• Title/Summary/Keyword: Cellular activity

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Protection of UV-derived Skin Cell Damage and Anti-irritation Effect of Juniperus chinensis Xylem Extract (향나무추출물의 광손상으로부터 피부세포 보호와 자극완화 효과에 대한 연구)

  • 김진화;박성민;심관섭;이범천;표형배
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.1
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    • pp.63-71
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    • 2004
  • The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98% at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30%. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.

Antioxidant and Antigenotoxic Effects of Shiitake Mushrooms Affected by Different Drying Methods (건조 방법에 따른 표고버섯의 항산화능과 항유전독성 효과)

  • Kim, Min-Jung;Chu, Won-Mi;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.8
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    • pp.1041-1048
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    • 2012
  • Shiitake mushroom (SM; Lentinus edodes) are cultivated and consumed in many Asian countries including Vietnam, China, Japan, Korea, and Thailand. In Asia, SM are mainly dried and used as flavoring. The aim of this study was to compare the effects of SM created with different drying processes, such as oven-dried and sun-dried, on the antioxidative and antigenotoxic effects. Raw and dried SM were extracted with acetone, ethanol, methanol, and hot water. The antioxidant effects of SM were evaluated by determining total phenolic content, DPPH radical scavenging activity (RSA), an ORAC assay, and a cellular antioxidant capacity (CAC) assay. The inhibitory effect of SM on oxidative stress-induced DNA damage in human leukocytes was evaluated by a Comet assay. The total phenolic content of raw SM extracted with methanol and of that extracted with water were significantly higher than the dried SM. Among the water extracts, the $IC_{50}$ for DPPH RSA of raw and sun-dried SM were significantly higher than that of oven-dried SM. Sun-dried SM showed the most potent ORAC value at 50 g/mL. The CAC against $AAPH^-$ induced oxidative stress in HepG2 cells, and $H_2O_2$ induced DNA damage were effectively protected against by all SM extracts. These results suggest that unprocessed SM are the best antioxidants, and that the sun-dried method would be the best option to use in terms of antioxidant activity and the antigenotoxic effect.

Antioxidant capacity and hepatoprotective effect of ethyl acetate fraction from shoot of Aralia elata on alcohol-induced cytotoxicity (두릅 아세트산 에틸 분획물의 산화방지 효과 및 알코올에 대한 간세포 보호효과)

  • Kwon, Bong Seok;Park, Seon Kyeong;Kim, Jong Min;Kang, Jin Yong;Park, Sang Hyun;Kang, Jeong Eun;Lee, Chang Jun;Park, Su Bin;Yoo, Seul Ki;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.216-224
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    • 2018
  • To evaluate physiological effect of Aralia elata, in vitro antioxidant activity and hepatic protective effects were investigated. Ethyl acetate fraction from Aralia elata (EFAE) had higher total phenolic content than other fractions (n-hexane, chloroform, and distilled water layers). EFAE also showed significantly greater radical scavenging activity against 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH), than other fractions. Moreover, EFAE showed dose-dependent inhibitory effect of malondialdehyde (MDA). Hepatoprotective effects of EFAE against ethanol- and $H_2O_2$-induced oxidative stress and cytotoxicity in H4IIE and HepG2 hepatic cells were examined using 2',7'-dichlorofluorescein diacetate (DCF-DA) and 3-[4,5-dimethythiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that EFAE reduced cellular oxidative stress, and increased hepatic cell viability. In addition, EFAE inhibited ethanol-induced lipid accumulation in HepG2 cells. Finally, physiological substances of EFAE were analyzed using high performance liquid chromatography (HPLC), and the major bioactive compounds identified were 3,5-dicaffeoylquinic acid and chlorogenic acid.

