Prema B. Rapuri

Smoking and bone metabolism in elderly women

Cigarette smoking has been implicated as a risk factor for osteoporosis. In the present study, the relationship between smoking and bone mineral density, calcitropic hormones, calcium absorption, and biochemical indices related to bone and mineral metabolism was examined at baseline, in subjects recruited for an osteoporotic study. The subjects included 489 elderly women, aged 65-77 years. After exclusions (thiazide users), 54 women constituted the smoking group and 390 women were classified as nonsmokers. The effect of frequency of smoking was also examined in this population (33 light smokers [<1 pack/day] and 21 heavy smokers [>1 pack/day]). Adjusted mean total body bone mineral density was 4% lower (0.968 +/- 0.019 vs. 1.009 +/- 0.004) and the total hip density was 6% lower (0.778 +/- 0.024 vs. 0.826 +/- 0.006) in heavy smokers compared with nonsmokers. At the other sites measured (spine, midradius, femoral neck, trochanter, and Wards triangle), a similar nonsignificant trend was observed. The adjusted mean calcium absorption corrected for weight was lower (13%) both in light and heavy smokers compared with nonsmokers, and serum 25-hydroxyvitamin D was significantly lower (16%) in heavy smokers than nonsmokers. Serum parathryroid hormone (PTH) was higher in heavy smokers, but was not significantly different from that of nonsmokers. A significant increase in bone remodeling markers, serum osteocalcin (4.35 +/- 0.271 vs. 3.79 +/- 0. 066) and urine N-telopeptide/creatinine (NTx/Cr) ratio (74.5 +/- 5. 75 vs. 49.8 +/- 1.4) was seen in heavy smokers compared with nonsmokers. These results suggest that smoking lowers bone mineral density, and is a result of decreased calcium absorption associated with secondary hyperparathyroidism and increased bone resorption.

Effect of Vitamins D2 and D3 Supplement Use on Serum 25OHD Concentration in Elderly Women in Summer and Winter

Vitamin D2 and D3 are generally considered equipotent in humans. A few studies have reported that serum 25OHD levels are higher in vitamin D3- compared with vitamin D2-supplemented subjects. As both vitamin D2 and D3 supplements are commonly used by elderly in United States, in the present study we determined the effect of self-reported vitamin D2 and vitamin D3 supplement use on serum total 25OHD levels according to season in elderly women aged 65–77 years. Serum total 25OHD levels were determined in winter and summer in unsupplemented women (N = 307) and in women who reported taking vitamin D2 (N = 56) and vitamin D3 (N = 55) supplements by competitive protein binding assay. In vitamin D2-supplemented women, the contribution of vitamin D2 and D3 to the mean serum total 25OHD level was assessed by HPLC. In summer, there were no significant differences in the mean total serum 25OHD levels (ng/ml) among the vitamin D2 (32 ± 2.1), vitamin D3 (36.7 ± 1.95), and unsupplemented (32.2 ± 0.95) groups. In winter, the mean serum total 25OHD levels were higher in women on vitamin D2 (33.6 ± 2.34, P < 0.05) and vitamin D3 (29.7 ± 1.76, NS) supplements compared with unsupplemented women (27.3 ± 0.72). In vitamin D2-supplemented women, about 25% of the mean serum total 25OHD was 25OHD2, in both summer and winter. Twelve percent of unsupplemented women and 3.6% of vitamin D-supplemented women had a mean serum total 25OHD level below 15 ng/ml in winter. In elderly subjects, both vitamin D2 and Vitamin D3 supplements may contribute equally to circulating 25OHD levels, with the role of vitamin D supplement use being more predominant during winter.

Caffeine decreases vitamin D receptor protein expression and 1,25(OH)2D3 stimulated alkaline phosphatase activity in human osteoblast cells

Of the various risk factors contributing to osteoporosis, dietary/lifestyle factors are important. In a clinical study we reported that women with caffeine intakes >300 mg/day had higher bone loss and women with vitamin D receptor (VDR) variant, tt were at a greater risk for this deleterious effect of caffeine. However, the mechanism of how caffeine effects bone metabolism is not clear. 1,25-Dihydroxy vitamin D(3) (1,25(OH)(2)D(3)) plays a critical role in regulating bone metabolism. The receptor for 1,25(OH)(2)D(3), VDR has been demonstrated in osteoblast cells and it belongs to the superfamily of nuclear hormone receptors. To understand the molecular mechanism of the role of caffeine in relation to bone, we tested the effect of caffeine on VDR expression and 1,25(OH)(2)D(3) mediated actions in bone. We therefore examined the effect of different doses of caffeine (0.2, 0.5, 1.0 and 10mM) on 1,25(OH)(2)D(3) induced VDR protein expression in human osteoblast cells. We also tested the effect of different doses of caffeine on 1,25(OH)(2)D(3) induced alkaline phosphatase (ALP) activity, a widely used marker of osteoblastic activity. Caffeine dose dependently decreased the 1,25(OH)(2)D(3) induced VDR expression and at concentrations of 1 and 10mM, VDR expression was decreased by about 50-70%, respectively. In addition, the 1,25(OH)(2)D(3) induced alkaline phosphatase activity was also reduced at similar doses thus affecting the osteoblastic function. The basal ALP activity was not affected with increasing doses of caffeine. Overall, our results suggest that caffeine affects 1,25(OH)(2)D(3) stimulated VDR protein expression and 1,25(OH)(2)D(3) mediated actions in human osteoblast cells.

