Bone and mineral最新文献

Carbonic anhydrase (CA) which catalyzes the reversible hydrolysis of carbon dioxide is known to be important in osteoclastic bone resorption, however, suggested roles in calcium phosphate mineral formation have not been previously demonstrated. Biochemical evidence is provided for the presence of CA in growth plate matrix vesicles (MV) and the level of activity determined by enzyme assay. Inhibition of CA activity with the specific inhibitor acetazolamide resulted in reduced rates of MV mineralization. Other inhibitor studies showed that MV mineralization was also impaired by 4,4-diisothiocyanatostilbene-2, 2-disulfonic acid (DIDS), a blocker of membrane bicarbonate channels. No evidence was found for the presence of any proton pumps or channels. When acetazolamide and DIDS were combined, their inhibitory effects on MV mineralization were additive. These findings suggest that MV posess a pH regulation system composed of carbonic anhydrase and a putative bicarbonate channel. This system may function in the MV by providing intraluminal buffering capacity. The control of intravesicular pH is important for the stabilization of the acid-labile nuclea-tional core complex and in preventing the build-up of protons during calcium phosphate phase transformations.

Previously we reported that intestinal cells contain estrogen receptors, and that 17β-estradiol enhanced calcium uptake by these cells in vitro. The current study was undertaken to examine the in vivo effects of 17β-estradiol on intestinal absorption of calcium and phosphorus. Three groups of rats were studied. Group 1 received solvent vehicle. Groups 2 and 3 received 5 μg and 40 μg 17β-estradiol/kg body weight/day, respectively, for 21 days. Hormone and solvent vehicle injections were given subcutaneously. Rats were fed a Teklad diet containing 0.4% Ca, 0.3% P and 3.0 U vitamin D/g during the study. Intestinal absorption of calcium and phosphorus was assessed over a 5-day period from day 15–19. Carmine red (25 mg/100 g diet) was added to the rat feed to mark the beginning and end of fecal collections. Administration of 17β-estradiol caused an increase in intestinal absorption of calcium and phosphorus. The increase was significant only for calcium, and in the animals that received high-dose 17β-estradiol (P < 0.05). Serum calcium and phosphorus levels were significantly greater in 17β-estradiol treated than in control animals. The urinary excretion of calcium and phosphorus was also increased in a dose-dependent manner by 17β-estradiol, and was significant for both calcium and phosphorus in animals that received high-dose 17β-estradiol (P < 0.05). In contrast, 17β-estradiol treatment did not significantly alter the serum levels of parathyroid hormone and l,25(OH)₂vitamin D. These findings indicate that estrogen administration promotes intestinal absorption of calcium in vivo. The enhanced calcium absorption, in spite of unaltered serum l,25(OH)₂vitamin D levels, suggests that estrogen does not promote calcium absorption mainly by increasing the circulating levels of l,25(OH)₂vitamin D.

A long-term investigation of bone mineral metabolism in a newly developed strain, the WBN/Kob rat, which spontaneously develops diabetes, possibly due in part to hemosiderin deposition, was conducted. WBN/Kob rats used in this study developed diabetes after 9 months of age. Bone mass peaked at 6 months or 8 months of age, and femoral breaking strength was maximal at 8 months of age, declining rapidly after the development of diabetes. In contrast, both the bone mass and the mechanical strength increased up to 14 months of age in controls. The serum osteocalcin (BGP) levels were lower at 4 months of age and serum l,25(OH)₂D levels were significantly lower throughout the study in WBN/Kob rats than in controls. These results suggest that abnormal bone and mineral metabolism is present in WBN/Kob rats before the onset of diabetes, and that bone strength and BMD decrease simultaneously with the development of diabetes. This strain can serve as a useful model, not only of hemosiderosis and diabetes, but also of osteopenia.

The Damara mole-rat, Cryptomys damarensis, has no access to obvious dietary or endogenous sources of vitamin D. We tested the hypotheses that mineral metabolism in these animals is independent of vitamin D status but rather is affected by dietary calcium (Ca) content. Furthermore, we questioned whether bone and teeth assist in plasma mineral homeostasis. Mole-rats increased Ca intake when dietary Ca content increased; however, mode of gastrointestinal uptake, vitamin D metabolite and plasma Ca concentrations were not altered. Similarly, oral vitamin D supplementation did not affect gastrointestinal Ca absorption or alter plasma Ca concentration, although significant increases in plasma concentrations of vitamin D were evident. Bone and teeth mineral (Ca and P_i) content were augmented with vitamin D supplementation. Mineral homeostasis was primarily maintained by manipulating mineral deposition in teeth, for mineral content in teeth increased significantly when dietary Ca content changed from 1.3 g/kg to 2.6 g/kg and higher. Mineral homeostasis in these subterranean rodents does not appear to be regulated at the level of the intestine, but rather by manipulating bone and teeth mineral reservoirs.

