Browsing by Subject "shilajit"

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    Determining The Effects of Fulvic acid on Biofilm/Planktonic Streptococcus Mutans Growth
    (Office of the Vice Chancellor for Research, 2014-04-11) Botros, Mark; Gregory, Richard L.
    Fulvic acid, a major organic compound extract of Shilajit has been the focus of dental research for the past few years. Shilajit, a sticky tar-like substance of dark brownish color, was used during the ancient times, thousands of years ago and continues to be the traditional method today in India to aid with curing bone/cartilage diseases. Shilajit has also been proven to have anti-inflammatory and pain suppressing effects. This experiment determined the minimum inhibitory concentration (MIC), which is the lowest concentration of fulvic acid, an active component of shilajit that inhibits the visible growth of S. mutans. This experiment also determined the minimum bactericidal concentration (MBC) which is the lowest concentration of fulvic acid that kills S. mutans. A 3-day procedure to determine the growth vs inhibition of the S. mutans was conducted and bacterial readings were recorded using a spectrophotometer after treating S. mutans with 10% formaldehyde, crystal violet stain, and iso-propanol with 30-45 minute incubations between each. The experiment determined that very high concentrations of fulvic acid killed S. mutans, while less concentrated fulvic acid inhibited the growth of S. mutans bacterial cells. A solution comprised of a 5% concentration of fulvic acid killed all of the S. mutans; 5.00%, 2.50%, and 1.25% fulvic acid concentrations had bacterial absorbance of 0.000, 0.009, and 0.027, respectively, as compared to the control group’s normal bacterial growth absorbance of 0.254. Additionally, solutions ranging from a two-fold dilution of fulvic acid to six-fold dilution of fulvic acid inhibited the growth of S. mutans. A similar trend was also observed in planktonic and biofilm formation. For all of the above, in the seventh and eighth dilution (0.078% and 0.039% respectively) of the fulvic acid, the growth of S. mutans bacteria was similar to the control group due to the level of dilution. Overall it was observed that fulvic acid is able to kill bacteria in strong concentrations. Additionally it is able to inhibit further growth of bacteria in lower concentrations, but once the solution becomes too dilute, it does not have an effect on bacterial growth. This contributes greatly to the field of oral health because this data can be utilized for further research on oral bacterial growth inhibitors. Furthermore, the data collected here is a significant starting point for research on the specific minimum concentrations necessary to inhibit oral bacteria growth, because this can be used to determine the smallest amounts of fulvic acid, the bacteria the human body can handle.
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    Determining the Minimum Inhibitory Concentration of Fulvic Acid on Early Streptococcus mutans Biofilm Formation
    (Office of the Vice Chancellor for Research, 2014-04-11) Tahir, Ali; Witcher, Phillip; Gregory, Richard L.
    Fulvic acid is an active ingredient in many homeopathic treatments. Shilajit is one such treatment that is mainly used in the Indian Subcontinent area. It is believed that fulvic acid has some remedial properties that can help with inhibition of many types of bacteria and various malignant diseases. In this experiment the effects of fulvic acid were analyzed on early Streptococcus mutans biofilm formation. S. mutans is an oral bacterium that contributes to the formation of dental caries forming bacterial biofilm on teeth. For the experiment, early S. mutans biofilm formation was treated with different concentrations of fulvic acid for 24 hours in sterile 96-well flat-bottom microtiter plates. The optical density (OD) of the S. mutans biofilm was then measured using a SpectraMax190, after staining with crystal violet. Data was analyzed on the knowledge that a greater OD is correlated to a greater bacterial biofilm. Results demonstrated that fulvic acid inhibited the growth of early S. mutans biofilm formation at fulvic acid concentrations greater than 5% (vol. %). After preliminary studies, different concentrations of fulvic acid closer to the estimated minimum inhibitory concentration (MIC) were applied to the S. mutans to find a more precise MIC of the fulvic acid on the biofilm growth. Upon completion of these various studies, fulvic acid was shown to inhibit early S. mutans biofilm formation and may show signs of oral health improvement if applied for human use.
