• Title/Summary/Keyword: Diffusion cell methods

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Antibacterial and Antibiotic Activity Enhancing Effect of Extract and Fractions from the Root of Rumex japonicus Houtt (참소리쟁이 뿌리 추출물 및 분획의 항균 활성과 항생제 증강 활성)

  • Yang, Sun A;Kim, A Young;Pyo, Byoung Sik;Kim, Sun Min
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.6
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    • pp.375-380
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    • 2017
  • Background: The objective of this study were to determine the antibacterial activity and antibiotic activity-enhancing effect of 70% ethanol extract of the root of Rumex japonicus Houtt. and its fractions when used in combination with gentamicin against aerobic skin flora. Methods and Results: The antibacterial activity and antibiotic (gentamicin) activity enhancing effect against aerobic skin flora were determined using the disc diffusion assay. Chloroform fraction (CF) and ethyl acetate fraction (EF) showed higher activities against Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. Regarding the antibiotic (gentamicin) activity-enhancing effect against aerobic skin flora, the n-hexane fraction (HF) and CF showed strong activity. The combination of HF and CF with gentamicin was evaluated using the broth dilution assay to determine the inhibitory effect on the growth of aerobic skin flora. The combination of CF with gentamicin exhibited the highest inhibitory effect on the growth of S. aureus and S. epdermidis. MTT assay performed to determine the viability of L929 cells revealed that EF treatment resulted in viability of 33.96 - 116.76% at the tested concentration. The combination of 70% ethanol extract and its other fractions with gentamicin showed low cell toxicity. Conclusions: Appropriate use of antimicrobial agents is important prior to the development of new antibiotics. The 70% ethanol extract of the root of R. japonicus Houtt. and its fractions showed significant synergism with gentamicin when used in combination against S. aureus and S. epdermidis. Thus, R. japonicus Houtt. could be used as a functional materials in antimicrobial-related fields.

Inhibitory Effects on Oral Microbial Activity and Production of Lipopolysaccharides-Induced Pro-Inflammatory Mediators in Raw264.7 Macrophages of Ethanol Extract of Perilla flutescens (L.) Britton

  • Jeong, Moon-Jin;Lim, Do-Seon;Lee, Myoung-Hwa;Heo, Kyungwon;Kim, Han-Hong;Jeong, Soon-Jeong
    • Journal of dental hygiene science
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    • v.20 no.4
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    • pp.213-220
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    • 2020
  • Background: The leaves of Perilla frutescens, commonly called perilla and used for food in Korea, contain components with a variety of biological effects and potential therapeutic applications. The purpose of this study was to identify the components of 70% ethanol extracted Perilla frutescens (EEPF) and determine its inhibitory effects on oral microbial activity and production of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharides (LPS)-stimulated Raw264.7 macrophages, consequently, to confirm the possibility of using EEPF as a functional component for improving the oral environment and preventing inflammation. Methods: One kg of P. frutescens leaves was extracted with 70% ethanol and dried at -70℃. EEPF was analyzed using high-performance liquid chromatography analysis, and antimicrobial activity against oral microorganisms was revealed using the disk diffusion test. Cell viability was elucidated using a methylthiazolydiphenyl-tetrazolium bromide assay, and the effect of EEPF on LPS-induced morphological variation was confirmed through microscopic observation. The effect of EEPF on LPS-induced production of pro-inflammatory mediators, NO and PGE2 was confirmed by the NO assay and PGE2 enzyme-linked immunosorbent assay. Results: The main component of EEPF was rosemarinic acid, and EEPF showed weak anti-bacterial and anti-fungal effects against microorganisms living in the oral cavity. EEPF did not show toxicity to Raw264.7 macrophages and had inhibitory effects on the morphological variations and production of pro-inflammatory mediators, NO and PGE2 in LPS-stimulated Raw264.7 macrophages. Conclusion: EEPF can be used as a functional material for improving the oral environment through the control of oral microorganisms and for modulating inflammation by inhibiting the production of inflammatory mediators.

