Immunoglobulin A response to SARS-CoV-2-N-protein potentially persists in oral fluids of patients with periodontitis six months after mRNA vaccine administration.

Few studies have investigated the mucosal immune response after BNT162b2-booster vaccination in individuals with periodontitis. In this study, we evaluated the persistence of IgA anti-SARS-CoV-2-N-protein in saliva and gingival crevicular fluid (GCF) of patients with periodontitis for at least six months post BNT162b2 vaccine booster. We included patients with moderate (n = 7) and severe (n = 7) periodontitis and participants without periodontitis (n = 7) as controls. The Bradford method measured the protein concentrations in the samples, and an enzyme-linked immunosorbent assay of the SARS-CoV-2 N protein was performed to analyze the targeted IgA level. For the tested SARS-CoV-2 antigen (N-protein), IgA levels in saliva and GCF showed a strong and significant correlation. Therefore, in patients with moderate or severe periodontitis, saliva and GCF can provide information regarding the IgA response against SARS-CoV-2-N-protein. The neutralizing activity of IgA against SARS-CoV-2 was not investigated in this study, necessitating further research.

Association between dysbiotic perio-pathogens and inflammatory initiators and mediators in COVID-19 patients with diabetes.

It has been suggested that a corona virus infection is linked to chronic periodontitis (COVID-19). Our objectives were to look at the expression of angiotensin-converting enzyme-2 (ACE2) in periodontal compartments containing periodontal infections to determine if ACE2 is directly or indirectly responsible for the inflammation in periodontal tissues getting worse. In this study, six non-COVID-19 periodontitis patients without diabetes served as controls, and 23 hospitalized periodontitis patients were admitted with PCR-confirmed COVID-19 with diabetes mellitus (Group 1/G1, n = 10), and without diabetes (Group 2/G2, n = 13). We evaluated the mRNA expression of ACE2, IL-6, IL-8, complement C3, and LL-37, as well as the relative proportion of Porphyromonas gingivalis, Fusobacterium nucleatum, and Veillonella parvula to represent the dysbiosis condition in periodontal microenvironment using subgingival plaque and gingival crevicular fluids (GCF) samples and quantitative real time PCR (qPCR). Every analysis was done to ascertain how they related to one another. The area under the curve (AUC) and receiver operating characteristic (ROC) curve were used to determine the sensitivity and specificity of inflammatory indicators. All the grouped patients had ACE2 detected, according to our findings, but only the G1 patients had a positive correlation (p < 0.05) between ACE2 expression and the inflammatory markers. The combination of IL-6 and C3 mRNAs was found to be 0.78 and 0.55 for the G1 group and the G2 group, respectively, based on the ROC and AUC values. According to our research, the relationship between complement C3 and IL-6 may be able to predict the degree of periodontal inflammation in COVID-19 patients who also have diabetes.

Changes in Interleukin-1ß, Tumor Necrosis Factor-α, and Interleukin-10 Cytokines in Older People with Periodontitis.

Aging can change the ability to respond to various stimuli and physical conditions. A decreased immune response is a form of deterioration of function in older people, who then become more vulnerable when exposed to pathogens. Periodontitis is an inflammatory disease of the periodontal tissues that often occurs in older people. This study aimed to clinically analyze the periodontal status and cytokine levels of IL-1ß, TNF-α, and IL-10 in older people and adults with periodontitis. This clinical study examined 20 persons in a group of older people and 20 persons in a group of adults. The clinical measurements of periodontal status included the Simplified Oral Hygiene Index (OHI-S), Plaque Index (PlI), and Papilla Bleeding Index (PBI). The cytokine levels in gingival crevicular fluid (GCF) were quantified by using ELISA kits. The OHI-S, PlI, and PBI were found to be higher in the older group. The mean values of cytokines were higher in the older group than in adults, although no statistically significant differences were found. A strong correlation was found between the clinical measurements and the cytokine levels in the GCF. There was an increasing tendency of pro-inflammatory and anti-inflammatory cytokines in the older group compared to the adult group.

