• Archives of Pharmacal Research
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• v.26 no.12
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• pp.1029-1035
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• 2003

Osteoporosis is recognized as one of the major hormonal deficiency diseases, especially in menopausal women and the elderly. When estrogen is reduced in the body, local factors such as IL-1 $\beta$ and IL-6, which are known to be related with bone resorption, are increased and promote osteoclastogenesis, which is responsible for bone resorption. In the present study, we investigated whether glucosidic isoflavones (Isocal, PIII) extracted from Sophorae fructus affect the proliferation of osteoblasts and prevent osteoclastogenesis in vitro by attenuating upstream cytokines such as IL-1$\beta$ and IL-6 in a human osteoblastic cell line (MG-63) and in a primary osteoblastic culture from SD rat femurs. Interestingly, IL-1$\beta$ and IL-6 mRNA were significantly suppressed in osteoblast-like cells treated with 17$\beta$-estradiol (E2) and PIII when compared to positive control (SDB), and this suppression was more effective at $10^{-8}$% than at the highest concentration of $10^{-4}$%. In addition, these were confirmed in protein levels using ELISA assay. In the cell line, the cells showed that E2 was the most effective in osteoblastic proliferation over the whole range of concentration ($10^{-4}%-10^{-12}$%), even though PIII also showed the second greatest effectiveness at $10^{-8}$%. Nitric oxide (NO) was significantly (p<0.05) upregulated in PIII and E2 over the concentration range $10^{-6}% to 10^{-8}$% when compared to SDB, without showing any dose dependency. In bone marrow primary culture, we found by TRAP assay that PIII effectively suppressed osteoclastogenesis next to E2 in comparison with SDB and culture media (control). In conclusion, these results suggest that local bone-resorbing cytokines can be regulated by PIII at lower concentrations and that, therefore, PIII may preferentially induce anti-osteoporosis response by attenuating osteoclastic differentiation and by upregulating NO.

• Archives of Pharmacal Research
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• v.27 no.1
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• pp.99-105
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• 2004

Isoflavones have been a central subject in research on the natural phytoestrogens found in Leguminosae. Their effects on bone formation and remodeling are important in that they can act like estrogen by binding on estrogen receptors on the target cell surface. We, therefore, believed that isoflavones may help in the treatment of patients with estrogen deficiency disease such as estrogen replacement therapy (ERT) for osteoporosis. As commonly known, osteoporosis is one of the hormonal deficiency diseases, especially in menopausal women. When estrogen is no longer produced in the body a remarkable bone remodeling process occurs, and the associated events are regulated by growth factors in the osteoblast lineage. In the present study, we investigated whether isoflavones (Isocal) extracted from Sophorae fructus affect the growth factors IGF-I and TGF-$\beta$ that have been known to be related with bone formation. In the study, we found that the active control (PIII) effectively enhanced the level of nitric oxide (NO) and growth factors, and thereby inhibited osteoclastogenesis. The most efficient concentration was $10^{-8}$% within five days, whereas the comparative control (soybean isoflavone) was not as effective even at a lower concentration. In conclusion, the products which contain enriched glucosidic isoflavone and nutrient supplements such as shark cartilage and calcium can be used for osteoporosis therapy by enhancing the production of IGF-I and TGF-$\beta$. Furthermore, the NO produced through endothelial constitutive NO synthase (ecNOS) may playa role in inhibiting bone reabsorption.

• The Korea Journal of Herbology
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• v.33 no.3
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• pp.45-53
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• 2018

