ScienceOpen: research and publishing network

For Researchers

Search
Advanced search

30

views

    

0

recommends

0

shares

Record: found
Abstract: found
Article: not found

Detecting microcalcifications in atherosclerotic plaques by a simple trichromic staining method for epoxy embedded carotid endarterectomies

review-article

Author(s): M. Relucenti ¹ , R. Heyn ¹ , L. Petruzziello ¹ , G. Pugliese ² , M. Taurino ³ , G. Familiari ¹

Publication date (Electronic): 26 September 2010

Journal: European Journal of Histochemistry : EJH

Publisher: PAGEPress Publications

Keywords: atherosclerosis, carotid endarterectomy, calcification, polychromatic stain, epoxy resin, transmission electron microscopy.

Read this article at

ScienceOpenPublisher PMC

There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

Abstract

Atherosclerotic plaques have a high probability of undergoing rapid progression to stenosis, becoming responsible of acute coronary syndrome or stroke. Microcalcifications may act as enhancers of atherosclerotic plaque vulnerability. Considering that calcifications with a diameter smalller than 10 µm in paraffin embedded tissue are rather difficult to detect, our aim was to analyze microcalcifications on semithin sections from epoxy resin embedded samples of carotid endarterectomies using an original trichromic stain (methylene blue-azur B - basic fuchsine - alizarin red). We have compared samples stained either with our method, methylene blue-azur B alone or with Von Kossa staining, and methylene blue-azur B -basic fuchsine alone or with Von Kossa staining.

Our method resulted to be simple and fast (ca. 2 min), it gives a sharp general contrast for all structures and allows to easy identify collagen and elastin. In addition, gray-green colour associated to intracellular lipid droplets evidences foam cells, which are particularly abundant in endarterectomies samples. Mast cells and their metachromatic granules are also well recognized. Calcifications over 0,5 µm are clearly recognizable. In conclusion, microcalcifications are clearly distinguished from the extracellular matrix in spite of their reduced dimensions. Methylene blue-azur B-basic fuchsine-alizarin red method is easy to use, reproducible, and is particularly suitable for the identification of microcalcifications in the morphological analysis of atherosclerotic plaques.

Related collections

Most cited references 25

Record: found
Abstract: found
Article: not found

From vulnerable plaque to vulnerable patient: a call for new definitions and risk assessment strategies: Part I.

Morteza Naghavi, Peter Libby, Erling Falk … (2003)

Atherosclerotic cardiovascular disease results in >19 million deaths annually, and coronary heart disease accounts for the majority of this toll. Despite major advances in treatment of coronary heart disease patients, a large number of victims of the disease who are apparently healthy die suddenly without prior symptoms. Available screening and diagnostic methods are insufficient to identify the victims before the event occurs. The recognition of the role of the vulnerable plaque has opened new avenues of opportunity in the field of cardiovascular medicine. This consensus document concludes the following. (1) Rupture-prone plaques are not the only vulnerable plaques. All types of atherosclerotic plaques with high likelihood of thrombotic complications and rapid progression should be considered as vulnerable plaques. We propose a classification for clinical as well as pathological evaluation of vulnerable plaques. (2) Vulnerable plaques are not the only culprit factors for the development of acute coronary syndromes, myocardial infarction, and sudden cardiac death. Vulnerable blood (prone to thrombosis) and vulnerable myocardium (prone to fatal arrhythmia) play an important role in the outcome. Therefore, the term "vulnerable patient" may be more appropriate and is proposed now for the identification of subjects with high likelihood of developing cardiac events in the near future. (3) A quantitative method for cumulative risk assessment of vulnerable patients needs to be developed that may include variables based on plaque, blood, and myocardial vulnerability. In Part I of this consensus document, we cover the new definition of vulnerable plaque and its relationship with vulnerable patients. Part II of this consensus document focuses on vulnerable blood and vulnerable myocardium and provide an outline of overall risk assessment of vulnerable patients. Parts I and II are meant to provide a general consensus and overviews the new field of vulnerable patient. Recently developed assays (eg, C-reactive protein), imaging techniques (eg, CT and MRI), noninvasive electrophysiological tests (for vulnerable myocardium), and emerging catheters (to localize and characterize vulnerable plaque) in combination with future genomic and proteomic techniques will guide us in the search for vulnerable patients. It will also lead to the development and deployment of new therapies and ultimately to reduce the incidence of acute coronary syndromes and sudden cardiac death. We encourage healthcare policy makers to promote translational research for screening and treatment of vulnerable patients.

0 comments Cited 446 times – based on 0 reviews      Review now

Record: found
Abstract: found
Article: not found

A hypothesis for vulnerable plaque rupture due to stress-induced debonding around cellular microcalcifications in thin fibrous caps.

