Effectiveness of Eugenia caryophyllus in toothpaste against oral microbial in the saliva of healthy subjects in Indonesia

Background: Toothpaste is essential in the process of oral care. One of the components of toothpaste, Eugenia caryophyllus, might have an anti-inflammatory and antibacterial effect. Objectives: The objective of this study is to analyze the effectiveness of E. caryophyllus in the toothpaste against Streptococcus mutans, Lactobacillus, and total microbial load in the saliva of healthy subjects after toothbrushing using E. caryophyllus toothpaste. Methods: Saliva (n = 10, aged 18–25 years) were collected before and 2 weeks after toothbrushing using toothpaste-containing E. caryophyllus (Antiplaque, Triple-Ace, Indonesia). The total microbial load of saliva was determined by the measurement of colony-forming unit (CFU) number on brain–heart infusion agar, at 37°C, for 24 h, in anaerobic-condition. Real-time polymerase chain reaction technique was used to quantify the S. mutans and Lactobacillus deoxyribonucleic acid using SYBR green and 16S-rRNA gene-specific primers for S. mutans and Lactobacillus. Primers were 5'-ATTCCCGCCGTTGGACCATTCC-3' (fwd); 5'-CCGACAAAGACCATTCCATCTC-3' (rvs) and 5'-CTTGTACACACCGCCC GT CA-3' (fwd); 5'-CTCAAAACTAAACAAAGTTTC-3' (rvs) for S. mutans and Lactobacillus, respectively. Data were analyzed using t-pair test with P < 0.05 set as the level of significance. Results: The result showed that there was a significant reduction of total microbial load, S. mutans, and Lactobacillus number in the saliva after toothbrushing. The total microbial number in the saliva was significantly decreased before (5.84 ± 0.43 log CFU/mL) and 2 weeks after toothbrushing (5.27 ± 0.61 log CFU/mL) (P < 0.05). The number of S. mutans and Lactobacillus was also significantly decreased before (8.27 ± 0.11 and 2.34 ± 0.71 log CFU/mL) and after toothbrushing (8.18 ± 0.11 and 1.91 ± 0.25 log CFU/mL) (P < 0.05). Conclusion: E. caryophyllus toothpaste may reduce the number of total microbial, S. mutans, and Lactobacillus in the saliva of healthy subjects. Further studies are needed to explore this result.