Objectives : In cases of osteoarthritis, the hypofunction of the cartilage and joint leads to a limited range of joint motion, swelling, and pain, which is generally treated using pharmaceutical drugs (e.g., anti-inflammatory agents, cartilage protectants, and nonsteroidal anti-inflammatory drugs) or replacement arthroplasty. However, long-term drug treatment is associated with adverse effects on the gastrointestinal systems. The present study aimed to evaluate the ability of Giheolsotong-hwan to treat of osteoarthritis symptoms in the MIA-induced rat model based on histological analysis, and factors that are associated with inflammation and bone mineral metabolism. Methods : Giheolsotong-hwan was administered orally at doses of 200 mg/kg/day or 400 mg/kg/day for 2 weeks before direct injection of monosodium iodoacetate ($3mg/50{\mu}{\ell}$ of 0.9% saline) into the intra-articular space of the rats' right knee. The rats subsequently received the same doses of oral Giheolsotong-hwan for another 4 weeks. We evaluated the treatment effects based on serum biomarkers and histopathological analysis of the knee joints. Results : Compared to those in control rats, the Giheolsotong-hwan treatments significantly decreased the serum concentration of inflammation factors (i.e., $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, $PGE_2$, and $LTB_4$), and bone degrade factors (i.e., MMP-9, CTX-II, and COMP). In addition, the Giheolsotong-hwan treatments significantly increased the concentration of glycosaminoglycans of bone defence factors, but no chage the TIMP-1. Furthermore, the Giheolsotong-hwan treatments effectively preserved the knee cartilage and proteoglycan. Conclusion : The results indicate that Giheolsotong-hwan treated osteoarthritis symptoms. Thus, Giheolsotong-hwan may be a novel oriental therapeutic option for the management of osteoarthritis.

• Anatomy and Cell Biology
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• v.56 no.4
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• pp.552-561
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• 2023

The endocrinology of type 2 diabetes (T2D) and its predisposing factors have been studied extensively while its skeletal effects have received negligible research despite this being a global disease. The cellular and molecular association between proximal humeral fractures and T2D has not been fully elucidated. We aimed to study bone cell quantities and immunolabel osteogenic and antiosteogenic cytokines. The study used 12-week-old rats (23 males) consisting of 8 Sprague Dawley (SD) and 15 Zucker Diabetic Sprague Dawley (ZDSD). Weekly mass measurements were taken while fasting blood glucose levels were recorded every 2 weeks with oral glucose tolerance tests conducted once every 4 weeks. Upon termination at the age of 28 weeks, humeri were fixed in 10% buffered formalin, prior to decalcification in ethylenediaminetetraacetic acid. The bone samples were then processed in ascending grades of alcohol using an automatic processor before embedding in paraffin wax. Sections were cut at 5 ㎛ thickness in a series for Haematoxylin and Eosin stain, and immunohistochemistry was performed with the anti-tartrate-resistant acid phosphatase (TRAP), anti-alkaline phosphatase (ALP), anti-bone morphogenetic protein 3 (BMP3), anti-transforming growth factor beta 1 (TGFβ1), anti-aged glycation end product (AGE) antibodies in the sequence. ZDSD rats had more adipocytes, BMP3 and AGEs expression with higher numbers of TRAP positive osteocytes and fewer ALP cells although no differences were found in TGFβ1 immunopositivity. We also found that T2D increases the number of AGEs immuno-positive cells, as well as its extracellular expression, thus providing a conducive environment for the interaction of the osteogenic cytokine and its antagonist to suppress osteoblastogenesis. ZDSD groups had higher adipocyte numbers therefore increased marrow adiposity in T2D.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.101-106
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• 2004

This study was conducted to examine the effect of bluefin tuna bone on the bone metabolism of the rats. Weaned 6-week old male rats were fed low-calcium diets for 2 weeks after the adjustment period. Rats were divided into 6 groups and were fed experimental diets for six weeks. Experimental groups were \circled1 Normal calcium: CC (0.5% CaCO$_3$; control) \circled2 TB (bluefin tuna bone powder) \circled3 CT (citrated bluefin tuna bone powder) \circled4 BB (bovine bone powder) \circled5 CL (calcium lactate) \circled6 Low calcium LC (0.15% CaCO$_3$). Low-calcium diet group (LC) showed the lowest calcium retention. There was no differences in calcium excretion in stool and calcium absorption among various calcium sources. Serum calcitonin levels were high in TB, CT and BB group compared to those in CC, CL LC group. Parathyroid hormone and osteocalcin levels showed no differences among experimental groups. Deoxypyridinoline (DPD) levels were significantly higher in LC group than in other groups. Wet weight of the femur were significantly high in TB and CT group, and dry weight of femur showed no differences among normal calcium groups. Bone density of femur in LC group was significantly lower than those of normal calcium feeding group, and TB group showed highest bone density among experimental groups. There was no differences in bone metabolism among various calcium sources. Therefore, it is pointed out that the amount of calcium intake is very important because there was significant differences between normal calcium diet and low calcium diet. According to the results of femur weight, ash, calcium and bone density, it is suggested that bluefin tuna bone have alternative effects to bovine bone powder on the maintenance of bone health.