Porphyromonas gingivalis association with inflammatory markers and exosomal miRNA-155 in saliva of periodontitis patients with and without diabetes diagnosed with COVID-19.

Background: /Purposes: Studies have indicated that salivary molecules from patients with periodontitis and diabetes are confounded with pathological conditions associated with SARS-CoV-2 infection. The study aimed to address whether the abundance of Porphyromonas gingivalis which causes periodontitis, differed compared with that of Aggregatibacter actinomycetemcomitans (used as control) and to analyze the correlation of periodontitis with the expression levels of severe acute respiratory syndrome coronavirus 2 receptor (ACE2) and periodontitis inflammatory markers (TLR-2/TLR-4, TNFα, and miR-155). Materials and Methods: A saliva sample (5 mL) was obtained from 23 hospitalized patients with COVID-19, categorized into two groups: diabetic (G1, n = 10) and non-diabetic (G2, n = 13). Saliva from patients with periodontitis without diabetes and coronavirus disease 2019 (COVID-19; n = 6) were included as control. The quantitative real-time polymerase chain reaction measured the levels of P. gingivalis and A. actinomycetemcomitans, as well as periodontitis markers in saliva. The obtained data were analyzed using one-way ANOVA and the Spearman correlation test. Results: The abundance of P. gingivalis was observed to be higher (p < 0.05) in saliva of patients with diabetes (G1) than in those without diabetes (G2). A contradictory trend was observed for A. actinomycetemcomitans. The transcription level of ACE2 was comparable in all groups tested, while the expression of periodontitis markers varied. The relationships and sensitivity/specificity among P. gingivalis infection ACE2 expression, and inflammatory markers were also evaluated. Conclusions: This study showed that the association between P. gingivalis infection and ACE2 expression might reflect the characteristics of saliva in COVID-19 patients with and without diabetes. However, the relationships between TLR-4 and miR-155 are more specific in discriminating against COVID-19 patients with and without diabetes.

The utility of salivary CRP and IL-6 as a non-invasive measurement evaluated in patients with COVID-19 with and without diabetes.

Background: The available evidence suggests that inflammatory responses, in both systemic and oral tissue, contribute to the pathology of COVID-19 disease. Hence, studies of inflammation biomarkers in oral fluids, such as saliva, might be useful to better specify COVID-19 features. Methods: In the current study, we performed quantitative real-time PCR to measure salivary levels of C-reactive protein (CRP) and interleukin-6 (IL-6) in saliva obtained from patients diagnosed with mild COVID-19, in a diabetic group (DG; n = 10) and a non-diabetic group (NDG; n = 13). All participants were diagnosed with periodontitis, while six participants with periodontitis but not diagnosed with COVID-19 were included as controls. Results: We found increases in salivary total protein levels in both the DG and NDG compared to control patients. In both groups, salivary CRP and IL-6 levels were comparable. Additionally, the levels of salivary CRP were significantly correlated with total proteins, in which a strong and moderate positive correlation was found between DG and NDG, respectively. A linear positive correlation was also noted in the relationship between salivary IL-6 level and total proteins, but the correlation was not significant. Interestingly, the association between salivary CRP and IL-6 levels was positive. However, a moderately significant correlation was only found in COVID-19 patients with diabetes, through which the association was validated by a receiver operating curve. Conclusions: These finding suggest that salivary CRP and IL-6 are particularly relevant as potential non-invasive biomarker for predicting diabetes risk in mild cases of COVID-19 accompanied with periodontitis.

Clinical and oral microbiome pattern of halitosis patients with periodontitis and gingivitis.

