• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.4
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• pp.331-339
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• 2007

Background: Distraction osteogenesis(DO) is a useful method for treating cases demanding the generation of new bone. During DO, the angiogenic activity is crucial factor in the new bone formation. The aim of this study was to detect the autocrine growth activity in the cellular components of the distracted bone with observation of the co-expression of vascular endothelial growth factor(VEGF) and its receptors following the mandibular DO. Materials and methods: Unilateral mandibular distraction(0.5 mm twice per day for 10 days) was performed in six mongrel dogs. Two animals were killed at 7, 14, and 28 days after completion of distraction, respectively. Immediately after the animals were killed, the right mandibles were harvested en block. Immunohistochemical staining was processed for observation of the VEGF expression, and double immunofluorescent staining was also processed for detection of the co-expression of osteocalcin and VEGF's two distinct receptors(VEGFR-1 and VEGFR-2). Results: At 7 and 14 days after distraction, the expressions of VEGF were significantly increased in the osteogenic cells of the distracted bone. Up to 28 days after distraction, VEGF was still expressed moderate in the osteoblastic cells of distracted bone. The co-expressions of osteocalcin/VEGFR-1 and osteocalcin/VEGFR-2 were observed in the distracted bone at 7 and 14 days after distraction. In the double immunofluorescent staining, the co-expression' s level of osteocalcin/VEGFR-1 was more than that of osteocalcin/VEGFR-2. Conclusion: Taken together, this study suggested that VEGF plays an important role in the osteogenesis, and these osteoblastic cell-derived VEGF might act as autocrine growth factor during distraction osteogenesis. In the other word, the cellular components, such as osteoblasts and immature fibroblast-like cellsor mesenchymal cells in the distracted bone, might have autocrine growth activity during distraction osteogenesis.

• Journal of Periodontal and Implant Science
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• v.36 no.4
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• pp.839-847
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• 2006

The aim of this study is to monitor reporter gene expression under osteocalcin gene promoter, using a real-time molecular imaging system, as tool to investigate osteoblast differentiation. The promoter region of mouse osteocalcin gene 2 (mOG2), the best-characterized osteoblast-specific gene, was inserted in promoterless luciferase reporter vector. Expression of reporter gene was confirmed and relationship between the reporter gene expression and osteoblastic differentiation was evaluated. Gene expression according to osteoblstic differentiation on biomaterials, utilizing a real-time molecular imaging system, was monitored. Luciferase was expressed at the only cells transduced with pGL4/mOGP and the level of expression was statistically higher at cells cultured in mineralization medium than cells in growth medium. CCCD camera detected the luciferase expression and was visible differentiation-dependent intensity of luminescence. The cells produced osteocalcin with time-dependent increment in BMP-2 treated cells and there was difference between BMP-2 treated cells and untreated cells at 14days. There was difference at the level of luciferase expression under pGL4/mOGP between BMP-2 treated cells and untreated cells at 3days. CCCD camera detected the luciferase expression at cells transduced with pGL4/mOGP on Ti disc and was visible differentiation-dependent intensity of luminescence This study shows that 1) expression of luciferase is regulated by the mouse OC promoter, 2) the CCCD detection system is a reliable quantitative gene detection tool for the osteoblast differentiation, 3) the dynamics of mouse OC promoter regulation during osteoblast differentiation is achieved in real time and quantitatively on biomaterial. The present system is a very reliable system for monitoring of osteoblast differentiation in real time and may be used for monitoring the effects of growth factors, drug, cytokines and biomaterials on osteoblast differentiation in animal.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.2
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• pp.309-317
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• 2000

Bisphosphonates inhibit bone resorption in vivo and in vitro. Currently proposed mechanism of action of bisphosphonates involves both direct effect on osteoclasts and indirect effect through the mediation of osteoblasts. Recent understanding of molecular mechanism of osteoclastogenesis indicates that osteoclast differentiation is quite tightly regulated by signaling molecules from differentiating osteoblasts. Therefore this investigation was designed to elucidate the effect of bisphosphonate on osteoblast differentation. For this purpose, in vitro effects of etidronate and alendronate on the expression of Cbfa1 a master control gene of osteoblast differentiation, several bone marker genes, and formation of calcified nodules were evaluated. To evaluate the effect of bisphosphonate on calcified nodule formation, osteoblasts isolated from rat calvaria were cultured in a-MEM containing $10^{-4},\;10^{-5},\;10^{-6}M$ of etidronate or $10^{-6},\;10^{-7},\;10^{-8}M$ of alendronate for 15 days, and then stained by alizarin red to determine mineralization. To evaluate the effect of bisphosphonate on osteoblast differentiation, osteoblast cells were cultured in a-MEM containing $10^{-4},\;10^{-5},\;10^{-6}M$ of etidronate or $10^{-6}$ M of alendronate for 8 days. And then total RNA was extracted and northern blot analysis was done to examine the expression of Cbfa1, type I collagen, alkaline phosphatase, osteopontin and osteocalcin. The results were as follows: 1. Etidronate suppressed the calcification of bone nodule in dose dependent manner, while alendronate didn't. 2. The expression of Cbfa1 was decreased dose dependently by etidronate, but increased by alendronate. 3. Etidronate suppressed the expression of type I collagen, osteopontin and osteocalcin in dose dependent manner however alendronate promote the expression of osteoblast marker gene. 4. The expression of alkaline phosphatase was not affected either etidronate nor alendronate. These results suggest that etidronate suppressed the expression of Cbfa1 in dose dependent manner, and consequently the expression of osteoblast marker genes, such as type I collagen, osteopontin and osteocalcin were also suppressed in similar manner. And finally this decreased expression of osteoblastic marker gene prevent calcined bone nodule formation.

