Immediately after removal, harvested endarterectomy specimens were placed in 10% formaldehyde. Representative parts of the specimen were cross-sectioned in approximately 4 mm thick samples. In further processing, the samples were decalcified by a hydrochloric acid solution and embedded in paraffin. The samples were cut into five-micron-thick tissue sections. Xylene was used as a deparaffinization agent, and graded alcohol was used for the hybridization of the tissue sections. For staining parallel sections, hematoxylin and eosin with the van Gieson/orcein method were used. The indirect immunohistologic method was used for the detection of endothelial cells (CD31 marker, primary mouse anti-human monoclonal antibody and clone JC70A) and macrophages (CD68 marker, primary mouse anti-human monoclonal antibody and clone PG-M1). All histological analyses were performed by one experienced pathologist (VM) using a bright-field optical microscope (Nikon Eclipse E 400).
Endarterectomy specimens were scanned for multiple histological features, including eccentricity, the presence of atheromatous or fibrous tissue, calcification, myxoid change, hemorrhage, thrombosis, inflammation, foamy macrophage, giant cell reaction, hemosiderin, neovascularisation or ossification (TAB 1). All specimens were divided into AHA groups IV/V, VIII, or VI, according to the AHA classification [9 (link)]. Plaques in the AHA VI group were gathered in the group of unstable plaques.
Endarterectomy specimens were scanned for multiple histological features, including eccentricity, the presence of atheromatous or fibrous tissue, calcification, myxoid change, hemorrhage, thrombosis, inflammation, foamy macrophage, giant cell reaction, hemosiderin, neovascularisation or ossification (TAB 1). All specimens were divided into AHA groups IV/V, VIII, or VI, according to the AHA classification [9 (link)]. Plaques in the AHA VI group were gathered in the group of unstable plaques.
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