• The Journal of Internal Korean Medicine
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• v.40 no.1
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• pp.58-69
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• 2019

Objective: This study investigated the effects of Hansu-Daebowon (HDW) on bone resorption in vitro and bone loss in vivo. Methods: Osteoclast differentiation was measured by counting TRAP (+) MNC formed from RAW 264.7 in the presence of RANKL. Bone pit formation was determined in an artificial bone slice loaded with RANKL-stimulated osteoclasts. To elucidate the mechanisms of the inhibitory effects of HDW on bone resorption and osteoclast differentiation, osteoclastogenic genes (i.e. TRAP, MMP-9, NFATc1, c-Fos, and Cathepsin K) were measured using real time PCR. Furthermore, bone loss was observed using micro-CT in an LPS-treated mammal model. Results: HDW inhibited the bone pit formation in vitro and inhibited bone loss in vivo. Moreover, HDW decreased the number of TRAP (+) MNCs in the presence of RANKL, and HDW inhibited the expressions of cathepsin K, MMP-9, TRAP, NFATc1, and c-Fos in the osteoclasts. Conclusion: HDW exerts inhibitory effects on bone loss and bone resorption resulting from the inhibitions of osteoclast differentiation and osteoclastogenic gene expression.

• Journal of Dental Rehabilitation and Applied Science
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• v.32 no.3
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• pp.158-168
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• 2016

This article is to review various methods used to investigate internal and marginal adaptation of fixed dental prostheses, and to summarize a merit, worth, and limitation of each method, using some results of previous studies. The methods of measuring internal and marginal gap are divided into two categories in this study; in vivo and in vitro. In vivo methods are clinical evaluations, including exploration, radiography, and impression technique. In vitro methods are laboratory evaluations such as direct view, cross-sectioning, and silicone replica technique using microscope. Measuring by micro computed tomography (CT) or profilometer is also in vitro methods. In recent years, the development of scanning systems is able to analyze 3-dimensional internal and marginal space in detail. As measuring and analyzing technology become more advanced, the ability to thoroughly examine crown adaptation is becoming both simpler and more efficient.

• The Korea Journal of Herbology
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• v.32 no.2
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• pp.41-47
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• 2017

Objective : The aim of this study is to investigate anti-arthritis activity using a korea's natural monument No. 265 designate 'Yeonsan-Ogye'. In this study, research by using extracts from different concentration of the Yeonsan-Ogye through an MIA-induced arthritis animal model was being conducted in vivo and scientifically verifying the efficacy of medicinal food. Methods : Yeonsan-Ogye was administered 500 mg/kg/day, 1000 mg/kg/day, 2000 mg/kg/day to SD-Rat, where arthritis was induced by monosodium iodoacetate (MIA) at $60mg/m{\ell}$. MMP-9, COMP, CTXII, calcitonin and glycosaminoglycan level in serum were measured by ELISA. The changes of relative hind paw weight bearing ratio by Incapacitance Test Meter and The cartilage of meniscus volume was examined and 3-D high-resolution reconstructions of the cartilage of meniscus were obtained using a Micro-CT system. Also, the histopathological analysis of knee was observed by H&E and safranin-O staining. Results : Production of MMP-9, COMP (all groups) and CTXII (500, 1000 mg/kg/day) level in serum was decreased, respectively, in comparison with control. The other way, production of calcitonin (500, 1000 mg/kg/day) and glycosaminoglycan (all groups) level in serum, Hind paw weight bearing ratio (all groups) was increased, espectively, in comparison with control. The cartilage of patella volume in micro-CT increased significantly. In addition, all groups showed a increase in the cartilage volume and proteoglycan. Conclusion : The results for Yeonsan-Ogye showed significant antiarthritis activity in serum and the cartilage. Therefore, it is thought to be that Yeonsan Ogye can be utilized as a variety of new korea medicie and health foods against arthritis diseases.

• Journal of Industrial and Engineering Chemistry
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• v.67
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• pp.244-254
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• 2018

Surface modified poly ${\text\tiny{L}}$-lactic acid (PLLA) samples with hydroxyapatite (HA), heparin and bone morphogenetic protein-2 (BMP-2) mediated by polydopamine (pDA) coating (PLLA/pDA/HA/Hep/BMP-2) were prepared, and their effects on the enhancements of bone formation and osseointegration were evaluated in vitro and in vivo as compared to PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. The changes in surface chemical compositions, morphologies and wettabilities were observed by X-ray photoelectron spectroscopy (XPS), field-emission scanning electron microscopy (FE-SEM), atomic force microscopy (AFM) and water contact angle measurements. Pre-coating of HA particles with pDA provided uniform and homogeneous anchoring of particles to PLLA surface. In addition, the strong ionic interaction between heparin and pDA led PLLA surface readily heparinized for loading of BMP-2. In vitro experiments revealed that the levels of alkaline phosphatase (ALP) activity, calcium deposition, and osteocalcin (OCN) gene expression were higher in MG-63 human osteosarcoma cell lines grown on PLLA/pDA/HA/Hep/BMP-2 than on control PLLA, PLLA/pDA/HA, and PLLA/pDA/Hep/BMP-2. In vivo studies using micro-computed tomography (micro-CT) also showed that PLLA/pDA/HA/Hep/BMP-2 screw exhibited greatest value of bone volume (BV) and bone volume/tissue volume (BV/TV) among samples. Histological evaluations with H&E and Von Kossa staining demonstrated that a combination of HA and BMP-2 contributed to the strong osseointegration.

