heart Valves/pathology , Heart Valves/ultrastructure , Humans , Microscopy, Electron, Scanning , Nanotechnology/methods , Vascular Calcification/pathologyABSTRACT
The lymphatic vasculature preserves tissue fluid balance by absorbing fluid and macromolecules and transporting them to the blood vessels for circulation. The stepwise process leading to the formation of the mammalian lymphatic vasculature starts by the expression of the gene Prox1 in a subpopulation of blood endothelial cells (BECs) on the cardinal vein (CV) at approximately E9.5. These Prox1-expressing lymphatic endothelial cells (LECs) will exit the CV to form lymph sacs, primitive structures from which the entire lymphatic network is derived. Until now, no conclusive information was available regarding the cellular processes by which these LEC progenitors exit the CV without compromising the vein's integrity. We determined that LECs leave the CV by an active budding mechanism. During this process, LEC progenitors are interconnected by VE-cadherin-expressing junctions. Surprisingly, we also found that Prox1-expressing LEC progenitors were present not only in the CV but also in the intersomitic vessels (ISVs). Furthermore, as LEC progenitors bud from the CV and ISVs into the surrounding mesenchyme, they begin expressing the lymphatic marker podoplanin, migrate away from the CV, and form the lymph sacs. Analyzing this process in Prox1-null embryos revealed that Prox1 activity is necessary for LEC progenitors to exit the CV.
Subject(s)
Cell Movement , Coronary Vessels/cytology , Embryo, Mammalian/blood supply , Embryo, Mammalian/cytology , Embryonic Stem Cells/cytology , Endothelium, Lymphatic/embryology , Homeodomain Proteins/metabolism , Tumor Suppressor Proteins/metabolism , Adherens Junctions/metabolism , Adherens Junctions/ultrastructure , Animals , Cadherins/metabolism , Coronary Vessels/embryology , Coronary Vessels/ultrastructure , Embryo, Mammalian/ultrastructure , Embryonic Development , Embryonic Stem Cells/ultrastructure , Endothelium, Lymphatic/ultrastructure , Homeodomain Proteins/genetics , Membrane Glycoproteins/metabolism , Mice , Mice, Knockout , Mice, Transgenic , Microscopy, Electron, Transmission , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Recombinant Fusion Proteins/metabolism , Tumor Suppressor Proteins/geneticsABSTRACT
: In contrast to cardiomyocytes, autophagy in cardiac microvascular endothelial cells (CMECs) during ischemia/reperfusion (I/R) injury has not been fully investigated. Tongxinluo (TXL), a traditional Chinese medicine, was shown to be vascular protective. We aimed to elucidate the role of autophagy and its regulatory mechanisms by TXL in CMECs subjected to I/R injury. CMECs were exposed to different treatments for 30 minutes and subjected to hypoxia/reoxygenation each for 2 hours. The results indicated that hypoxia/reoxygenation significantly induced autophagy, as identified by an increased number of monodansylcadaverine-positive CMECs, increased autophagosome formation, and a higher type II/type I of light chain 3 ratio, but not Beclin-1 expression. Autophagy inhibition using 3-methyladenine was proapoptotic, but rapamycin-induced autophagy was antiapoptotic. TXL enhanced autophagy and decreased apoptosis in a dose-dependent manner, reaching its largest effect at 800 µg/mL. 3-methyladenine attenuated the TXL-promoted autophagy and antiapoptotic effects, whereas rapamycin had no additional effects compared with TXL alone. TXL upregulated mitogen-activated protein kinase and extracellular signal-regulated kinase (ERK) phosphorylation; however, PD98059 abrogated ERK phosphorylation and decreased autophagy and increased apoptosis compared with TXL alone. These results suggest that autophagy is a protective mechanism in CMECs subjected to I/R injury and that TXL can promote autophagy through activation of the mitogen-activated protein kinase/ERK pathway.