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Intensive morphometric analysis of enormous alterations in skeletal bone system with micro-CT for AHNAK−/− mice

2020 , Kim, I.Y. , Yi, S.S. , Shin, J.H. , Kim, Y.N. , Ko, C.-Y. , Kim, H.S. , Lee, S.Y. , Bae, Y.S. , Seong, J.K.

AHNAK has been reported to be involved in actin cytoskeleton rearrangement of some cell types, calcium homeostasis, and activation of T cells. Although the functional role of AHNAK in muscle cells, epidermis, and brain has been determined, its association with apparent clinical impairment has not been found yet. During phenotypic analysis of AHNAK knock out (KO) mice for many years, we observed that AHNAK KO mice showed very slow growth. Snouts of these animals were very short, and their bones were easily broken compared to normal mice. It is known that AHNAK is closely related to calcium. However, intensive morphological studies on phenotypes of bone have yet been reported for AHNAK. Thus, the objective of the present study was to analyze the morphology of skull, mandibular, limbs, and caudal bones of AHNAK KO mice intensively using micro-CT with many factors for various ages of these mice (6 weeks, 18 weeks, and 40 weeks). As a result, it was found that the facial region of AHNAK KO mouse showed a large difference in mandible than skull. Their both femur and tibia were shortened, and bone strength was also significantly decreased compared to normal mice. Particularly, the tail bone of AHNAK KO mice exhibited morphological abnormality by age. Taken together, these results suggest that AHNAK plays an important role in bone shape, development, and metabolism. Although our results demonstrated that AHNAK has a distinct role in bone, further investigations are needed to determine various features of bone metabolism related to AHNAK in the future.

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Bilateral asymmetry in microarchitecture of trabecular bone in male c57bl/6 mouse tibia: Implication for experimental sample size estimations

2013 , Ko, C.-Y. , Jung, Y.-J. , Seo, D.H. , Kim, H.S.

This study aimed to determine whether there is bilateral asymmetry between the left and right tibiae in the microarchitectural characteristics of the trabecular bone and estimate a moderate sample size for detections of such bilateral differences. The left and right tibiae of 20 C57BL/6 mice (12 weeks old) were scanned by micro-computed tomography, and the structural parameters of the trabecular bone were measured. There were significant differences in the structural parameters between the left and right tibiae (p<0.05); specifically, the microarchitecture was greater in quantity and quality in the left tibia than in the right tibia. Moreover, the percent of bilateral differences between the left and right tibiae ranged from 2.26% to 22.52%. For most of the structural parameters except for trabecular bone thickness (Tb.Th) and trabecular bone separation (Tb.Sp), the 20 mice involved in this study were enough to detect differences. These results show that the directional left-right asymmetry in the microarchitecture of the trabecular bone may exist despite the samples being from an inbred strain. Furthermore, we estimated a sample size for detections of such differences between the left and right tibiae.

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Enhancement of bone quality and longitudinal growth due to free-fall motion in growing rats

2015 , Cho, S. , Eom, S. , Seo, D.-H. , Park, J. , Ko, C.-Y. , Kim, H.S.

Objectives: This study is to investigate the synchronous phenomena between bone quality and longitudinal length in a same subject affected by landing exercise. Physical exercise on the ground induces external loading to human body due to resistance from ground which can activate bone generation or remodeling. Especially, when the impact stimulation is applied to bone, it may improve bone quality and lengthening. Methods: 6-week-old male Wistar rats were randomly allocated to one of two conditions: free fall from 40 cm-height (I40; n = 7), and control (IC; n = 7). The impact stimulations were administered to the free fall groups, 10 times/day, and 5 days/week for 8 weeks. Structural parameters and longitudinal length of tibia were measured to quantitatively evaluate the variation in morphological characteristics and bone length with maturing. Results: The landing impact seems to be commonly effective on the enhancement of bone quality as well as longitudinal growth. Howe ver, the extent of enhancement may be more dominant in bone quality than longitudinal growth. On the other hand, the ratio of longitudinal growth seems to be dependent on the duration of stimuli whereas the enhancement of bone quality does not. Conclusions: This study verified that free-falls exercise can be effective on the enhancement of bone qualities and promotion of vertical growth in long bones. We expect that it might be possible for the moderate impact stimulation to be proposed as an aid for prevention of bone loss and promotion of bone lengthening. © 2015, Korean Society of Medical and Biological Engineering and Springer.

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The effects of minimally invasive laser needle system on suppression of trabecular bone loss induced by skeletal unloading

2013 , Ko, C.-Y. , Kang, H. , Ryu, Y. , Jung, B. , Kim, H. , Jeong, D. , Shin, H.-I. , Lim, D. , Kim, H.S.

This study was aimed to evaluate the effects of low-level laser therapy (LLLT) in the treatment of trabecular bone loss induced by skeletal unloading. Twelve mice have taken denervation operation. At 2 weeks after denervation, LLLT (wavelength, 660 nm; energy density, 3 J/cm2) was applied to the right tibiae of six mice (LASER) for 5 days/week over 2 weeks by using a minimally invasive laser needle system (MILNS) which consists of a 100 m optical fiber in a fine needle (diameter, 130 µm). Structural parameters and histograms of bone mineralization density distribution (BMDD) were obtained before LLLT and at 2 weeks after LLLT. In addition, osteocyte, osteoblast, and osteoclast populations were counted. Two weeks after LLLT, bone volume fraction, trabeculae number, and trabeculae thickness were significantly increased and trabecular separations, trabecular bone pattern factor, and structure model index were significantly decreased in LASER than SHAM (rho < 0.05). BMDD in LASER was maintained while that in SHAM was shifted to lower mineralization. Osteocyte and osteoblast populations were significantly increased but osteoclast population was significantly decreased in LASER when compared with those in SHAM (rho < 0.05). The results indicate that LLLT with the MILNS may enhance bone quality and bone homeostasis associated with enhancement of bone formation and suppression of bone resorption.