Analysis of plaque microbiota and salivary proteins adhering to dental materials.

OBJECTIVES: Plaque causes oral diseases and aspiration-pneumonia in the elderly. It is not known whether pellicle-like attached salivary proteins and microbiota on dental materials are identical to those on teeth. The purpose of this study was to determine the properties of salivary proteins and microbiota that attach to dental materials. METHODS: Eight subjects wore removable oral splints with pieces of pure-titanium, cobalt-chromium alloy, silver-palladium-copper-gold-alloy, denture-base-resin, and hydroxyapatite for 24 h. The bacteria that adhered to each material were analyzed using 16S rRNA sequencing simultaneously. Each material sample was then immersed in pooled saliva, and the attached proteins were collected. Salivary proteins were analyzed using MALDI-TOF/MS, and high molecular weight proteins were identified using peptide mass fingerprinting. RESULTS: Among the dental materials, the α- and ß-diversity of adherent flora were similar. The bacterial species that adhered easily to materials were Streptococcus sp. oral taxon 058, Neisseria mucosa, Gemella haemolysans, and Rothia dentocariosa. Regardless of material, the peaks or spots of attached salivary proteins had similar patterns, containing functioning proteins such as anchoring receptors for early colonizers. CONCLUSIONS: There were no significant differences in microbiota and protein adherence in hydroxyapatite compared to the dental materials. Therefore, similar microbiota was determined to have formed on the similar pellicle-like proteins. In our study, the characteristics of plaque adhesion on both hydroxyapatite and dental materials were clarified. Based on this study, the creation of new methods of inhibiting plaque adhesion to prevent aspiration-pneumonia and oral infections can be undertaken.

Analysis of oral microbiota in Japanese oral cancer patients using 16S rRNA sequencing.

OBJECTIVES: It is important to determine the cause of increasing oral cancer occurrence and mortality rates in Japan, because the mortality rate has recently decreased in other developed countries. The impact of microbiota in carcinogenesis, especially in the digestive tract has been reported. This study aimed to clarify the relationship between oral cancer and oral microbiota in Japanese patients. METHODS: DNA was extracted from salivary samples of 60 oral cancer patients and 80 non-cancer individuals as controls. We performed metagenomic analysis using 16S rRNA amplicon sequencing. Statistical analysis in this study was performed using R (version 3.5.0). RESULTS: Oral cancer patients showed higher α-diversity compared to the control group, and the ß-diversity between the two groups differed significantly. Further, there was a significant difference in the abundance ratio of bacterial genera between the two groups. Peptostreptococcus, Fusobacterium, Alloprevotella, and Capnocytophaga were more abundant in the cancer group compared to the control, whereas Rothia and Haemophilus were less abundant (p < 0.01). A negative correlation in the microbiota composition was confirmed between the operational taxonomic units (OTU) of genus Rothia and T-stage progression using the TNM classification method. We performed logistic regression analysis to investigate the impact factor for the oral cancer group, and the result showed that Chao 1 index and sex are statistically significant variables. CONCLUSIONS: In this study, we observed an increased bacterial diversity in oral cancer patients and found distribution changes for some bacteria.