Effects of carvacrol on osteoblasts and common oral pathogens

doi:10.12307/2023.124

Abstract

Abstract: BACKGROUND: The natural plant extract carvacrol has the characteristics of broad-spectrum antibacterial activity, low drug resistance and low cytotoxicity, which has become a research hotspot of antibiotic substitutes with a good prospect in recent years.
OBJECTIVE: To determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of carvacrol against Streptococcus mutans and Staphylococcus aureus, two common oral pathogens and to explore the antibacterial activity of carvacrol against these two bacteria and its effect on osteoblast proliferation, thereby providing a new choice for oral disease prevention and treatment.
METHODS: A microplate reader was used to determine the MIC and MBC values of carvacrol against Streptococcus mutans and Staphylococcus aureus. The antibacterial activity of carvacrol was tested by a zone of inhibition test. Cell counting kit-8 was used to detect the proliferation of MC3T3-E1 cells induced by different concentrations of carvacrol at 1, 4, and 7 days. Alkaline phosphatase activity and Runx2 and osteoprotegerin gene expression levels were detected by alkaline phosphatase and real-time fluorescence quantitative PCR kits at 1, 7, and 14 days, respectively.
RESULTS AND CONCLUSION: The MIC values of carvacrol against Streptococcus mutans and Staphylococcus aureus were 0.20% and 0.04%, and the MBC values were 0.60% and 0.12%, respectively. In the inhibition zone test, carvacrol showed an obvious antibacterial zone, and the difference was statistically significant (P < 0.05). Cell counting kit-8 assay showed that the proliferation activity of cells treated with carvacrol was significantly increased (P < 0.05). The activity of alkaline phosphatase increased in all carvacrol concentration groups. The increase of alkaline phosphatase activity was more obvious on day 7 and the effect of 1MIC group was the most obvious (P < 0.05). Results from the real-time fluorescence quantitative PCR showed that different genes were highly expressed at the mRNA level after treated with carvacrol and all of them occurred in the 1MIC group (P < 0.05). To conclude, carvacrol has strong antibacterial activity against two common oral pathogens, Streptococcus mutans and Staphylococcus aureus, can improve the osteogenic activity of MC3T3-E1 cells, and promote their proliferation and differentiation.

Key words: carvacrol, minimum inhibitory concentration, minimum bactericidal concentration, osteogenesis, bone tissue engineering

CLC Number:

Zhang Zihan, Wang Wenli, Li Jinnuo, Li Yourui. Effects of carvacrol on osteoblasts and common oral pathogens[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(11): 1765-1771.

