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      Validating Intravascular Imaging with Serial Optical Coherence Tomography and Confocal Fluorescence Microscopy

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          Abstract

          Atherosclerotic cardiovascular diseases are characterized by the formation of a plaque in the arterial wall. Intravascular ultrasound (IVUS) provides high-resolution images allowing delineation of atherosclerotic plaques. When combined with near infrared fluorescence (NIRF), the plaque can also be studied at a molecular level with a large variety of biomarkers. In this work, we present a system enabling automated volumetric histology imaging of excised aortas that can spatially correlate results with combined IVUS/NIRF imaging of lipid-rich atheroma in cholesterol-fed rabbits. Pullbacks in the rabbit aortas were performed with a dual modality IVUS/NIRF catheter developed by our group. Ex vivo three-dimensional (3D) histology was performed combining optical coherence tomography (OCT) and confocal fluorescence microscopy, providing high-resolution anatomical and molecular information, respectively, to validate in vivo findings. The microscope was combined with a serial slicer allowing for the imaging of the whole vessel automatically. Colocalization of in vivo and ex vivo results is demonstrated. Slices can then be recovered to be tested in conventional histology.

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          Most cited references15

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          Inflammation in atherosclerosis: transition from theory to practice.

          Inflammation drives the formation, progression, and rupture of atherosclerotic plaques. Experimental studies have demonstrated that an inflammatory subset of monocytes/macrophages preferentially accumulate in atherosclerotic plaque and produce proinflammatory cytokines. T lymphocytes can contribute to inflammatory processes that promote thrombosis by stimulating production of collagen-degrading proteinases and the potent procoagulant tissue factor. Recent data link obesity, inflammation, and modifiers of atherosclerotic events, a nexus of growing clinical concern given the worldwide increase in the prevalence of obesity. Modulators of inflammation derived from visceral adipose tissue evoke production of acute phase reactants in the liver, implicated in thrombogenesis and clot stability. Additionally, C-reactive protein levels rise with increasing levels of visceral adipose tissue. Adipose tissue in obese mice contains increased numbers of macrophages and T lymphocytes, increased T lymphocyte activation, and increased interferon-gamma (IFN-gamma) expression. IFN-gamma deficiency in mice reduces production of inflammatory cytokines and inflammatory cell accumulation in adipose tissue. Another series of in vitro and in vivo mouse experiments affirmed that adiponectin, an adipocytokine, the plasma levels of which drop with obesity, acts as an endogenous antiinflammatory modulator of both innate and adaptive immunity in atherogenesis. Thus, accumulating experimental evidence supports a key role for inflammation as a link between risk factors for atherosclerosis and the biology that underlies the complications of this disease. The recent JUPITER trial supports the clinical utility of an assessment of inflammatory status in guiding intervention to limit cardiovascular events. Inflammation is thus moving from a theoretical concept to a tool that provides practical clinical utility in risk assessment and targeting of therapy.
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            Atherosclerosis in angiographically "normal" coronary artery reference segments: an intravascular ultrasound study with clinical correlations.

