J Dent Hyg Sci.  2020 Dec;20(4):221-229. 10.17135/jdhs.2020.20.4.221.

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

• Jeong MJ
• Lim DS
• Heo K
• Jeong SJ

• KMID: 2510018
• DOI: http://doi.org/10.17135/jdhs.2020.20.4.221

Abstract

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 mg/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.

Keyword

Anti-inflammatory effect; Anti-microbial activity; MC3T3-E1 osteoblastic cell; Rosmarinic acid; Titanium