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.

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.

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.

An in vitro evaluation of cytotoxicity of curcumin against human periodontal ligament fibroblasts.

INTRODUCTION: Curcumin, a component of turmeric (Curcuma longa L.), is a molecule of multitude of medicinal properties. Although curcumin has found a place in the treatment of gingival and periodontal diseases, there are no reported cytotoxicity studies on the cells of clinical significance (i.e., periodontal ligament [PDL] fibroblasts). AIMS: The objective of this research was to assess the in vitro cytotoxicity of curcumin against human PDL fibroblasts. MATERIALS AND METHODS: Human PDL fibroblasts from premolar teeth were cultured and used for cytotoxicity tests from healthy children presented for orthodontic extractions. Test concentrations of curcumin (100%, 50%, and 25%) were prepared by diluting 95% curcumin with di­methyl­sulfoxide and added to 96­well microtiter plate (in triplicate) containing the fibroblast culture (approximately 2 × 104 cells/well). Fibroblast cells without treatment (without curcumin) acted as a control group. The viability of cells after 48 h of incubation at 37°C in a humidified atmosphere of 5% CO2 and 95% air was ascertained by the 3­(4, 5­dimethyl­thiazol­2­yl)­2, 5­diphenyl­tetrazolium bromide (MTT) assay. The viability of PDL fibroblast cells of experimental wells was expressed relative to that of control, in terms of change in the color intensity. Absorbencies were recorded at 450 nm on a microplate reader with background subtraction at 620 nm. The cell viability at various concentrations of curcumin against the PDL fibroblasts was calculated as mean absorbance (optical density) and percentage values. RESULTS: Cell viability of PDL fibroblasts to 100%, 50%, and 25% curcumin concentration was 111.75%, 112.50%, and 114.40%, respectively. CONCLUSIONS: No in vitro cytotoxicity was detected for curcumin against human PDL fibroblasts, at any of the concentrations used (100%, 50%, and 25%) by MTT assay at the end of 48 h.

Evaluation of Physical Parameters of Novel Licorice Varnish Versus Fluoride and Combination Varnish: An In-Vitro Study.

OBJECTIVES: The aim of this study was to evaluate the physical properties of locally prepared Licorice varnish (LV), commercially available Fluoride varnish (FV) and a Combination of both Varnishes (CV). MATERIAL AND METHODS: LV was prepared using authenticated licorice roots. Commercially available FV (Bifluorid 12) was used as a positive control and CV was prepared in six different concentrations of both varnishes. Conventional antibacterial activity assessment, employing disc diffusion and broth dilution methods, was inconclusive. Therefore a novel assessment method was used, whereby the varnish was directly added to a mixture of Brain Heart Infusion broth with Streptococcus mutans and incubated. Physical parameters such as pH, rate of evaporation, viscosity, film forming ability, and cost incurred for preparation were assessed and compared. RESULTS: FV, LV and CV (except the combination of LV 80% + FV 20%) showed antibacterial activity against Streptococcus mutans. All three varnishes formed films on the tooth surface as confirmed by Scanning Electron Microscopy. Mean pH was in the range of 4-4.5, viscosity 48-52 centipoise (cP), rate of evaporation was 150-160 seconds. They were comparable to each other in the physical parameters tested, except for the shelf life of LV. CONCLUSION: All three varnishes showed antibacterial activity against Streptococcus mutans which was established using an innovative method of antibacterial activity assessment. LV was most economical of all but had a shorter shelf life. The results of this study need to be evaluated through an in vivo study.

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.

Design and Synthesis of Some Novel Fluorobenzimidazoles Substituted with Structural Motifs Present in Physiologically Active Natural Products for Antitubercular Activity.

Keeping in view the drawbacks associated with research on anti-TB drugs based on plant extracts and the non-availability of fluorinated natural products with antitubercular activity has prompted us to make an effort towards the synthesis and characterization of a novel series of fifteen substituted fluorobenzimidazoles. The newly synthesized compounds were characterized by I.R, 1H-NMR, 13C-NMR, Mass, and elemental analysis. The synthesized compounds 4(a-f) and 5(b-j) have been evaluated for their in-vitro antimycobacterial activity against H37Rv strain (ATCC 27294) by MABA method. Incorporation of methylenedioxyphenyl moiety at 2- and 6-position of the benzimidazole ring furnished compounds 4d and 5i with antitubercular activity comparable or more potent than the naturally occurring compounds with reported antitubercular activity. Among the fifteen tested compounds, 4d and 5i emerged as promising hits characterized by MIC lower than that determined for sesamin against the pathogenic H37Rv strain. Antitubercular activity results indicate that these compounds may be suitable for further lead optimization. The cytotoxic effect of these active compounds on THP-1 cell line was assessed by MTT assay and the results suggest that these two molecules are potential candidates for further development as antitubercular agents.

Comparative evaluation of the efficacy of a herbal mouthwash and chlorhexidine mouthwash on select periodontal pathogens: An in vitro and ex vivo study.

