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Objectives: The COVID-19 vaccine is currently being administered worldwide to address the ongoing pandemic. Although these vaccines have proven effective in preventing severe disease, the level of immunity required to prevent respiratory mucosal infection remains less well understood. Therefore, it is desirable to develop a noninvasive screening strategy such as oral fluid to monitor secreted antibodies longitudinally as potential surrogates of mucosal immunity., Methods: We evaluated the anti-spike protein antibodies in gingival crevicular fluid (GCF) and saliva and compared them to immune responses in the blood of 50 healthy health care workers following 2 doses of intramuscular Pfizer/BioNTech-BNT162b2 vaccine., Results: The antibodies to SARS-CoV-2 spike and subdomain proteins (RBD, S1, S2, and NTD) were significantly higher in serum than oral fluids but showed a greater detection rate and higher median titres in GCF than saliva. For all tested SARS-CoV-2 antigens, IgG in GCF (as opposed to saliva) showed a more significant and stronger correlation with IgG in serum. Serum-neutralising antibodies (Nab) titres also displayed a significant and stronger correlation with anti-spike protein and their subdomains in GCF than saliva. Interestingly, the time post-second dose of vaccine and sex had a similar influence on IgG in serum and GCF. However, interferon (IFN)-γ-producing T-cell responses showed no association with SARS-Cov-2 IgG antibodies in serum, GCF, or saliva and neutralisation antibodies in serum. The correlation matrix of all measured parameters grouped serum and GCF IgG parameters separately from salivary IgG parameters indicating that GCF better represents the humoural response in serum than saliva., Conclusions: Within limitations, we propose that GCF could be a less invasive alternative to serum and more appropriate than saliva to detect antibody responses by current COVID-19 vaccines if the GCF collection procedure could be standardised. Further research is needed to investigate the suitability of GCF for community immune surveillance for vaccines., Competing Interests: Conflict of interest None disclosed., (Copyright © 2022. Published by Elsevier Inc.)

Objective: This prospective clinical study aim was to analyze the effect of the probiotic Lactobacillus reuteri Prodentis lozenges on salivary microbiome of subjects wearing fixed orthodontic appliances., Methods: Saliva samples were collected prior to consumption and 14th-day post probiotic lozenges consumption (n=40, age 18-23). Oral hygiene index-score (OHI-S) and papilla bleeding index (PBI) were recorded. The salivary microbiome was profiled by next-generation sequencing using the V3-V4 region of 16S-rRNA. Microbial composition, diversity and taxonomic biomarkers were analysed in comparison to probiotic intervention and the clinical characteristics of the cohort using standard bioinformatics tools., Results: The diversity and bacterial community structures did not change significantly in salivary microbiome of periodontally healthy subjects during short-term probiotic intervention. Probiotic consumption correlated with reduction of OHI and PBI scores (50% reduction of scores, P<0.001). The reduction of clinical indices was evident in conjunction with significantly reduced abundance of oral pathogens, such as Porphyromonas pasteri, Treponema sp., Fretibacterium fastidiosum, Kingella oralis and Propionibacterium acnes., Conclusion: Short-term probiotic intervention helped maintaining good oral health in patients undergoing fixed orthodontic therapy. Although overall oral microbiome structure remained largely unchanged, a significant alteration in the abundance of health and disease-associated species highlighted the beneficial effect of probiotic., Competing Interests: No potential conflict of interest was reported by the author(s)., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

Pregnancy gingivitis peaks during mid-pregnancy and resolves transiently towards the postpartum period. However, the role of maternal immune response in orchestrating gingival inflammation has not yet been fully understood. Hence, in this study, we examined the salivary protein profile during the three trimesters of pregnancy, in context to pregnancy gingivitis, employing iTRAQ-based quantitative proteomics. Unstimulated saliva was collected from 10 subjects in each trimester of pregnancy and postpartum period. Samples were analysed using iTRAQ analysis and ELISA and SEM was performed to validate results. Neutrophil mediated immune response was overrepresented in all three trimesters of pregnancy, despite the decrease in phagocytic responses during the second and third trimesters. ELISA showed a significantly higher Neutrophil Extracellular Traps (NETs) formation in the third trimester of pregnancy coinciding with the resolution of pregnancy gingivitis. The NETs-associated proteins (neutrophil elastase and myeloperoxidase) showed a positive correlation with estrogen hormones, which was also highest during the third trimester. Sex hormone-driven NETs formation could be the mainstay of defence that contributes to the remission of pregnancy gingivitis. This study has provided a new insight into the role of immune-modulation in pregnancy gingivitis, which will aid development of new therapeutics for managing pregnancy gingivitis in future.

