Evaluation of antimicrobial efficacy of ethanolic extract of Cuminium Cyminium as intracanal medicament on common bacteria of endodontic infections: An in vitro study.

Context: Antimicrobial intracanal medicaments play a vital role in successful outcome of any endodontic procedure. One such plant extract Cuminium cyminium, as intracanal medicaments needs to be researched. Aims: The purpose of this study was in vitro assessment of the antibacterial activity of ethanol extract of C. Cyminium in comparison to Calcium hydroxide (Ca[OH]2) as intracanal medicament against the pathogens of endodontic infection, at an interval 1 h, 24 h, 48 h, and 72 h. Settings and Design: The study was conducted in the central research laboratory of our institute. Freshly prepared C. cyminium extract was procured from AYUSH approved laboratory and direct contact test (DCT) was utilized, which is based on turbidometric determination of microbial growth in a 96-well microplate, carrying 6 times for each bacteria. Methodology: Three groups were assigned for each material in a 96 microwell plate for DCT. Bacterial growth kinetics was monitored at intervals of 1 h, 24 h, 48 h, and 72 h using spectrophotometer at 595 nm. The optical density of T2 (Test group), P2 (Positive control), and N2 (Negative control) was considered. Statistical Analysis Used: After compiling the data, based on the normality of data, further statistical analysis was conducted using Kolmogorov-Smirnov test, Paired t-test, and pairwise comparisons by Turkey's multiple post hoc procedures. The level of statistical significance was set at P = 0.05. Results: The comparison of mean optical density values of C. cyminium in comparison with Ca(OH)2 against the microorganisms of endodontic origin showed a statically significant decrease in bacterial viability at the end of 24 h, 48 h, and 72 h. Conclusion: Based on the results of the study, it can be concluded that C. cyminium has significant antibacterial action against endodontic origin, at interval of 24 h, 48 h, and 72 h.

Simultaneous detection and evaluation of Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, and Prevotella melaninogenica in subgingival plaque samples of chronic periodontitis and healthy individuals through multiplex polymerase chain reaction.

Background: Prevotella is a Gram-negative anaerobic bacilli. The phenotypic characteristics of the various species of Prevotella are similar, which often makes it difficult in routine differentiation and identification of all the species. Aim: The purpose of the study was to detect and compare presence of Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica, and Prevotella loescheii in subgingival plaque samples of chronic periodontitis and healthy individuals. Materials and Methods: Two hundred and thirty-six subjects were considered consisting of chronic periodontitis (128) and healthy (108) individuals. Subgingival plaque sample was collected in reduced transport fluid and analyzed. DNA extraction and polymerase chain reaction (PCR) were performed for genus Prevotella followed by positive samples were considered for the detection of selected species through multiplex PCR using specific primers. Results: Out of 236 samples, 94.1% were positive for genus Prevotella. Out of 222 cases P. nigrescens showed the highest number of cases positive (59.5%) followed by P. melaninogenica (57.2%), P. intermedia (55.4%), and P. loescheii (40.1%). Species were analyzed individually between chronic periodontitis and healthy, P. intermedia, P. nigrescens, and P. loescheii showed greater positivity in healthy compared to chronic periodontitis. Positivity for P. melaninogenica was high in chronic periodontitis compared to healthy. Conclusion: The number of positive cases for species, when correlated with clinical parameters showed an increase in mean score for all clinical parameters assessed, suggesting the presence of variation in the prevalence of Prevotella species and geographic variation do exist in oral microflora. Findings suggest that they can be normal commensals and opportunistic.

Regeneration of periodontal ligament fibers around mini dental implants and their attachment to the bone in an animal model: A radiographic and histological study.

Background: Tissue-engineered periodontal ligament (PDL) around a dental implant by using PDL stem cells (PDLSCs) may be useful in periodontal regeneration and can reduce or eliminate certain shortcomings of dental implants. Materials and Methods: PDLSCs were isolated from extracted human PDL cells and cultured in a bioreactor. They were identified using markers CD45, CD73, CD90, CD105, and CD146. After the formation of multiple cellular layers, they were then attached on titanium mini dental implants and placed in rabbit tibia. The rabbits were sacrificed after 9 months, and the implants were analyzed histologically and radiographically by Cone beam computed tomography (CBCT). Results: Isolated PDLSCs obtained from human premolars showed a colony-forming ability on the 7th day and 14th day. Immunocytochemistry revealed that cells had taken up the adequate positive stains for primary antibodies CD73, CD90, CD105, and CD146 and negative staining for CD45. The histological sections obtained from sacrificed rabbits, when viewed under the light microscope, clearly showed the presence of PDL around dental implants. CBCT examination showed that the implant was well within the bone and did not migrate. The site appeared to be normal without any lytic changes in the bone. Conclusion: It can safely be postulated from the present study that tissue engineering of PDL can be achieved around dental implants using PDLSCs. Important inter-tissue interactions like the formation of a functional PDL around the implantation site, and induction of bone formation in the vicinity of the implants may be possible. Future research in humans is required for further research.

