Bone Mineral Analysis through Dual Energy X-Ray Absorptiometry in Laboratory Animals

Tsujio, Masashi; Mizorogi, Toshihiro; Kitamura, Itsuko; Maeda, Yasuhiro; Nishijima, Ken‐ichi; Kuwahara, Sachi; Ohno, Tamio; Niida, Shunpei; Nagaya, Masahiro; Saito, Ryoichi; Tanaka, Shin

doi:10.1292/jvms.001493

Cited by 19 publications

(16 citation statements)

References 13 publications

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“…9,24 Based on our review of current literature, the studies that used broad ROIs to measure BMD via DXA in mice specified ROIs that contained whole bones (e.g., whole femur). [17][18][19][20] These ROIs were not specific to trabecular bone-rich regions ( Fig. 2A).…”

Section: Discussionmentioning

confidence: 90%

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Guidelines for Dual Energy X-Ray Absorptiometry Analysis of Trabecular Bone-Rich Regions in Mice: Improved Precision, Accuracy, and Sensitivity for Assessing Longitudinal Bone Changes

Shi

Lee

Uyeda

et al. 2016

Tissue Engineering Part C: Methods

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Trabecular bone is frequently studied in osteoporosis research because changes in trabecular bone are the most common cause of osteoporotic fractures. Dual energy X-ray absorptiometry (DXA) analysis specific to trabecular bone-rich regions is crucial to longitudinal osteoporosis research. The purpose of this study is to define a novel method for accurately analyzing trabecular bone-rich regions in mice via DXA. This method will be utilized to analyze scans obtained from the International Space Station in an upcoming study of microgravity-induced bone loss. Thirty 12-week-old BALB/c mice were studied. The novel method was developed by preanalyzing trabecular bone-rich sites in the distal femur, proximal tibia, and lumbar vertebrae via high-resolution X-ray imaging followed by DXA and micro-computed tomography (micro-CT) analyses. The key DXA steps described by the novel method were (1) proper mouse positioning, (2) region of interest (ROI) sizing, and (3) ROI positioning. The precision of the new method was assessed by reliability tests and a 14-week longitudinal study. The bone mineral content (BMC) data from DXA was then compared to the BMC data from micro-CT to assess accuracy. Bone mineral density (BMD) intra-class correlation coefficients of the new method ranging from 0.743 to 0.945 and Levene's test showing that there was significantly lower variances of data generated by new method both verified its consistency. By new method, a Bland-Altman plot displayed good agreement between DXA BMC and micro-CT BMC for all sites and they were strongly correlated at the distal femur and proximal tibia (r = 0.846, p < 0.01; r = 0.879, p < 0.01, respectively). The results suggest that the novel method for site-specific analysis of trabecular bone-rich regions in mice via DXA yields more precise, accurate, and repeatable BMD measurements than the conventional method.

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Section: Discussionmentioning

confidence: 90%

“…2B). [17][18][19][20] The conventional method did not contain specific animal positioning instructions for DXA, with one exception: the PIXImus 2 densitometer user manual stated that mice were to be laid prone with heads placed in the tray headrest.…”

Section: Animals and Experimental Designmentioning

confidence: 99%

Guidelines for Dual Energy X-Ray Absorptiometry Analysis of Trabecular Bone-Rich Regions in Mice: Improved Precision, Accuracy, and Sensitivity for Assessing Longitudinal Bone Changes

Shi

Lee

Uyeda

et al. 2016

Tissue Engineering Part C: Methods

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“…Once exsanguinated, excise both femurs. Pay special care to remove as much soft tissue as possible, for this has been shown to affect density measurements 11 . Wrap the femurs with water soaked gauze and store at -20C until you are ready to analyze them.…”

Section: Post-mortem Analysis Of Bone Densitymentioning

confidence: 99%

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, <em>in vivo</em>, Methodology