Halitosis is caused by a bacterial proteolytic process that induces the production of volatile sulfur compounds, odor-causing gases. The aim of this study was to determine the clinical oral hygiene state and oral microbiome pattern of halitosis patients with periodontitis and gingivitis. The oral hygiene state of halitosis patients with periodontitis and gingivitis was assessed using the oral hygiene index simplified (OHI-S), decay missing filled teeth (DMFT), and tongue biofilm. The dorsum of the tongue and subgingival swabs were cultured for bacteria, and bacterial morphology was evaluated using Gram staining. Evaluation of the bacterial genus using the Bergey's systematic bacteriology diagram as a guide. A total of ten patients with periodontitis and gingivitis were included. Our data indicated that the scores of OHI-S and DMFT were different significantly between halitosis patients with periodontitis and gingivitis (both had p<0.001) while tongue biofilm score was not different between groups. On the dorsum of the tongue, periodontitis patients had a significant higher oral microbiome population (85.65x106 CFU/mL) compared to those with gingivitis (0.047x106 CFU/mL) with p=0.002. In contrast, the number of microbiomes in the subgingival had no significant different between periodontitis and gingivitis. On the dorsum of the tongue, six bacterial genera were isolated from periodontitis cases and seven genera were detected from gingivitis patients. On subgingival, 10 and 15 genera were identified from periodontitis and gingivitis, respectively. Fusobacterium, Propionibacterium, Eubacterium and Lactobacillus were the most prevalent among periodontitis cases while Porphyromonas was the most prevalent in gingivitis patients. In conclusion, although OHI-S and DMFT are different between periodontitis and gingivitis, overlapping of bacterial genera was detected between periodontitis and gingivitis cases.

ACE2 expression in saliva of patients with COVID-19 and its association with Candida albicans and Aggregatibacter actinomycetemcomitans.

Background: A relationship between oral microbiota and susceptibility to SARS-CoV-2 infection has been extensively studied. However, the relationship between oral commensal flora and expression of angiotensin-converting enzyme 2 ( ACE2) remains to be established. In this observational study, we collected saliva from patients with COVID-19 and evaluated the relationship between ACE2 expression and Candida albicans as well as with selected gram-negative bacteria ( Aggregatibacter actin o mycetemcomitans, Fusobacterium nucleatum, and Veillonella parvula). We investigated how this may be directly or indirectly involved in oral dysbiosis in patients with COVID-19. Methods: We included 23 hospitalized patients admitted to Universitas Indonesia Hospital with PCR-confirmed COVID-19, with six healthy participants serving as controls. Saliva and tongue surface swabs were collected from patients with diabetes (DG) and without diabetes (NDG) and subject controls. Using quantitative PCR (qPCR) we assessed the mRNA expression of ACE2, the abundance of C. albicans, and the transcription levels of its biofilm-associated genes, agglutinin-like protein 3 ( ALS3), hyphal wall protein 1 ( HWP1), and yeast-form wall protein 1 ( YWP1). We also counted the relative proportion of the three selected gram-negative oral bacteria in saliva. All analyses were performed to determine the relationship between ACE2 expression and C. albicans and gram-negative bacteria. Results: ACE2 mRNA expression was significantly higher in tongue swab samples than in saliva. However, no significant difference was observed between the patient groups. Conversely, DG patients had a significantly higher abundance of C. albicans in saliva compared to NDG patients and control group patients. The correlation and sensitivity/specificity relationship between ACE2 expression and C. albicans or the selected oral bacteria were also observed. Conclusions: The data show that ACE2 expression can be detected in saliva of patients with COVID-19 and its association with C. albicans and gram-negative oral bacteria might contribute toward developing an oral dysbiosis based predictor for prognosis of COVID-19 severity.

The Red and Orange Complex Subgingival Microbiome of Cognitive Impairment and Cognitively Normal Elderly with Periodontitis.