• Journal of Biomedical Engineering Research
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• v.41 no.6
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• pp.235-246
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• 2020

This study aimed to evaluate the inhibitory effect of micro-current stimulation(MCS) on adipogenesis regarding with Wnt/β-catenin pathway using the ob/ob mouse and 3T3-L1 cell line. 6-week old ob/ob male mice were equally assigned to four groups: obese group(ob), obese with MCS groups(50 μA, 200 μA, and 400 μA). 6-week old C57BL/6J male mice were assigned to the control group(CON). We analyzed abdominal adipose tissue volume by using in vivo micro-CT and measured the body weight, feed intake, liver weight and triglycerides in serum. All the MCS groups showed that significantly reduced body weight and triglycerides in serum. In the case of liver weight and abdominal adipose tissue volume, the inhibitory effect of adipogenesis was shown in the 200 μA and 400 μA groups. To elucidate the anti-obesity effect of MCS, β-catenin, C/EBPα and FAS protein expressions were analyzed by western blotting. β-catenin expression was upregulated, C/EBPα and FAS expression were down-regulated in the relatively high-intensity groups(200 μA and 400 μA). Thus, the 200 μA and 400 μA for the intensity of MCS were chosen for cell experiments. In the 3T3-L1 cell line, Wnt/β-catenin pathway including Wnt10b, Wnt3a, β-catenin and Cyclin D1 was activated in all MCS groups. Accordingly, the expression level of C/EBPα was decreased during the differentiation and lipid droplet was significantly reduced in Oil red O staining results. These results suggest that the Wnt/β-catenin signaling might be activated by MCS with current intensities between 200-400 μA and it may lead to anti-obesity effects.

• Biomolecules & Therapeutics
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• v.28 no.4
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• pp.337-343
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• 2020

Activation of osteoclast and inactivation of osteoblast result in loss of bone mass with bone resorption, leading to the pathological progression of osteoporosis. The receptor activator of NF-κB ligand (RANKL) is a member of the TNF superfamily, and is a key mediator of osteoclast differentiation. A flavanone glycoside isolated from the fruit of Poncirus trifoliata, poncirin has anti-allergic, hypocholesterolemic, anti-inflammatory and anti-platelet activities. The present study investigates the effect of poncirin on osteoclast differentiation of RANKL-stimulated RAW264.7 cells. We observed reduced formation of RANKL-stimulated TRAP-positive multinucleated cells (a morphological feature of osteoclasts) after poncirin exposure. Real-time qPCR analysis showed suppression of the RANKL-mediated induction of key osteoclastogenic molecules such as NFATc1, TRAP, c-Fos, MMP9 and cathepsin K after poncirin treatment. Poncirin also inhibited the RANKL-mediated activation of NF-κB and, notably, JNK, without changes in ERK and p38 expression in RAW264.7 cells. Furthermore, we assessed the in vivo efficacy of poncirin in the lipopolysaccharide (LPS)-induced bone erosion model. Evaluating the micro-CT of femurs revealed that bone erosion in poncirin treated mice was markedly attenuated. Our results indicate that poncirin exerts anti-osteoclastic effects in vitro and in vivo by suppressing osteoclast differentiation. We believe that poncirin is a promising candidate for inflammatory bone loss therapeutics.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.34 no.1
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• pp.1-11
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• 2012

