Effects of Multi-Deficiencies-Diet on Bone Parameters of Peripheral Bone in Ovariectomized Mature Rat

Abstract

Many postmenopausal women have vitamin D and calcium deficiency. Therefore, vitamin D and calcium supplementation is recommended for all patients with osteopenia and osteoporosis. We used an experimental rat model to test the hypothesis that induction of osteoporosis is more efficiently achieved in peripheral bone through combining ovariectomy with a unique multi-deficiencies diet (vitamin D depletion and deficient calcium, vitamin K and phosphorus). 14-week-old Sprague-Dawley rats served as controls to examine the initial bone status. 11 rats were bilaterally ovariectomized (OVX) and fed with multi-deficiencies diet. Three months later the treated group and the Sham group (n = 8) were euthanized. Bone biomechanical competence of the diaphyseal bone was examined on both, tibia and femur. Image analysis was performed on tibia via µCT, and on femur via histological analysis. Lower torsional stiffness indicated inferior mechanical competence of the tibia in 3 month OVX+Diet. Proximal metaphyseal region of the tibia showed a diminished bone tissue portion to total tissue in the µCT despite the increased total area as evaluated in both µCT and histology. Cortical bone showed higher porosity and smaller cross sectional thickness of the tibial diaphysis in the OVX+Diet rats. A lower ALP positive area and elevated serum level of RANKL exhibited the unbalanced cellular interaction in bone remodeling in the OVX+Diet rat after 3 month of treatment. Interestingly, more adipose tissue area in bone marrow indicated an effect of bone loss similar to that observed in osteoporotic patients. Nonetheless, the presence of osteoid and elevated serum level of PTH, BGP and Opn suggest the development of osteomalacia rather than an osteoporosis. As the treatment and fracture management of both osteoporotic and osteomalacia patients are clinically overlapping, this study provides a preclinical animal model to be utilized in local supplementation of minerals, drugs and growth factors in future fracture healing studies.

Effects of chronic mild stress on parameters of bone assessment in adult male and female rats

“…The mild bone loss or sometimes non-significant differences between CMS and control groups may be due not only to the induction mechanism but also to the time of observation. El Khassawna et al (2013), for example, have shown that tibia BV/TV of ovariectomized rats fed with multideficient diet, assessed by microcomputed tomography, can achieve less than 10% after three months of induction. Feik et al (1997) showed that in humans, cortical area of cross-sections from femoral midshaft increases in both sexes until the 7 th decade and then it starts to decline, more pronounced in women.…”

Quantification of calcium content in bone by using ToF-SIMS–a first approach

“…The non-mineralised collagen matrix with small remains of Ca in the centre of the trabecular has been described in more detail in our previous study [11]. After histological and serological investigations El Khassawna et al also conclude the development of osteomalacia in this rat model [26]. In consequence the Ca quantification might become more complicated due to measurement of non-mineralised regions which might result in too low Ca values for the OVX and diet group.…”

Vitamin K Supplementation Modulates Bone Metabolism and Ultra-Structure of Ovariectomized Mice

“…Results were expressed in fold of amplification. Better description of methodology were scrutinized in the following references [18,19].…”

“…As long as the effects of VK in bone metabolisms are still inconclusive, we decided to investigate its effects in a well-established model for osteoporosis, the ovariectomized (OVX) mice [17][18][19]. Surgical removal of the ovaries mimics menopause resulting in a suitable model to investigate mechanisms of VK action in bone as well as its interaction with estrogen either on BMD or microarchitecture [19].…”