Increasing evidence has shown an association between periodontitis and cognitive impairment. Subgingival microbiota play a great role in periodontitis pathogenesis. However, the correlation between the subgingival microbiome and cognitive impairment remains unclear. This study aimed to evaluate the red and orange complex subgingival microbiome of cognitively impaired and cognitively normal elderly Indonesian subjects with periodontitis. Twenty-eight elderly subjects diagnosed with periodontitis underwent two cognitive examinations using the Hopkins Verbal Learning Test and the Mini-Mental State Examination. Gingival crevicular fluid taken from the periodontal pocket, at a depth between 5 and 7 mm, using a paper point was used as the subgingival samples. The subgingival microbiome in the cognitive impairment group (n = 14) and cognitively normal group (n = 14) was compared using the 16S rRNA Metagenomic iSeq™ 100 Sequencing System. There was ß-diversity in the subgingival microbiota between the cognitively impaired and cognitively normal subjects. The metagenomic analysis showed a higher abundance of Porphyromonas and Treponema bacteria in the cognitive impairment group than in the normal cognitive group (p < 0.05). The abundance of Porphyromonas gingivalis and Treponema denticola was higher in the cognitively impaired elderly subjects. The role of P. gingivalis and T. denticola in the pathogenesis of cognitive impairment needs further investigation.

Validation of RNA Aptamer Probes to Image Candida albicans in Paraffin-Embedded Sections of Wistar Rat Tongue.

OBJECTIVE: This study aimed to validate the use of Ca-apt-1, an RNA aptamer, that we generated previously as a probe for immunostaining of Candida albicans in rat tongue paraffin-fixed tissue sections MATERIAL AND METHODS: The performance of Ca-apt-1 as a detector molecule was compared with that of anti-C. albicans polyclonal antibody (PcAb), which was used as a positive control. Immunostaining images were visualized by light microscopy and were analyzed by using ImageJ software. RESULTS: Microscopic results demonstrated that Ca-apt-1 specifically recognized and immunostained C. albicans cells of rat tongue candidiasis, with a specificity comparable to that of PcAb. ImageJ analysis showed that the area (pixels) detected by Ca-apt-1 was wider than that detected by the antibody. This indicates that the binding affinity of Ca-apt-1 toward C. albicans was better than that of PcAb on paraffin-embedded tissues. CONCLUSION: This study demonstrates that Ca-apt-1 can be used as a probe for immunostaining of fixed tissue sections for oral candidiasis diagnosis.

Correlation between the extent of smoking, salivary protein profiles, and dental caries in young adult smokers.

Context.Proteins in the saliva are one of the defense mechanism factors that can protect the oral cavity from disease. However, smoking might affect the properties of saliva. AIM: To determine the differences in salivary protein profiles and total concentrations in smokers and non-smokers and their correlation with dental caries severity as indicated by the Decayed, Missing, Filled-Teeth (DMF-T) scores. METHODS AND MATERIAL: This cross-sectional study included 25 smokers and 25 non-smokers. The DMF-T scores were recorded. The total salivary protein was measured by the Bradford method, and the profile proteins were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The average of salivary protein concentration in smokers was lower than that in non-smokers (551.486 µg/mL versus 765.361 µg/mL), but the difference was not statistically significant (P > 0.05). Further correlation analyses showed a negative correlation between the concentration of proteins based on the extent of smoking. A weak negative correlation was found between protein concentration and DMF-T scores (r = -0.239). Dominant salivary protein bands of 11.6 kDa and 54.5 kDa were found in smokers and 27 kDa, 60 kDa, and 94.5 kDa were found in non-smokers. CONCLUSION: Different protein bands appeared in smokers and non-smokers. There was a weak correlation between protein concentration, DMF-T scores, and the extent of smoking.

A pilot study of red complex and three genera subgingival microbiome in periodontitis subjects with and without diabetes, evaluated by MinION platform.

