• The Journal of Korean Medicine
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• v.38 no.3
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• pp.59-72
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• 2017

Objectives: This study was performed to evaluate effects of Sochungryong-tang Extract(SRE) on osteoclast differentiation and bone resorptionin order to find out the possibility for clinical use in preventing and treating osteoporosis. Methods: To evaluate the effect of SRE on osteoclast differentiation, we induced RAW 264. 7 cells to be differentiated to osteoclasts by RANKL (receptor activator of nuclear $factor-{\kappa}B$ ligand). We measured effect on TRAP (Tartrate-resistant acid phosphatase), NFATc, cathepsin K, MMP-9, inflammation related factors, histogenesis factors and bone resorption. Results: SRE decreased osteoclast differentiation, and also decreased expression of bone resorbing factors such as MMP-9, cathepsin K, TRAP, NFATc1, MITF, c-Fos, osteoclast stimulatory transmembrane protein, calcitonin receptor in RANKL-induced osteoclast. SRE also decreased Cyclooxygenase-2, indusible nitric oxide synthase, $TNF-{\alpha}$, which are thought to be related with the inflammatory bone destruction. Conclusion: SRE inhibits osteoclast differentiation and bone resorption. The results indicate that the BHT extract can potentially be applied for preventing and treating osteoporosis.

• The Journal of Internal Korean Medicine
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• v.24 no.2
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• pp.203-212
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• 2003

Objective : As a link in chain of research to confirm the oriental medical prescription which has the anti-atherosclerosis effects, this research evaluated the effects on the macrophage-related factors by using KanghwalSokdantang(KS). Methods : In order to perform this research, we have evaluated the effects on the oxLDL formation from the macrophages, the nitric oxide formation, and the oxidation of macrophages. Thus, with this evaluation, we have investigated the applicapability on the artherosclerosis. Results : KanghwalSokdantang has showed a noticeable reduction of protein oxidation in the process of oxLDL formation, has remarkably restrained phospholipid peroxidation, an index to estimated the phospholipid oxidation and reduction that are formed in the process of macrophage's oxLDL formation, and has increased the nitrite concentration noticeably in the LDL-dealing macrophages. By increasing the survival rate of macrophages, KanghwalSokdantang has restrained the cellular damages. KanghwalSokdantang is ineffective on the LDH outflow from damaged cells. $1{\mu}g/ml$ KanghwalSokdantang sample has increased acid phosphatase activity remarkably. Conclusion : KanghwalSokdantang has the possibility to be used in the prevention and treatment of atherosclerosis, which is formed by the oxLDL formation of macrophages.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.5
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• pp.411-417
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• 2019

Humanin (HN) is a mitochondrial peptide that exhibits cytoprotective actions against various stresses and diseases. HN has been shown to induce the phosphorylation of AMP-activated protein kinase (AMPK), which is a negative regulator of receptor activator of nuclear factor-${\kappa}B$ ligand (RANKL). However, the role of HN in osteoclastogenesis or other skeletal disorders remains unknown. Here, we examined whether HN regulates osteoclastogenesis via AMPK activation using bone marrow-derived macrophage (BMM) cultures. Our results show that HN inhibited RANKL-induced osteoclast formation and reduced the expression of genes involved in osteoclastogenesis, including nuclear factor of activated T-cells cytoplasmic 1, osteoclastassociated receptor, cathepsin K, and tartrate-resistant acid phosphatase. Moreover, HN increased the levels of phosphorylated AMPK protein; compound C, an AMPK inhibitor, recovered HN-induced osteoclast differentiation. In addition, we found that HN significantly decreased the levels of RANKL-induced reactive oxygen species in BMMs. Therefore, these results indicate that HN plays an important role in osteoclastogenesis and may function as an inhibitor of bone disorders via AMPK activation.

• International Journal of Oral Biology
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• v.44 no.3
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• pp.89-95
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• 2019

Piperlongumine (PL) is a natural product found in long pepper (Piper longum). The pharmacological effects of PL are well known, and it has been used for pain, hepatoprotection, and asthma in Oriental medicine. No studies have examined the effects of PL on bone tissue or bone-related diseases, including osteoporosis. The current study investigated for the first time the inhibitory effects of PL on osteoclast differentiation, bone resorption, and osteoclastogenesis-related factors in RAW264.7 macrophages stimulated by the receptor activator for nuclear factor-${\kappa}B$ ligand (RANKL). Cytotoxicity was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and osteoclast differentiation and bone resorption were confirmed by tartrate-resistant acid phosphatase (TRAP) staining and pit formation analysis. Osteoclast differentiation factors were confirmed by western blotting. PL exhibited toxicity in RAW264.7 macrophages, inhibiting osteoclast formation and bone resorption, in addition to inhibiting the expression of osteoclastogenesis-related factors, such as tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Fos, and NFATc1, in RANKL-stimulated RAW264.7 macrophages. These findings suggest that PL is suitable for the treatment of osteoporosis, and it serves as a potential therapeutic agent for various bone diseases.