• International Journal of Industrial Entomology and Biomaterials
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• v.30 no.1
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• pp.17-25
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• 2015

This study aimed to investigate the effects of silk fibroin on bone metabolism in ovariectomized rats. A total of 30 Sprague-Dawley rats were randomized into sham-operated (SHAM), ovariectomized control (OVX), alendronate (OVX+ALEN, 10 mg/kg body weight/d), low silk fibroin (OVX+SF100, 100 mg/kg body weight/d), and high silk fibroin (OVX+SF300, 300 mg/kg body weight/d) groups. All the rats were fed by gavage for 12 wk. At the end of 12 wk, blood and urine were collected for analysis of bone turnover markers, and bone mineral density (BMD) was measured by micro-computed tomography. The results show that the OVX group (p < 0.05) displayed the highest mean body weight gain. Among the five groups, serum levels of bone alkaline phosphatase (ALP) and urine levels of deoxypyridinoline (DPD) were highest in the OVX group (p < 0.05). Bone ALP levels in the ALEN group were significantly lower than that of the silk-treated groups. On the other hand, DPD levels were not significantly different between the ALEN and silk-fibroin-treated groups (p < 0.05). The trabecular BMD was significantly higher in the ALEN and silk-treated groups compared to the OVX group (p < 0.05). In conclusion, this study showed that silk fibroin has similar effects as alendronate, which is used in osteoporosis medication. Therefore silk fibroin might be a new candidate for the prevention and treatment of osteoporosis in patients.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.2
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• pp.139-147
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• 1998

This study has been performed to evaluate the relationship between the remained mineral components in a decalcified bone matrix and an ectopic bone formation efficiency. The freezed rat diaphyseal cortical bones measuring 0.5cm in length were demineralized in heated 0.6N HCl at $60^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 minutes, respectively, using a controlled heat ultrasonic cleaner. Each 1cc of decalcifying solution taken during decalcification procedure was used to calculate calcium content using calcium dignostics kit under 600nm of spectrophotomer. After decalcification, each specimen was also weighed. Then each prepared specimen was implanted into the dorsal pouch of 24 Sprague-Dawley rats divided into 8 groups by time course. The implants were harvested at 1, 2, and 3 weeks and prepared for routine H-E stain specimens to evaluate osteogenic activity. The results are as follows: 1. There was statistical significant difference in change of calcium concentration up to demineralization of 30 minutes and each allogenic bones decalcifed up to 20 minutes revealed 99.65% of decalcification in average. 2. There was statistical significant difference in change of weight in demineralized allogenic bone up to 20 minutes treatment but, no significant change was noted after that time. 3. The histologic analysis revealed active ectopic bone formation in the implanted allografts demineralized for 20, 25, 30 minutes, respectively. However, the other groups of allografts showed relatively poor osteoinductive activity. These findings suggest that complete decalcification with a minimized degeneration of collagen matrix is necessary to induce maximal osteogenesis by decalcified bone allograft.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.44 no.1
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• pp.25-30
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• 2011

We investigated the effect of a lactic acid extract of Sargassum horneri (ExSL) as a calcium supplement on bone formation in 48 female Sprague-Dawley rats for 4 weeks of their growth phase. The rats were divided into four groups based on diet: two calcium-sufficient and two calcium-deficient diets. The normal control group (NC) was fed AIN-93G; the NCS group was fed the same diet containing 1% extract; the calcium-deficient control (DC) diet was based on AIN-93G; and the DCS group received the same calcium-deficient diet plus 1% extract. Bone formation in the rats was evaluated using the wet weight, length, diameter, and bone mineral density (BMD) of the femur. Serum parameters were also examined. The food intake among the groups did not differ significantly (P<0.05). The NCS group gained the most body weight, while the DC group gained much less weight than the other groups. The feeding efficiencies of the groups that received the extract (NCS and DCS) were slightly higher than those of the control groups (NC and DC). The calcium intakes of all groups depended on the amount of calcium in the feed; the NCS and DCS diets contained 12.15 mg more calcium than the NC and DC diets. The calcium absorption was lower in NCS than in DC and DCS, but significantly higher than in NC (P<0.05). The BMDs in the calcium-sufficient groups were not significantly different (P<0.05), while in the calcium-deficient groups the BMD was significantly higher in DCS than in DC (P<0.05). The serum calcium and phosphorus levels in all groups were not associated with markers of bone growth related to the extract. The osteocalcin content and alkaline phosphatase (ALPase) activity were higher in the calcium-deficient groups than in the normal groups (P<0.05). Ultimately, the osteocalcin content and ALPase activity were lower in DCS compared to DC. These results suggest that the addition of ExSL promotes bone formation and calcium absorption in growing rats.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.501-521
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• 2002