Background: Subgingival niche is one biofilm habitat containing rich microbiota, which plays an active role in maintaining the health of periodontal tissue and determining host response. As such, a study of changing subgingival biofilms is important for understanding the effect of a systemic condition. In this study, we compared the occurrence of six bacteria cohabiting in the subgingival area of periodontitis subjects, with (DP, n = 8) and without (NDP, n = 4) diabetes. Methods: The six genus and species of targeted bacteria were confirmed by 16S rRNA amplicon sequencing on MinION nanopore platform. Descriptive statistic was used to describe the obtained data. Results: We found that the six genus and species of targeted bacteria were detected but in different quantities in either group's periodontal pocket. Our data showed that Tannerella forsythia was the most abundant species in subgingival biofilms of the DP group of the red complex bacteria. In contrast, Aggregatibacter sp., which belongs to the phylum of proteobacteria, was present at a relatively lower level. In contrast, Fusobacterium sp., which belongs to orange complex bacteria, showed relative similarities in subgingival biofilms of both groups tested, while Veillonella sp., were abundant in the DP groups.  Conclusions: Our data show that the diversity of classic periodontopathogens increased in the subgingival niche of periodontitis subjects with diabetes. It is the first study in Indonesia to apply MinION-based, full-length 16S rRNA amplicon sequencing in periodontitis patients with and without diabetes.

Diversity of Oral Microbiome of Women From Urban and Rural Areas of Indonesia: A Pilot Study.

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.

COVID-19 Awareness Among Dental Professionals in Indonesia.

Background: The COVID-19 pandemic caused by SARS-CoV-2 has claimed thousands of lives worldwide. To enhance knowledge and awareness of COVID-19, considerable online resources have been made available, including aspects related to the dental profession. The study aim was to examine the knowledge, perception, and attitude of dental professionals in Indonesia toward COVID-19. We conducted a survey via a questionnaire created using Google docs and distributed to 632 members of the Indonesian Dental Association in the context of a webinar hosted by the Indonesian Oral Biology Association on first June, 2020. Materials and Methods: The questionnaire consisted of 17 items pertaining to demographic data, knowledge and virus identification, awareness regarding drugs commonly used in dentistry during pandemic and research opportunities. Participants were asked to complete the questionnaire after the webinar by choosing one answer to each question. For the analysis, participants were divided into three groups according to their professional background i.e., employment at national hospital, private hospital, or academic faculty. Data were analyzed using descriptive statistics and expressed as frequencies and percentages. The chi-square test was used to investigate the association between professional activity and the level of knowledge, perceptions, and attitudes about COVID-19. Results: Sixty percent of the participants correctly identified the pathogenesis of the disease. This knowledge was not associated with their professional affiliation (p = 0.95). Sixty-seven percentage had comprehensive knowledge about virus detection methods. This knowledge was not associated with their affiliation either (p = 0.54). Questions regarding drugs of choice, prevention, and the spread of COVID-19 were correctly answered by 89, 96, and 82% of the participants, respectively. Knowledge of these aspects were significantly associated with the professional affiliation (p < 0.05). All respondents were optimistic regarding research opportunities (p < 0.01). Respondents from academics were more interested in joining COVID-19-related research projects with governmental institutions (p < 0.01). Conclusion: Knowledge and awareness of COVID-19 among Indonesian dentists are reasonably good. However, further improvement would be beneficial to manage patients during this pandemic. As the number of COVID-19 cases continue to rise in Indonesia, it is important that dentists keep abreast of the updated knowledge on this moving field. Dentist knowledge on infection control should be strengthened through continuous educational programs.

Effect of cell-free spent media prepared from Aggregatibacter actinomycetemcomitans on the growth of Candida albicans and Streptococcus mutans in co-species biofilms.

This study explored the influence of cell-free spent media prepared from Aggregatibacter actinomycetemcomitans LuxS mutant (Aa-LuxS), its wild type strain (Aa-WT), and the laboratory strain (Aa-Y4), on the interaction between Candida albicans and Streptococcus mutans while growing in co-species biofilm for 48 h. By analyzing the results of crystal violet staining, [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide] (MTT) assays, and quantitative real-time polymerase chain reaction (qPCR), we found that the presence of Aa-LuxS in treated biofilms did not affect biofilm development, while added Aa-WT or Aa-Y4 resulted in a significant decrease in both biofilm mass and the number of cells. The inhibitory effect of Aa-WT or Aa-Y4 was not dependent on the protein concentration in the spent media tested (1 and 10%). Gene transcription analyses indicated that Aa-WT/Aa-Y4 exhibits comparable inhibitory effects on the expression of hyphal-associated genes (ALS3 and HWP1), but not on the expression of YWP1, which encodes a yeast form of C. albicans. In contrast, except for gtfD, the expression of S. mutans gtfB/C genes encoding glucosyltransferase was not affected in Aa-WT and Aa-Y4 treated biofilms compared to the levels found in Aa-LuxS treated biofilms. Our results indicate that AI-2-containing spent media derived from Aa can reduce biofilm biomass without significantly inhibiting the survival rate of S. mutans.