Objective: The studies on the influence of geographical and socio-economic factors on the oral microbiome remain underrepresented. The Indonesia basic health research (RISKESDAS) 2018, showed an increasing trend in non-communicable diseases compared with the previous report in 2013. The prevalence of diabetes, heart disease, hypertension, and obesity are reported to be higher in urban areas than in rural areas. Interestingly, non-communicable diseases were found to be more prevalent in women than men. This pilot study aimed to examine the oral health and oral microbiome derived from tongue samples of healthy Indonesian women from urban and rural areas. Methods: Twenty women aged 21-47 years old from West Jakarta, residents of DKI Jakarta ( n = 10) as representative of the urban area, and residents of Ende, Nangapanda, East Nusa Tenggara ( n = 10) as representative of the rural area were recruited for this pilot study. The participants were evaluated by the Simplified Oral Hygiene Index (OHI-S) according to the criteria of Greene and Vermillion and divided into three groups. High-throughput DNA sequencing was performed on an Illumina iSeq 100 platform. Results: The principal component analysis displayed a marked difference in the bacterial community profiles between the urban and rural localities. The presence of manifest was associated with increased diversity and an altered oral bacterial community profile in the urban women. Two bacterial taxa were present at significantly higher levels (adjusted p < 0.01) in the urban oral microflora (Genus Prevotella and Leptotricia ) could account for this difference irrespective of the individual oral hygiene status. The linear discriminant analysis effect size (LEfSe) analysis revealed several distinct urban biomarkers. At the species level, Leptotrichia wadei, Prevotella melaninogenica, Prevotella jejuni , and P. histicola , show an excellent discriminatory potential for distinguishing the oral microflora in women between urban and rural areas. Further, using SparCC co-occurrence network analysis, the co-occurrence pattern in the dominant core oral microbiome assembly was observed to be specific to its ecological niche between two populations. Conclusions: This is the first pilot study demonstrating the characterization of the oral microbiome in Indonesian women in urban and rural areas. We found that the oral microbiome in women displays distinct patterns consistent with geographic locality. The specific characterization of the microbiota of Indonesian women is likely linked to geographical specific dietary habits, cultural habits, and socio-economic status or the population studied., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Widyarman, Theodorea, Udawatte, Drestia, Bachtiar, Astoeti and Bachtiar.)

Purpose: One of the key approaches to minimize the risk of COVID-19 transmission would be to reduce the titres of SARS-CoV-2 in the saliva of infected COVID-19 patients. This is particularly important in high-risk procedures like dental treatment. The present randomized control trial evaluated the efficacy of three commercial mouth-rinse viz. povidone-iodine (PI), chlorhexidine gluconate (CHX) and cetylpyridinium chloride (CPC), in reducing the salivary SARS-CoV-2 viral load in COVID-19 patients compared with water., Methods: A total of 36 SARS-CoV-2-positive patients were recruited, of which 16 patients were randomly assigned to four groups-PI group (n = 4), CHX group (n = 6), CPC group (n = 4) and water as control group (n = 2). Saliva samples were collected from all patients at baseline and at 5 min, 3 h and 6 h post-application of mouth-rinses/water. The samples were subjected to SARS-CoV-2 RT-PCR analysis., Results: Comparison of salivary Ct values of patients within each group of PI, CHX, CPC and water at 5 min, 3 h and 6 h time points did not show any significant differences. However, when the Ct value fold change of each of the mouth-rinse group patients were compared with the fold change of water group patients at the respective time points, a significant increase was observed in the CPC group patients at 5 min and 6 h and in the PI group patients at 6 h., Conclusion: The effect of decreasing salivary load with CPC and PI mouth-rinsing was observed to be sustained at 6 h time point. Within the limitation of the current study, as number of the samples analyzed, the use of CPC and PI formulated that commercial mouth-rinses may be useful as a pre-procedural rinse to help reduce the transmission of COVID-19. ISRCTN (ISRCTN95933274), 09/09/20, retrospectively registered.

