Determining the best anti-microbial properties of dental cements used for pulp capping procedures using deep dentinal carious material.

Introduction: The exposure of the healthy pulp in cases of deep dentinal caries (DDC) that contain carious microorganisms can be prevented by placing a layer of pulp capping agent on the affected dentin. The cements used for pulp-capping should also ensure good anti-microbial properties. The present study was carried out to detect the antimicrobial efficacy of the commonly used cements by culturing the samples directly from DDC. Aims: To determine the efficacy of dental cements in the growth inhibition of microorganisms involved in DDC using direct contact anaerobic culture test. Method: 100 samples of DDC were collected in RTF. Ten microliters of the specimen containing RTF was incubated in thioglycolate broth consisting of 1 mm3 cement blocks of GIC, CaOH2, ZnOE and MTA anaerobically for 24 hours. This was further sub-cultured using selective media for streptococcus mutans, lactobacillus and bifidobacterium. Growth inhibition was measured by calculating the number of CFUs and statistically analysed using ANOVA and Tukey's post-hoc tests. Results: Tests showed variation in the anti-microbial effects of the cements and was highly significant at P < 0.001. Bifidobacterium showed most number of CFUs. MTA was the most effective pulp capping agent exhibiting 87.13% reduction in microbial growth, followed closely by ZnOE (84.6%). Conclusion: A conservative approach to treat DDC is the need of the hour which calls for the use of pulp capping cements of good antimicrobial efficacy. The current study revealed bifidobacterium to be the most prevalent in DDC and the cement that would best inhibit the mixed culture growth was MTA followed closely by ZnOE.

Prediction and validation of microbial community function from normal pulp to pulpitis caused by deep dentinal caries.

INTRODUCTION: Microbial function changes may be responsible for dental pulp transformation from normal to diseased. However, studies on the prediction and verification of the function of the microbial community in the deep dentine and pulp of caries-induced pulpitis are lacking. METHODS: This study included 171 cases of deep dentinal caries divided into normal pulp (NP), reversible pulpitis (RP), and irreversible pulpitis (IRP). In Experiment I, the microbial community composition was identified in 111 samples using 16S ribosomal DNA. Function prediction was performed through phylogenetic investigation of communities by Phylogenetic Investigation of Communities by Reconstruction of Unobserved States prediction and qPCR. In Experiment II, different microbiome functions were confirmed in 60 samples using liquid chromatography-tandem mass spectrometry. RESULTS: In Experiment I, microbial abundance significantly differed in the IRP group compared to the other two groups. The RP and NP groups had the same microbiome composition, but the predicted functional difference between the RP and NP groups pertained to membrane transport (p < .010). The predicted functional difference between the IRP and NP groups pertained to amino-acid, co-factor, and vitamin metabolism (p < .010). In Experiment II, Kyoto Encyclopedia of Genes and Genomes functional annotation revealed that the differential metabolites between the RP and NP groups did not participate in membrane transport; however, the differential metabolites between the IRP and NP groups participated in amino-acid metabolism. CONCLUSIONS: The near-pulp microbiome in RP and NP with deep dentinal caries had the same differential function. However, amino acid metabolism in near the pulp microbial community differed between IRP and NP with deep dentinal caries.

Study of microbial diversity in saliva and plaque samples from caries-free and caries-affected children using denaturing gradient gel electrophoresis.

BACKGROUND: Recent investigations have shown the possible involvement of bacteria other than mutans group and Lactobacilli in the etiology of caries. Molecular methods have been used to study the microbial diversity in caries-active (CA) and caries-free (CF) children. Among them, denaturing gradient gel electrophoresis (DGGE) is more popular and has been used in the present study. AIMS: The aim of the present study was to investigate the difference in bacterial diversity in saliva and plaque samples from CF and CA children using DGGE. MATERIALS AND METHODS: The study involved saliva and plaque samples from 56 children of which 28 were CF, 20 with CA, and 8 with white spot lesions (WSP). DNA was extracted and subjected to polymerase chain reaction amplification with universal primers. It was then run in polyacrylamide gel electrophoresis with gradients of urea and formamide and stained with SYBR green. Multiple bands were produced in each sample lane and each band represents one organism. STATISTICAL ANALYSIS: A dendrogram was generated using Phoretix software and similarity index was calculated using a specific formula. RESULTS: Samples in each group formed several clusters indicating a specific pattern of the bacterial profile. Similarity coefficient was calculated based on the number of bands, intensity, and location. The diversity was less in the saliva and plaque samples of CA group as compared to those of CF and WSP groups. CONCLUSIONS: DGGE can be used to study distinctive bacterial profiles in healthy and caries-affected sites. DGGE can be further developed as a pattern recognition tool with which to identify specific groups of bacteria. Saliva may be used to study bacterial diversity in dental caries.