Anti-Oral Microbial Activity and Anti-Inflammatory Effects of Rosmarinic Acid in Lipopolysaccharide-Stimulated MC3T3-E1 Osteoblastic Cells on a Titanium Surface원문보기
Jeong, Moon-Jin
(Department of Oral Histology and Developmental Biology, School of Dentistry, Chosun University)
,
Lim, Do-Seon
(Department of Dental Hygiene, Graduate School of Public Health Science, Eulji University)
,
Heo, Kyungwon
(Department of Oral Histology and Developmental Biology, School of Dentistry, Chosun University)
,
Jeong, Soon-Jeong
(Department of Dental Hygiene & Institute of Basic Science for Well-Aging, Youngsan University)
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 usi...
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.
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.
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제안 방법
) for 1 hour. The development was performed using an X-ray film (FUJI FILM, Tokyo, Japan) after detection using an ECL solution (Merck Millipore). The intensity of bands was measured using a Science Lab Image Gauge (FUJI FILM, Tokyo, Japan).
The various studies on the antimicrobial activities of RA against oral microorganisms causing oral diseases, including PI, are insufficient, and there are no studies on the antiinflammatory effect of RA in the lipopolysaccharide (LPS)-stimulated osteoblasts on the Ti surface to enable the elucidation of its potential for the regulation of PI. The purpose of this study was to investigate the antimicrobial activity of RA against oral microorganisms and the anti-inflammatory effect of RA on LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of PI in Ti-based dental implants.
대상 데이터
All the experiments were carried out in triplicate. All the data were expressed as means±standard deviations.
Microbial strains to confirm the antimicrobial activity of RA against oral microorganisms were purchased from the Korea Microbial Conservation Center (KCCM) and the Gene Bank (KCTC) and used in the experiment (Table 1). S.
) agar and broth. The fungus, C. albicans was cultured in potato dextrose agar (MB cell Ltd.) and broth (MB cell Ltd.) and used in the experiment.
데이터처리
, Chicago, IL, USA). Statistically significant differences were determined using the Student’s t-test. The significance level for determining statistical significance was set at 0.
이론/모형
Total RNA was extracted by processing according to the manufacturer’s method using RiboEXTM reagent (GeneAll, Seoul, Korea). Complementary DNA (cDNA) was synthesized using 1 g of isolated total RNA using RT Premix (GeNet Bio, Daejeon, Korea).
성능/효과
This means that the control of pro-inflammatory mediators and cytokines can also control osteoclast formation and bone resorption. This study showed that RA reduced the production of pro-inflammatory mediators, NO and PGE2 and pro-inflammatory cytokines, TNF- and IL-1 in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels (Fig. 2∼4). This implies that RA plays a role at the transcriptional level.
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