Yuliya Vengrenyuk, S Carlier, P Ganatos … (2006)

In this article, we advance a hypothesis for the rupture of thin fibrous cap atheroma, namely that minute (10-mum-diameter) cellular-level microcalcifications in the cap, which heretofore have gone undetected because they lie below the visibility of current in vivo imaging techniques, cause local stress concentrations that lead to interfacial debonding. New theoretical solutions are presented for the local stress concentration around these minute spherical inclusions that predict a nearly 2-fold increase in interfacial stress that is relatively insensitive to the location of the hypothesized microinclusions in the cap. To experimentally confirm the existence of the hypothesized cellular-level microcalcifications, we examined autopsy specimens of coronary atheromatous lesions using in vitro imaging techniques whose resolution far exceeds conventional magnetic resonance imaging, intravascular ultrasound, and optical coherence tomography approaches. These high-resolution imaging modalities, which include confocal microscopy with calcium-specific staining and micro-computed tomography imaging, provide images of cellular-level calcifications within the cap proper. As anticipated, the minute inclusions in the cap are very rare compared with the numerous calcified macrophages observed in the necrotic core. Our mathematical model predicts that inclusions located in an area of high circumferential stress (>300 kPa) in the cap can intensify this stress to nearly 600 kPa when the cap thickness is <65 microm. The most likely candidates for the inclusions are either calcified macrophages or smooth muscle cells that have undergone apoptosis.

0 comments Cited 123 times – based on 0 reviews      Review now

Record: found
Abstract: found
Article: not found

von Kossa staining alone is not sufficient to confirm that mineralization in vitro represents bone formation.

N Camacho, M Dallas, W Dallas … (2003)

Numerous techniques are currently used to characterize biological mineralization in intact tissues and cell cultures; the von Kossa staining method, electron microscopic analysis (EM), X-ray diffraction, and Fourier transform infrared spectroscopy (FTIR) are among the most common. In this study, we utilized three of these methods to compare the mineralization of cultured fetal rat calvarial cells (FRC) and the osteoblast cell lines 2T3 and MC3T3-E1 with the in vivo mineral of rat calvarial bone. The cells were cultured with or without ascorbic acid (100 microg/ml) and beta-glycerophosphate (2.5, 5, or 10 mM betaGP), and harvested between 16 and 21 days (FRC cells and 2T3 cells) or at 30 days of culture (MC3T3-E1 cells). In the FRC cultures, maximal von Kossa staining was observed with 2.5 and 5 mM betaGP in the presence of 100 microg/ml ascorbate. FRC cells also showed some von Kossa staining when cultured with bGP alone. In contrast, maximal von Kossa staining for MC3T3-E1 cells was observed with 10 mM betaGP. Only the cultures of MC3T3-E1 cells that received both ascorbate and betaGP produced von Kossa positive structures. The 2T3 cultures produced von Kossa positive staining only upon treatment with ascorbic acid and betaGP, which was greatly accelerated by bone morphogenic protein-2 (BMP-2). FTIR was performed on the mineral and matrix generated in FRC, MC3T3, and 2T3 cultures, and the results were compared with spectra derived from 16-day-old rat calvaria. The mineral-to-matrix ratios calculated from FTIR spectra for rat calvaria ranged from 2.97 to 7.44. FRC cells made a bonelike, poorly crystalline apatite, and, with increasing betaGP, there was a statistically significant (P

0 comments Cited 59 times – based on 0 reviews      Review now

Author and article information

Journal

Journal ID (nlm-ta): Eur J Histochem

Journal ID (pmc): EJH

Journal ID (publisher-id): EJH

Title: European Journal of Histochemistry : EJH

Publisher: PAGEPress Publications (Pavia, Italy )

ISSN (Print): 1121-760X

ISSN (Electronic): 2038-8306

Publication date (Electronic): 26 September 2010

Publication date Collection: 26 September 2010

Volume: 54

Issue: 3

Electronic Location Identifier: e33

Affiliations

[1 ]Laboratory of Electron Microscopy Pietro M. Motta, Department of Human Anatomic, Histologic, Forensic and Locomotor Apparatus Sciences Sapienza University of Rome, Italy

[2 ]Department of Clinical Sciences, II Medical School, S. Andrea Hospital, Sapienza University of Rome, Italy

[3 ]Department of Vascular Surgery, II Medical School, S. Andrea Hospital, Sapienza University of Rome, Italy

Author notes

Correspondence: Dr. Michela Relucenti, Laboratory of Electron Microscopy Pietro M. Motta, Department of Human Anatomy, Sapienza University of Rome, via Alfonso Borelli 50, 00161 Rome, Italy. Tel. +39.06.49918012 - Fax +39.06.49918070. E-mail: michela.relucenti@ 123456uniroma1.it

Authors' contribution: all authors have contributed equally to this work.

Article

Publisher ID: ejh.2010.e33

DOI: 10.4081/ejh.2010.e33

PMC ID: 3167311

PubMed ID: 20819772

SO-VID: 140b8ed9-532c-4ec2-b2cb-faa48028ba00

Copyright © ©Copyright M. Relucenti et al., 2010

License:

This work is licensed under a Creative Commons Attribution 3.0 License (by-nc 3.0).

Licensee PAGEPress, Italy

History

Date received : 18 March 2010

Date accepted : 3 June 2010

Categories

Subject: Technical Report

ScienceOpen disciplines: Clinical chemistry

Keywords: polychromatic stain,calcification,atherosclerosis,transmission electron microscopy.,epoxy resin,carotid endarterectomy

Data availability:

ScienceOpen disciplines: Clinical chemistry

Keywords: polychromatic stain, calcification, atherosclerosis, transmission electron microscopy., epoxy resin, carotid endarterectomy

Comments

Comment on this article

scite_

Similar content 407

See all similar

Cited by 6

See all cited by

Most referenced authors 210

See all reference authors