Evaluation of Presence of Enterococcus faecalis in Root Canals of Deciduous Molars with Necrotic Pulp by Agar Culture and Polymerase Chain Reaction.

Objective: To assess the presence of Enterococcus faecalis in root canals of deciduous molars with necrotic pulp by agar culture and polymerase chain reaction (PCR) assay. Materials and methods: This is an experimental study, where a total of 120 endodontic samples were taken from deciduous molars with necrotic pulps. The presence of Enterococcus faecalis was assessed by culture, using Enterococcus confirmatory agar, and by PCR assay. Statistical analysis of the results was performed using McNemar's test. Results: The presence of Enterococcus faecalis was detected in 20 samples (16.67% of total) by microbial culture and in 45 samples (37.5% of total) by PCR assay, with a statistically significant difference between the two methods (p < 0.001). Microbial culture and PCR both detect Enterococcus faecalis, with the latter detecting an additional 25 positive samples. Conclusion: In this study, PCR assay was significantly more sensitive than agar culture method in detecting the presence of Enterococcus faecalis in root canals of deciduous molars with necrotic pulp, that is, 37.5% of all samples. Clinical significance: Importance of presence of Enterococcus faecalis in necrotic pulps of deciduous teeth, as it is primarily responsible for failure of endodontic treatment, thus helping clinicians to advocate the use of local drug delivery in primary teeth endodontics and also aids clinicians in choosing the most effective intracanal medication. How to cite this article: Nalawade TM, Bhat KG, Kale AD, et al. Evaluation of Presence of Enterococcus faecalis in Root Canals of Deciduous Molars with Necrotic Pulp by Agar Culture and Polymerase Chain Reaction. Int J Clin Pediatr Dent 2023;16(6):816-819.

Detection and comparison of prevalence of Porphyromonas gingivalis through culture and Real Time-polymerase chain reaction in subgingival plaque samples of chronic periodontitis and healthy individuals.

Introduction: The micro-flora of oral cavity is a myriad of micro-organism. Any infection of oral cavity leads to diseased condition which is a transitional transformation of the micro-organism in a specific paradigm depending upon the diseased condition. Periodontitis is one of the predominant chronic diseases which is a multifactorial infection. Porphyromonas gingivalis is a key etiological agent in causing periodontitis. To study the predominance of these bacteria in the diseased condition is important to detect, quantify and to find its efficacy by comparing different methods for identification. Aim and Objectives: The aim of the study is to determine the prevalence of P. gingivalis by anerobic culture and by real-time polymerase chain reaction (PCR) from subgingival plaque samples of chronic periodontitis and healthy individual and to compare efficacy of two methods. Materials and Methods: A total of 400 subjects were considered, and subgingival plaque was collected using paper points. Individual were equally divided into two groups: chronic periodontitis (200) and healthy individuals (200). Each plaque sample collected was divided into two aliquots of which the first aliquot was subjected for anerobic culture to isolate P. gingivalis. Phenotypical identification was done morphologically and biochemically further quantification of P. gingivalis was done by colony-forming unit. The second aliquot was subjected for DNA extraction and real-time PCR was conducted to detect and quantify P. gingivalis using specific primer. Results: Out of 400 samples, 73% showed detection of P. gingivalis by culture method and through reverse transcription-PCR (RT-PCR), the detection was 75%. Individual detection of P. gingivalis by culture in chronic periodontitis was 89.5% and 54.4% in healthy individuals, while detection by RT-PCR was found to be 91.5% in chronic periodontitis and 58% in healthy individuals. However, comparison between two techniques in detection of P. gingivalis was statistically insignificant. Conclusion: When we compared RT-PCR with culture RT-PCR showed higher positivity. RT-PCR is more sensitive and requires less time to detect. However, in the present study, culture also showed good positivity, suggesting proper dilution and with extended incubation, the specificity of culture can be improved to a great extent.

Green tea catechins showed antibacterial activity on streptococcus mutans -An in vitro study.

