• Journal of Nutrition and Health
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• v.44 no.6
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• pp.481-487
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• 2011

Obesity not only reduces bone mineral density but also increases inflammatory markers. Therefore, we examined the change in inflammatory markers and morphological microstructure of the bones using a mouse model fed a high-fat diet. C57BL/6J 4-week-old male mice were divided into a control group (n = 6) and a experimental group (n = 6); the control group was provided with 10% Kcal fat diet, and the high-fat diet group was provided with 45% Kcal fat diet for 12 weeks using the free provision method. Blood was analyzed for inflammatory markers, and micro-computed tomography was used to measure the morphological microstructure of the femoral bone. The weight increases in the control group and high-fat diet group were $5.85{\pm}1.84g$ and $16.06{\pm}5.64g$, respectively (p < 0.01), glucose was $115.00{\pm}16.88mg/dL$ and $188.33{\pm}13.29mg/dL$ (p < 0.01), and triglycerides were $65.00{\pm}6.19mg/dL$ and $103.33{\pm}8.02mg/dL$ (p < 0.05) respectively. Leptin and interleukin (IL)-6 were significantly higher in the high-fat diet group than that in the control group (p < 0.01). As a result of a biochemical index analysis of bone metabolism, osteocalcin tended to be lower in the high-fat diet group, whereas CTx was significantly higher in the high-fat diet group compared to that in the control group (p < 0.01). The thickness of the bony trabecula was significantly narrower in the high-fat diet group than that in the control group (p < 0.05), and the gap in the bony trabecula was significantly wider in the high-fat diet group than that in the control group (p < 0.05). IL-6 and the gap in the bone trabecula, which was a morphological microstructure of the bones, showed a positive correlation (p < 0.05). Taken together, inducing obesity through a high-fat diet in mice during the growth phase caused a change in bone microstructure and was correlated with the inflammation index. Accordingly, restriction of excessive fat intake may be needed to suppress the inflammatory reactions and promote normal bone formation.

• Journal of Acupuncture Research
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• v.29 no.6
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• pp.11-21
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• 2012

Objectives : BHH10 is traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify BHH10 extract induces osteogenic activity in human osteoblast-like MC3T3-E1 cells. Methods : MC3T3-E1, pre-osteoblast cell line, were treated with BHH10 of various concentrations($0.1{\mu}g/mL$, $1{\mu}g/mL$, $10{\mu}g/mL$). And then, the effect of BHH10 on osteoblast differentiation was examined by alkaline phosphatase(ALP) activity, von Kossa staining and RT-PCR for osteoblast differentiation markers such as osteocalcin(OCN), osteopontin(OPN). Results : BHH10 had dose-dependent effect on the viability of osteoblastic cells, and dose-dependently increased alkaline phosphatase(ALP) activity. BHH10 markedly increased mRNA expression for OCN, OPN in MC3T3-E1 cells. Also, BHH10 significantly induced mineralization in the culture of MC3T3-E1 cells. Conclusions : In conclusion, these results propose that BHH10 can play an important role in osteoblastic bone formation, osteogenesis, and may possibly lead to the development of bone-forming drugs.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.6
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• pp.713-722
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• 2013

The purpose of this study was to examine the effects of dietary green tea powder supplementation on bone metabolism in streptozotocin-induced diabetic rats. Thirty-two male Sprague-Dawley rats (body weight $210{\pm}3g$) were divided into two groups, diabetic and non-diabetic groups. Each group was randomly divided into two subgroups which were fed with the control and 1% green tea powder diets. The serum and urine concentrations of calcium and phosphorus were determined. Serum osteocalcin and ALP and urinary DPD crosslinks value were measured in order to monitor bone formation and resorption. Bone mineral density (BMD) and bone mineral content (BMC) were estimated using PIXImus in the spine and femur. Body weight gain and FER were lower in the diabetic group than in the non-diabetic group regardless of diets. The serum concentration of calcium and phosphorus were not changed among all groups. Urinary calcium and phosphorus excretion were higher in the diabetic group than in the non-diabetic group regardless of diets; however, they were not significantly different by green tea powder intake. Serum alkaline phosphatase (ALP) was increased in the diabetic group than in thenon-diabetic group. Further, there were no significant differences in serum osteocalcin and urinary deoxypyridinoline crosslinks value among all groups. The levels of spine and femur bone mineral density of the diabetic group were significantly lower than that of the non-diabetic group. Within the diabetic group, spine BMD was significantly higher in rats fed with the green tea powder diet than in rats fed the control diet. Therefore, this study suggests that green tea powder has a beneficial effect on bone health, although it is not directly applicable to humans.