Effects of Potassium-Channel Opener on Thallium-201 Kinetics: In-vitro Study in Rat Myocyte Preparations and In-vivo Mice Biodistribution Study (K-통로개방제가 배양심근세포와 생쥐 체내의 Thallium-201역동학에 미치는 영향)

  • Lee, Jae-Tae;Kim, Eun-Ji;Ahn, Byeong-Cheol;Sohn, Kang-Kyun;Lee, Kyu-Bo;Ha, Jeoung-Hee;Kim, Chun-K.
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.4
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    • pp.507-515
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    • 1996
  • Background : Potassium channel opener (K-opener) opens ATP-sensitive K'-channel located at cell membrane and induces potassium efflux from cytosol, resulting in intracellular hyperpolarization. Newly synthesized K-opener is currently examined for pharmacologic potency by means of rubidium release test from smooth muscle strip pre-incubated with Rb-86. Since in-vivo behavior of thallium is similar to that of rubidium, we hypothesized that K-opener can alter T1-201 kinetics in vivo. Purpose : This study was prepared to investigate the effects of pinacidil (one of potent K-openers) on the T1-201 uptake and clearance in cultured myocyte, and in-vivo biodistribution in mice. Methods : Spontaneous contracting myocytes were prepared to imitate in-vivo condition from 20 hearts of 3-5 days old Sprague-Dawley rat and cultured for 3-5 days before use ($5{\times}10^5$ cells/ml). Pinacidil was dissolved in 10% DMSO solution at a final concentration of 100nM or l0uM and was co-incubated with T1-201 in HBSS buffer for 20-min to evaluate its effect on cellular T1-uptake, or challenged to cell preparation pre-incubated with T1-201 for washout study. Two, 40 or $100{\mu}g$ of pinacidil was injected intravenously into ICR mice at 10 min after $5{\mu}Ci$ T1-201 injection, and organ uptake and whole body retention rate were measured at different time points. Results : Co-incubation of pinacidil with T1-201 resulted in a decrease in T1-201 uptake into cultured myocyte by 1.6 to 2.5 times, depending on pinacidil concentration and activity of T1-201 used. Pinacidil enhanced T1-201 washout by 1.6-3.1 times from myocyte preparations pre-incubated with T1-201. Pinacidil treatment appears to be resulted in mild decreases in blood and liver activity in normal mice, in contrast, renal and cardiac uptake were mildly decreased in a dose dependent manner. Whole body retention ratios of T1-201 were lower at 24 hour after injection with $100{\mu}g$ of pinacidil than control. Conclusion : These results suggest that treatment with K-opener may affect the interpretation of T1-201 myocardial images, due to decreasing thallium accumulation and enhancing washout from myocardium.

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Paenibacillus kimchicus sp. nov., an antimicrobial bacterium isolated from Kimchi (김치로부터 분리된 항균 활성 세균 Paenibacillus kimchicus sp. nov.)

  • Park, A-rum;Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.319-326
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    • 2016
  • An antimicrobial bacterium to pathogenic microorganisms, strain $W5-1^T$ was isolated from Korean fermented-food Kimchi. The isolate was Gram-staining-variable, strictly aerobic, rod-shaped, endospore-forming, and motile with peritrichous flagella. It grew at $15-40^{\circ}C$, at pH 6.0-10.0, and in the presence of 0-4% NaCl. Strain $W5-1^T$ could hydrolyze esculin and xylan, and assimilate $\small{D}$-mannose, but not $\small{D}$-mannitol. Strain $W5-1^T$ showed antimicrobial activity against Listeria monocytogens, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhi. The G+C content of the DNA of strains $W5-1^T$ was 52.6 mol%. The predominant respiratory quinone was menaquinone-7 (MK-7) and the major cellular fatty acids were $C_{16:0}$, antieiso-$C_{15:0}$, $C_{18:0}$, and $C_{12:0}$. The strain contained meso-diaminopimelic acid in cell-wall peptidoglycan. On the basis of 16S rRNA gene sequence and phylogenetic analysis, the strain W5-1 was shown to belong to the family Paenibacillaceae and was most closely related to Paenibacillus pinihumi $S23^T$ (98.4% similarity) and Paenibacillus tarimensis $SA-7-6^T$ (96.4%). The DNA-DNA relatedness between the isolate and Paenibacillus pinihumi $S23^T$ was 8.5%, indicating that strain $W5-1^T$ represented a species in the genus Paenibacillus. On the basis of the evidence from this polyphasic study, it is proposed that strain $W5-1^T$ is considered to represent a novel species of the genus Paenibacillus, for which the name Paenibacillus kimchicus sp. nov. is proposed. The type strain is $W5-1^T$ (=KACC $15046^T$ = $LMG 25970^T$).