BACKGROUND: The main bacterial aetiological agents in caries formation are the α-haemolytic Streptococcal species Streptococcus mutans, which has been found to be the initiator of most dental caries. The leaves of Camellia sinensis known as green tea, has properties, such as antibacterial and anti-cariogenic. Epigallocatechin-3-gallate (EGCG) one of the most abundant catechins found in green tea is known to contribute to these effects. AIM: To evaluate the antibacterial effect of green tea catechins namely EGCG on S. mutans with two different methods at different concentrations. OBJECTIVES: 1) To assess the antimicrobial efficacy of EGCG by disc diffusion test at concentrations of 100, 75, and 50 µg/mL. 2) To assess the antimicrobial efficacy of EGCG by Minimum inhibitory concentration (MIC) test at concentrations ranging from 0.2 to 100 µg/mL. METHODOLOGY: Commercially available purest form of green tea polyphenol EGCG was used in the study. Disc diffusion test on agar medium and MIC test was used to determine the susceptibility of the S. mutans to green tea catechins EGCG. RESULTS: The results of the agar well diffusion method showed that the EGCG extract has shown zones of inhibition against S. mutans at concentrations of 100 µg/mL (28.67 mm), 75 µg/mL (15.33 mm), 50 µg/mL (10.33 mm) while that of MIC test of EGCG extract of concentrations ranging from 0.2 to 100 µg/mL against S. mutans shows that the mean MIC value was 1.07. CONCLUSION: Catechins in the tea are potentially anti-cariogenic agents which can reduce bacterial presence in the oral cavity and have the potential to be further used for the preparation of dentifrice and mouthwash.

Cancer Stem Cell Traits in Tumor Spheres Derived from Primary Laryngeal Carcinoma Cell Lines.

OBJECTIVE: Cancer stem cells (CSCs) belong to a subpopulation of undifferentiated cells present within tumors that have the potential to regenerate, differentiate, maintenance of pluripotency, drug resistance, and tumorigenicity when transplanted into an innate host. These can influence the growth and behavior of these tumors and are used to investigate the initiation, progression, and treatment strategies of laryngeal cancer. Research on CSC science and targeted therapies were hinge on their isolation and/or enrichment procedures. The object of the study is to isolate cancer stem cells from primary laryngeal carcinoma (CSCPLC) by tumor spheres enrichment. We checked the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance. MATERIALS AND METHODS: We performed tumor sphere formation assay (primary, secondary, and tertiary) chemotherapy resistance by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay were performed to evaluate the CSC cells. Immunofluorescence for stem cell markers (CD133+, CD44+) and gene expression of stem cell markers for CD133+, CD44+, OCT4, SOX2, and NANOG was done using the real-time polymerase chain reaction technique. RESULTS: We were able to isolated CSC subpopulations from PLC cell lines by the tumor sphere method. These cells exhibited good primary, secondary, and tertiary tumor sphere formation efficiency and also disclosed a resistant index of more than 2. Immunofluorescence for stem cell markers (CD133+ and CD44+) confirms the presence of CSC. There was significantly higher mRNA expression of stem cell markers in CSC enriched subpopulations compared to the parental cell lines. CONCLUSION: We conclude that tumor spheres enrichment is an efficient, economical, and reliable approach for the isolation and characterization of CSC from PLC cell lines. These cells demonstrated the properties of self-renewal, stemness, clonogenicity, and chemotherapeutic resistance.

Solitary Median Maxillary Central Incisor: A Case Report with 3-Year Follow-Up and Literature Review.

A solitary median maxillary central incisor (SMMCI) is a rare anomaly that can occur alone or be associated with other systemic abnormalities. Early diagnosis of SMMCI is crucial as it might indicate the presence of an associated congenital or developmental abnormality. The prevalence of live-born children with SMMCI is determined to be 1:50,000 and is more common among females. The purpose of this paper was to report an unusual case of a 9-year-old girl with SMMCI who had no growth deficiency or any other systemic involvement. Since pediatricians and dentists are the first professionals to evaluate an SMMCI's patient in most cases, it is important that they be aware of the possibility of other related systemic problems that require systemic care. Appropriate treatment, diagnosis, and referral should also include neuropediatric evaluation, genetic testing, and craniofacial profile analysis along with multidisciplinary approach.

Coculture method for in vitro cultivation of uncultured oral bacteria.