A novel antimicrobial-containing nanocellulose scaffold for regenerative endodontics

  • Victoria Kichler ;Lucas Soares Teixeira ;Maick Meneguzzo Prado ;Guilherme Colla ;Daniela Peressoni Vieira Schuldt ;Beatriz Serrato Coelho ;Luismar Marques Porto ;Josiane de Almeida
    • Restorative Dentistry and Endodontics
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    • v.46 no.2
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    • pp.20.1-20.11
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    • 2021
  • Objectives: The aim of this study was to evaluate bacterial nanocellulose (BNC) membranes incorporated with antimicrobial agents regarding cytotoxicity in fibroblasts of the periodontal ligament (PDLF), antimicrobial activity, and inhibition of multispecies biofilm formation. Materials and Methods: The tested BNC membranes were BNC + 1% clindamycin (BNC/CLI); BNC + 0.12% chlorhexidine (BNC/CHX); BNC + nitric oxide (BNC/NO); and conventional BNC (BNC; control). After PDLF culture, the BNC membranes were positioned in the wells and maintained for 24 hours. Cell viability was then evaluated using the MTS calorimetric test. Antimicrobial activity against Enterococcus faecalis, Actinomyces naeslundii, and Streptococcus sanguinis (S. sanguinis) was evaluated using the agar diffusion test. To assess the antibiofilm activity, BNC membranes were exposed for 24 hours to the mixed culture. After sonicating the BNC membranes to remove the remaining biofilm and plating the suspension on agar, the number of colony-forming units (CFU)/mL was determined. Data were analyzed by 1-way analysis of variance and the Tukey, Kruskal-Wallis, and Dunn tests (α = 5%). Results: PDLF metabolic activity after contact with BNC/CHX, BNC/CLI, and BNC/NO was 35%, 61% and 97%, respectively, compared to BNC. BNC/NO showed biocompatibility similar to that of BNC (p = 0.78). BNC/CLI showed the largest inhibition halos, and was superior to the other BNC membranes against S. sanguinis (p < 0.05). The experimental BNC membranes inhibited biofilm formation, with about a 3-fold log CFU reduction compared to BNC (p < 0.05). Conclusions: BNC/NO showed excellent biocompatibility and inhibited multispecies biofilm formation, similarly to BNC/CLI and BNC/CHX.

Early Response of Cardiopulmonary Exercise Test in Patients with Locally Advanced Non-Small Cell Lung Cancer Treated with Systemic Chemotherapy (국소진행성 폐암환자에 대한 전신항암화학요법이 운동부하 심.폐기능에 미치는 영향 : 초기변화를 중심으로)

  • Shin, Kyeong-Cheol;Chung, Jin-Hong;Lee, Kwan-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.4
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    • pp.369-378
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    • 2002
  • Background : The effects of chemotherapy on pulmonary function are mainly a reduced diffusion capacity and a restrictive ventilatory impairment. Exercise can expose cardiovascular and pulmonary abnormalities not evident at rest. Exercise related cardiopulmonary function is important in patients with malignant disease as a determinant of quality of life. We performed this study to evaluate the changes of body composition and cadiopulmonary exercise perfoemance of patients with locally advanced, non-small cell, lung cancer (NSCLC) before and after chemotherapy. Methods : We evaluated resting pulmonary function, body composition, physiologic performance status, and cardiopulmonary exercise function in 11 patients with locally advanced NSCLC, at diagnosis and prior to the fourth cycle of chemotherapy. Results : After chemotherapy, 4 patients (36.4%) showed partial response and 7 (63.4%) had stable disease. After chemotherapy, diffusion capacity of the lung for carbon monoxide was reduced ($89.7{\pm}34.1%$, vs. $71.9{\pm}20.5%$) but not significantly. There were no significant changes in body composition or the state of physiologic performance after chemotherapy. There was a significant impairment of cardiopulmonary exercise tolerance in patients with NSCLC, evidenced by a reduction of maximal oxygen uptake ($VO_2$max, ml/kg/min, $17.9{\pm}2.6$ : $12.6{\pm}6.1$, <0.05) and $O_2$pulse ($O_2$ pulse, ml/beat, $7.0{\pm}1.7$, $5.2{\pm}2.1$, <0.05). Conclusion : Systemic chemotherapy resulted in a loss of cardiopulmonary exercise function in patients with locally advanced NSCLC within the short-term period, but not a physiologic change of body composition within the same period.

Engraftment of Human Mesenchymal Stem Cells in a Rat Photothrombotic Cerebral Infarction Model : Comparison of Intra-Arterial and Intravenous Infusion Using MRI and Histological Analysis

