The effect of methotrexate on mouse bone cells in culture

Arthritis and Rheumatism
K P MayS G West

Abstract

We have recently shown that administration of long-term, low-dose methotrexate (MTX) causes severe osteopenia in female rats. This osteopenia is characterized both by decreased osteoblast function without a decrease in osteoblast numbers, and by increased bone resorption that is believed to represent a physiologic remodeling response by osteoclasts. The present study investigates the effects of varying doses of MTX on mouse bone cells in culture. Cells were obtained by sequential digestion of neonatal mouse calvariae, and cultured with fetal calf serum (10% for osteoblast-like cells and 2% for osteoclast-like cells). After 1 week, MTX was added to each culture in concentrations of 0.6 microM, 0.4 microM, 0.2 microM, 0.1 microM, 1 nM, and 0.5 nM. All experiments were done on 24 wells for each MTX concentration and for the controls. The effect on osteoblastic cells was assessed, at 7 days, by cell counts and by measurement of lysate alkaline phosphatase and supernatant osteocalcin levels, and, at 21 days, by analysis of the calcified matrix production, which was cultured with ascorbic acid and beta-glycerophosphate. For osteoclastic cells, cell count and lysate acid phosphatase levels were determined. Levels of osteoblastic cells...Continue Reading

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Citations

Sep 8, 2012·Osteoporosis International : a Journal Established As Result of Cooperation Between the European Foundation for Osteoporosis and the National Osteoporosis Foundation of the USA·B F RicciardiJ M Lane
Feb 18, 2004·Clinica Chimica Acta; International Journal of Clinical Chemistry·Samia I GirgisRobert J Shiner
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Jan 22, 2013·The Journal of Foot and Ankle Surgery : Official Publication of the American College of Foot and Ankle Surgeons·Maria J BischoffTom M van Raaij
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Dec 18, 2001·Biopharmaceutics & Drug Disposition·M P IqbalS Pervez
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