Proteomics approach for biomarkers and diagnosis of periodontitis: systematic review.

Quantitative proteomic workflow based on mass spectrometry (MS) is recently developed by the researchers to screen for biomarkers in periodontal diseases comprising periodontitis. Periodontitis is known for chronic inflammatory disease characterized by progressive destruction of the tooth-supporting apparatus, yet has a lack of clear pathobiology based on a discrepancy between specified categories and diagnostic vagueness. The objective of this review was to outlined the accessible information related to proteomics studies on periodontitis. The Preferred Reporting Items for Systematical Reviews and Meta-Analysis (PRISMA) statement guides to acquaint proteomic analysis on periodontal diseases was applied. Three databases were used in this study, such as Pubmed, ScienceDirect and Biomed Central from 2009 up to November 2019. Proteomics analysis platforms that used in the studies were outlined. Upregulated and downregulated proteins findings data were found, in which could be suitable as candidate biomarkers for this disease.

Quantification and Pathogenicity of Candida albicans in Denture-Wearing and Nondenture-Wearing Elderly.

OBJECTIVE: The primary purpose of this study was to evaluate and compare the microbial loads and pathogenicity traits of oral Candida albicans in denture-wearing (DW; n = 15) and nondenture-wearing (NDW; n = 15) elderly persons. MATERIALS AND METHODS: The fungal counts of the saliva, tongue dorsa, and prosthesis-fitting surfaces of the participants were assessed using real-time polymerase chain reaction to compare the quantity and expression of selected C. albicans biofilm-associated genes (ALS3, HWP1, and YWP1). STATISTICAL ANALYSIS: The obtained data were analyzed by one-way analysis of variance, followed by Bartlett's test. When appropriate, the Student's t-test was also used; a value of p < 0.05 was considered statistically significant. RESULTS: In both groups, the count of C. albicans was found to be significantly higher in saliva than in other oral samples. The expression of the hypha-specific genes (ALS3 and HWP1) in the tongue dorsa was higher in the DW group (p < 0.05), whereas the transcription level of the yeast-specific gene (YWP1) was significantly higher in the NDW group. CONCLUSION: Both tongue dorsa and dentures appear to be sharing factors that are important for C. albicans biofilm growth in abiotic and biotic oral surfaces of the elderly.

Expression of TNF, IL1B, and iNOS2 in the neural cell after induced by Porphyromonas gingivalis with and without coating antibody anti -Porphyromonas gingivalis.

Porphyromonas gingivalis has virulence factors such as gingipain and lipopolysaccharide, causing bacteremia to reach the brain and activate neuroinflammatory release cytokines. This study analyzed the effect of the co-culture of neuron cells with P. gingivalis coated with anti -P. gingivalis antibodies against cytokines produced by neuron cells. The gene expressions of the TNF, IL1B, NOS2 in neurons was evaluated using RT-qPCR. The results showed that P. gingivalis coated with anti -P. gingivalis antibody before co-culture with neuron cells could decrease the gene expression of TNF, IL1B, and NOS2 of neuron cells.

RNA aptamers selected against yeast cells inhibit Candida albicans biofilm formation in vitro.