PURPOSE: The purpose of the study is to culture uncultured oral bacteria with helper strains using the coculture method from the subgingival plaque samples of chronic periodontitis patients. MATERIALS AND METHODS: The samples were processed and inoculated on a blood agar medium enriched with hemin and Vitamin K. A helper strain Propionibacterium acnes (ATCC 6919) was cross-streaked across the inoculums to facilitate coculture. The plates were then incubated for 7 days with subsequent subculturing and further incubation. RESULTS: Satellite colonies around helper strain showed one colony type of Porphyromonas gingivalis, one was of nonpigmented Prevotella, three were of Fusobacterium nucleatum and five isolates remained unidentified. CONCLUSIONS: Coculture could be used effectively as one of the methods in the isolation and in vitro cultivation of oral bacteria. Incubation using the anaerobic jar technique was found to be economical and efficient for the growth of anaerobic oral bacteria.

Antimicrobial susceptibility pattern of oral gram negative anaerobes from Indian subjects.

OBJECTIVES: There is paucity of information on the antimicrobial susceptibility pattern of oral anaerobic bacteria. In this study, an attempt has been made to evaluate the antimicrobial susceptibility/resistance trend of oral Gram negative bacteria from Indian subjects. METHODS: Minimum inhibitory concentrations (MIC) of 304 isolates against twelve different antibiotics were determined using gradient diffusion MIC strips. The organisms were isolated and identified based on phenotypic characteristics and included Porphyromonas gingivalis, Prevotella species, Tannerella forsythia, Fusobacterium nucleatum, Aggregatibacter actinomycetemcoitans, Eickenella corrodens and Capnocytophaga species. For each antimicrobial agent, MIC50 and MIC90 were calculated and expressed. RESULTS: Resistance to azithromycin, clindamycin, and amoxicillin was observed in most of the anaerobic bacterial species studied. High degree of susceptibility was observed to amoxillin-clavulanic acid, doxycycline and moxifloxacin. A single strain of P. melaninogenica was resistant to moxifloxacin. The susceptibility pattern varied with cephalosporins among species. Ceftriaxone showed highest and cefazolin least efficacy among cephalosporins. All anaerobic bacteria tested were susceptible to metronidazole. Strains of T. forsythia were more resistant to several antibiotics than other anaerobic bacteria. All three species of capnophilic bacteria displayed high degree of resistance to metronidazole and significant resistance to amoxicillin, azithromycin, clindamycin, cefazolin and cefuroxime. CONCLUSIONS: Amoxicillin-clavulanic acid, doxycycline, moxifloxacin and metronidazole appeared to be the most effective drugs against gram negative anaerobic bacteria. However, the MIC50 and MIC90 values against metronidazole were on the higher side of the normal indicating a potential for developing resistance.

Effect of curcumin on growth, biofilm formation and virulence factor gene expression of Porphyromonas gingivalis.

Porphyromonas gingivalis is a keystone pathogen and major colonizer in host tissue which plays a pivotal role in periodontitis among the other polymicrobial infections. Increasing facts demonstrate that curcumin has antibacterial activity and anti-biofilm effect against the periodontopathogens through diverse mechanisms that have a positive impact on periodontal health. The present study was aimed to elucidate the effect of curcumin on biofilm formation and virulence factor gene expression of P. gingivalis. By using gene expression studies, we exploited the mechanism of anti-biofilm effects of curcumin on P. gingivalis. The minimum inhibitory concentration and minimum bactericidal concentration of curcumin for both ATCC and clinical strains of P. gingivalis were found to be 62.5 and 125 µg ml-1 respectively. Curcumin prevented bacterial adhesion and biofilm formation in a dose-dependent manner. Further, curcumin attenuated the virulence of P. gingivalis by reducing the expression of genes coding for major virulence factors, including adhesions (fimA, hagA, and hagB) and proteinases (rgpA, rgpB, and kgp). The results indicated that curcumin has shown anti-biofilm as well as antibacterial activity against P. gingivalis. Further, curcumin because of its pleiotropic actions could be a simple and inexpensive therapeutic strategy in the treatment of periodontal disease.

Adjunctive systemic antimicrobials for the non-surgical treatment of periodontitis.