Objectives: The dysbiotic oral microbiome plays a key role in the pathogenesis of caries in children. Topical application of casein phosphopeptide-amorphous calcium phosphate containing fluoride (CPP-ACP/F) is an effective treatment modality for children with caries (CC). Hitherto the mechanism by which CPP-ACP/F modules the oral microbiome in CC has not been investigated. The study aimed to examine the CPP-ACP/F effect on the dental plaque microbiome of children group with caries., Methods: This preliminary prospective clinical cohort included 10 children with caries. The children received topical fluoride CPP-ACP/F once-a-week for one month. Plaque samples were collected before and after treatment and subjected to 16S rDNA-based next-generation-sequencing. Microbial composition, diversity and functional roles were analyzed in comparison to the clinical characteristics of cohort using standard bioinformatics tools., Results: CPP-ACP/F treatment modulated dysbiotic oral microbiome towards healthier community as the higher proportion of Proteobacteria and certain microbial protective species were enriched following CPP-ACP/F treatment. Despite overall uniformity of community structure in children with caries between the groups, some bacterial species were differentially represented in a statistically significant manner between pre- and post- treatments. Three bacterial species were found to be predictive of strongly sensitive to the CPP-ACP/F treatment, marked by decreased abundance of Lautropia mirabalis and increased abundance of Gemella haemolysans and Schwartzia succinivorans., Conclusion: Within the limits of the current study, it could be concluded that the CPP-ACP/F varnish treatment modulated the microbial composition of the dental plaque microbiome towards symbiosis. These symbiotic changes may demonstrate the potential clinical significance of CPP-ACP/F varnish treatment., (Copyright © 2021. Published by Elsevier Ltd.)

Background and Purpose: Candida albicans is the major fungal species associated with superficial mucosal infections such as oral candidiasis as well as systemic mycoses with high morbidity and mortality. On top of the rising drug resistance, currently available antifungal agents have significant adverse effects. Nephrotoxicity is the major treatment complication associated with antifungal agents.Recently, we discovered a novel antifungal small molecule SM21 with promising antifungal activity. The present study aimed to comparatively evaluate the in vivo and in vitro nephrotoxicity of SM21 comparing with Amphotericin B and voriconazole., Experimental Approach: Nephrotoxicity of SM21 and its analogue were comparatively evaluated with Amphotericin B (AmB) and voriconazole. Immortalized human kidney proximal tubule epithelial cells (HK-2) were used for in vitro analysis of nephrotoxicity using cytotoxicity assays and qPCR gene expression analysis (Kim-1/ HAVcr-1, CASP3 ). Sprague Dawley (SD) rat model was used to evaluate the nephrotoxicity in vivo using classical (SCr and BUN) and next-generation kidney injury urinary biomarkers (Kim-1, CLU, ALB, NGAL, β2M, and Cys C) alongside histopathological and immunohistochemical standards., Key Results: AmB treatment showed a stronger cytotoxic impact on HK-2 viability and gene expression of cell death markers (Kim-1/ HAVcr-1, CASP3 ) compared with SM21 and SM21 analogue in vitro (P < 0.01). In vivo data further demonstrated that SM21 did not significantly increase classical as well as novel nephrotoxic biomarkers, and minimal renal tubular necrosis and abnormalities were observed (15 mg kg -1 BW/day)., Conclusions and Implications: SM21 had a significantly better safety profile in terms of nephrotoxicity with no major tubular epithelial abnormalities observed in kidney cells and no augmentation of kidney injury biomarkers compared to AmB. Kim-1 and CLU were the most sensitive biomarkers for detection of AmB-induced kidney damage. Future clinical trials should consider inclusion of these novel biomarkers as early indicators of acute kidney injury in antifungal-induced nephrotoxicity., (Copyright © 2020 Udawatte, Kang, Wang, Arumugam and Seneviratne.)