• Journal of Yeungnam Medical Science
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• v.25 no.1
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• pp.19-30
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• 2008

Osteoporosis, a disease characterized by low bone mass and microarchitectural deterioration of bone tissue leading to enhanced bone fragility and fracture risk, is a major public health problem. The diagnostic methods for osteoporosis include simple radiography, bone scan, DXA (Dual energy X-ray Absortiometry) and biochemical markers of bone turnover. Optimal treatment and prevention of osteoporosis require modification of risk factors, particularly smoking cessation, adequate physical activity, and attention to diet, in addition to pharmacologic intervention. The estrogens and raloxifene both prevent bone loss in postmenopausal women, and the estrogens probably also decrease the risk of first fracture. There is good evidence that raloxifene prevents further fractures in postmenopausal women who already have had fractures and some evidence that estrogen does as well. Bisphosphonate prevents bone loss and reduces fractures in healthy and osteoporotic postmenopausal women and in osteoporotic men as well. Risedronate is more potent and has fewer side effects than alendronate and reduces the incidence of fractures in osteoporotic women. Calcitonin increases bone mineral density in early postmenopausal women and men with idiopathic osteoporosis, and also reduces the risk of new fractures in osteoporotic women. All of the agents discussed above prevent bone resorption, whereas teriparatide and strontium increase bone formation and are effective in the treatment of osteoporotic women and men. New avenues for targeting osteoporosis will emerge as our knowledge of the regulatory mechanisms of bone remodeling increases, although issues of tissue specificity may remain to be addressed.

• Korean Journal of Community Nutrition
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• v.7 no.1
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• pp.121-129
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• 2002

Osteoporosis, the typical metabolic bone disease of the elderly, is characterized by a reduction in bone mineral density (BMD) and increased fracture risk. Genetic and environmental factors are known to play a key role in bone metabolism, and diet is also considered to be one of the important factors. The purpose of the present study was to investigate the relationship among the factors affecting BMD, including stature, body weight, age, time period since onset of menopause, and biochemical markers of bone turnover in postmenopausal women. Seventy-eight postmenopausal women who visited health promotion center for health examinations volunteered to participate in this study and they were divided into two groups according to the time period since onset of menopause : women with a time period since onset of menopause of less than 5 years (Group 1) and women with a time period since onset of menopause of 5 years or more (Group 2). The demographic characteristics and dietary intake were surveyed using a questionnaire. BMDs of the lumbar spine and femoral neck of subjects were measured by dual energy X-ray absorptiometry. Serum levels of 25-hydroxy-vitamin D and parathyroid hormone (PTH), known to be indicators of bone related hormone status, were anlyzed. Serum samples were measured for calcium, phosphorus, alkaline phosphatase, and osteocalcin as bone formation indicators, and urine was analysed for deoxypyridinoline, creatinine, calcium, and sodium as bone resorption indicators. The results are as follow : The mean BMDs of the lumbar spin and femoral neck were $1.02 \pm 0.02 g/cm^2 and 0.81 \pm 0.02 g/cm^2 respectively, and the BMD level of Group 2 was significantly lower than tat of Group 1 (p<0.01, p<0.05, respectively). The mean daily intake of energy was 1838 $\pm$ 55 kcal. When nutrient intake was compared with the recommended dietary allowances (RDA) of the subjects, only calcium, vitamin A and riboflavin intake showed means lower than the RDA. The nutrient intake did not show any significant differences between Group 1 and 2 Serum and urine levels of biochemical markers of bone turnover did not show any significant differences between Group 1 and 2, and all were within the normal range. However, the PTH and deoxypyridinoline levels showed a tendency to be higher, and the osteocalcin level to be lower in Group 2 than in Group 1. Although age and years after menopause (YAM) showed negative correlations with lumbar spine bone mineral density (LBMD) (r= -0.38, p<0.001, and r= -0.26, p< 0.05, respectively), no correlation was found with femoral neck bone mineral density (NBMD). While height, body weight and body mass index (BMI) showed a positive correlation with LBMD (r= 0.32, p<0.001, r= 0.38, p<0.001, r= 0.22, p= 0.05, respectively), only body weight and BMI showed a positive correlation with NBMD (r= 0.30, p<0.01, and r= 0.27, p<0.05, respectivley). There was no significant corealtion between BMDs and the nutrient intake of subjects, except in the case of carbohydrates (r= 0.22, p<0.05). Also, serum and urine levels of bone turnover markers showed no significant correlation with nutrient intake. On the other hand, serum osteocalcin had a positive correlation with vitamin C intake (r= 0.22, p= 0.05), and urine deoxypyridinolin showed a negative correlation with niacin intake (r= -0.22, p= 0.05). Urinary na was negatively correlated with protein intake(r= -0.23, p= 0.05). The results suggested that it is difficult to prevent the decrease in bone mass among postmenopausal women eating the usual Korean diet. However, the BMDs of the lumbar spine and femoral neck were positively related to body weight ad BMI in postmenopausal women. Therefore, this study confirmed that one of the most effective ways to minimize bone loss in postmenopausal women would be to maintain an adequate body weight with balanced nutrient intake and activity in the pre-and postmenopausal periods.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.2
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• pp.97-106
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• 2010

Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.4
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• pp.306-312
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• 2010

Purpose: The purpose of this experiment was to evaluate the clinical effect of cultured autoglogous osteoblasts as a way to treat the defect of mandible in rabbits. Materials and Methods: Twelve rabbits were used to determine the rate of osteogenesis. The osteoblasts were obtained from the iliac crest of rabbits using aspiration. They were then cultured in Dulbecco's Modified Eagles's Medium (DMEM) with beta-glycerophosophatate, L-ascorbicacid, and dexamethasone to proliferate and differentiate osteoprogenitor cells. The expression of osteogenic markers were detected by reverse transcription-polymerase chain reaction (RT-PCR) and silver nitrate staining techniques. Five, 10-mm holes were placed in each rabbit mandible to simulate defective regions with the use of a low speed trephine bur. In the experimental group, the previously cited defects were grafted with both activated osteoblastic and autogenous bone. The control group, however, was only grafted with autogenous bone. Both groups were then analyzed at 2, 4, and 8-week intervals using bone histomorphometric analysis. Results: According to histomorphologic analysis, the rates of new bone formation at the 2, 4, and 8-week intervals were 36%, 51%, and 23% for the control group, respectively; 52%, 39%, and 28%, for the experimental group, respectively. The experimental group showed higher rates of new bone formation compared to the control group at both the 2-week and 8-week interval. Conclusion: Bone marrow-derived osteoblasts seems to be a promising bone graft material.

• The Korean Journal of Nuclear Medicine
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• v.24 no.2
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• pp.222-228
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• 1990

The development of histomorphometric and histodynamic investigations has permitted the description of a specific and complex osteopathy in hyperthyroidism. The increased bone turnover rate in hyperthyroid patients may be accompanied by a considerable bone loss. These features are associated with both inclosed osteoclastic bone resorption and increased osteoblastric bone formation, with an accelerated calcification rate. Conventional biochemical markers of bone metabolism, i.e. serum calcium and alkaline phosphatase and urinary hydroxyproline and calcium are normal in most patients with hyperthyroidism. However, the correlation between serum BGP and serum concentration of thyroid hormon suggests that serum BGP may be a sensitive marker of increased bone formation due to the hypersecretion of thyroid hormones. Any increase in bone turnover, whether focal or diffuse, will result in an increase in $^{99m}Tc-methylenediphosphonate$ uptake (MDP). The measurement of this uptake in hyperthyroid patients by bone provides a sensitive and objective means of quantifying skeletal metabolism. Using a standard shadow-shield whole-body monitor and radioimmunoassay kit, we have measured whole-body retention of $^{99m}Tc-MDP$ up to 24hr and concentration of serum Osteocalcin in 20 patients with hyperthyroidism and in 42 normals. The results were as follows; 1) The average of serum Osteocalcin level in 42 patients with normals was $9.90{\pm}4.87(ng/ml)$ and in 20 patients with hyperthyroidism was $19.54{\pm}5.7(ng/ml)$. Both the averages of serum Osteocalcin and 24hr $^{99m}Tc-MDP$ uptakes in hyperthyroid patients were higher than those in normals. 2) $^{99m}Tc-MDP$ uptakes in skeletal system increased in proportion to normal ageing after 40 yrs old in 42 patients with normals. The average of $^{99m}Tc-MDP$ uptakes in hyperthyroid patients were higher than those in normals without related ageing. 3) A significant relationships between the $^{99m}Tc-MDP$ uptakes and serum Osteocalcin level were peformed (r=0.55, $y=17.58+6.7\times$). From the above results we concluded that the measurement of serum Osteocalcin and 24hr $^{99m}Tc-MDP$ uptakes can be used for evaluation of bone turnover as a specific marker in hyperthyroid patients.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.13-23
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• 2010

