Dataset Information

Bone development in growing female mice fed calcium and vitamin D at lower levels than is present in the AIN-93G reference diet.

ABSTRACT:

Background

The AIN-93G reference (REF) diet is used to allow the comparison within and between studies of different research groups but its levels of vitamin D (vit D) and calcium (Ca) may be higher than required for healthy bone structure and bone mineral density (BMD).

Objective

To determine if lower dietary levels of Ca (3.5, 3 or 2.5?g?Ca/kg diet) at 1 of 2 levels of vit D (100 or 400?IU/kg diet) supports similar development of bone structure and BMD compared to AIN-93G reference (REF) diet in female CD-1 mice at 2 and 4?months of age.

Methods

Within a trial, weanling female mice (n?=?12-15/group) were randomized to 1 of 4 diets until necropsy at 4?months of age: Trial 1: 100?IU?vit D/kg?+?3.5, 3 or 2.5?g?Ca/kg diet or 1000?IU?vit D/kg?+?5?g?Ca/kg diet (REF); and Trial 2: 400?IU?vit D/kg?+?3.5, 3 or 2.5?g?Ca/kg diet or 1000?IU?vit D/kg?+?5?g/kg diet (REF). At age 2 and 4?months, in vivo bone structure and BMD were assessed using micro-computed tomography (?CT) at the proximal and midpoint tibia. At age 4?months, lumbar vertebra 4 (L4) and mandible structure were analyzed ex vivo, femur strength at midpoint and neck was assessed and serum 25(OH)D₃ and PTH were quantified.

Results

For Trial 1 (100?IU?vit D/kg), there were no differences in tibia structure at age 2 and 4?months nor L4 or mandible structure or femur strength at the midpoint or neck at 4?months of age despite lower serum 25(OH)D₃ among all groups compared to REF. For Trial 2 (400?IU?vit D/kg), mice fed 2.5?g?Ca/kg diet had lower (p?p?3 or femur strength at the midpoint or neck. Serum PTH was not elevated compared to REF in either Trial.

Conclusion

Lowering both dietary vit D (100?IU/kg) and Ca (2.5?g/kg) in AIN-93G diet did not result in differences in bone development of female CD-1 mice at early adulthood. Translational relevance of bone studies conducted using the AIN-93G diet may be affected by its high vit D and Ca content.

SUBMITTER: Yumol JL

PROVIDER: S-EPMC6020397 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Publications

Bone development in growing female mice fed calcium and vitamin D at lower levels than is present in the AIN-93G reference diet.

Yumol Jenalyn L JL Wakefield C Brent CB Sacco Sandra M SM Sullivan Philip J PJ Comelli Elena M EM Ward Wendy E WE

Bone reports 20180519

<h4>Background</h4>The AIN-93G reference (REF) diet is used to allow the comparison within and between studies of different research groups but its levels of vitamin D (vit D) and calcium (Ca) may be higher than required for healthy bone structure and bone mineral density (BMD).<h4>Objective</h4>To determine if lower dietary levels of Ca (3.5, 3 or 2.5 g Ca/kg diet) at 1 of 2 levels of vit D (100 or 400 IU/kg diet) supports similar development of bone structure and BMD compared to AIN-93G refere ...[more]

