CC BY 4.0 · TH Open 2021; 05(02): e113-e124
DOI: 10.1055/s-0041-1725042
Original Article

Colocalization of Erythrocytes and Vascular Calcification in Human Atherosclerosis: A Systematic Histomorphometric Analysis

Elsa Wilma Böhm
1  Department of Cardiology, University Medical Center, Mainz, Germany
,
Maria Pavlaki
2  Department of Cardiology, Democritus University of Thrace, Alexandroupolis, Greece
,
Georgios Chalikias
2  Department of Cardiology, Democritus University of Thrace, Alexandroupolis, Greece
,
Dimitrios Mikroulis
3  Department of Cardiothoracic Surgery, Democritus University of Thrace, Alexandroupolis, Greece
,
George S. Georgiadis
4  Department of Vascular Surgery, Democritus University of Thrace, Alexandroupolis, Greece
,
Dimitrios N. Tziakas
2  Department of Cardiology, Democritus University of Thrace, Alexandroupolis, Greece
,
Stavros Konstantinides
2  Department of Cardiology, Democritus University of Thrace, Alexandroupolis, Greece
,
Katrin Schäfer
1  Department of Cardiology, University Medical Center, Mainz, Germany
› Author Affiliations
Funding This research was funded by the German Research Foundation (Deutsche Forschungsgemeinschaft, SCHA 808/9–1 and 9–2 to K.S.).

Abstract

Background Intimal calcification typically develops in advanced atherosclerosis, and microcalcification may promote plaque progression and instability. Conversely, intraplaque hemorrhage and erythrocyte extravasation may stimulate osteoblastic differentiation and intralesional calcium phosphate deposition. The presence of erythrocytes and their main cellular components (membranes, hemoglobin, and iron) and colocalization with calcification has never been systematically studied.

Methods and Results We examined three types of diseased vascular tissue specimens, namely, degenerative aortic valve stenosis (n = 46), atherosclerotic carotid artery plaques (n = 9), and abdominal aortic aneurysms (n = 14). Biomaterial was obtained from symptomatic patients undergoing elective aortic valve replacement, carotid artery endatherectomy, or aortic aneurysm repair, respectively. Serial sections were stained using Masson–Goldner trichrome, Alizarin red S, and Perl's iron stain to visualize erythrocytes, extracelluar matrix and osteoid, calcium phosphate deposition, or the presence of iron and hemosiderin, respectively. Immunohistochemistry was employed to detect erythrocyte membranes (CD235a), hemoglobin or the hemoglobin scavenger receptor (CD163), endothelial cells (CD31), myofibroblasts (SMA), mesenchymal cells (osteopontin), or osteoblasts (periostin). Our analyses revealed a varying degree of intraplaque hemorrhage and that the majority of extravasated erythrocytes were lysed. Osteoid and calcifications also were frequently present, and erythrocyte membranes were significantly more prevalent in areas with calcification. Areas with extravasated erythrocytes frequently contained CD163-positive cells, although calcification also occurred in areas without CD163 immunosignals.

Conclusion Our findings underline the presence of extravasated erythrocytes and their membranes in different types of vascular lesions, and their association with areas of calcification suggests an active role of erythrocytes in vascular disease processes.



Publication History

Received: 29 October 2020

Accepted: 18 January 2021

Publication Date:
14 April 2021 (online)

© 2021. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

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