Identification of apoptotic changes in osteocytes in normal and pathological human bone

Abstract

Previous work on bone growth and biomechanics suggests that osteocytes might sense the requirement for bone remodeling and signal to cells in the basic multicellular unit that undertake this function. The present study looked for evidence of apoptosis in human osteocytes in adult, pediatric, and pathological bone to compare these situations of differing levels of turnover and considered the possibility of a functional role for this death mechanism in bone modeling and remodeling. Apoptosis was identified in bone tissue by agarose gel electrophoresis of DNA (to demonstrate DNA ladders). In cryostat sections it was possible to visualize individual cells with fragmented DNA in situ using a modified nick translation technique (NT). In addition, visualization of apoptotic morphology was undertaken using light and electron microscopy. Adult femoral head and iliac crest bone showed no evidence of DNA ladders and very small numbers of osteocytes with DNA fragmentation using NT. In contrast, samples of pediatric calvaria, adult heterotopic bone, and osteophytes all displayed characteristic laddering of extracted DNA and showed evidence of potentially apoptotic osteocytes in situ using NT. In agreement with these findings, transmission electron microscopy showed numbers of osteocytes in infant calvaria with advanced chromatin condensation and cell shrinkage indicative of apoptosis. Since all three types of positive bone are involved in rapid matrix turnover, apoptotic changes in human osteocytes in vivo might be related in general terms to the modeling and remodeling activity level of the bone sampled. It was further found that the distribution of potentially apoptotic cells in the infant and pathological bone was anatomically nonuniform, raising the intriguing possibility of a functional relationship between bone turnover and the controlled cell death of osteocytes.