Biochemical Characterization of Phospholipase C$\delta$from liver of Mud loach (Misgurnus mizolepis) (미꾸라지 간으로부터 포스포리파아제 C델타 단백질의 생화학적 특성)

  • Seo, Jung-Soo;Lim, Sang-Uk;Kim, Na-Young;Lee, Sang-Hwan;Oh, Hyun-Suk;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.67-80
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    • 2005
  • Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.

The difference of photosynthetic efficiency and electron transport rate by control of the red tide organism using algicidal substance and yellow clay (살조물질과 황토를 이용한 적조생물 제어에 따른광합성 효율 및 전자전달율의 차이)

  • Son, Moonho;Baek, Seung Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.4
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    • pp.2951-2957
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    • 2015
  • The development of worldwide harmful algal blooms(HAB) is a serious problem for public health and fisheries industries. To evaluate the algicidal impact on the HAB species, algicide thiazolidinedione derivative (TD49) and yellow clay were examined, which is focus on assess the algicidal effects and inhibition to photosynthesis of HAB species. To obtain the detailed information, we analyzed the viability of target species related to activity Chl. a, photosynthetic efficiency($F_v/F_m$), and electron transport rate(ETR). Culture experiment was conducted to evaluate the algicidal effects of three harmful species(raphidophyceae Heterosigma akashiwo, Chattonella marina, and dinophyceae Heterocapsa circularisquama) and one non-harmful species (cryptophyceae Rhodomonas salina). Our experiments revealed that three HAB species were easily destroyed of the cell walls after TD49 dosing. Also, they had significantly reducing values of active Chl. a, $F_v/F_m$, and ETR, due to the damage of photosystem II by inter-cellular disturbance. As a result, the algicidal effect(%) for the three HABs were as follows, in the order of greatest to the least: H. circularisquama> C. marina> H. akashiwo. However, the algicidal effect for yellow clay remained to be <30% (p>0.01), implying that it may not have damaged the photosystem II. On the other hand, non-HAB R. salina was promoted at both TD49 and yellow clay treatments. Our results demonstrated that the TD49 is a good agent for the control of HABs H. akashiwo, C. marina, and H. circularisquama, whereas the yellow clay would not be suitable for the field application based on our experimental results.

Effect of NADPH Oxidase Inhibition on Heme Oxygenase-1 Expression in Human Hepatoma Cell Line HepG2 (인간 간암세포주 HepG2에서 NADPH oxidase 활성 억제에 의한 heme oxygenase-1 발현의 조절)

  • Lee, Sang-Kwon;Kim, Kang-Mi;Park, Kwang-Hoon;Park, Young-Chul
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1625-1630
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    • 2011
  • Heme oxygenase-1 (HO-1) is a stress-responsive protein that is known to regulate cellular functions such as cell proliferation, inflammation, and apoptosis. In this study, we investigated the role of NADPH oxidase on the expression of HO-1 in human liver hepatoma cell line HepG2. Diphenyleneiodonium (DPI), an NADPH oxidase inhibitor, markedly inhibited HO-1 expression and the nuclear translocation of transcription factor Nrf2 in cobalt protoporphyrin (CoPP) or hemin-treated HepG2 cells. Similarly, the knockdown of $p47^{phox}$, a cytosolic factor for NADPH oxidase activity, by siRNA inhibited the CoPP-induced expression of HO-1. In addition, GSHmee, an intracellular antioxidant, blocked the expression of HO-1 in CoPP-treated cells. Based on these results, we conclude that the blockage of NADPH oxidase with DPI or $p47^{phox}$ siRNA inhibits CoPP-induced HO-1 expression in HepG2 cells, and also suggest that the expression of HO-1 in CoPP-induced HepG2 cells is associated with increase of intracellular ROS by NADPH oxidase activity.