BACKGROUND: Systemic antimicrobials can be used as an adjunct to mechanical debridement (scaling and root planing (SRP)) as a non-surgical treatment approach to manage periodontitis. A range of antibiotics with different dosage and combinations are documented in the literature. The review follows the previous classification of periodontitis as all included studies used this classification. OBJECTIVES: To assess the effects of systemic antimicrobials as an adjunct to SRP for the non-surgical treatment of patients with periodontitis. SEARCH METHODS: Cochrane Oral Health's Information Specialist searched the following databases to 9 March 2020: Cochrane Oral Health's Trials Register, CENTRAL, MEDLINE, and Embase. The US National Institutes of Health Ongoing Trials Register ClinicalTrials.gov and the World Health Organization International Clinical Trials Registry Platform were searched for ongoing trials. SELECTION CRITERIA: We included randomized controlled trials (RCTs) which involved individuals with clinically diagnosed untreated periodontitis. Trials compared SRP with systemic antibiotics versus SRP alone/placebo, or with other systemic antibiotics. DATA COLLECTION AND ANALYSIS: We selected trials, extracted data, and assessed risk of bias in duplicate. We estimated mean differences (MDs) for continuous data, with 95% confidence intervals (CIs). We assessed the certainty of the evidence using GRADE. MAIN RESULTS: We included 45 trials conducted worldwide involving 2664 adult participants. 14 studies were at low, 8 at high, and the remaining 23 at unclear overall risk of bias. Seven trials did not contribute data to the analysis. We assessed the certainty of the evidence for the 10 comparisons which reported long-term follow-up (≥ 1 year). None of the studies reported data on antimicrobial resistance and patient-reported quality of life changes. Amoxicillin + metronidazole + SRP versus SRP in chronic/aggressive periodontitis: the evidence for percentage of closed pockets (MD -16.20%, 95% CI -25.87 to -6.53; 1 study, 44 participants); clinical attachment level (CAL) (MD -0.47 mm, 95% CI -0.90 to -0.05; 2 studies, 389 participants); probing pocket depth (PD) (MD -0.30 mm, 95% CI -0.42 to -0.18; 2 studies, 389 participants); and percentage of bleeding on probing (BOP) (MD -8.06%, 95% CI -14.26 to -1.85; 2 studies, 389 participants) was of very low certainty. Only the results for closed pockets and BOP showed a minimally important clinical difference (MICD) favouring amoxicillin + metronidazole + SRP. Metronidazole + SRP versus SRP in chronic/aggressive periodontitis: the evidence for percentage of closed pockets (MD -12.20%, 95% CI -29.23 to 4.83; 1 study, 22 participants); CAL (MD -1.12 mm, 95% CI -2.24 to 0; 3 studies, 71 participants); PD (MD -1.11 mm, 95% CI -2.84 to 0.61; 2 studies, 47 participants); and percentage of BOP (MD -6.90%, 95% CI -22.10 to 8.30; 1 study, 22 participants) was of very low certainty. Only the results for CAL and PD showed an MICD favouring the MTZ + SRP group. Azithromycin + SRP versus SRP for chronic/aggressive periodontitis: we found no evidence of a difference in percentage of closed pockets (MD 2.50%, 95% CI -10.19 to 15.19; 1 study, 40 participants); CAL (MD -0.59 mm, 95% CI -1.27 to 0.08; 2 studies, 110 participants); PD (MD -0.77 mm, 95% CI -2.33 to 0.79; 2 studies, 110 participants); and percentage of BOP (MD -1.28%, 95% CI -4.32 to 1.76; 2 studies, 110 participants) (very low-certainty evidence for all outcomes). Amoxicillin + clavulanate + SRP versus SRP for chronic periodontitis: the evidence from 1 study, 21 participants for CAL (MD 0.10 mm, 95% CI -0.51 to 0.71); PD (MD 0.10 mm, 95% CI -0.17 to 0.37); and BOP (MD 0%, 95% CI -0.09 to 0.09) was of very low certainty and did not show a difference between the groups. Doxycycline + SRP versus SRP in aggressive periodontitis: the evidence from 1 study, 22 participants for CAL (MD -0.80 mm, 95% CI -1.49 to -0.11); and PD (MD -1.00 mm, 95% CI -1.78 to -0.22) was of very low certainty, with the doxycycline + SRP group showing an MICD in PD only. Tetracycline + SRP versus SRP for aggressive periodontitis: we found very low-certainty evidence of a difference in long-term improvement in CAL for the tetracycline group (MD -2.30 mm, 95% CI -2.50 to -2.10; 1 study, 26 participants). Clindamycin + SRP versus SRP in aggressive periodontitis: we found very low-certainty evidence from 1 study, 21 participants of a difference in long-term improvement in CAL (MD -1.70 mm, 95% CI -2.40 to -1.00); and PD (MD -1.80 mm, 95% CI -2.47 to -1.13) favouring clindamycin + SRP. Doxycycline + SRP versus metronidazole + SRP for aggressive periodontitis: there was very low-certainty evidence from 1 study, 27 participants of a difference in long-term CAL (MD 1.10 mm, 95% CI 0.36 to 1.84); and PD (MD 1.00 mm, 95% CI 0.30 to 1.70) favouring metronidazole + SRP. Clindamycin + SRP versus metronidazole + SRP for aggressive periodontitis: the evidence from 1 study, 26 participants for CAL (MD 0.20 mm, 95% CI -0.55 to 0.95); and PD (MD 0.20 mm, 95% CI -0.38 to 0.78) was of very low certainty and did not show a difference between the groups. Clindamycin + SRP versus doxycycline + SRP for aggressive periodontitis: the evidence from 1 study, 23 participants for CAL (MD -0.90 mm, 95% CI -1.62 to -0.18); and PD (MD -0.80 mm, 95% CI -1.58 to -0.02) was of very low certainty and did not show a difference between the groups. Most trials testing amoxicillin, metronidazole, and azithromycin reported adverse events such as nausea, vomiting, diarrhoea, mild gastrointestinal disturbances, and metallic taste. No serious adverse events were reported. AUTHORS' CONCLUSIONS: There is very low-certainty evidence (for long-term follow-up) to inform clinicians and patients if adjunctive systemic antimicrobials are of any help for the non-surgical treatment of periodontitis. There is insufficient evidence to decide whether some antibiotics are better than others when used alongside SRP. None of the trials reported serious adverse events but patients should be made aware of the common adverse events related to these drugs. Well-planned RCTs need to be conducted clearly defining the minimally important clinical difference for the outcomes closed pockets, CAL, PD, and BOP.