PMID: 29955642

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Project description:UnlabelledBackgroundPrevious studies examining the metabolic consequences of dietary iron deficiency have reported elevated serum glucose concentrations in iron-deficient animals. Importantly, the majority of these findings were observed using an earlier version of a laboratory animal diet (AIN-76A) in which the primary carbohydrate source was sucrose - a disaccharide known to negatively impact both glucose and lipid homeostasis. The AIN-76A diet formula was improved in 1993 (AIN-93) to optimize animal nutrition with a major change being the substitution of cornstarch for sucrose. Therefore, we sought to examine the effects of iron deficiency on steady-state glucose homeostasis and the hepatic expression of glucose- and lipid-related genes in rats fed an iron-deficient diet based on either an AIN-76A or AIN-93 diet.MethodsThe study design consisted of 6 treatment groups: control (C; 40 mg Fe/kg diet), iron deficient (ID; ≤ 3 mg Fe/kg diet), or pair-fed (PF; 40 mg Fe/kg) fed either an AIN-76A or AIN-93 diet for 21 d. Hemoglobin and hematocrit were measured in whole blood. Serum insulin and cortisol were measure by ELISA. Serum glucose and triacylglycerols were measured by standard colorimetric enzyme assays. Alterations in hepatic gene expression were determined by real-time qPCR.ResultsHemoglobin and hematocrit were significantly reduced in both ID groups compared to the C and PF groups. Similarly, animals in the both ID groups exhibited elevated steady-state levels of blood glucose and insulin, and significantly decreased levels of circulating cortisol compared to their respective PF controls. Serum triacyglycerols were only increased in ID animals consuming the AIN-76A diet. Hepatic gene expression analyses revealed a ~4- and 3-fold increase in the expression of glucokinase and pyruvate dehydrogenase kinase-4 mRNA, respectively, in the ID group on either diet compared to their respective PF counterparts. In contrast, the expression of lipogenic genes was significantly elevated in the AIN-76 ID group, while expression of these genes was unaffected by iron status in the AIN-93 ID group.ConclusionsThese results indicate that an impaired iron status is sufficient to alter glucose homeostasis, though alterations in lipid metabolism associated with ID are only observed in animals receiving the AIN-76A diet.

Project description:A study of 959 black and white women shows that the normal range for 24-h urine calcium excretion in white women is 30-300 mg and 10-285 mg in black women. Clinical laboratories use a normal range for 24-h urine calcium excretion of 100-300 mg; there is a need for them to update their age- and race-specific ranges.IntroductionRecommendations for a normal range for 24-h urine calcium vary from a low of 50 mg to a high of 400 mg; most "laboratory normal ranges" based on older literature are incorrect. The objective of this analysis is to define a normal range for young women age 25-45 years and older women age 55-90 years, white and black, for 24-h urine calcium and calcium/creatinine ratio, and to examine the relationship between 24-h urine calcium, calcium absorption, and vitamin D metabolites.MethodsData from 3 studies was collected on 959 normal black and white women, ages 25-87 years, for 24-h urine calcium, creatinine, calcium intake, serum 25-hydroxyvitamin D (25OHD),1,25 dihydroxyvitamin D (1,25(OH)2D), and calcium absorption. Urine calcium and creatinine were measured on an auto-analyzer. Calcium absorption was measured by a single isotope method. Serum 25OHD and 1,25(OH)2D were measured by competitive protein binding assay or radioimmunoassay. Because 24-h urine calcium did not fit a normal distribution, non-parametric methods were used to determine the 95% reference interval (RI) and 90% non-parametric confidence intervals (CI) calculated for groups.ResultsThe 95% reference intervals for 24-h urine calcium for women were as follows: black older 7-225 mg, black younger 8-285 mg; white older 37-275 mg, and white younger 23-287 mg. In older white women, 24-h urine calcium was significantly correlated with serum 1,25(OH)2 D, r = 0.23, p < 0.001, calcium intake r = 0.12, p = 0.001, and calcium absorption r = 0.18, p = 0.003, but not serum 25OHD r = 0.07, p = 0.06.ConclusionsThe normal reference interval for 24-h urine calcium for black women is lower than white women. Twenty-four-hour urine calcium was correlated with serum 1,25(OH)2 D calcium intake and calcium absorption, but not serum 25OHD. This range will be useful clinically for defining hypercalciuria and for following patients on vitamin D and calcium treatment.

Dataset Information

Bone development in growing female mice fed calcium and vitamin D at lower levels than is present in the AIN-93G reference diet.

Background

Objective

Methods

Results

Conclusion

Publications

Bone development in growing female mice fed calcium and vitamin D at lower levels than is present in the AIN-93G reference diet.

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