BACKGROUND: Several herbal mouthwash and herbal extracts have been tested in vitro and in vivo in search of a suitable adjunct to mechanical therapy for long-term use. In this study, we aimed to look at the antimicrobial effect of the herbal mouthwash and chlorhexidine (CHX) mouthwash on select organisms in in vitro test and an ex vivo model. MATERIALS AND METHODS: The antimicrobial effects were determined against standard strains of bacteria that are involved in different stages of periodontal diseases. The in vitro tests included determination of minimum inhibitory concentration (MIC) using broth dilution and agar diffusion. In the ex vivo part of the study supragingival dental plaque were obtained from 20 periodontally healthy adult volunteers. Descriptive analysis was done for the entire quantitative and qualitative variable recorded. RESULTS: The MIC by broth dilution method found no statistically significant difference between the mouthwashes. The agar dilution method showed CHX was more effective as compared to the herbal mouthwash against standard strains of Streptococcus mutans, Streptococcus sanguinis, and Aggregatibacter actinomycetemcomitans. However, no difference was observed between the mouthwashes for Porphyromonas, Pseudomonas aeruginosa, and Fusobacterium nucleatum. The ex vivo results conclude that none of the selected mouthwashes were statistically significantly different from each other. CONCLUSION: In the present study, CHX showed higher levels of antimicrobial action than the herbal mouthwash against bacterial species. The results reinforce the earlier findings that the in vitro testing is sensitive to methods and due diligence is needed when extrapolating the data for further use. However, long-term use and in vivo effectiveness against the periopathogens need to be tested in well-planned clinical trials.

In vitro antibacterial activity of ethanolic extract of Morus alba leaf against periodontal pathogens.

CONTEXT: Antibiotic resistance is a major problem with inadvertent usage. Thus, there is a need to search for new antimicrobial agents of herbal origin to combat antibiotic resistance. One such plant is Morus alba which has a long history of medicinal use in traditional Chinese medicine. AIM: To compare the antibacterial activity of ethanolic extract of M. alba leaves with chlorhexidine gluconate against Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia. SETTINGS AND DESIGN: Experimental in vitro study. METHODOLOGY: Crude extract from the leaves of M. alba were prepared by Soxhlet extraction method by using ethanol as a solvent. Minimum inhibitory concentration (MIC) of the extract was assessed against A. actinomycetemcomitans, P. gingivalis and T. forsythia, and compared with that of chlorhexidine gluconate by broth dilution method. RESULTS: P. gingivalis was the most sensitive organism against the M. alba extract with an MIC value of 1.95 mg/ml; while T. forsythia and P. gingivalis both were most sensitive organisms against chlorhexidine gluconate with MIC values of 0.00781 mg/ml. CONCLUSION: M. alba possess good antibacterial activity against A. actinomycetemcomitans, P. gingivalis and T. forsythia and thus would be beneficial for the prevention and treatment of periodontal disease. However, chlorhexidine gluconate was found to be more effective when compared to M. alba.

Effect of Ocimum sanctum on Oral Cancer Cell Line: An in vitro Study.

BACKGROUND: Cancer till today remains the leading cause of death in both developed and developing countries. Plants have been beacon of therapeutic sources for curing diseases from times immemorial. Hence, the present study aimed at evaluating the antiproliferative activity of extract of Ocimum sanctum leaves on oral cancer cell line. OBJECTIVES: To evaluate the antiproliferative effect and to analyze dose dependent cytotoxic activity of aqueous extract of O. sanctum leaves on KB mouth cell line. To compare the effectiveness among different variety of O. sanctum. MATERIALS AND METHODS: KB cells (Mouth Epidermal Carcinoma Cells) were used for the present study. Aqueous and dry extract of O. sanctum with both dark (Krishna Tulsi) and light (Rama Tulsi) leaves were prepared in the institution. The antiproliferative and cytotoxic activity on KB cell line was evaluated by MTT assay. Statistical analysis with Mann-Whitney U-test and Wilcoxon matched pairs test was carried out. RESULTS: The aqueous extract of O. sanctum of both the leaves exhibited significant cytotoxic effect against oral cancer cell line. CONCLUSION: Aqueous extract of O. sanctum leaves was effective as an antiproliferative agent which caused apoptosis in oral cancer cell line. CLINICAL SIGNIFICANCE: Ocimum sanctum herb which is abundantly grown in India can be used for its anticancer properties for treating oral cancer. This will not only be cost-effective but will also have less or no side effects.

Effect of Azadirachta indica (Neem) and Aloe vera as compared to subantimicrobial dose doxycycline on matrix metalloproteinases (MMP)-2 and MMP-9: An in-vitro study.

BACKGROUND: A critical outcome of periodontal diseases is degradation of collagen in the periodontal tissues, by enzymes such as Matrix Metallo-Proteinases (MMPs). Doxycycline is known to down-regulate the activity of MMPs. Azadirachta indica (Neem) and Aloe vera are herbs known to have an anti-inflammatory effect. The present study was designed to evaluate the anti-inflammatory effect of Neem and Aloe vera by way of its inhibitory effect on MMP-2 and MMP-9 activity in cases of chronic periodontitis and compare it with doxcycline. MATERIALS AND METHODS: A total of 30 subjects were enrolled in this study. Gingival tissue samples were obtained from patients diagnosed with the chronic periodontitis. The tissue extracts were treated with the said drug solutions and inhibition of MMP-2 and MMP-9 was analyzed. Enzymatic activity was detected by electrophoresis. The data was subjected to Student's paired t-test. RESULTS: The results showed that the activity of MMP-2 and MMP-9 was significantly decreased by the use of doxycycline, Neem and Aloe vera. A 53.5% reduction in the MMP-2 and 52.5% reduction in the MMP-9 activity was seen when samples were subjected to Neem treatment at the concentration of 1500 µg/ml. Tissues treated with Aloe vera in the concentration of 2000 µg/ml showed a 20.09% reduction in the MMP-2 and 20.4% reduction in the MMP-9 activity. Doxycycline in the concentration of 300 µg/ml, showed an 82.1% reduction in the MMP-2 and 82.6% reduction in the MMP-9 activity. CONCLUSION: The present study demonstrated an inhibitory effect of Neem and Aloe vera on MMP-2 and MMP-9, which are involved in the extracellular matrix degradation during periodontitis.