Figures/Tables 8

References

[1] 薛元泰,吴世芳,李义恒,等.8种精油对乳酸菌及致病菌的选择抑制性筛选及其应用研究[J].食品与发酵工业:1-11[2022-04-25].
[2] FACHINI-QUEIROZ FC, KUMMER R, ESTEVÃO-SILVA CF, et al. Effects of Thymol and Carvacrol, Constituents of Thymus vulgaris L. Essential Oil, on the Inflammatory Response. Evid Based Complement Alternat Med. 2012;2012:657026.
[3] 许璐,喻毅,曹宇,等.牛至油及其主要成分的体外抑菌实验[J].世界中医药,2020,15(14):2072-2075.
[4] BNYAN IA, ABID AT, OBIED HN. Antibacterial Activity of Carvacrol against Different Types of Bacteria. J Nat Sci Res. 2014;4(9):12-16.
[5] ČABARKAPA IS, ŠKRINJAR MM, LEVIĆ JD, et al. Influence of thymol and carvacrol on initial cell attachment and biofilm of Candida albicans. Food Feed Res. 2015;42(1):23-30.
[6] MILADI H, ZMANTAR T, KOUIDHI B, et al. Synergistic effect of eugenol, carvacrol, thymol, p-cymene and γ-terpinene on inhibition of drug resistance and biofilm formation of oral bacteria. Microb Pathog. 2017; 112:156-163.
[7] KOUIDHI B, ZMANTAR T, HENTATI H, et al. Cell surface hydrophobicity, biofilm formation, adhesives properties and molecular detection of adhesins genes in Staphylococcus aureus associated to dental caries. Microb Pathog. 2010;49(1-2):14-22.
[8] KOUKOS G, SAKELLARI D, ARSENAKIS M, et al. Prevalence of Staphylococcus aureus and methicillin resistant Staphylococcus aureus (MRSA) in the oral cavity. Arch Oral Biol. 2015;60(9):1410-1415.
[9] MERGHNI A, BEN NEJMA M, DALLEL I, et al. High potential of adhesion to biotic and abiotic surfaces by opportunistic Staphylococcus aureus strains isolated from orthodontic appliances. Microb Pathog. 2016; 91: 61-67.
[10] CIANDRINI E, CAMPANA R, FEDERICI S, et al. In vitro activity of Carvacrol against titanium-adherent oral biofilms and planktonic cultures. Clin Oral Investig. 2014;18(8):2001-2013.
[11] VERARDI G, CENCI MS, MASKE TT, et al. Antiseptics and microcosm biofilm formation on titanium surfaces. Braz Oral Res. 2016;30:S1806-83242016000100220.
[12] VU AA, BOSE S. Natural Antibiotic Oregano in Hydroxyapatite-Coated Titanium Reduces Osteoclastic Bone Resorption for Orthopedic and Dental Applications. ACS Appl Mater Interfaces. 2020;12(47):52383-52392.
[13] DEEPAK V, KASONGA A, KRUGER MC, et al. Carvacrol Inhibits Osteoclastogenesis and Negatively Regulates the Survival of Mature Osteoclasts. Biol Pharm Bull. 2016;39(7):1150-1158.
[14] KUO PJ, HUNG TF, LIN CY, et al. Carvacrol Ameliorates Ligation-Induced Periodontitis in Rats. J Periodontol. 2017;88(7):e120-e128.
[15] 邢承宇,王璐,陈星光,等.香芹酚-壳聚糖纳米颗粒的制备及抑菌活性研究[J].食品与发酵工业:1-10[2022-04-25].
[16] DE L GUIMARÃES LG, DA SILVA ML, REIS PC, et al. General Characteristics, Phytochemistry and Pharmacognosy of Lippia sidoides. Nat Prod Commun. 2015;10(11):1861-1867.
[17] TUFEKCI E, CASAGRANDE ZA, LLNDAUER SJ, et al. Effectiveness of an essential oil mouthrinse in improving oral health in orthodontic patients. Angle Orthod. 2008;78:294-298.
[18] LANA-RUIZ-CABELLO M, MAISANABA S, PUERTO M, et al. Evaluation of the mutagenicity and genotoxic potential of carvacrol and thymol using the Ames Salmonella test and alkaline, Endo III- and FPG-modified comet assays with the human cell line Caco-2. Food Chem Toxicol. 2014;72:122-128.
[19] AMINI P, ARAUJO MW, WU MM, et al. Comparative antiplaque and antigingivitis efficacy of three antiseptic mouthrinses: a two week ran-domized clinical trial. Braz Oral Res. 2019;23:319-325.
[20] GIANNELLI M, CHELLINI F, MARGHERI M, et al. Effect of chlorhexidine digluconate on different cell types: a molecular and ultrastructural investigation. Toxicol In Vitro. 2008;22(2):308-317.
[21] RODRIGUEZ HERRERO E, BOON N, PAUWELS M, et al. Necrotrophic growth of periodontopathogens is a novel virulence factor in oral biofilms. Sci Rep. 2017;7(1):1107.
[22] AHMAD A, KHAN A, AKHTAR F, et al. Fungicidal activity of thymol and carvacrol by disrupting ergosterol biosynthesis and membrane integrity against Candida. Eur J Clin Microbiol Infect Dis. 2011;30(1):41-50.
[23] 甘盈盈,袁中伟,张天翼,等.香芹酚对耐甲氧西林金黄色葡萄球菌(USA300)抑菌机制的研究[J].四川农业大学学报,2019,37(2): 276-282.
[24] BEN ARFA A, COMBES S, PREZIOSI-BELLOY L, et al. Antimicrobial activity of carvacrol related to its chemical structure. Lett Appl Microbiol. 2006;43(2):149-154.
[25] MILADI H, ZMANTAR T, KOUIDHI B, et al. Use of carvacrol, thymol, and eugenol for biofilm eradication and resistance modifying susceptibility of Salmonella enterica serovar Typhimurium strains to nalidixic acid. Microb Pathoge. 2017;104:56-63.
[26] 戴雨芸,李超,袁中伟,等.香芹酚抑制金黄色葡萄球菌生物被膜的形成[J].微生物学通报,2020,47(3):813-820.
[27] GARVEY MI, RAHMAN MM, GIBBONS S, et al. Medicinal plant extracts with efflux inhibitory activity against Gram-negative bacteria. Int J Antimicrob Agents. 2011;37(2):145-151.
[28] CHOUHAN S, SHARMA K, GULERIA S. Antimicrobial Activity of Some Essential Oils-Present Status and Future Perspectives. Medicines (Basel, Switzerland). 2017;4(3):E58.