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Shaking Stimuli Can Retard Accelerated Decline of Bone Strength of a Mouse Model Assumed to Represent a Postmenopausal Woman

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Shaking Stimuli Can Retard Accelerated Decline of Bone Strength of a Mouse Model Assumed to Represent a Postmenopausal Woman Journal of Analytical Bio-Science Vol. 33, No 4 (2010) 〈Original article〉 Shaking stimuli can retard accelerated decline of bone strength of a mouse model assumed to represent a postmenopausal woman Kouji Yamada*, Kazuhiro Nishii and Takehiko Hida Summary In contrast in elderly people, if bone formation and resorption are out of balance, bone mineral density (BMD) diminishes leading to progressive development of osteoporosis, a cause of easy bone fracture. Standing still for prolonged periods leads to fatigue. Humans tend to avoid this by shifting the body weight unconsciously or consciously left and right or back and forth. Standing in an erect posture on a board that continuously shakes horizontally causes all the skeletal muscles of the body as well as the entire skeleton to act to restore balance. As a result of the application of such shaking–induced stimuli to a reduced–BMD mouse model of postmenopausal osteoporosis, rapid decline in the bone strength of the femur was suppressed. A shaking stimulus such as was used in this study could easily be applied to human beings in future once further study is conducted to validate the results. Key words: Osteogenesis, Bone resorption, Shaking, Physical therapy 1. Introduction (BMD) continues to increase for a while after cessation of linear growth. Generally, bone mass in A rise in average longevity along with improved humans reaches a peak in the late twenties to thirties, treatment for organ disorders and various diseases remaining in a static state for some time thereafter. has rendered age-associated bone mass reduction a With further aging, bone volume is maintained but great clinical issue. This problem needs to be solved mineral content and strength both progressively by elucidation of its pathogenesis and establishment of decrease. After reaching peak bone mass, women appropriate therapeutic options as well as preventive lose cortical bone by 35% and cancellous bone by methods. Postmenopausal women rapidly lose bone 50% until the end of life. Women rapidly lose bone mass within several years of menopause due to mineral content, especially in cancellous component- reduced estrogen action. In human beings, bone dominant bone, immediately after menopause and grows in synchrony with body growth during the bone mass loss continues slowly thereafter for life1). growth period and thereafter bone mineral density In humans, bone loss can be either acute or School of Health Sciences, Fujita Health University, *Corresponding author; Kouji Yamada, Ph.D. 1-98 Kutsukake-cho, Toyoake, Aichi 470-1192, Japan School of Health Sciences, Fujita Health University, 1-98, Kutsukake-cho, Toyoake, Aichi 470-1192, Japan Received for Publication April 9, 2010 Accepted for Publication April 28, 2010 － 355 － Journal of Analytical Bio-Science chronic. For example, about 1% of bone mass is lost firm down to the highest level of being able to keep a after one week of immobilization, while 10 - 20% standing posture. of bone mass is lost after several months2). Bone Bone histomorphometry is cited as one means of mass is reduced by Ca, 1 - 2% over a year in assessing bone metabolism. We prepared decalci- postmenopausal osteoporosis. In rats, the rear legs of fied thin sections of mouse femurs to measure cancel- which were unloaded by tail suspension, their femoral lous bone volume, trabecular thickness, and trabecular bone mass was reduced within 14 days irrespective of number which are parameters of bone structure. BMD whether the cancellous bone-rich metaphysis or the is maintained through balanced bone resorption and cortical bone-rich diaphysis was measured3). In the bone formation. Loss of this balance leads to osteo- case of mice subjected to tail-suspension, cancellous porosis. We measured osteoid surface/bone surface, bone mass was lost by up to 44% after 7 days of osteoblast surface/bone surface, mineralizing such treatment as compared to control. A further 7 surface/bone surface and mineral apposition rate as days of such treatment did not lead to a further parameters of bone formation and eroded surface/bone decrease, however4). surface and osteoclast number/bone perimeter as In this study, we tested exercise therapy using a parameters of bone resorption. Increasing numbers of physical stimulus imposed by a shaker in an attempt to osteoclasts and their increased lifespan depend on prevent the accelerated reduction of BMD observed in animal species, bone site, and age. In rats, osteo- a mouse model of postmenopausal women. The clast formation decreases with increasing age, while in stimulus was provided as follows: A shaking board