Quantitative assessment of Scardovia wiggsiae from dental plaque samples of children suffering from severe early childhood caries and caries free children.

Scardovia wiggsiae has recently been identified as a potential pathogen associated with dental caries. The aim of the present study was to detect and quantify S. wiggsiae from dental plaque samples of children suffering from severe early childhood caries and children who were caries free by employing a real time DNA polymerase chain reaction (Real-time PCR) method. Dental plaque samples were collected from children suffering from severe early childhood caries (S-ECC) (n = 30) and caries free children (CF) (n = 30) reporting to the out-patient clinics of the department of paediatric and preventive dentistry. Plaque samples from each group were subjected to real-time PCR, post DNA extraction. Both the groups showed the presence of the organism S. wiggsiae, however there was a significant difference in its quantification between groups, with the median number being 1.49 × 108 cells per ml in caries free samples compared to 1.40 × 109 cells per ml in S-ECC samples. S. wiggsiae were isolated from nearly all samples of children, both caries free and those suffering from S-ECC. However, their numbers differ drastically in both groups with the scales tipping towards the S-ECC group, proving their association with the disease process in a significant manner. The present study shows significant association of S. wiggsiae in severe early childhood caries.

Roles of the matricellular protein Tenascin-C in T-lymphocyte trafficking and etiopathogenesis of Oral Lichen Planus.

OBJECTIVE: This study was aimed at examining the role of Tenascin-C in T cell trafficking in Oral Lichen Planus (OLP). DESIGN: For the in vivo immunohistochemical analyses, 115 OLP samples were collected from patients and immunostaining was performed. The intensity and distribution of TN-C expression were quantified and correlated with histological analyses of basement membrane integrity and presence of inflammatory infiltrate. For the in vitro study, TN-C and collagen were coated on culture plates and migration of T lymphocytes was assessed. RESULTS: TN-C immunoexpression was increased in terms of both distribution and intensity along the basement membrane zone. These changes were significantly associated with basement membrane duplication (distribution p < 0.002 and intensity p < 0.001) and bands of inflammation (distribution p < 0.002 and intensity p < 0.001) assessed by Chi-square test. T lymphocytes demonstrated significant migration towards TN-C as compared to collagen (n = 3, p < 0.05). CONCLUSIONS: These findings indicate TN-C may have a key role in promoting T cell migration at the epithelial-mesenchymal junction in OLP. These observations suggest TN-C could be a good target for therapeutic intervention, either in itself or synergistically with anti-inflammatory directed strategies in this chronic disease management.

In vitro evaluation of cytotoxicity of Emblica officinalis (amla) on cultured human primary dental pulp fibroblasts.

