Involvement of hydrogen peroxide in the differentiation of clonal HD-11EM cells into osteoclast-like cells.

The present study uses the osteoclast precursor clonal line, HD-11EM, to study the potential of hydrogen peroxide (H2O2) in mediating the differentiation of HD-11EM into osteoclast-like cells. HD-11EM cells are a newly established clonal cell line that, in response to 1alpha,25-(OH)2D3, differentiate into osteoclast-like cells that are multinucleated (more than three nuclei), express tartrate-resistant acid phosphatase (TRAP), and excavate resorption pits when cultured on dentin slices in the presence of osteoblasts (Hsia et al., 1995, J. Bone Miner. Res., 10(Suppl 1):S424; Hsia, and Hauschka, 1997, unpublished data). Here we demonstrate that HD-11EM express the reduced nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase specific cytochrome b558 subunits, and that stimulation of HD-11EM with 1 or 10 nM 1alpha,25-(OH)2D3 increases the extracellular release of H2O2 within 5-10 min. Ours is the first report that stimulation of a cell with 1alpha,25-(OH)2D3 enhances the activation of NADPH-oxidase and increases the basal release of superoxide and the formation of its dismutation product, H2O2. To determine the possible involvement of H2O2 in the differentiation of HD-11EM, these cells were exposed to glucose/glucose oxidase. This enzyme system was used to deliver a pure and continuous source of H2O2 in nanomole amounts consistent with quantities produced by HD-11EM in response to 1alpha,25-(OH)2D3. Both 1alpha,25-(OH)2D3 and the exogenously generated H2O2 stimulated a dose- and time-dependent increase in TRAP activity/cell and the number of multinucleated cells 24-48 hr after treatment. Northern analysis confirmed an increase in expression of TRAP mRNA in response to either 1alpha,25-(OH)2D3 or H2O2. Decreases in cell proliferation and v-myc mRNA were also observed in response to these agents. Taken together, our findings indicate that production of H2O2 by HD-11EM is an important local factor involved in differentiation of HD-11EM into osteoclast-like cells, and suggest that H2O2 may play a role in native osteoclast differentiation.

Hypertension--unexpected research results and program redirection.

The California Department of Health Services (DHS) has conducted two statewide surveys to evaluate the effectiveness of a coordinated hypertension control program. Unfortunately, the expected reduction in the prevalence of elevated blood pressure did not occur. Public knowledge about hypertension did increase, but among hypertensive persons awareness, treatment, and control did not improve. The responses to several survey questions were examined to determine why there was no improvement in the rate of control of elevated blood pressure. Obviously, many persons with hypertension have not complied with medical advice at various points in the hypertension control system. Consequently, the DHS is redirecting its Hypertension Control Program. Less emphasis is being placed on the coordination of blood pressure control resources, including public education, while more emphasis is being placed on increasing compliance with treatment recommendations, focusing on patient tracking as described by the National High Blood Pressure Education Program and long-term adherence to therapy as discussed in the 1984 Report of the Joint National Committee on Detection, Evaluation, and Treatment of High Blood Pressure.

Using the results of hypertension research in program redirection.

This is one of a series of articles from western state public health departments.