  • Byun, Jun Soo;Kwak, Byung Kook;Kim, Jae Kyun;Jung, Jisung;Ha, Bon Chul;Park, Serah
    • Journal of Korean Neurosurgical Society
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    • v.54 no.6
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    • pp.467-476
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    • 2013
  • Objective : This study aimed to evaluate the hypotheses that administration routes [intra-arterial (IA) vs. intravenous (IV)] affect the early stage migration of transplanted human bone marrow-derived mesenchymal stem cells (hBM-MSCs) in acute brain infarction. Methods : Male Sprague-Dawley rats (n=40) were subjected to photothrombotic infarction. Three days after photothrombotic infarction, rats were randomly allocated to one of four experimental groups [IA group : n=12, IV group : n=12, superparamagnetic iron oxide (SPIO) group : n=8, control group : n=8]. All groups were subdivided into 1, 6, 24, and 48 hours groups according to time point of sacrifice. Magnetic resonance imaging (MRI) consisting of T2 weighted image (T2WI), $T2^*$ weighted image ($T2^*WI$), susceptibility weighted image (SWI), and diffusion weighted image of rat brain were obtained prior to and at 1, 6, 24, and 48 hours post-implantation. After final MRI, rats were sacrificed and grafted cells were analyzed in brain and lung specimen using Prussian blue and immunohistochemical staining. Results : Grafted cells appeared as dark signal intensity regions at the peri-lesional zone. In IA group, dark signals in peri-lesional zone were more prominent compared with IV group. SWI showed largest dark signal followed by $T2^*WI$ and T2WI in both IA and IV groups. On Prussian blue staining, IA administration showed substantially increased migration and a large number of transplanted hBM-MSCs in the target brain than IV administration. The Prussian blue-positive cells were not detected in SPIO and control groups. Conclusion : In a rat photothrombotic model of ischemic stroke, selective IA administration of human mesenchymal stem cells is more effective than IV administration. MRI and histological analyses revealed the time course of cell migration, and the numbers and distribution of hBM-MSCs delivered into the brain.

Antibacterial Effect of Haedokgeumhwa-san against Methicillin-Resistant Staphylococcus aureus (해독금화산(解毒金花散)의 Methicillin-Resistant Staphylococcus aureus에 대한 항균효과)

  • Lee, Ha-Il;Lee, Su-Kyung;Kwon, Young-Mi;Song, Yung-Sun
    • Journal of Korean Medicine Rehabilitation
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    • v.25 no.2
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    • pp.1-13
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    • 2015
  • Objectives Methicillin-Resistant Staphylococcus aureus (MRSA) is a human pathogen and a major cause of hospital-acquired infections. New antibacterial agents that have not been compromised by bacterial resistance are needed to treat MRSA-related infections. In this study, we investigated the antimicrobial activity ofethanol extract of Haedokgeumhwa-san (HGH) which prescription is composed of korean medicine against MRSA. Methods The antibacterial activity of HGH extract was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method (minimal inhibitory concentration; MIC), checkerboard dilution test, and time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors. The checkerboard dilution test was used to examined synergistic effect of ampicillin, oxacillin, ciprofloxacin, vancomycin, gentamicin and norfloxacin in combination with HGH ethanol extract. A time-kill assay was performed a survival curve which was obtained by plotting viable colony counts depending on time on bacterial growth. Results The minimum inhibitory concentration (MIC) of ethanol extract (HGH) ranged from 1,000 to $2,000{\mu}g/mL$ against all the tested bacterial strains, respectively. We are able to confirm that HGH extract has potentially strong antibacterial activity. In the checkerboard dilution test, fractional inhibitory concentration index of HGH in combination with antibiotics indicated synergy or partial synergism against S. aureus. A time-kill study showed that the growth of the tested bacteria was considerably inhibited after 8 hr of treatment with the combination of HGH with selected antibiotics. For measurement of cell membrane permeability, HGH $250{\sim}1,000{\mu}g/mL$ along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (Tris) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide ($NaN_3$) was used as an inhibitor of ATPase. TX, Tris, DCCD and $NaN_3$ cooperation against S. aureus showed synergistic action. Accordingly, antimicrobial activity of HGH was affected by cell membrane and inhibitor of ATPase. Conclusions These results suggest that Haedokgeumhwa-san extract has antibacterial activity, and that HGH extract offers a potential as a natural antibiotic against MRSA.

Characterization of ginsenoside compound K loaded ionically cross-linked carboxymethyl chitosan-calcium nanoparticles and its cytotoxic potential against prostate cancer cells