CONTEXT: The dental pulp tissue is capable of healing after surgical amputation of infected/inflamed tissue during vital pulp therapy, when in contact with a suitable medicament. Emblica officinalis (amla), a traditional medicine, is one such medicament which has never been evaluated for its healing potential in pulp therapy. AIMS: The aim of the study was to evaluate the cytotoxicity of E. officinalis (amla) against human primary dental pulp fibroblasts. SETTINGS AND DESIGN: This was in vitro study. SUBJECTS AND METHODS: Human dental pulp fibroblasts were obtained from dental pulp tissue of extracted over-retained primary incisors. The primary cells were cultured using the Dulbecco's Modified Eagle's Medium and used for the study after the fourth passage. The test medicament was E. officinalis with mineral trioxide aggregate (MTA) (100%) and untreated cells as positive and negative controls, respectively. Methyl-thiazol-diphenyl-tetrazolium (MTT) cytotoxicity assay was performed, and the cell survival was observed and analyzed at intervals of 24, 48, and 72 h. STATISTICAL ANALYSIS USED: Cell survival within groups was compared with Wilcoxon matched-paired t-test and in between groups at each point interval was analyzed with the Kruskal-Wallis ANOVA test. The level of significance was set at 0.05. RESULTS: Within the groups, across the time periods of evaluation, there was a decline in cell survival in both the groups but was statistically significant in the MTA group. On interval-wise comparison, the decline in cell survival was statistically significant between the three groups at 72 h (P = 0.001). CONCLUSIONS: E. officinalis preserved the vitality of the human primary dental pulp fibroblasts and has the potential to be developed into vital pulp therapy medicament.

Antimicrobial susceptibility pattern of oral isolates of Aggregatibacter actinomycetemcomitans.

BACKGROUND: Aggregatibacter actinomycetemcomitans is involved in the etiology of localized aggressive periodontitis (LAP), a condition that frequently requires supplemental antibiotic therapy. Information on antimicrobial susceptibility pattern and guidelines for oral antibiotic therapy are not available on Indian patients. AIM: The main aim of the present study was to screen clinical isolates on a panel of antibiotics commonly used for oral/systemic therapy. MATERIALS AND METHODS: The study included 40 strains of A. actinomycetemcomitans isolated from patients with LAP. The subgingival plaque was plated onto Trypticase Soy Serum Bacitracin Vancomycin Agar medium and incubated for 72 h, and suspected colonies were confirmed by phenotypic tests. Each isolate was tested against a panel of 12 antibiotics using MIC gradient strip test. ATCC strains of A. actinomycetemcomitans serotype A and C were used as standards. Performance and interpretation of the test were done according to the manufacturers' instructions. Distribution of MICs among isolates (n = 40) were used to calculate concentrations inhibiting 50% (MIC50) and 90% (MIC90) of strains. RESULTS: Moxifloxacin, cefotaxime and ceftriaxone showed excellent activity with 100% growth inhibition followed by amoxicillin, amoxiclav and doxycycline (>90% activity). The bacterial strains were moderately susceptible to cefuroxime, cefazolin and tetracycline but displayed poor susceptibility to clindamycin and azithromycin. All isolates were resistant to metronidazole. CONCLUSION: The isolates of A. actinomycetemcomitans displayed a high level of resistance to azithromycin and clindamycin. Development of resistance against tetracycline also appears to be significant. Variable resistance among the different members of the cephalosporin group is a factor to be investigated further since susceptibility profile against these antibiotics and interpretative criteria for oral bacteria are not available.

Effect of Resveratrol on biofilm formation and virulence factor gene expression of Porphyromonas gingivalis in periodontal disease.

Periodontal disease is an oral inflammatory disease that destroys the tooth supporting periodontal tissues resulting in tooth loss. Porphyromonas gingivalis is a keystone pathogen that plays a significant role in periodontitis. In previous studies, resveratrol has shown significant results by targeting inflammatory and adhesive markers. Virulence factors of P. gingivalis play an important role in the bacterial adhesion and colonization. In this study, we aimed to demonstrate the anti-biofilm and anti-bacterial activity of resveratrol and also study the effect of resveratrol on the expression of virulence factor genes of P. gingivalis using reverse transcriptase polymerase chain reaction (RT-PCR). The anti-microbial and anti-biofilm activity of resveratrol on P. gingivalis was carried out by broth microdilution assay and biofilm adhesion reduction-crystal violet assay, respectively. We carried out the gene expression analysis by RT-PCR with the P. gingivalis treated compound to analyze the change in the expression of virulence factors: fimbriae and gingipain. Minimal inhibitory concentrations (MIC) of resveratrol against P. gingivalis and other clinical strains are in the range of 78.12-156.25 µg/mL. Resveratrol dose-dependently prevented the biofilm formation and also attenuated the virulence of P. gingivalis by reducing the expression of virulence factor genes such as fimbriae (type II and IV) and proteinases (kgp and rgpA). Resveratrol demonstrated superior anti-bacterial and anti-biofilm activity against P. gingivalis. There was significant reduction in the expression of fimbriae and gingipain with the resveratrol-treated compound. The results suggest that resveratrol, due to its multiple actions, may become a simple and inexpensive therapeutic strategy for treating periodontal disease.

