In vitro Evaluation of Immunomodulatory Effects of the Test Formulation by the Estimation of Natural Killer Cells and Phagocytosis Activities

Although the immunomodulatory effects of vitamins, minerals, and many herbs per se have been extensively studied, while research related to possible immunomodulatory effects of a combined formulation is relatively scarce. Here, the potential immunomodulatory effects of the combination of eight components viz. zinc chloride, ferrous sulfate, sodium selenate, nanocurcumin, copper chloride, magnesium gluconate, vitamin C, and vitamin D3 were investigated. These components are widely used in therapeutic, cosmetics and dietary supplements. The current study investigated the in vitro immunomodulatory effect of a Consciousness Energy Healing (The Trivedi Effect®) Treated nanocurcumin-based test formulation in lipopolysaccharide (LPS)-induced natural killer (NK) cells activity in splenocytes cell-line co-incubated with mouse lymphoma cell line (Yac-1). The formulation was divided into two parts, in which one part of the formulation was received the Biofield Treatment by a renowned Biofield Energy Healer, Mr. Mahendra Kumar Trivedi and was defined as the Biofield Energy Treated sample, while the other part was denoted as the untreated test sample. Cell viability using MTT assay in mouse splenocytes showed more than 98% cells were viable upto 10.4 µg/mL of the tested concentrations, indicating that the test formulation was safe and nontoxic. The NK cells activity was measured in cell supernatants using ELISA. The NK cells activity was significantly altered by 26.73%, 11.82%, 7.64%, 0.36% and 36.36% in the untreated formulation group at 0.1, 0.5, 1, 5.2, and 10.4 µg/mL, respectively compared to the vehicle control (VC) group. Further, Biofield Energy Treated test formulation exhibited 39.64%, 8.18%, 17.27%, 51.93% and 42.55% alteration of the NK cells activity at the concentrations of 0.1, 0.5, 1, 5.2, and 10.4 µg/mL, respectively compared to the VC group. Moreover, the cell viability of the test formulation was assessed in RAW 264.7 (Mouse Macrophage Cell Line) for phagocytosis activity was safe upto 13.3 µg/mL with >90% on cell viability. The phagocytosis assay data showed that at two concentrations (1.3 and 7.6 µg/mL) the Biofield Energy Treated test formulation improved extent of phagocytosis by 6.31% and 5.91%, respectively compared to the untreated test formulation group. In summary, the Biofield Energy Treated test formulation may have indeed immunomodulatory effect by enhanced NK cells activity and extent of phagocytosis responses. These observations indicated that the Biofield Energy Treated test formulation has the potential effects through modulating the expression of NK cells function and phagocytosis and might be developed as a useful anti-inflammatory product for various inflammatory disorders.

The cell viability of the nanocurcumin-based test formulation was found as safe at the tested concentrations upto 10.4 µg/mL in mouse splenocytes. Besides, the untreated test formulation altered NK cells activity by 26.73%, 11.82%, 7.64%, 0.36% and 36.36% at 0.1, 0.5, 1, 5.2, and 10.4 µg/mL, respectively compared to the vehicle control (VC) group. However, the NK cells activity was significantly altered by 39.64%, 8.18%, 17.27%, 51.93% and 42.55% at 0.1, 0.5, 1, 5.2, and 10.4 µg/mL, respectively compared to the VC group. Further, the cell viability of the test formulation was assessed in RAW 264.7 (Mouse Macrophage Cell Line) for phagocytosis activity was safe upto 13.3 µg/mL with >90% on cell viability. The phagocytosis assay data showed that at two concentrations (1.3 and 6.7 µg/mL) the Biofield Energy Treated test formulation improved extent of phagocytosis by 6.31% and 5.91%, respectively compared to the untreated test formulation group. The study results summarized that the nanocurcumin-based formulation showed significant modulation of NK cells expression and extent of phagocytosis responses in mouse splenocyte cells co-incubate with mouse lymphoma cell line (Yac-1) and mouse macrophage cell line (RAW264.7), respectively. Overall, data indicated a significant improvement of natural killer cells activity and phagocytosis upon exposure with the nanocurcumin-based test product on tested cell-lines. Thus, The Trivedi Effect® - Consciousness Energy Healing Treatment has the significant antioxidant and immunomodulatory activities in vitro. It is then anticipatated that the Biofield Energy Treated herbomineral formulation could be a more useful as an immunomodulatory formulation for healthy human and in patients in the near future. The immunomodulatory activity of the Biofield Treated novel formulation could be expanded to other therapeutic arena such as organs (kidney, liver, and heart) transplant, autoimmunedisorders (Graves’ Disease, Addison Disease, Multiple Sclerosis, Myasthenia Gravis, Rheumatoid Arthritis, Systemic Lupus Erythematosus, etc.), inflammatory disorders (Crohn’s Disease, Irritable Bowel Syndrome, Ulcerative Colitis, Vasculitis, etc.), aging, and strees.