mice these parameters depend on strain. In the case of was constructed of a rotating horizontal plate, the long bones, human postmenopausal bone mass pivot of which made circular motions. When a mouse reduction follows different courses between the was placed on the shaking board, the animal tried diaphyses and metaphyses6). The lumbar spine rapidly hard to keep its footing; this posture provided a kind of loses bone mass by about 15% in 5 - 6 years after exercise to distort bone and accelerated reduction of menopause and more slowly thereafter7). In contrast, BMD was thereby slowed down. However, real the distal end of the forearm bone loses bone mass by phenomena occurring in the living body seem to be as much as about 25% in a linear manner in 15 - 16 due to indirect influences on the signal transduction years post-menopause. Whole body bone mass pathways exerted by mechanical load-induced flows of decreases exponentially in the first 12 years after extracellular fluid rather than by direct physical distor- menopause, slowing down thereafter6). A lowered tion of the bone. level of estrogen greatly affects primarily cancellous The cells in bone that sense mechanical load are bone. The long bones of the limbs are predominantly presumably osteoblasts and osteocytes. Reportedly, composed of cortical bone, while their epiphyseal mechanical signals that cells recognize in vivo are portions are relatively rich in cancellous bone. This is distortion of bone tissues, changes in hydrostatic the reason for easy fracture occurring in the femoral pressure, liquid current inside the bone canaliculus and neck or in the distal end of the radius1). Estrogen electromagnetic fields generated by liquid currents5). elevates the calcitonin concentration that suppresses Seemingly, these mechanical loads are generated by bone resorption. This hormone, furthermore, also movement of skeletal muscles attached to the bones. stimulates vitamin D action that enhances intestinal Such movements are conducted by tendons, which absorption of calcium8). passively convey stimuli to bones and these stimuli Bone fulfils several roles in the body, acting as a then affect bone dynamics. In this context, since we store of calcium and phosphorous, providing structural assumed that the maximum stimulus should be close support and acting as a passive locomotive organ. to but under the threshold for falling, we determined Maximum locomotive potential requires maintenance the appropriate stimulus strength by gradually reducing of bone strength as well as muscle. Bone responds to the level of stimulus from the critical level of standing an applied mechanical load by adjusting the signals － 356 － Journal of Analytical Bio-Science Vol. 33, No 4 (2010) governing remodeling. When bone cells sense a stimulus from a mechanical load, bone formation is 2. Materials and Method stimulated on the surface of trabeculae, leading to an increase in bone mass. However if such stimuli are 1. Mouse Model below the threshold level, bone resorption is enhanced, Ovaries were removed from 9-week-old female resulting in loss of bone mass. In adults, these ICR mice under nembutal anesthesia. Estrogen processes, occur continuously so that bone mass is secretion is decreased by ovariectomy, therefore, these stable. In contrast in elderly people, if bone formation mice represent an osteopenic model (OVX mouse). and resorption are out of balance, BMD diminishes Animals in this study were kept in the vivarium of leading to progressive development of osteoporosis, a Fujita Health University, and studies were conducted cause of easy bone fracture. under Animal Use Committee regulations. There is a relatively new loading modality in which dynamic loads are to induce bone formation in 2. Shaking stimulus the femur. The method of knee loading that the left Twelve ovariectomized mice (OV group) and 12 knee of mice was loaded with 0.5 N force at 5, 10, or non-ovariectomized mice (i.e., wild type: WT group) 15 Hz for 3 min/day for 3 consecutive days9). We are were further subdivided into stimulated and non- interested in that evaluate the effects of loading stimulated groups, to create a total of 4 groups: frequencies (in Hz) on bone formation at the site Shaking stimulus/ovariectomy (+/+), no away from the loading site on the knee. However, our stimulus/ovariectomy (-/+), shaking stimulus/intact stimulation method is the method that is almost ovaries (+/-), and no stimulus/intact ovaries (-/-). The exercise therapy by stimulation using shaker. shaking stimulus was provided by a platform, shaking In this study, mice were bilaterally ovariectomized at a rate of 150 horizontal rotations per minute for 30 to create a mouse model (OVX mice) with suppressed minutes (NR-3; TAITEC Co. Ltd Japan). This secretion of estrogen. These mice were assumed to stimulus load was repeated 6 times a week for 10 mimic postmenopausal women. We developed a weeks.
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