Detection of antibodies against Aggregatibacter actinomycetemcomitans in serum and saliva through ELISA in periodontally healthy individuals and individuals with chronic periodontitis.

BACKGROUND: Periodontitis is a persistent polymicrobial infection, which leads to chronic inflammation in the tooth supporting tissues. Aggregatibacter actinomycetemcomitans are normal commensals of oral cavity but are low in number in periodontally healthy subjects. They are one of the major pathogens aetiologically linked to periodontal disease. Plasma and salivary antibody measurement may be useful to support diagnosis, disease activity, classification and prognosis of periodontitis. The aim of the present study was to investigate the association between the serum and salivary antibody levels to A. actinomycetemcomitans and therefore, to find whether this association was varying in different grades of periodontitis. METHOD: Total of 50 periodontally healthy and 50 chronic periodontitis subjects (35-65 years) of both sexes were included for the study. 2 ml of un-stimulated saliva and 5 ml of venous blood was collected under sterile conditions. The detection of antibodies against A. actinomycetemcomitans in periodontally healthy individuals and individuals with chronic periodontitis was performed using indirect ELISA. RESULTS: Results showed serum IgG, IgA mean levels against A. actinomycetemcomitans were higher in chronic periodontitis subjects compared to mean levels in periodontally healthy subjects. Similarly, salivary IgG, IgA levels were also raised in chronic periodontitis patients as compared in healthy subjects. Also the mean levels of serum IgG and salivary IgA were increased as the severity of disease increased. CONCLUSION: Antibody titer using saliva and serum could be useful tool for screening of patients with chronic periodontitis. Further, monitoring the various phases of treatment outcome using saliva could be a useful, non-invasive, prognostic indicator.

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.

Comparative evaluation of the antimicrobial susceptibility and cytotoxicity of husk extract of Cocos nucifera and chlorhexidine as irrigating solutions against Enterococcus Faecalis, Prevotella Intermedia and Porphyromonas Gingivalis - An in-vitro study.

AIM AND BACKGROUND: The aim of the present study is to evaluate and compare the antimicrobial susceptibility and cytotoxicity of Cocos nucifera and chlorhexidine (CHX) as irrigating solutions against Enterococcus faecalis, Prevotella intermedia, and Porphyromonas gingivalis. MATERIALS AND METHODS: The ethanolic extract of husk of C. nucifera was prepared. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the extract were determined using the serial broth dilution method and its cytotoxicity was evaluated against human periodontal fibroblasts using 3-(4,5-dimethyl-thiazole-2-yl)-2,5-diphenyl tetrazolium bromide assay. Antibacterial susceptibility for two irrigating solutions, namely 2% CHX gluconate irrigant (Group I) and 1.5% C. nucifera husk irrigant (Group II), was tested against P. gingivalis, P. intermedia, and E. faecalis. RESULTS: The MIC and MBC of C. nucifera husk extract for P. gingivalis were 468.75 µg/ml and 1562.5 µg/ml, for P. intermedia were 48.8 µg/ml and 1875 µg/ml, and for E. faecalis were 1562.5 µg/ml and 3750 µg/ml, respectively. The extract was nontoxic to the human periodontal fibroblast. Both the materials have shown similar antibacterial susceptibility and no difference was observed at baseline, 10, 30, and 60 min using two-way repeated measures of ANOVA. However, a statistically significant difference was observed between different time points for P. gingivalis and P. intermedia using Bonferroni multiple comparison test (f = 826.1390, P ≤ 0.05). CONCLUSION: 1.5% of ethanolic husk extract of C. nucifera has a significant antibacterial action against polymicrobial dental biofilm and its activity is comparable to that of 2% CHX which validates its use as a future irrigating solution for overcoming bacterial resistance with synthetic agents.