• Applied Microscopy
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• v.24 no.3
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• pp.10-22
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• 1994

To investigate the morphology of the synovial lining cells, synovitis was induced by carrageenin injection into the rat knee joint cavities. Synovial membranes were excised at 1, 3, 5, 7 and 14 days, and histologic, electron microscopic, histochemical (periodic acid Schiff: PAS, toluidine blue), and enzyme histochemical (acid phosphatase: ACP, nonspecific esterase: NSE and endogenous peroxidase) studies were performed. The results are as follows: Carrageenin induced synovial membrane hypertrophy with synovial cell proliferation and granuloma formation. The proliferated synovial lining cells and macrophages in the granulomatous lesion had round to oval nuclei and large, plump cytoplasm with many phagocytotic materials and vacuoles. Electron microscopically, these cells had small number of granular endoplasmic reticulum and many lysosomes, phagosomes and vaculoes. Mitotic figures were observed at early stage of experiment. PAS and toluidine blue stains showed strongly positive reaction in the cytoplasm of the proliferated lining cells and macrophages in granulomatous lesion. ACP and NSE activities were strong positive in the cytoplasm of the proliferated synovial lining cells and macrophages in the granulomatous lesion. But endogenous peroxidase stains were negative in all prolifeative lining cells and macrophages in granulomatous lesion. Conclusively, carrageenin-induced synovitis showed proliferation of synovial lining cells and granuloma formation in deep layer. The macrophages, which consisted of the lesions and have active phagocytic function, were speculated to proliferate by mitosis of superficial synovial A cells and histiocytes in the deep layer of the synovial membrane.

• Korean Journal of Veterinary Research
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• v.47 no.3
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• pp.265-271
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• 2007

This study investigated the protective effects of ethylene glycol-bis(${\beta}$-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), an extracellular calcium chelator, and ethylenediaminetetraacetic acid (EDTA), which chelates calcium and most metal ions, against carbon tetrachloride ($CCl_4$)-induced acute hepatotoxicity in mice. Mice were treated with EGTA or EDTA at a dose of 20 (low) or 100 mg/kg (high) subcutaneously 1h before $CCl_4$ administration. The mice were fasted and sacrificed 18h after $CCl_4$ treatment. Blood samples were collected from the carotid artery by decapitation under light ether anesthesia. Serum alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglyceride (TG), and cholesterol levels were measured. Malondialdehyde (MDA) production was determined as an index of lipid peroxidation in the liver. The liver, kidneys, and spleen were weighed. We also evaluated the histopathological changes in the liver in each group. The relative weights of the liver were significantly higher in the $CCl_4$-treatment group than in the normal group, except in the high-EDTA treatment group. EGTA and EDTA treatment caused a significant decrease in serum ALP, ALT, and AST levels. Of all of the doses of EGTA and EDTA tested, the high-EDTA dose resulted in the most remarkable inhibitory action. The protective effect in the high-EDTA-treatment group was confirmed histopathologically. The low-EGTA-treatment group showed a significant decrease in serum TG and cholesterol levels. Liver MDA levels were significantly decreased in the EGTA (20 mg/kg) and EDTA (20, 100 mg/kg) groups. These results suggest that EDTA, which chelates both calcium and metal ions, confers better protection in $CCl_4$-induced acute liver damage than does EGTA, a calcium chelator.

• The Korean Journal of Health Service Management
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• v.7 no.3
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• pp.71-82
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• 2013

Arterial stiffness(AS) is an important pathologic state of vascular injury. This study was carried out to elucidate the effect of physiological variables on brachial-ankle pulse wave velocity(BAPWV), index of AS. Four hundred adults(volunteers) participated in this study. Body indices, biochemical, cardiac and inflammatory markers, and right(Rt)- and left(Lt)-BAPWV were measured. Body mass index(BMI), Rt- and Lt-BAPWV, glucose, triglyceride, alkaline phosphatase(ALP), gamma-glutamyl transferase(GGT), creatinine, uric acid, troponin-I(TNI), NT-proBNP and high sensitivity C-reactive protein(hs-CRP) levels were higher than the reference value of each variable. Rt- and Lt-BAPWV were directly correlated with age, body weight, BMI, glucose, ketone, aspartate aminotransferase, alanine aminotransferase, ALP, GGT, total cholesterol, low density lipoprotein, lipoprotein(a), apolipoprotein-B, blood urea nitrogen, heart rate, TNI, creatine kinase, CK-MB, lactic dehydrogenase, myoglobin, hs-CRP, lipase, reumatoid factor, fibrinogen and D-dimer (P<0.05, P<0.01, P<0.001 or P<0.000, respectively), but inversely associated with total bilirubin, uric acid, apolipoprotein-A1 and GFR (P<0.05). These observations suggest that a variety of physiological variables may influence BAPWV, resulting in increased risk or prevention of cardiovascular and/or cerebrovascular attacks. Therefore, physiological variables affecting BAPWV should be regularly controlled.

