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.

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.

Conditioned-medium of stem cells from human exfoliated deciduous teeth prevent apoptosis of neural progenitors.

Purpose: This study aimed to evaluate the neuroprotective ability of the conditioned medium of stem cells from human exfoliated deciduous teeth (CM-SHED) to prevent glutamate-induced apoptosis of neural progenitors. Materials and methods: Neural progenitors were isolated from two-day-old rat brains, and the conditioned medium was obtained from a mesenchymal stem cell SHED. Four groups were examined: neural progenitor cells cultured in neurobasal medium with (N + ) and without (N-) glutamate and glycine, and neural progenitor cells cultured in CM-SHED with (K + ) and without (K-) glutamate and glycine. Results: The expression of GABA A1 receptor (GABAAR1) messenger RNA (mRNA) in neural progenitor measured by real-time quantitative PCR. GABA contents were measured by enzyme-linked immunosorbent assay, whereas the apoptosis markers caspase-3 and 7-aminoactinomycin D were analysed with a Muse® cell analyzer. The viability of neural progenitor cells in the K + group (78.05 %) was higher than the control group N- (73.22 %) and lower in the N + group (68.90 %) than in the control group. The K + group showed the highest GABA content, which significantly differed from that in the other groups, whereas the lowest content was observed in the N + group. The expression level of GABAAR1 mRNA in the K + group was the highest compared to that in the other groups. CM-SHED potently protected the neural progenitors from apoptosis. Conclusions: CM-SHED may effectively prevent glutamate-induced apoptosis of neural progenitors.

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.

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.

Chitosan gel prevents the growth of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola in mini-implant during orthodontic treatment.

AIMS: We evaluated the effect of chitosan gel on total oral bacteria, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, during orthodontic treatment with mini-implants. MATERIAL AND METHODS: Thirty subjects with 52 orthodontic mini-implants were divided into three groups: one group was treated with chitosan gel, the other group with chlorhexidine gel, and the control group with placebo. The plaque of the orthodontic peri-mini-implant area was collected before and after gel treatment. The total oral bacteria and red-complex bacteria of P. pingivalis, T. forsythia, and T. denticola were determined with reverse transcription-quantitative PCR. RESULTS: Thirty-four orthodontic mini-implants (65.38%) appeared as healthy and showed no clinical signs of inflammation. The total number of bacteria was reduced after chitosan gel application. The highest decrease in the proportion of P. gingivalis was observed in the chlorhexidine gel application group, which showed a value of 70.86%, whereas the chitosan gel application showed a reduction of only 26.59%, and the control gel application showed the lowest reduction effect of only 2.55%. The difference in the reduction between gel application groups was significant (P < 0.05) for T. denticola and T. forsythia. CONCLUSION: The gel containing chitosan reduced the levels of total oral bacteria and red-complex bacteria.

Possible Role of Porphyromonas gingivalis in the Regulation of E2F1, CDK11, and iNOS Gene Expression in Neuronal Cell Cycle: A Preliminary Study.

OBJECTIVE: This study aimed at evaluating the in vitro effect of Porphyromonas gingivalis exposure in gene expression of E2F1 (family of transcription factors), cyclin-dependent kinase-1 (CDK11), and inducible nitric oxide synthase (iNOS) of the neuronal cell cycle. MATERIALS AND METHODS: The culture of neuronal cell line SH-SY5Y was exposed to P. gingivalis ATCC 33277, and the gene expression of E2F1, CDK11, and iNOS was analyzed by using a real-time polymerase chain reaction. RESULTS: It was shown that E2F1, a G1 phase biomarker and transcription factor, was upregulated in neuronal cells exposed to P. gingivalis compared with that in control cells. However, CDK11, a biomarker of G2/M checkpoint and iNOS, was downregulated in neuronal cells exposed to P. gingivalis compared with that in control cells. CONCLUSIONS: P. gingivalis can regulate the neuronal cell cycle, as indicated in the E2F1, CDK11, and iNOS gene expression.

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.

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.

Establishing methods for isolation of stem cells from human exfoliated deciduous from carious deciduous teeth.