  • Zhang, Jianmei;Zhou, Jinyi;Yuan, Qiaoyun;Zhan, Changyi;Shang, Zhi;Gu, Qian;Zhang, Ji;Fu, Guangbo;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.228-235
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    • 2021
  • Backgroud: Ginsenoside compound K (GK) is a major metabolite of protopanaxadiol-type ginsenosides and has remarkable anticancer activities in vitro and in vivo. This work used an ionic cross-linking method to entrap GK within O-carboxymethyl chitosan (OCMC) nanoparticles (Nps) to form GK-loaded OCMC Nps (GK-OCMC Nps), which enhance the aqueous solubility and stability of GK. Methods: The GK-OCMC Nps were characterized using several physicochemical techniques, including x-ray diffraction, transmission electron microscopy, zeta potential analysis, and particle size analysis via dynamic light scattering. GK was released from GK-OCMC Nps and was conducted using the dialysis bag diffusion method. The effects of GK and GK-OCMC Nps on PC3 cell viability were measured by using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Fluorescent technology based on Cy5.5-labeled probes was used to explore the cellular uptake of GK-OCMC Nps. Results: The GK-OCMC NPs had a suitable particle size and zeta potential; they were spherical with good dispersion. In vitro drug release from GK-OCMC NPs was pH dependent. Moreover, the in vitro cytotoxicity study and cellular uptake assays indicated that the GK-OCMC Nps significantly enhanced the cytotoxicity and cellular uptake of GK toward the PC3 cells. GK-OCMC Nps also significantly promoted the activities of both caspase-3 and caspase-9. Conclusion: GK-OCMC Nps are potential nanocarriers for delivering hydrophobic drugs, thereby enhancing water solubility and permeability and improving the antiproliferative effects of GK.

Iontophoretic Delivery of Levodopa: Permeation Enhancement by Oleic Acid Microemulsion and Ethanol (Levodopa의 이온토포레시스 경피전달: 올레인산 아이크로에멀젼 및 에탄올의 투과증진)

  • Jung, Shin-Ae;Gwak, Hye-Sun;Chun, In-Koo;Oh, Seaung-Youl
    • Journal of Pharmaceutical Investigation
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    • v.38 no.6
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    • pp.373-380
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    • 2008
  • In order to develop optimal formulation and iontophoresis condition for the transdermal delivery of levodopa, we have evaluated the effect of two permeation enhancers, ethanol and oleic acid in microemulsion, on transdermal delivery of levodopa. In vitro flux studies were performed at $33^{\circ}C$, using side-by-side diffusion cell and full thickness hairless mouse skin. Current density applied was $0.4\;mA/cm^2$ and current was off after 6 hours application. Levodopa was analysed by HPLC at 280 nm. The o/w microemulsions of oleic acid in buffer solution (pH 2.5 & 4.5) were prepared using oleic acid, Tween 80 and ethanol. The existence of microemulsion regions were investigated in pseudo-ternary phase diagrams. Contrary to our expectation, cumulative amount of levodopa transported from microemulsion (pH 2.5) for 10 hours was similar to that from aqueous solution in all delivery methods (passive, anodal and cathodal). When pH of the micro-emulsion was pH 4.5, cumulative amount of levodopa transported for 10 hours increased about 40% (anodal) to 50% (cathodal), when compared to that from aqueous solution. Flux from pH 4.5 microemulsion showed higher value than that from pH 2.5 in all delivery methods. These results seem to indicate that electroosmosis plays more dominant role than electrorepulsion in the flux of levodopa at pH 2.5. The effect of ethanol on iontophoretic flux was studied using pH 2.5 phosphate buffer solution containing 3% or 5% (v/v) ethanol. Flux enhancement was observed in passive and anodal delivery as the concentration of the ethanol increased. Without ethanol, cathodal delivery showed higher flux than anodal delivery. Anodal delivery increased the cumulative amount of levodopa transported 1.6 fold by 5% ethanol after 10 hours. However, in cathodal delivery, no flux enhancement of levodopa was observed during current application and only marginal increase in cumulative amount transported after 10 hours was observed by 5% ethanol. These results seem to be related to the decrease in dielectric constant of the medium and the lipid extraction of the ethanol, which decrease the electroosmotic flow, and thus decrease the flux. Overall, the results provide important insights into the role of electroosmosis and electrorepulsion in the transport of levodopa through skin, and provide some useful informations for optimal formulation for levodopa.

Treatment test for bovine mastitis by the determination of ATP based on firefly bioluminescence (Bioluminescence 반응에 의한 ATP측정을 이용한 젖소 유방염 치료에 관한 연구)

  • Kim, Tae-jong;Kim, Jong-bae;Lee, Seong-bae;Jeon, Young-soo
    • Korean Journal of Veterinary Research
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    • v.29 no.3
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    • pp.393-405
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    • 1989
  • This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

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Anti-Oral Microbial Activity and Anti-Inflammatory Effects of Rosmarinic Acid in Lipopolysaccharide-Stimulated MC3T3-E1 Osteoblastic Cells on a Titanium Surface

  • Jeong, Moon-Jin;Lim, Do-Seon;Heo, Kyungwon;Jeong, Soon-Jeong
    • Journal of dental hygiene science
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    • v.20 no.4
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    • pp.221-229
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    • 2020
  • Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.