• Asian-Australasian Journal of Animal Sciences
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• v.16 no.6
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• pp.889-893
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• 2003

Immunomodulatory and therapeutic potential of Enrofloxacin was studied in bovine sub clinical mastitis (SCM). The therapeutic efficacy was adjudged by Somatic cell count and Total bacterial count of the milk, whereas, the immuno modulatory potential of the drug was assessed by measuring myeloperoxidase (MPO) and acid phosphates (ACP) enzyme level in the milk leukocytes. Forty-five cows were divided into three equal groups. Gr I consisting 15 cows served as healthy control, whereas, 30 cows (SCM), Gr II and Gr III, selected on the basis of California Mastitis Test (CMT) positive reaction. Gr II cows received 150 mg of Enrofloxacin, once a day for three days and Gr.III received sterile 5 ml PBS (pH 7.4) for 7days, both the treatment were given by intramammary route. The observation was made up to 30 days post-treatment (PT). The CMT of the healthy milk was negative (0), whereas, it ranged between 1 point score and 2 point score in SCM. The Somatic cell count (SCC) and Total bacterial count (TBC) decreased significantly (p<0.05) on day 3 PT in GrII cows in Enrofloxacin treated group, however, such changes were insignificant in PBS treated group. Traces of MPO and ACP enzyme were found in the healthy milk. The mean ACP level enhanced by 70% on day 3 PT in GrII and only 18.7% in Gr. III cows. The mean MPO level enhanced to 32% in Gr. II and 18 % in Gr. III cows on day 3 PT. Concomitant use of Enrofloxacin in SCM at sub optimal dose was found to reduce the bacterial load by increasing the bactericidal enzyme level in the milk polymorphonuclear cells (PMNs) in bovine SCM, which indicates its immunomodulatory potential in mastitis.

• Macromolecular Research
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• v.14 no.5
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• pp.565-572
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• 2006

Recombinant human bone morphogenic protein-2 (rhBMP-2), which is known as one of the major local stimuli for osteogenic differentiation, was immobilized on the surface of hyaluronic acid (HA)-modified poly$(\varepsilon-caprolactone)$ (PCL) (HA-PCL) scaffolds to improve the attachment, proliferation, and differentiation of human bone marrow stem cells (hBMSCs) for bone tissue engineering. The rhBMP-2 proteins were directly immobilized onto the HA-modified PCL scaffolds by the chemical grafting the amine groups of proteins to carboxylic acid groups of HA. The amount of covalently bounded rhBMP-2 was measured to 1.6 pg/mg (rhBMP/HA-PCL scaffold) by using a sandwich enzyme-linked immunosorbant assay. The rhBMP-2 immobilized HA-modified-PCL scaffold exhibited the good colonization, by the newly differentiated osteoblasts, with a statistically significant increase of the rhBMP-2 release and alkaline phosphatase activity as compared with the control groups both PCL and HA-PCL scaffolds. We also found enhanced mineralization and elevated osteocalcin detection for the rhBMP-2 immobilized HA-PCL scaffolds, in vitro.

• The Journal of Advanced Prosthodontics
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• v.2 no.1
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• pp.18-24
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• 2010

PURPOSE. The aim of this pilot study was to investigate the effect of etched microgrooves on the hydrophilicity of Ti and osteoblast responses. MATERIAL AND METHODS. Microgrooves were applied on Ti to have 15 and $60{\mu}m$ width, and 3.5 and $10{\mu}m$ depth by photolithography, respectively. Further acid etching was applied to create Ti surfaces with etched microgrooves. Both smooth- and acid-etched Ti were used as the controls. The hydrophilicity of Ti was analyzed by determining contact angles. Cell proliferation and osteogenic activity of MC3T3 mouse preosteoblasts were analyzed by bromodeoxyuridine assay and alkaline phosphatase (ALP) activity test, respectively. One-way ANOVA, Pearson's correlation analysis and multiple regression analysis were used for statistics. RESULTS. Etched microgrooves significantly increased the hydrophilicity of Ti compared to the smooth Ti. $60{\mu}m$-wide etched microgrooves significantly enhanced cell proliferation, whereas the osteogenic activity showed statistically non-significant differences between groups. Result of the osteogenic activity significantly correlated with those of hydrophilicity and cell proliferation. Hydrophilicity was determined to be an influential factor on osteogenic activity. CONCLUSION. This study indicates that increase in hydrophilicity of Ti caused by etched microgrooves acts as an influential factor on osteogenic activity. However, statistically non-significant increase in the ALP activity suggests further investigation.