AIM: This study aims to establish the isolation method of stem cells from pulp tissue of carious deciduous teeth. METHODS: The teeth were soaked in 1% povidone-iodine solution for about 1 min followed by washing in PBS with 1% antibiotic-antimycotic thrice. Dental pulp tissue was removed by extirpation, and then cultivated in the culture medium. Characterization of mesenchymal stem cell (MSC) was carried out using human MSC analysis kit with positive markers CD90, CD73, and CD105, but negative for expressions of CD45, CD34, CD11b, CD19, and HLA-DR. Differentiation capacity of stem cells from human exfoliated deciduous (SHED) was determined by staining with Alizarin S, Alcian Blue, and Oil Red O. RESULTS: There is no contamination after 3 days of culture. SHED derived from dental pulp were expressions of 99.2% of positive marker and 0.3% of the negative marker. At passage 5, SHED was differentiated into osteocyte, chondrocyte, and adipocyte types of cells in the induction medium. CONCLUSION: SHED derived from carious deciduous teeth can be used as a source of stem cell for regenerative medicine.

Relationship between Candida albicans and Streptococcus mutans in early childhood caries, evaluated by quantitative PCR.

Background: The aim of this study was to analyze the synergistic relationship between Candida albicans and Streptococcus mutans in children with early childhood caries (ECC) experience. Methods: Dental plaque and unstimulated saliva samples were taken from 30 subjects aged 3-5 years old, half with (n=15, dmft > 4) and half without (n=15) ECC. The abundance of C. albicans and S. mutans and relative to total bacteria load were quantify by real-time PCR (qPCR). This method was also employed to investigate the mRNA expression of glycosyltransferase ( gtfB) gene in dental plaque. Student's t-test and Pearson's correlation were used to perform statistical analysis. Results: Within the ECC group, the quantity of both microorganisms were higher in the saliva than in dental plaque. The ratio of C. albicans to total bacteria was higher in saliva than in plaque samples (p < 0.05). We observed the opposite for S. mutans (p < 0.05). The different value of C. albicans and S. mutans in saliva was positively correlated, and negatively correlated in dental plaque. Transcription level of S. mutans gtfB showed a positive correlation with C. albicans concentration in dental plaque.  Conclusion:C. albicans has a positive correlation with cariogenic traits of S. mutans in ECC-related biofilm of young children.

Mutans Streptococci counts from saliva and its protein profile in early childhood caries.

AIM: This study aims to analyze the number Mutans Streptococci (MS) and its protein profile from the saliva of early childhood caries (ECC) and caries-free subjects. METHODS: MS counts were cultured from saliva samples, and the protein profile of MS was determined from ECC and caries-free subjects. The number of colonies were counted, and the protein bands with the molecular weight of 13, 29, 39, 41.3, 74, and 95 kDa were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis method. RESULTS: We found that the number of colonies from saliva of ECC patients was higher than those caries-free (22.20 × 106 CFU/ml vs. 19.16 × 106 CFU/ml, p < 0.05). There are higher expression frequencies in protein 29, 39, 41.3, and 74 kDa of MS in ECC than caries-free subjects. CONCLUSIONS: There is the higher number of MS colonies and difference of MS protein profile isolated from saliva among children with ECC and caries-free counterparts.

Proinflammatory MG-63 cells response infection with Enterococcus faecalis cps2 evaluated by the expression of TLR-2, IL-1ß, and iNOS mRNA.

OBJECTIVE: We have previously demonstrated that unencapsulated Enterococcus faecalis cps2 inhibits biofilm formation of Candida albicans, a fungus commonly found with E. faecalis in periapical lesion. In this study, we compared encapsulated and unencapsulated E. faecalis cps2 strains relationship with osteoblastic (MG-63) cells, whereas E. faecalis ATCC 29212 were used as a reference strain. RESULTS: The binding capacity of E. faecalis to MG-63 cells as shown by each tested strain was comparable, but the unencapsulated strain was less invasive compared to the encapsulated and the reference strains. Moreover, quantitative real time-PCR (qPCR) results showed that infecting unencapsulated E. faecalis cps2 is a stronger stimulator for toll like receptor 2 (TLR2) and interleukin-1ß (IL-1ß) mRNAs, but not for inducible nitric oxide synthase (iNOS) mRNA in osteoblastic cells. In conclusion, the performance of unencapsulated E. faecalis cps2 when the bacterium interacts with osteoblastic cells is quite different from that of encapsulated E. faecalis cps2 and reference strains. It appears that the unencapsulated strain might contribute to the persistence of the periapical inflammatory response, depending on down-regulation of iNOS mRNA expression.