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Calcification of decalcified bone grafts in rat: a histochemical study.

When EDTA-decalcified bone of rat was subcutaneously implanted to allogeneic hosts, calcification of osseous and cartilage matrices proceeded as an accumulation of oval spheres of calcium deposits. The spheres were PAS-positive but not strongly basophilic, and surrounded by fine net of collagen fibrils. Treat-ments of decalcified grafts with toluidine blue, cetyl pyridinium chloride or cetyltrimethylammonium bromide, all of which easily bind with acid mucopoly-saccharides, inhibited exposure of fibrils and suppressed calcification. Subsequent bathing in a high concentration of CaCl2 restored the original calcifiability. Bathing of decalcified grafts in low concentration of CaCl2 rendered tendons, ligaments, chondrocytes and muscles calcifiable. The calcified loci were also PAS-positive. The results mentioned above that mucopolysaccharides have a close relationship in the process of calcification.