Antibiotics solution (penicillin-streptomycin) was procured from HiMedia, India. DMEM and FBS were procured from GIBCO, USA. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium) (MTT), quercetin, trypsin, and ethylenediaminetetraacetic acid (EDTA) were purchased from Sigma Chemical Co. St. Louis, MO, USA. H₂O₂ and DMSO were obtained from Fisher Scientific, India. All the other chemicals used in this experiment were analytical grade procured from India.

Human hepatoma cell line (HepG2) was procured from National Centre for Cell Science (NCCS), Pune, India, used as a test system in the present study. HepG2 cell line was maintained under DMEM medium supplemented with 10% fetal bovine serum (FBS) for routine culture. Growth conditions were maintained at 37°C, 5% CO₂, and 95% humidity and sub-cultured by tapping the flask and splitting the cell suspension into fresh flasks and supplementing with fresh cell growth medium.

The DMEM growth medium and HepG2 cells were subjected to the Biofield Energy Treatment (The Trivedi Effect^®) by Mahendra Kumar Trivedi under standard laboratory conditions for ~3 minutes from a distance of ~25 cm. The energy transmission was done without touching the medium and HepG2 cells. Following the Biofield Energy Treatment, the DMEM and HepG2 cells were used for this experiment.

The experiment was performed with four different groups. Group (G) 1 contained untreated HepG2 cells and untreated DMEM supplemented with 1% FBS and served as a baseline control. G2 served as negative control includes untreated HepG2 cells, untreated DMEM supplemented with 1% FBS, and 1 mM H₂O₂. G3 assigned as positive control consisted of untreated HepG2 cells, untreated DMEM supplemented with 1% FBS, H₂O₂ (1 mM), and quercetin at two different concentrations viz. 10 µM and 50 µM. G4 defined as the Biofield Energy Treated DMEM along with the untreated HepG2 cells supplemented with 1% FBS and 1 mM H₂O₂. Moreover, G5 referred as the Biofield Energy Treated HepG2 cells in addition to the untreated DMEM supplemented with 1% FBS and H₂O₂ (1 mM).

Human hepatoma cell line (HepG2) was used in this experiment for the assessment of cell proliferation. The DMEM and HepG2 cells were trypsinized, counted, and plated in wells of flat bottom 96-well plates at the density corresponding to 10 X 103 cells/well/180 µL of growth medium. Following respective treatments, the cells in the 96-well plates were incubated for 16 hours in a CO₂ incubator at 37°C, 5% CO₂, and 95% humidity. After 16 hours of incubation, the plates were taken out and 20 µL of 5 mg/mL of MTT 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide solution was added to all the wells followed by additional incubation for 3 hours at 37°C. The supernatant was aspirated and 150 µL of DMSO was added to each well to dissolve formazan crystals. The absorbance of each well was then read at 540 nm using Synergy HT microplate reader. Concentrations were determined and the experiment was done in triplicates. The percentage cell viability was calculated using formula (1):

% Cell viability = (X/R)*100................................(1)

Where,X = Absorbance of cells corresponding to positive control and test group

R = Absorbance of cells corresponding to baseline control

The data were expressed as the mean ± standard deviation (SD) of three independent experiments. The analysis was performed with SigmaPlot Statistical Software (Version 11.0). Differences between means were assessed for the statistical differences using Student’s t-test (between two groups) and for multiple comparison one-way analysis of variance (ANOVA) and post-hoc analysis was done by Dunnett’s test. p≤0.05 was considered as statistically significant.

The effect of the Biofield Energy Treated DMEM on cell viability (as absorbance) after challenged with hydrogen peroxide in HepG2 cells is represented in Table 1. The cell viability was evaluated with the help of MTT assay. The effect of the Biofield Energy Treatment on the percent cell viability in HepG2 cells and DMEM after challenged with H₂O₂ is shown in Figure 1. The percent cell viability in the baseline control group was defined as 100%. After challenged with H₂O₂ the percent cell viability was significantly (p≤0.001) decreased by 81.3% in the negative control group due to generation of oxidative stress compared to the baseline control group. The reference item, quercetin showed a significant (p≤0.001) increment of cell viability by 14.2% and 52.7% at the concentration of 10 and 50 µM, respectively compared to the negative control group. Zhang et al. 2011, demonstrated that the antioxidant properties of quercetin, a plant-derived aglycone has been used as a nutritional supplement and may be beneficial against a variety of diseases, including cancer 32. Besides, the percent cell viability was significantly (p≤0.001) increased by 13.6% in the Biofield Energy Treated DMEM group; while decreased by 3.2% in the Biofield Energy Treated HepG2 cells group compared to the negative control group.

Overall, the Consciousness Energy Treated DMEM possess better protection against oxidative stress, which was induced by H₂O₂. On the other hand, the Biofield Energy Treated HepG2 cells altered the protection against oxidative stress as compared to the negative control group. Hence, the study results demonstrated that The Trivedi Effect^® - Consciousness Energy Healing Treatment on DMEM showed a significant antioxidant activity rather than Biofield Energy Treated HepG2 cells. Bioenergy Therapy is considered as an important supplement to conventional cancer treatment. A lot of evidence exists on the benefits of Bioenergy Therapies for stress management and boosting the immune system. From literature, it has been stated that the Biofield Energy Treatment was free from side-effects and it helps cancer patients both physically and emotionally 33. Numerous literatures reported the beneficial effects of Bioenergy Therapies on stress management, reducing fatigue and anxiety, and increasing the quality of life and immune system among cancer patients 343536. In this experiment, authors found a significant impact of Consciousness Energy Healing Treatment (The Trivedi Effect^®) against oxidative stress. It is assumed that The Trivedi Effect^® - Consciousness Energy Healing could be use as potential antioxidant activity to protect various vital organs disorders.

DMEM: Dulbecco's Modified Eagle's Medium

FBS: Fetal bovine serum

ROS: Reactive oxygen species

MTT:3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

SOD: Super oxide dismutase

RNS: Reactive nitrogen species

BHA: Butylated hydroxyanisole

BHT: Butylated hydroxytoluene

Authors gratefully acknowledged to Trivedi Global, Inc., Trivedi Science, Trivedi testimonials and Trivedi master wellness for their support. In addition to, authors are thankful for the support of Dabur Research Foundation for conducting this study.