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Von Kossa stain, immunohistochemistry staining for osteopontin and anti-CD68 stains in diet, control, and reversibility group valves. Von Kossa stain (A) for calcium aggregates (brown) demonstrates calcification only in diet group (white arrow). Staining for osteopontin (B) in orange (white arrow) is expressed only in diet group. Staining using anti-CD68 antibody (C) for macrophages (in green) and marked with white arrow is demonstrated only in diet group.