McManus

Grill

2011

JoVE

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Spinal cord injury (SCI) is often accompanied by osteoporosis in the sublesional regions of the pelvis and lower extremities, leading to a higher frequency of fractures 1 . As these fractures often occur in regions that have lost normal sensory function, the patient is at a greater risk of fracture-dependent pathologies, including death. SCI-dependent loss in both bone mineral density (BMD, grams/cm 2 ) and bone mineral content (BMC, grams) has been attributed to mechanical disuse . The use of rodent models of SCI-induced osteoporosis can provide invaluable information regarding the mechanisms underlying the development of osteoporosis following SCI as well as a test environment for the generation of new therapies 5-7 (and reviewed in 8 ). Mouse models of SCI are of great interest as they permit a reductionist approach to mechanism-based assessment through the use of null and transgenic mice. While such models have provided important data, there is still a need for minimally-invasive, reliable, reproducible, and quantifiable methods in determining the extent of bone loss following SCI, particularly over time and within the same cohort of experimental animals, to improve diagnosis, treatment methods, and/or prevention of SCI-induced osteoporosis.An ideal method for measuring bone density in rodents would allow multiple, sequential (over time) exposures to low-levels of X-ray radiation. This study describes the use of a new whole-animal scanner, the IVIS Lumina XR (Caliper Instruments) that can be used to provide low-energy (1-3 milligray (mGy)) high-resolution, high-magnification X-ray images of mouse hind limb bones over time following SCI. Significant bone density loss was seen in the tibiae of mice by 10 days post-spinal transection when compared to uninjured, age-matched control (naïve) mice (13% decrease, p<0.0005). Loss of bone density in the distal femur was also detectable by day 10 post-SCI, while a loss of density in the proximal femur was not detectable until 40 days post injury (7% decrease, p<0.05). SCI-dependent loss of mouse femur density was confirmed postmortem through the use of Dual-energy X-ray Absorptiometry (DXA), the current "gold standard" for bone density measurements. We detect a 12% loss of BMC in the femurs of mice at 40 days post-SCI using the IVIS Lumina XR. This compares favorably with a previously reported BMC loss of 13.5% by Picard and colleagues who used DXA analysis on mouse femurs post-mortem 30 days post-SCI 9 . Our results suggest that the IVIS Lumina XR provides a novel, high-resolution/high-magnification method for performing long-term, longitudinal measurements of hind limb bone density in the mouse following SCI. Video LinkThe video component of this article can be found at https://www.jove.com/video/3246/ Protocol 1. Mouse spinal transection model 1. Adult, male, C57BL6 mice (approximately 20-25 g) are anesthetized using a combination of ketamine (200 mg/kg) and xylazine (10 mg/kg).All surgical procedures are performed in an Institutional, IACUC-approved su...

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“…to detect initial appearances of ossification centers; in clinical practice, DXA is widely used for the diagnosis and evaluation of osteoporosis and a new generation of DXA scanners offer software for performing vertebral morphometric analysis (Blake & Fogelman, 1997). Furthermore, numerous bone analyses have been performed on experimental animals using DXA (Tsujio et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Densitometric Analysis of the Human Ulna

Aguado-Henche¹,

Arriba²,

Carrascosa-Sánchez³

2015

Int. J. Morphol.

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SUMMARY: This study describes the distribution of the normal bone mass in the mature human ulna, as knowledge of distribution may provide information to orthopaedic surgeons for the treatment of such fractures while advancing the understanding of the responses of osteosynthesis elements during bone repair. A random sample of 41 dry ulnas was studied with a DXA system using a Norland XR-26 densitometer. Two projections were performed, lateral and antero-posterior. In both projections five regions of interest (ROI) were selected: proximal (ROI 1), proximal-intermediate (ROI 2), intermediate (ROI 3), distal-intermediate (ROI 4) and distal (ROI 5). All statistical calculations were performed using Stat graphics Plus (version 5.1) and SPSS (version 15.0). Bone mineral density (BMD) and bone mineral content (BMC) were calculated. Paired data were analysed using parametric Student t-Test to compare BMD and BMC in every region in both projections. DXA show that the higher BMD is in the proximal-intermediate region (ROI 2). The higher BMC is found in the proximal region (ROI 1). Lower BMD and BMC are found in the distal region (ROI 5). The total BMD shows significant statistical differences (p≤0.001), which indicates the heterogeneous nature of the distribution of bone mass in the studied bone. Bone densitometry, measured by the DXA technique, is useful for assessing trabecular architecture of the human ulna.

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Bone Mineral Analysis through Dual Energy X-Ray Absorptiometry in Laboratory Animals

Cited by 19 publications

References 13 publications

Guidelines for Dual Energy X-Ray Absorptiometry Analysis of Trabecular Bone-Rich Regions in Mice: Improved Precision, Accuracy, and Sensitivity for Assessing Longitudinal Bone Changes

Guidelines for Dual Energy X-Ray Absorptiometry Analysis of Trabecular Bone-Rich Regions in Mice: Improved Precision, Accuracy, and Sensitivity for Assessing Longitudinal Bone Changes

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, <em>in vivo</em>, Methodology

Densitometric Analysis of the Human Ulna

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