Aptamers that bind live bacterial cells have been widely investigated, but their potential to inhibit Candida albicans biofilm formation needs to be further explored. The aims of this study were to evaluate the binding of C. albicans to RNA aptamers and to examine the potential of aptamers to inhibit C. albicans biofilm formation in vitro. In this study, RNA aptamers selected against yeast cells of C. albicans ATCC 10231 were developed using the systematic evolution of ligands by exponential enrichment (SELEX) technique. The binding affinity of the resulting aptamers was then determined by an aptamer-linked immobilized sorbent assay (ALISA), and a colorimetric (MTT) assay was used to measure the metabolic activity of Candida biofilms. After 11 rounds of SELEX, two candidate aptamers, Ca-apt-1 and Ca-apt-12, were identified. The Ca-apt-1 aptamer also recognized C. albicans isolated from clinical specimens but did not recognize other oral microorganisms (i.e., Streptococcus mutans and Saccharomyces cerevisiae). The ALISA results showed that the binding affinity of these aptamers was comparable to that of an anti-C. albicans monoclonal antibody. In addition, Ca-apt-1 could inhibit biofilm and hyphal formation of C. albicans in vitro, as demonstrated using biofilm assays. This study shows that RNA aptamers could potentially be used in diagnostic and therapeutic applications for C. albicans-related disease in the future.

Scaling and Root Planing Effect to mRNA Expression of Matrix Metalloproteinase-9 and Periodontal Clinical Parameters on Chronic Periodontitis

Abstract Objective: To evaluate the relationship between the mRNA transcription level of Matrix Metalloproteinase-9 (MMP-9) and the selected clinical periodontal healing at one month of scaling and root planing. Material and Methods: A total of six chronic periodontitis patients and one periodontally healthy subject were recruited. The gingival crevicular fluid was collected from all subjects, and the expression level of MMP-9 mRNA was measured by quantitative real-time PCR. Pocket depth, papilla bleeding index, and clinical attachment loss were measured on day 1 at baseline and day 30. Scaling and root planing was performed on day 1. Data were analyzed using SPSS 22.0 software Results: In comparison to the control, periodontal clinical parameters in the treatment group were significantly reduced after scaling and root planing. MMP-9 mRNA expression did not show a significant change after the 30th day. A weak correlation was noted between the MMP-9 mRNA transcription level and the changed PBI measurement Conclusion: Scaling and root planing is clinically effective for chronic periodontitis with a 4-6 mm pocket, whereas the expression of MMP-9 mRNA was not altered. Further studies with a more extended observation period are needed to confirm or reject the present findings.

The Anti-inflammatory Effects of Glycerol-supplemented Probiotic Lactobacillus reuteri on Infected Epithelial cells In vitro.

BACKGROUND: One of the most interesting effects of probiotics is their ability to modulate the immune system through the induction of cytokines and to enhance the host immune response. AIMS: The purpose of this study was to evaluate the anti-inflammatory effect of glycerol-supplemented Lactobacillus reuteri on the transcription level of interleukin (IL)-8 and human-beta-defensin (hBD)-2 expressed by epithelial cells after exposure to bacteria. MATERIALS AND METHODS: The confluent-cultured HaCat cell line (105 cells/mL) was exposed to Streptococcus mutans ATCC-25175 and Porphyromonas gingivalis ATCC-33277 (107 colony-forming units [CFU]/mL) for 24 h and challenged with probiotic L. reuteri ATCC-55730 (107 CFU/mL) supplemented with glycerol. Subsequently, the transcription levels of IL-8 and hBD-2 in HaCat cells were analyzed using reverse-transcription polymerase chain reaction (RT-PCR). In addition, cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. All the obtained data were statistically analyzed using the one-way analysis of variance test, with P < 0.05 set as the level of significance. RESULTS: The MTT assays confirmed no cytotoxic effects of glycerol-supplemented L. reuteri on HaCat cells (viability >90%). mRNA expression of IL-8 and hBD-2 increased after exposure to both bacteria. The presence of glycerol-supplemented L. reuteri significantly reduced the expression of IL-8 and hBD-2 on HaCat cells (P < 0.05). CONCLUSION: Glycerol-supplemented L. reuteri reduced the expression of IL-8 and hBD-2, and the results may be proof of principle for a probiotic approach to combating inflammation. However, further studies are needed to validate this probiotic effect.