The purpose of this study is to evaluate the influences of the bone morphogenetic protein (BMP) on the healing of periodontal ligament and alveolar bone after replantation of tooth, and to examine the possibility of its clinical application. 45 Sprague Dawley rats weighted about 100 gram were divided into 3 experimental groups by different dose of BMP. All the upper right and left 1st molar were extracted after 5 days feeding of 0.4% ${\beta}$-aminopropionitrile, and right molar were used as experimental group and left molar were used as control group. The root surface of experimental molar were treated with 25,50 and l00ng/ml of human recombinant Bone morphogenetic protein-4 (rh-BMP-4) with micropipet, and 1M Sodium hypochloride were used on control root surface. All the experimental animals were sacrificed as 1, 2, 4, 7 and 14 days after autoreplantation of upper 1st molar into their own position. The maxilla were disected included both side of 1st molar. The collected tissue were processed from demineralization to paraffin embeding as usual procedure, and the specimens were prepared with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows : 1. There was no significant differences between control and experimental site on 1 and 2 days after replantation of tooth. In the case of 4th days, the evidence of tissue regeneration were observed on experimental site to compare the controls. New osteoid were revealed on high concentration of BMP at 7 days after replantation, and it became more obvious at 14 days, 2. The effect of the rh-BMP-4 coated on root surface was revealed slight influences for the prolifertion of cells of periodontium and tissue regeneration as dose-dependent pattern. 3. Bony ankylosis was observed between alveolar bone and root surface due to the remarkable amount of osteoid formation on the 14 days after replantation of root. In the conclusion, it was suggested that topical application of the rhBMP-4 on the root surface has influence on the periodontal ligament and alveolar bone. The application method of BMP on the root should be designed with calculation of proper concentration.

• International Journal of Industrial Entomology and Biomaterials
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• v.32 no.2
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• pp.80-89
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• 2016

The effects of Cudrania tricuspidata (CT) extract on markers of osteoporosis were examined in ovariectomized rats. We classified 26 rats into five groups and provided a pellet chow diet and tap water throughout the 27-wk experimental period. During the last 15 wk, we added oral injections to each group as follows: sham-operated (SHAM, n=4) and ovariectomized-control (OVX, n=5) with distilled water, alendronate with 10 mg/kg/d of alendronate sodium (ALEN, n=5), CT (CT100, n=6) with 100 mg/kg/d of CT, and CT (CT300, n=6) with 300 mg/kg/d of CT. After the experimental period, blood, urine, and micro-CT images were assessed. The CT100 and OVX groups did not show any significant differences in urinary n-terminal telopeptide (NTx) (p<0.05 ), but with increases in CT concentration, the NTx level was slightly reduced. Serum osteocalcin was significantly higher in the CT groups than in all other groups (p<0.05 ). Notably, the serum calcium levels of all groups were within the normal range, but urinary calcium levels in the CT groups were significantly lower than the OVX group (p<0.05 ). In addition, the CT groups exhibited higher trabecular BMD than the OVX groups while showing similar BMD to the ALEN group (p<0.05 ). The Tb.Th of the ALEN group was lower than all other groups. Based on the overall analysis of results, CT prevented bone loss by inhibiting bone resorption and enhancing bone formation. Although alendronate showed a similar effect in preventing bone loss, it did so by solely inhibiting bone resorption, and its long-term use reportedly causes paradoxical effects such as hip fractures. Thus, for osteoporosis induced by ovariectomy, we conclude that CT extract is an effective natural treatment without severe side effects.