An enzyme histochemical study of human salivary duct calculi

An investigation was undertaken to study the distribution of enzymes associated with submandibular gland salivary calculi. Ten calculi were freeze-sectioned and incubated for acid and alkaline phosphatases and for lactate, succinate and maleate dehydrogenases. All calculi were partly covered by a 50-210 micrometers wide zone of organic material consisting of connective tissue and metaplastic squamous epithelium facing the mineralized calculus, or of a structureless substance attached to the mineralized calculus. The epithelium showed an intense staining reaction for acid phosphatase and lactate dehydrogenase and a moderate reaction for succinate dehydrogenase throughout all levels of the epithelium. The structureless peripheral zone exhibited a moderate activity of acid phosphatase and succinate dehydrogenase located to an area close to the mineralized matrix. Also alkaline phosphatase and lactate dehydrogenase were found in a special pattern in the structureless zone. Sodium fluoride and sodium vanadate added to the incubation medium inhibited acid phosphatase activity whereas cupric chloride only lowered the staining reaction. Enzyme activity was found only within the peripheral zone of organic material with one exception. The results suggest that the calcification process of salivary calculi is not a passive calcification of necrotic material or mucin but rather an active process promoted by enzymes in the surrounding organic substances.