            This study evaluated the magnitude, patterns and clinical correlates of atherosclerosis in angiographically "normal" reference segments in patients undergoing transcatheter therapy for symptomatic coronary artery disease. Pathologic studies indicate that the extent of coronary atherosclerosis is underestimated by visual analysis of angiographically normal coronary artery segments. Intravascular ultrasound allows detailed, high quality cross-sectional imaging of the coronary arteries in vivo. Intravascular ultrasound was used to study angiographically normal coronary artery reference segments in 884 patients evaluated for transcatheter therapy for symptomatic native coronary artery disease. The reference segment was the most visually normal intravascular ultrasound cross section within 10 mm proximal to the target lesion but distal to any major side branch. Results are presented as mean value +/- 1 SD. Only 60 (6.8%) of 884 angiographically normal reference segments were normal by intravascular ultrasound. Reference segment percent cross-sectional narrowing measured 51 +/- 13% and correlated poorly with the target lesion percent cross-sectional narrowing (r = 0.166, p < 0.0001). Reference segments contained less calcific and dense fibrotic plaque and proportionately more soft plaque elements. Independent predictors of reference segment percent cross-sectional narrowing were male gender, patient age, diabetes mellitus, hypercholesterolemia and presence of multivessel disease. Independent predictors of reference segment calcification were patient age and serum creatinine levels. Reference segment percent cross-sectional narrowing in 723 patients undergoing transcatheter therapy was similar to that in patients studied for diagnostic purposes; however, reference segment arc of calcium was greater in treated patients (43 +/- 81 vs. 25 +/- 57, p = 0.015). Reference segment disease was not an independent predictor of subsequent angiographic restenosis or clinical events within 12 months of follow-up. Atherosclerosis is ubiquitous in angiographically normal coronary artery reference segments. Reference segment disease parallels the severity of target lesion disease and is associated with many of the conventional risk factors for coronary artery disease. Because of its sensitivity in detecting atherosclerosis in angiographically normal reference segments, intravascular ultrasound should enhance the study of risk factors for atherosclerosis and the results of therapies to control disease progression.
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              Quantitative measurement of attenuation coefficients of weakly scattering media using optical coherence tomography.

              From calibrated, weakly scattering tissue phantoms (2-6 mm-1), we extract the attenuation coefficient with an accuracy of 0.8 mm-1 from OCT data in the clinically relevant 'fixed focus' geometry. The data are analyzed using a single scattering model and a recently developed description of the confocal point spread function (PSF). We verify the validity of the single scattering model by a quantitative comparison with a multiple scattering model, and validate the use of the PSF on the calibrated samples. Implementation of this model for existing OCT systems will be straightforward. Localized quantitative measurement of the attenuation coefficient of different tissues can significantly improve the clinical value of OCT.
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                Author and article information

                Contributors
                Role: Academic Editor
                Role: Academic Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                15 December 2016
                December 2016
                : 17
                : 12
                : 2110
                Affiliations
                [1 ]Département de Génie Électrique et Institut de Génie Biomédical, École Polytechnique de Montréal, Montreal, QC H3T 1J4, Canada; maxime.abran@ 123456gmail.com (M.A.); alexandre.castonguay87@ 123456gmail.com (A.C.); joel.lefebvre@ 123456gmail.com (J.L.)
                [2 ]Montreal Heart Institute, Montreal, QC H1T 1C8, Canada; mariejeanne.bertrand@ 123456gmail.com (M.-J.B.); barbarastaehli@ 123456hotmail.com (B.E.S.); nolwenn.merlet@ 123456gmail.com (N.M.); Teodora.Mihalache-Avram@ 123456icm-mhi.org (T.M.-A.); Pascale.Geoffroy@ 123456icm-mhi.org (P.G.); melanie.mecteau@ 123456icm-mhi.org (M.M.); david.busseuil@ 123456icm-biobanque.org (D.B.); Eric.Rheaume@ 123456icm-mhi.org (É.R.); philippe.l_lallier@ 123456icloud.com (P.L.); jean-claude.tardif@ 123456icm-mhi.org (J.-C.T.)
                [3 ]Département de Médecine, Université de Montréal, Montreal, QC H3C 3J7, Canada
                [4 ]National Research Council Canada (NRCC), Montreal, QC H3A 1A3, Canada; Feng.Ni@ 123456cnrc-nrc.gc.ca (F.N.); abedelnasser.abulrob@ 123456nrc-cnrc.gc.ca (A.A.)
                Author notes
                [* ]Correspondence: pierluctardif@ 123456gmail.com (P.-L.T.); frederic.lesage@ 123456polymtl.ca (F.L.); Tel.: +1-514-340-4711 (ext. 7542) (F.L.)
                Article
                ijms-17-02110
                10.3390/ijms17122110
                5187910
                27983695
                13bae4d3-125f-4ca4-bfa2-aca37234b247
                © 2016 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 31 August 2016
                : 09 December 2016
                Categories
                Article

                Molecular biology
                intravascular ultrasound (ivus),near-infrared fluorescence (nirf),atherosclerosis,ex vivo three-dimensional (3d) histology,optical coherence tomography (oct),confocal fluorescence microscopy

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