Fat cells contribute not only to systemic lipid metabolism, but also to osteogenesis and hemopoiesis in the bone marrow. The present study represents the first attempt to culture mature unilocular fat cells of the bone marrow. Two methods devised in our laboratory were employed: one is the "ceiling culture method" that utilizes the floating property of the cells; the other, a three-dimensional collagen gel matrix culture that captures unilocular fat cells in the gel matrix. Using these methods, the proliferation of unilocular fat cells from the bovine metacarpal bone marrow was demonstrated. First, we confirmed the proliferative ability of unilocular fat cells derived from the bone marrow, using autoradiography to study 3H-thymidine incorporation into the nuclei. The unilocular fat cells de-differentiated into fibroblast-like fat cells and then proceeded to proliferate. When they underwent a contact inhibition of growth, re-differentiation from fibroblast-like fat cells into unilocular fat cells occurred at a high rate. A specific enzymatic marker of the fat cell, alpha-glycerophosphate dehydrogenase activity related to lipogenesis, was then demonstrated in the cultured fat cells. We examined the functional reactivity of the fat cells by treatment with insulin and cyclic-AMP, and both lipogenesis and lipolysis were also confirmed in them. We concluded that unilocular fat cells from the bone marrow de-differentiated, proliferated and re-differentiated in culture. The present results may help to clarify the various functions of fat cells in the bone marrow.
We compared histological and functional findings in rapidly destructive coxarthrosis (RDC) and slowly progressive osteoarthritis (OA) to investigate whether osteoclasts contribute to the extensive bone destruction observed in RDC. A histological analysis of tissue specimens from the synovium obtained from 10 cases of RDC and 40 cases with OA of the hip was performed after staining for tartrate-resistant acid phosphatase (TRAP). The cells isolated from these tissue specimens from the synovium were cultured for 24 h, and the numbers of TRAP-positive giant cells were counted. Thereafter, we performed a resorption pit formation assay by isolated cells cultured on dentine slices for 7 days. The number of TRAP-positive multinuclear giant cells present in the synovial membrane obtained from RDC patients was significantly larger than that obtained from OA patients. Large lacunar resorption pits were only seen on the dentin slices in a culture of isolated cells from RDC patients without any stimulators. This is the first report, to our knowledge, to reveal that mature and activated osteoclasts exist only in the synovium of RDC and not in the OA synovium. This result might suggest that the underlying mechanism of RDC is therefore associated with osteoclastogenesis in the synovium.
The pathogenesis of corticosteroid-induced femoral head necrosis is assumed to be related to lipid metabolism. Mature fat cells are believed to play a central role in lipid metabolism. The purpose of this study was to investigate the size of mature fat cells in the human femoral head after steroid treatment. Cancerous bone tissue was obtained from the femoral heads of 20 women who had undergone total hip arthroplasty. This bone tissue was subsequently incubated in a medium containing 10(-7) or 10(-5) M dexamethasone for 5 days. Mature fat cells from the bone marrow were observed by scanning electron microscopy, and the largest diameter of individual fat cells was measured. The size of the mature fat cells in human bone marrow increased after high-dose steroid treatment. The largest fat cell volume after steroid treatment was one and one-half times larger than that observed in the control. Steroid-induced osteonecrosis is known to sometimes occur after high-dose steroid treatment. These findings may indicate the pathogenetic factors in the early stage of steroid-induced osteonecrosis.
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