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Chronobiol Int. 2003 Mar;20(2):325-36.

Circadian rhythms in serum bone markers and their relation to the effect of etidronate in rats.

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Division of Pharmacology, Department of Oral Biology, Tohoku University Graduate School of Dentistry, Aoba-ku, Sendai, Japan.


Circadian rhythmicity is an essential feature of bone metabolism. The present study was undertaken to (Aoshima et al., 1998) determine the changes in bone resorption and formation in rats over 24h, (Black et al., 1999) evaluate the effect of the consecutive administration of etidronate on circadian rhythms of serum bone markers, and (Blumsohn et al., 1994) determine whether the effect of etidronate on bone metabolism is circadian time-dependent. One hundred twenty male Wistar rats, which had been adapted to a 12/12h light/dark cycle, were injected subcutaneously once daily with either 0.5 mgP/kg etidronate or 0.9% NaCl (control group) at 0090, 1300, 1700, 2100, 0100, or 0500h for 10d. Serum was collected and tibiae were dissected 24h after the last injection. Serum pyridinoline (Pyd), tartrate-resistant acid phosphatase (TRAP), osteocalcin (OC), alkaline phosphatase (ALP), calcium (Ca), phosphorus (Pi), calcitonin (CT), and parathyroid hormone (PTH) were determined. Bone mineral density (BMD) in the proximal tibia, and the rate of formation of longitudinal trabecular bone over the past 48h were also determined using a chronological labeling method with NTA-Pb. The results showed characteristic circadian rhythms in serum bone markers in rats, with peaks in both bone resorption and bone formation during the animals' rest span. The administration of etidronate at the different times of the day decreased the level of bone-resorption markers (Pyd and TRAP) without affecting the circadian patterns of markers of bone formation (OC and ALP). However, the magnitude of the decrease due to etidronate was not uniform throughout the day, and was greatest during the daytime. Etidronate increased the BMD in the tibial metaphysis in all of the time-treatment groups, but the magnitude of the increase did not vary with the time of etidronate administration. The present data provide a physiological basis for future studies on bone metabolism and may be important in the design of future experiments and in the interpretation of experimental data.

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