Background: Alcohol has been shown to be associated with
osteoporosis and osteonecrosis in patients and in animal models. Recent
studies have demonstrated that alcohol contributes to abnormal lipid
metabolism in the stromal cells of bone marrow, but the mechanisms have not
been defined. The purpose of this study was to evaluate the effects of alcohol
on the differentiation of a stem cell that was cloned from bone marrow.
Methods: D1 cells (cloned bone-marrow stem cells from a BALB/c
mouse) were treated either with increasing concentrations of ethanol (0.09,
0.15, and 0.21 mol/L) or without alcohol to serve as controls. Morphologic
features of the cells were monitored with use of a phase-contrast microscope.
Alkaline phosphatase activity was determined with use of a colorimetric assay.
The expression of genes that are indicators of adipogenesis [422(aP2),
PPAR?] and osteogenesis (osteocalcin) was evaluated using Northern blot
and reverse transcription-polymerase chain reaction assays.
Results: The cells treated with ethanol started to accumulate
triglyceride vesicles at day seven. The number of adipocytes and the
percentage of the area that contained the cells with fat vesicles increased
significantly (p < 0.05), and the level of alkaline phosphatase activity
diminished with longer durations of exposure to ethanol and with higher
concentrations. Analysis of gene expression showed diminished expression of
osteocalcin. This occurred without a significant increase in the expression of
either the fat-cell-specific gene 422(aP2) or PPAR? in cells treated
with ethanol, suggesting that adipogenesis may occur at a point downstream in
the fatty-acid-metabolism pathway.
Conclusions: Alcohol treatment decreases osteogenesis while
enhancing adipogenesis in a cloned bone-marrow stem cell, indicating that
alcohol abuse may be one of the mechanisms leading to osteoporosis and
osteonecrosis. This finding explains the clinical observation that there is
increased adipogenesis in alcohol-induced osteoporosis and osteonecrosis.
Clinical Relevance: The inhibition of bone-marrow adipogenesis and
the concomitant enhancement of osteogenesis may provide a novel approach to
the prevention or treatment of osteonecrosis and osteoporosis.