A semi-anthropomorphic ‘distal radius like’ phantom, developed by Kalender and Ruegsegger for use in peripheral bone densitometry using single photon (DPA) dual X-ray (DXA) and quantitative computed tomography (QCT) machines, has been studied with a view to cross-calibrating different types and brands of densitometers in current use. In the context of an EU ‘Concerted Action’ (second Framework Programme) the phantom was repeatedly measured on six SPA machines, three DXA machines and nine QCT machines (545 measurements). Linear regression equations were derived, individual to each machine, which allowed the derivation of ‘standardized densities’. In this way we converted measurements made by machines of the same modality to a common scale of measurements. Two machines (one DXA, one SPA) showed statistically significant instability over time emphasising the need for rigorous quality control in the application of densitometry. In other respects these results provide an encouraging basis for the derivation of standardized normative ranges and the more effective use of peripheral densitometry in future clinical and epidemiological studies.

Interleukin-4 (IL-4) potently inhibits bone resorption by preventing the differentiation of osteoclast precursors to osteoclasts. To elucidate the role of IL-4 in bone formation, we studied the effects of human IL-4 on human osteoblast-like cells obtained from trabecular bone, which showed increased osteocalcin production in response to l,25-(OH)₂D₃ in more than 10 passages. IL-4 stimulated the proliferation of osteoblast-like cells in a concentration-dependent manner, showing the minimal and maximal stimulatory effects at 10 pg/ml and 100–1000 pg/ml, respectively. IL-4 also stimulated the expression of alkaline phosphatase mRNA (1.7-fold) and the enzyme activity to the same extent at 10–100 pg/ml. Furthermore, IL-4 stimulated collagen type I mRNA expression in human osteoblast-like cells. The cytokine did not affect osteocalcin production in a short culture period (3 days). These in vitro findings suggest that IL-4, a bone-resorption-inhibitory cytokine produced by activated T cells in bone marrow, may exert an anabolic effect on osteoblast-like cells in trabecular bone through a paracrine mechanism.

Osteoporosis is a well-known side-effect of chronic treatment with glucocorticoids. We have studied vertebral bone mineral density (BMD) and biochemical markers of bone metabolism in 30 patients diagnosed of Addison's disease (AD) to determine the effect of long-term replacement treatment with hydrocortisone (30 mg/day) or prednisone (7.5 mg/day). Lumbar bone mineral density was measured with dual energy X-ray absorptiometry in L-1–4 in two occasions, separated by 12 months. BMD in premenopausal women and men with AD was similar to healthy controls and postmenopausal women had slightly lower results. Rate of change of bone density followed up over a period of 12 months was −0.82%. Bone loss was not influenced by duration or type of steroid treatment. Biochemical parameters, serum calcium, alkaline phosphatase, osteocalcin, procollagen type I, PTH and 25(OH)vitamin D were within normal limits. Our results show that in patients with AD, after replacement with low doses of glucocorticoids there is no significative trabecular bone loss neither modifications in bone formation markers.

A specific HPLC system was developed to assess urinary excretion of collagen crosslinks (pyridinoline (Pyr) and deoxypyridinoline (D.Pyr)) in two models of osteopenia in rats, ovariectomy and adjuvant polyarthritis. The sensitivity of this method was in the picomolar range. In ovariectomized rats, a specific model of bone resorption, Pyr and D.Pyr levels rose early, reaching a peak 2 weeks after surgery. Both levels remained raised during the whole observation period (6 weeks) with no change in the Pyr/D.Pyr ratio. So, in this high bone turnover model, hyperresorption is reflected by the parallel increase of both crosslinks resulting in a significant decrease of bone mineral density (BMD) at 6 weeks (−7.3% vs. control). In polyar-thritic rats, in the 2 post-adjuvant weeks, Pyr levels increased in parallel with inflammatory parameters, whereas D.Pyr levels remained unchanged. This is in agreement with our previous report that at the end of the 2nd week after adjuvant there is no change in bone resorption. From the 3rd week, both Pyr and D.Pyr increased. The Pyr/D.Pyr ratio was always significantly higher in polyarthritic rats. These results suggest that the early increase of Pyr level reflects non-osseous collagen breakdown and that bone resorption occurs at a later stage when D.Pyr rises, leading to a dramatic decrease of BMD at 4 weeks (−17.7% vs. control). Taken together, our results suggest that in rat as in human, urinary Pyr is a marker of bone and cartilage breakdown, whereas D.Pyr is a specific marker of bone loss. This automated method described may constitute a very useful tool to evaluate bone and/or cartilage breakdown in rats and for the assessment of protective treatments.