Histochemical and biochemical determination of calcium in pleomorphic adenoma

Although calcification seldom occurs in pleomorphic adenoma, it often occurs in salivary glands, and so we decided to investigate the possible role of calcium in this difference. A histochemical method using glyoxal bis(2-hydroxyanil) demonstrated a small amount of calcium outlining lumina and separated cells of epithelial structures and associated with cells of myxoid and chondroid regions in pleomorphic adenoma, and a conspicuous amount in the acini of the associated salivary glands. A biochemical method using dry ashing demonstrated a significantly higher level of calcium in the glands than in pleomorphic adenoma. The results indicate that the calcium is mainly associated with secretory granules, which are scarce in pleomorphic adenoma, and with proteoglycan present intercellularly and in stromal regions of pleomorphic adenoma. The calcium in secretory granules is of possible importance in calcification in lumina and epithelium, and that bound to proteoglycan is possibly released following necrosis to be of importance in stromal calcification. However, the overall low level of calcium in pleomorphic adenoma is the likely explanation for the usual lack of calcification.