Purpose: The purpose of this study was to evaluate the bone regeneration capacity of silk fibroin (SF) when combined with beta tricalcium phosphate (${\beta}$-tricalcium phosphate [TCP]) and rh-bone morphogenetic protein (BMP) in vivo by micro-computed tomography (CT), soft x-ray, and histological analysis. Methods: A total of 56 critical size defects formed by a trephine bur made on 28 adult female Spague-Dawley rats were used for this study and the defect size was 5.0 mm in diameter. The defects were transplanted with (1) no graft material (raw defect), (2) autogenous bone, (3) SF ($10{\mu}g$), (4) SF-BMP ($10{\mu}g$, $0.8{\mu}g$ each), and (5) SF+${\beta}$-TCP ($10{\mu}g$). At 4 and 8 weeks after operation, the experimental animals were sacrificed. Samples were evaluated with soft x-ray, histological examinations and 3-dimensional micro-CT analysis. Results: In the 3-dimensional micro-CT evaluation, bone volume and bone surface data were higher in the SF-BMP ($12.8{\pm}1.5$, $138.6{\pm}45.0$ each) (P<0.05) and SF-TCP ($12.3{\pm}1.5$, $144.9{\pm}30.9$ each) group than in the SF group ($6.1{\pm}3.3$, $77.2{\pm}37.3$ each) (P<0.05), except for the autogenous group ($15.0{\pm}3.0$, $190.7{\pm}41.4$ each) at 4 weeks. At 8 weeks, SF-BMP ($16.8{\pm}3.5$, $173.9{\pm}34.2$ each) still revealed higher (P<0.05) bone volum and surface, but SF-TCP ($11.3{\pm}1.5$, $1132.9{\pm}52.1$ each) (P=0.5, P=0.2) revealed the same or lower amount compared with the SF group ($13.8{\pm}2.7$, $127.5{\pm}44.8$ each). The % of bone area determined by radiodensity was higher in the SF-TCP ($31.4{\pm}9.1%$) and SF-BMP ($36.2{\pm}16.2%$) groups than in the SF ($19.0{\pm}10.4$) group at the period of 4 weeks. Also, in the histological evaluation, the SF-BMP group revealed lower inflammation reaction, lower foreign body reaction and higher bone healing than the SF group at postoperative 4 weeks and 8 weeks. The SF-TCP group revealed lower inflammation at 4 weeks, but accordingly, as the TCP membrane was absorbed, inflammatory and foreign body reaction are increased at 8 weeks. Conclusion: The current study provides evidence that the silk fibrin can be used as an effective grafted material for tissue engineering bone generation through a combination of growth factor or surface treatment.

• Journal of Biomedical Engineering Research
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• v.38 no.1
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• pp.49-55
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• 2017

The prevalence of obesity has noticeably increased worldwide over several decades with various complication. Even though anti-obesity drug treatments have been spotlighted by resulting in effective mean weight losses, its adverse effects cannot be overlooked. Thus, this study aimed to evaluate the effects of multi-frequency whole body vibration, one of the mechanical stimulus, as a countermeasure against obesity. Thirty-two-6-week-old C57BL/6J male mice were equally assigned to four groups: the Control group (CON, n = 8), the Sham group (Sham, n = 8), the sham with single frequency whole body vibration (S+V, n = 8), and the sham with multi frequency whole body vibration (S+MV, n = 8). After 4 weeks, morphologic changes in the adipose tissue were evaluated from three-dimensional images using in vivo micro-computed tomography. At 4 weeks, the volume of the abdominal adipose tissue, which had the highest value in Sham group, noticeably reduced in S+MV group compared to it in S+V group. These results implied that the accumulation of abdominal adipose tissue can be effectively reduced through applying multi-frequency whole body vibration.

• Journal of the Korean Society for Precision Engineering
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• v.29 no.5
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• pp.578-583
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• 2012

The pharmacological therapies and whole body vibration as non-pharmacological therapies were known to have adverse side effects. Therefore, partial stimulation was suggested and its effects were evaluated. This study aimed to evaluate the site-specific effects of partial stimulator for treatment of osteoporosis induced by estrogen deficiency. Sixteen virginal Sprague-Dawley rats (12 weeks old) were divided into 2 groups(no stimulation, stimulation groups). All rats were ovariectomised to induce osteoporosis. After 3 weeks of operation, the right tibiae in rats of stimulation group (frequency: 10Hz, cycle: 1500, strain on bone surface: $2000{\mu}{\varepsilon}$) were stimulated perpendicularly at right tibia by using partial stimulator for 6 weeks (3days/week). The right tibiae in rats were scanned, before stimulation (0 week) and at 6 weeks after stimulation by using in-vivo micro computed tomography. For investigation of changes in morphological characteristics, structural parameters were measured and calculated. At 6 weeks the morphological characteristics (relative value) in stimulation group were significantly enhanced than those in no stimulation group (p<0.05). In this study, we find that after 6 weeks of partial stimulation, the morphological characteristics of tibia trabecular bone were enhanced. Thus, we concluded that partial stimulation could be used to treat osteoporosis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.40
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• pp.5.1-5.8
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• 2018

Background: The objective of this study was to evaluate the influence of masticatory muscle injection of botulinum toxin type A (BTX-A) on the growth of the mandibular bone in vivo. Methods: Eleven Sprague-Dawley rats were used, and BTX-A (n = 6) or saline (n = 5) was injected at 13 days of age. All injections were given to the right masseter muscle, and the BTX-A dose was 0.5 units. All of the rats were euthanized at 60 days of age. The skulls of the rats were separated and fixed with 10% formalin for micro-computed tomography (micro-CT) analysis. Results: The anthropometric analysis found that the ramus heights and bigonial widths of the BTX-A-injected group were significantly smaller than those of the saline-injected group (P < 0.05), and the mandibular plane angle of the BTX-A-injected group was significantly greater than in the saline-injected group (P < 0.001). In the BTX-A-injected group, the ramus heights II and III and the mandibular plane angles I and II showed significant differences between the injected and non-injected sides (P < 0.05). The BTX-A-injected side of the mandible in the masseter group showed significantly lower mandibular bone growth compared with the non-injected side. Conclusion: BTX-A injection into the masseter muscle influences mandibular bone growth.