Falls are associated with decreased renal function and insufficient calcitriol production by the kidney

In a double blind placebo controlled 3-year osteoporosis study in elderly women, we collected prospective data on falls. The study population comprised 489 normal elderly women aged 65-77 years randomized to four groups: placebo, calcitriol 0.25 mcg b.i.d., conjugated equine estrogens (0.625 mg/day) and calcitriol+estrogen. Falls occurred in 57% of all women. Using a Poisson regression model, the placebo group with low GFR-creatinine clearance (CrCl<60 ml/min) had 60% more falls compared to the group with CrCl> or =60 ml/min. Further sub group analyses showed that there is no increased risk of falls with CrCl 60-70, 70-80 and >90 ml/min. Calcitriol treatment significantly reduced the number of falls by 50% (OR=0.5, 95% CI: 0.4-0.9, p=0.010) compared to placebo in the low CrCl group. The group with lower CrCl had lower calcium absorption (p<0.001), lower serum 1,25-dihydroxyvitamin D (1,25(OH)(2)D) (p<0.001) and normal serum 25OHD suggesting that there is decreased conversion of 25OHD to 1,25(OH)(2)D by the aging kidney. It is postulated that the decrease in falls on calcitriol therapy is related to an increase in serum 1,25(OH)(2)D, upregulation of VDR and improvement in muscle strength although one cannot exclude an effect on the central nervous system.

Association between Vitamin D receptor polymorphisms and the rate of bone loss in elderly women - Importance of adjusting for dietary and lifestyle factors

The association between the restriction length polymorphisms of the Vitamin D receptor (VDR) gene and the bone mineral density (BMD) or the rate of bone loss is still under debate. In a longitudinal study of untreated postmenopausal elderly women, we evaluated the relationship between the VDR gene polymorphisms (BsmI, TaqI, ApaI, and FokI) and the rate of bone loss over a 3-year period. We also examined the effect of adjustments for dietary and lifestyle factors on these associations. Before adjustments, the rate of femoral neck bone loss was - 3.76 +/- 1.58% in women with BB genotype and 0.45 +/- 0.65% in women with bb genotype, which was not significantly different. Upon adjustment for dietary and lifestyle factors, statistically significant (P = 0.03) bone loss was observed at femoral neck in women with BB genotype (- 3.66 +/- 2.44%) compared to that of bb genotype (2.39 +/- 1.32%). Similar results were observed with TaqI genotypes. The rates of bone loss at other skeletal sites were not different between VDR genotypes defined by BsmI and TaqI. VDR gene polymorphisms defined by ApaI and FokI were not related to the rate of bone loss.

Effect of vitamin D supplement use on serum concentrations of total 25OHD levels in elderly women

Vitamin D(2) and D(3) are generally considered equipotent in humans. As Vitamin D(2) supplements are commonly used by elderly in United States, we determined the contribution of 25OHD(2) to the total serum 25OHD levels by HPLC in elderly women who reported taking Vitamin D(2) supplements (n = 56) and also in a group of randomly selected unsupplemented women (n = 60). In addition, we compared the total serum 25OHD measured by HPLC with competitive protein-binding assay (CPBA), a method routinely employed to measure Vitamin D status. A correlation of 0.91 (P < 0.001) was observed between the two methods for the serum total 25OHD measurement. The mean serum 25OHD level in Vitamin D(2) supplemented group was significantly higher than in unsupplemented group measured by HPLC (32 versus 28 ng/ml) and marginally higher measured by CPBA (33 vs. 31 ng/ml). Seventy eight percent of women taking Vitamin D(2) supplements had appreciable amounts of circulating 25OHD(2,) which constituted about 25 percent of their total serum 25OHD. It is also interesting to note that Vitamin D deficiency was less prevalent in elderly women taking Vitamin D(2) supplements (1.8%) compared to women not taking any supplements (12%).