Zinc mapping in bone tissues by histochemistry and synchrotron radiation–induced x-ray emission: correlation with the distribution of alkaline phosphatase

Zinc distribution in osteons was mapped by synchrotron radiation-induced X-ray emission analysis in both human and porcine adult bone, as well as in porcine bone by histochemistry using Timm’s method. Both procedures showed that zinc is not uniformly distributed, being in its highest concentration on haversian bone surfaces. When Timm’s method was applied in conjunction with a procedure leading to partial zinc extraction, three zinc pools were specifically detected: a loose one, found in the mineralizable osteoid; a mineral one, bound to the bone mineral; and a tenacious one, firmly bound to an organic component located in the osteoid and mineralizing organic matrix. The alkaline phosphatase distribution was also mapped in porcine adult bone by histochemistry and immunohistochemistry and it was found codistributed with tenacious zinc mainly at the calcification front. The data suggest that alkaline phosphatase is buried as a bone matrix protein during initial mineralization.