"Efficacy of cytotoxic effect of green tea catechins on the human periodontal fibroblasts and human dental pulp fibroblasts -An in vitro study".

Background: Inflammation of tooth-supporting tissue and the pulp tissue is followed by wound healing and regeneration process that involves the specific type of connective tissue cells, the fibroblasts. During periodontitis and pulpitis, the inflammation of the tissue causes damage to the fibroblasts. These fibroblasts secrete collagen proteins and maintain the structural framework; along with this the inflammatory process moves toward healing where in the specific cells such as the fibroblast cells play important roles. Green tea catechins epigallocatechin-3-gallate (EGCG) being one of the major catechins is known to have multiple beneficial effects on human fibroblasts. Objective: To assess the in vitro cytotoxicity of green tea catechins on the human periodontal ligament (PDL) fibroblasts and human dental pulp fibroblasts. Materials and Methods: Human PDL fibroblasts (hPDLFs) and human dental pulp fibroblasts were isolated from the two extracted premolar teeth that were indicated for orthodontic treatment. The fibroblasts were then seeded in 96 well tissue culture plate for cell viability study. EGCG was used at different concentration to treat the cells. After 48 h; (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) (MTT) assay was performed to determine the cell viability. Results: The vitality of hPDLFs and human dental pulp fibroblasts was found to be inversely proportional to EGCG concentrations. Conclusions: hPDLFs have shown 37% proliferation at lowest concentration of EGCG used and human dental pulp fibroblasts have shown 99% viability at lowest concentration of EGCG used.

Assessment of the effects of hydrogen water on human gingival fibroblast cell culture in patients with chronic periodontitis.

Background: Activated inflammatory cells produce reactive oxygen species (ROS) to eliminate pathogens. Under normal conditions, the pathogens are taken care of, and tissues are repaired. However, in periodontal disease, persistent inflammation causes increased ROS release and impaired healing. Therefore, removal of overproduced ROS using antioxidants is necessary. Hydrogen water has an antioxidative effect on cells and impedes oxidative stress-related disorders. Aim: To study the effect of hydrogen water on cell viability, migration, and its antioxidative potential in fibroblasts obtained from chronic periodontitis patients. Materials and Methods: The gingival tissue samples were obtained from 26 subjects (13 periodontally healthy individuals and 13 chronic periodontitis patients) and processed. The human gingival fibroblasts were cultured and the assays were commenced once adequate growth was detected. The effect of hydrogen water on cell viability was checked by neutral red assay, while the migration potential was assessed by transwell migration assay. The antioxidative potential of hydrogen water was evaluated by CUPRAC assay. Statistical Analysis: Intergroup comparison was done using Mann-Whitney U-test. Intragroup comparison was done using Wilcoxon signed-rank test. Results: Hydrogen water was nontoxic to the fibroblasts at 24 h and 48 h. The intergroup comparison of the cell viability between hydrogen water-treated periodontally healthy gingival fibroblasts (HF) and fibroblasts from patients with chronic periodontitis (CF) showed a statistically significant (P = 0.00) difference at 24 h and 48 h. Hydrogen water also positively influenced the migratory capacity. Hydrogen water-treated fibroblasts obtained from chronic periodontitis patients showed more migration in comparison to the healthy group (P = 0.00). Hydrogen water showed an antioxidative potential. The maximum potential was seen in relation to the fibroblasts obtained from chronic periodontitis patients at 48 h. Conclusion: Hydrogen water was nontoxic, increased the migratory capacity, and showed an antioxidative potential on human fibroblasts obtained from periodontally healthy individuals and patients with chronic periodontitis.

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.

Response of stem cells from human exfoliated deciduous teeth (SHED) to three bioinductive materials - An in vitro experimental study.

INTRODUCTION: Stem cells have unmatched capacity and potential for regeneration and when used alone or in combination with scaffolds to replace or repair damaged cells, can differentiate into any mature cell. AIM: To evaluate the functional differentiation potential of EMD (Enamel Matrix Derivative), MTA (Mineral Trioxide Aggregate) and Biodentine on Stem Cells from Human Exfoliated Deciduous teeth (SHED). OBJECTIVE: To determine functional differentiation potential (osteogenic/odontogenic) of various biomaterials on SHED. MATERIAL AND METHOD: SHED derived from 5th linear passage after sub-culturing were treated with EMD, MTA and Biodentine individually and their effect on cell viability was compared and evaluated by MTT (3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide) assay for 7 days. Alizarin red S staining was used to assess mineralization potential of these materials by the staining calcium deposits for 14 days. The results were analyzed using One-way ANOVA, Post hoc Tukey's test for multiple comparisons. RESULTS: It was observed that EMD imparted the highest cell viability at the end of 7 days (p < 0.001) followed by Biodentine and MTA. Likewise EMD showed highest potential to enhanced mineralization and expression of dentine sialoprotein (p < 0.001) followed by Biodentine and MTA at the end of 14 days (p<0.001). CONCLUSION: It can be concluded that all the tested materials are bioinductive to SHED. EMD can be used for various vital pulp therapies as that of Biodentine and MTA with predictable as well as enhanced success rate.

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.