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    Evaluating the Effect of Fulvic Acid on Oral Bacteria and Cancerous Oral Cells
    (Office of the Vice Chancellor for Research, 2014-04-11) Witcher, Phillip; Gregory, Richard L.; Windsor, L. Jack
    Shilajit is a homeopathic treatment used by local inhabitants of India and Pakistan. It may have specific components that inhibit the formation of cavities and the growth of cancer cells. This experiment analyzed the effects of fulvic acid, an active component of shilajit, on the growth of oral bacteria and squamous cell carcinoma. The effect of fulvic acid was evaluated on early Streptococcus mutans (S. mutans) biofilm formation and established S. mutans biofilm by treating each group with different concentrations of fulvic acid for 24 hours in sterile 96-well flat-bottom microtiter plates. S. mutans was used because it is a common cause of dental caries. The optical density (OD) of the S. mutans biofilm was measured after crystal violet staining using a SpectraMax190; greater growth correlated to greater OD. It was determined that fulvic acid inhibits the growth of newly forming S. mutans biofilm at fulvic acid concentrations greater than 1.25% (vol. %) and established S. mutans biofilm at fulvic acid concentrations greater than 5% (vol. %). To evaluate the effect of fulvic acid on squamous cell carcinoma (SCC-25) cells, six-well plates seeded with SCC-25 cells (1*105 cells/well) were exposed to different concentrations of fulvic acid (buffered to a pH of 7.5) for 72 hours. The cytotoxicity and cell proliferation were measured using a cytotoxicity detection kit and a water soluble tetrazolium kit (Roche Applied Science), respectively. It was determined that fulvic acid inhibits the growth of SCC-25 cells at concentrations of fulvic acid above 2% (volume %). The effects of fulvic acid (0.5%) on matrix metalloproteinase expression and collagen degradation ability of SCC-25 cells is being analyzed. The suppressive mechanisms observed by fulvic acid on both S. mutans and SCC-25 cells could improve overall oral health.
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    Study of Fulvic Acid: A Natural Dietary Supplement
    (Office of the Vice Chancellor for Research, 2014-04-11) Syed, Ghayasul I.; Gregory, Richard L.; Windsor, L. Jack; Song, Fengyu
    Shilajit is a substance found in parts of Asia. Although there have been no clinical studies, it is used by the locals and is marketed because it is thought to have antiseptic, anti-inflammatory and pain suppressing effects. Fulvic acid (F-A) is a major constituent of shilajit and was used in the analysis of the anti-pathogenic tendencies of shilajit and cytotoxic effects on human cells of the oral cavity. The bacterial study was performed on Streptococcus mutans, a normal flora of the oral cavity. The idea was to test the metabolic activity of the bacteria in F-A-containing media. Menadione-XTT reagent was used for this. The bacterial biofilm was allowed to grow in TSBS in a microtiter plate of 96 wells. The F-A solution of different concentration were introduced into each well in a gradually decreasing amount and the last control wells had a zero concentration. The XTT reagent was introduced and after incubation the biofilm of S. mutans reduced the XTT to an orange color, the change in color was detected by measuring the absorbance at 490nm. Between 2.5% to 5.0% of F-A the wells showed signs of decreased activity. The numbers indicated that absorbance of the wells with concentrated F-A was lower compared to the wells with more diluted F-A solutions. From this it can be concluded that F-A had a negative effect on the growth and metabolic activity of S. mutans. For human testing, pulp and fibroblast cells were subjected to different concentrations of F-A. The cytotoxicity was measured by the amount of Lactate Dehydrogenase released from the treated cells (sign of damage). Overall, the experiment validates the potency of F.A as an effective antibacterial. Further testing is needed but the compound shows promise and can be employed as an effective ingredient of mouthwash and other such antiseptic products.