Antioxidative Effect and Tyrosinase Inhibitory Activity of the Unripened Fruit Extract of Rubus coreanus Miquel (미성숙 복분자 과실의 항산화 효능 및 타이로시네이즈 저해 활성 평가)

  • Han, Saet Byeol;Kwon, Soon Sik;Kong, Bong Ju;Kim, Kyeong Jin;Park, Soo Nam
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.39 no.4
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    • pp.295-302
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    • 2013
  • In this study, the antioxidative effects and inhibitory activities of unripened fruit extract of Rubus coreanus Miquel (R. coreanus Miquel) on tyrosinase were investigated and the potential applicability as a cosmeceutical ingredients was evaluated. All experiments were performed with 50% ethanol extract, ethyl acetate fraction and aglycone fraction of unripened fruit extract of R. coreanus Miquel. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activites ($FSC_{50}$) of 50% ethanol extract (6.56 ${\mu}g/mL$) and ethyl acetate fraction (6.14 ${\mu}g/mL$) of unripened fruit extract of R. coreanus Miquel were higher than (+)-${\alpha}$-tocopherol (8.98 ${\mu}g/mL$), which is known as a typical hydrophobic antioxidant. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of 50% ethanol extract (0.83 ${\mu}g/mL$), ethyl acetate fraction (0.84 ${\mu}g/mL$) and aglycone fraction (1.13 ${\mu}g/mL$) of R. coreanus Miquel on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were higher than L-ascorbic acid (1.5 ${\mu}g/mL$), which is known as s typical hydrophilic antioxidant. The cellular protective effect of extract and fraction of unripened fruit extract of R. coreanus Miquel on the rose bengal sensitized photohemolysis of human erythrocytes was increased in a concentration dependent manner (1 ~ 50 ${\mu}g/mL$). And 50% ethanol extract in 50 ${\mu}g/mL$ showed the most protective effect among extracts (${\tau}_{50}$ = 296.3 min). The inhibitory effects on tyrosinase of ethyl acetate and agylcone fractions were higher than arbutin. These results indicate that unripened fruit extracts of R. coreanus Miquel can be applied to antioxidant scavenging ROS including radical as an alternative whitening agent to replace arbutin.

Effects of bisphosphonate and indomethacin on alveolar bone remodeling in rats (Bisphosphonate와 Indomethacin이 백서 치조골의 골개조에 미치는 영향)

  • Cho, Myeong-Sook;Kim, Jong-Chul
    • The korean journal of orthodontics
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    • v.26 no.2 s.55
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    • pp.163-174
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    • 1996
  • The purpose of this study was to examine the effects of bisphosphonate and indomethacin, blockers of bone resorption with different mechanisms, on alveolar bone remodeling. Male rats were divided into control, bisphosphonate and indomethacin groups, and then each group was divided info an experimental side and a control side according to the force application. Bisphosphonate(6.3mg/kg,$2.52x10^{-2}mol/L$) and indomethacin (9mg/kg, $2.52x10^{-2}mol/L$) were injected 6 hours and 1 hour before or 24 hours after the force application. The rats were killed 72 hours after the force application and histologic examination was perfomed. The values of serum acid phosphatase and lactate dehydrogenase were also measured in the control md experimental groups treated with bisphosphonate or indomethacin 1 hour before the force application. In the experimental side, the least number of osteoclasts was noted in the groups treated 1 hour before the force application with indomethacin or bisphosphonate, while there were no differences between the control and the groups treated with drugs 6 hours before or 24 hours after the force application. In the control side, the number of osteoclasts was not inecreased with no differences among the groups. Histologic examination revealed a severe alveolar bone resorption in the control group and the groups treated with indomethacin 6 hours before or 24 hours after the force application. Indomethacin treatment 1 hour before the force application and bisphosphonate treatment at any time significantly attenuated the bone resorption. Electron microscopically, ruffled border and clear zone of osteoclasts were observed in the control and indomethacin groups, while some osteoclasts were detached from the bone surface and exhibited dull cellular projections in the bisphosphonate groups. The bisphosphonate and indomethacin groups showed lower values of acid phosphatase and lactate dehydrogenase than the control group. The acid phosphatase value in the bisphosphonate group was lower than that in the indomethacin group, whereas there was no difference in the lactate dehydrogenase value between the groups. These results suggest that bisphosphonate reduces the activity of osteoclasts as well as the number of osteoclasts and that indomethacin reduces the number of osteoclasts without affecting the activity of osteoclasts. Bisphosphonate has a larger inhibitory effect on bone resorption md thus less limitation in the application time than indomethacin.

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