Objective: The present study was carried out to induce differentiation of human embryonic stem cells (hESCs) into germ cells and to establish a culture condition for single hESCs dissociated by enzyme. Methods: Embryonic body (EB) was formed by hanging drop culture for 3 days from hESCs colony. The EBs were cultured in the medium supplemented with retionic acid (RA) or/and bone morphogenetic protein-4 (BMP4) for 14 days to differentiate into germ cells. Germ cell specific markers, c-kit and VASA were used for immunohistochemistry of EB. Human ESCs colonies were dissociated into single cells by Collagenase, Tryple and Accutase, and then colony formation rate of the single cells was examined. Rho-associated kinase inhibitor (ROCK inhibitor, Y27632) was added into the culture medium of single cells to reduce the apoptotic damage during the dissociation. Results: Single cells dissociated with Tryple or Accutase showed higher colony formation rates compared to the cells dissociated with Collagenase. Seeding of $5{\times}10^3$ cells/well (4 well dish) was efficient to obtain high colony formation rate compared to other concentrations of seeding cell. Addition of Y27632 significantly increased the colony formation rate of the single cells dissociated by Tryple. Immunohistochemistry of EB with c-kit and VASA markers showed a weak fluorescence signals compared to the signals from the testicular tissue. Conclusion: Dissociation with Tryple was useful to obtain healthy single cells and addition of Y27632 was beneficial for survival and colony formation of the single cells. Unlike other studies, we just observed a dim fluorescence staining of the germ cell markers, probably caused by the short-term culture for the differentiation of EB compared to other studies.

• Journal of Nutrition and Health
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• v.31 no.3
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• pp.279-288
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• 1998

This study explored the effect of calcium levels and/or ovariectomy on bone metabolism using female Sprague-Dawley weanling rats as a model . Rats received a low (0.1%) calcium diet for 8 weeks. The rats were then divided into three subgroups that were fed 0.1% ,0.5% and 1.5% calcium diets for 8 weeks after operation. The results of this experiment indicate that body weight gin was higher in ovariectomy groups than in sham groups regardless of calcium level and food intake. Serum Ca and P concentrations were of normal level regardless of calcium level and ovariectomy. Estrogen concentration was low in the ovariectomized group. Serum alkaline phophatase activity and urinary hydroxyproline have been used as markers of bone formation and resorption. These values were increased in ovariectomized groups. The weight, length and breaking force of femur were not significantly different between the groups. Ash, Ca, P and total lipid contents in femur and lumbar were decreased in the groups fed low calcium . Mg content was decreased in the ovariectomy and total protein content was not affected by calcium level and ovariectomy. The effect groups of ovarectomy on calcium contents of bone was more prominent in lumbar than in femur. In conclusion, though low calcium intakes during growth period may retard the attainment of peak bone mass, calcium supplementation after this period increased bone growth and mineral contents, but not significant effect in three calcium levels. Furthermore, calcium intake was shown to have a greater influence on the mineral contents of femur than of lumbar, and removal of endogenous estrogen production by ovariectomy was shown to be more deleterious on the ash and calcium contents of the lumbar than of femur.