• Proceedings of the Korean Nutrition Society Conference
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• 2001.05a
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• pp.181.1-181
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• 2001

• Journal of Acupuncture Research
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• v.25 no.2
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• pp.243-257
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• 2008

Objective & Methods : The purpose of this study is to observe the effects of Cyperi Rhizoma Herbal-acupuncture infusion solution at Umgok($KI_{10}$) on Osteoporotic Rats Induced by Ovariectomy. The author performed several experimental items to analyze cytotoxic, osteoporosis evaluation, change of ALP, osteocalcin, Ca and histological change of tibia. Result : 1. BMD were increased non-meaningly in CR-HA than control group. 2. ALP was not inhibited by CR-HA, osteocalcin was decreased meaningly in CR-HA than control group. 3. Osteoclast like cell in tibia was increased meaningly in control group, decreased meaningly in CR-HA. 4. In the histological study in tibia, TBV was significantly increased, GPL was significantly decreased in the CR-HA than control group. Conclusions : From the result above, the results suggest that CR-HA at $KI_{10}$ has significant effect on osteoporosis, and to be put to practical use in the future osteoporosis clinic.

• The Journal of Korean Medicine
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• v.23 no.3
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• pp.54-62
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• 2002

Objectives : This study was undertaken to investigate the effects of Amydae Carapax (AC) on parameter related to bone metabolism in ovariectomized rats. Methods : We measured alkaline phosphatase activity and contents of estrogen, calcium, hydroxyproline, osteocalcin, calcitonin and parathyroid hormone after the ovariectomized rats were treated with AC for 30 days. Results : Serum estrogen, calcium and calcitonin contents in ovariectomized rats significantly decreased, but increased after AC treatment. [Significant increase of serum alkaline phosphatase activity, parathyroid hormone activity and osteocalcin content in ovariectomized rats was remarkably decreased by AC treatment. Increase of urinary calcium and hydroxyproline content in ovariectomized rats was decreased by AC treatment.] Conclusions : These results shows that AC has the ability to counteract abnormal calcium metabolic processes due to sex hormone inequality, promoting bone absorption and inhibiting bone formation.

• The Journal of Korean Medicine
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• v.40 no.4
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• pp.61-71
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• 2019

Objectives: The purpose of this study is to determine the effect of Yuhyangjeongtong-san on the recovery of tibial fractures in rats. Methods: In this study, osteocalcin and Calcitonin, CTX-2, TGF-β and BMP-2, which are used as indicators of bone formation, were analyzed after hematologic fractures using experimental rats. In addition, the fracture union process was confirmed using X-rays. Results: Osteocalcin, Calcitonin and BMP-2 showed a significant increase compared with the control at 4 weeks. CTX-2 and TGF-β showed a significant increase compared with the control at 3 weeks. On X-ray, YJS treated group, as the experiment progressed, the boundary line became blurred, the bone outline was clearly visible, and the fracture recover was progressing. Conclusion: The findings suggest that YJS can play a significant role in the repair of fractures. Therefore YJS is likely to be used to treat fractures.

• The Journal of Korean Medicine
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• v.22 no.4
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• pp.1-9
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• 2001

Objectives : To investigate the effect of Taxilli Ramulus (TR) extract on bone metabolism of ethanol-treated animal model. Methods : The changes of serum calcium, calcitonin, estrogen level, a1ka1ine phosphatase activity, osteocalcin, parathyroid hormone content and urine calcium level were observed with ethanol treatment for 60 days. The results were compared with an ethanol- TR extract double treatment group. Results : We observed increment of serum osteocalcin, parathyroid hormone content, alkaline phosphatase activity and urine calcium level by chronic ethanol feed and they were recovered to near normal level with Taxilli Ramulus extract treatment. Weight gain, serum calcium level, calcitonin and estrogen content were remarkably reduced with ethanol treatment and their levels were normalized by Taxilli Ramulus extract. Conclusions : These results showed that Taxilli Ramulus extract have the ability to recover to normal in the body an abnormal calcium metabolism process due to external factors. These results suggested that Taxilli Ramulus extract have preventive effects on calcium concentration loss and osteoporosis.

• YAKHAK HOEJI
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• v.43 no.4
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• pp.526-534
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• 1999

We investigated the effect of safflower-seed on fracture healing of fracture model in rat. Fracture healing was evaluated by examining the degree of wound healing macroscopically, radiography, bone histomorphometry and biochemical examination. After 1, 3, 5, 7 days, the would healing was accelerate in safflower-seed diet group. Radiography does not reveal the difference in fracture healing between two group. After 2 weeks, safflower-seed had a significant, stimulatory effect on external callus formation (p<0.05). But after 4, 6, 8 weeks, no difference was observed between normal and safflower-seed dietgroup in callus size. Urinary hydroxyproline, osteocalcin and total alkaline phosphatase decreased significantly (p<0.05) in safflower-seed treated group at 2 week after tracture.

• The Journal of Internal Korean Medicine
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• v.22 no.2
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• pp.175-181
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• 2001

Objectives : This study was undertaken to study the action mechanism of Rhizoma Davalliae (RD) at parameter related to regulating bone density. Methods : We measured alkaline phosphatase activity and the contents : calcium, hydroxyproline, osteocalcin, calcitonin, and parathyroid hormone after the ovariectomized rats had been treated with RD for 30 days. Results : The following changes occurred: First, the serum calcium content and the calcitonin content, which decreased in ovariectomized rats, increased with RD treatment. Second, the serum alkaline phosphatase activity, the parathyroid hormone ativity, the osteocalcin content, the urinary calcium content, and the hydroxyprolin content-which increased in ovariectomized rats-decreased with RD treatment. Conclusions : These results show that RD treatment can recover abnormal calcium metabolic process by sex hormone inequality, promoting bone formation and inhibiting bone absorption.