Published: Feb 15, 2019 Volume: 2 Issue: 4 Page Number: 1-12
DOI: 10.14302/issn.2329-9487.jhc-19-2582 ISSN: 2329-9487
Authors: Mahendra Kumar Trivedi, Snehasis Jana
Citation: Mahendra Kumar Trivedi, Snehasis Jana (2019) In vitro Assessment of the Biofield Treated Test Item on Cardiac Function Using Rat Cardiomyocytes Cell Line (H9c2) via Multiparametric Analysis. Journal Of Hypertension And Cardiology - 2(4):1-12.
Introduction: Heart disorders are the major concern of population health worldwide. According to WHO estimates 2018, 17.9 million peoples were died due to cardiovascular disorders.
Aim: The aim of this study was to investigate the cardioprotective activity of Biofield Energy Treated test item, Dulbecco's Modified Eagle Medium (DMEM) using rat cardiomyocytes (H9c2).
Methods: The test item (DMEM) was divided into three parts, first part received one-time Biofield Energy Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and was labeled as the one-time Biofield Energy Treated (BT-I) DMEM, while second part received the two-times Biofield Energy Treatment and is denoted as BT-II DMEM. The third part did not receive any treatment and defined as the untreated DMEM group.
Results: Cell viability of the test samples by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay showed 89.03% and 98.49% in the BT-I and BT-II groups, respectively suggested a nontoxic and safe in nature of the tested test item. The BT-I group showed 16.01% restoration of cell viability. The level of lactate dehydrogenase (LDH) was significantly inhibited by 50.37% and 49.35% in the BT-I and BT-II groups, respectively compared to the untreated DMEM group. Moreover, percent protection of creatine kinase-myocardial band (CK-MB) by 49.48% and 59.79% in the BT-I and BT-II groups, respectively, compared to the untreated DMEM group. Reactive oxygen species (ROS) level in terms of mean fluorescence unit (FU) was reduced by 6.64% in the BT-I group than untreated DMEM. Besides, BT-I and BT-II groups significantly increased the level of % apoptotic cells by 63.16% and 97.37% (p≤0.05), respectively than untreated DMEM.
Conclusion: Allover, results envisaged that Biofield Treatment significantly improved different cardiac parameters. Thus, Biofield Energy Treatment (The Trivedi Effect®) could be utilized as a cardio-protectant against several cardiac disorders such as coronary artery disease, heart attack, arrhythmias, heart failure, congenital heart disease, cardiomyopathy, etc.
The Trivedi Effect®, Biofield Energy Treatment, H9c2, Cardiac health, CK-MB, Apoptosis, LDH
Heart disorders are the major concern of population health worldwide. About 6 lakh peoples die due to heart disease in the United States every year; that’s 1/4 deaths . Cardiovascular disease (CVD) and stroke produce an immense health and economic burdens in the United States and globally. According to WHO estimates, in 2016, 17.9 million people around the globe die of cardiovascular diseases each year. This represents about 1/3 of all deaths globally [2, 3]. CVD is the leading cause of death in Europe, accounting for over 4 million deaths each year. It has been projected that by 2020, CVD would be more numerous in India and China than in all economically developed countries in the world . Three main criteria to keep a healthy heart like opening blood vessels, strengthening the heart muscle, and controlling free radical damage by antioxidants . Apart from animal models and primary cardiac myocytes derived animal, even recent work has been done to develop human cardiomyocyte model systems for the screening of cardioprotective activity of substances . The use of in vitro test model for the prediction of heart damages provides several advantages over in vivo assessment. As this model require few animals, test material, and give high accuracy data . Rat cardiomyocytes cell line (H9c2) have been widely used as an alternative model to human cardiomyocytes in vitro for the assessment of cardio-protectant properties of any test substances .
Various study data suggested the effect of Energy Therapy in cancer patients through therapeutic touch , massage therapy , etc. Complementary and Alternative Medicine (CAM) therapies are preferred model of treatment, among which Biofield Therapy (or Healing Modalities) is one approach to enhance emotional, mental, physical, and human wellness. The National Center of Complementary and Integrative Health (NCCIH) has recognized and allowed Biofield Energy Healing as a CAM approach in addition to other therapies and medicines such as natural products, chiropractic/osteopathic manipulation, Qi Gong, deep breathing, Tai Chi, yoga, meditation, massage, special diets, healing touch, relaxation techniques, traditional Chinese herbs and medicines, naturopathy, movement therapy, homeopathy, progressive relaxation, guided imagery, pilates, acupuncture, acupressure, Reiki, rolfing structural integration, hypnotherapy, Ayurvedic medicine, mindfulness, essential oils, aromatherapy, and cranial sacral therapy. Human Biofield Energy has subtle energy that has the capacity to work in an effective manner . CAM therapies have been practiced worldwide with reported clinical benefits in different health disease profiles . This energy can be harnessed and transmitted by the experts into living and non-living things via the process of Biofield Energy Healing. Biofield Energy Treatment has been reported with a significant revolution in the field of cancer research [13,14], materials science [15-17], microbiology [18-20], agriculture [21,22], nutraceuticals [23, 24], and biotechnology [25,26]. Besides, The Trivedi Effect® also significantly improved bioavailability of various low bioavailable compounds [27-29], an improved overall skin health [30, 31], bone health [32-34], human health and wellness. Based on the excellent contribution of Biofield Energy in wide spectrum of areas, authors intend to extend the treatment modality to study the impact of the Biofield Energy Healing Treatment (The Trivedi Effect®) on the test item (DMEM) for cardiomyocytes cell line (H9c2).
Materials and Methods
Chemicals and Reagents
N-acetyl cysteine (NAC), 2′,7′-Dichlorofluorescin diacetate (DCFDA), 3-(4,5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide (MTT), and ethylenediaminetetraacetic acid (EDTA) were obtained from Sigma Chemical Co. (St. Louis, MO). Antibiotics solution (penicillin-streptomycin) was purchased from HiMedia, India. Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco, India. Creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) kits were obtained from Biovision, USA. Annexin-V kit was purchase from Guava Technologies, USA. The positive control, trimetazidine (TMZ) was procured from Zliesher Nobel, USA. All the other chemicals used in this experiment were analytical grade procured from India.
Biofield Energy Healing Strategy
The test item (DMEM) was used in this experiment and one portion was considered as the untreated group, where no Biofield Treatment was provided. Further, the untreated group was treated with “sham” healer for comparison purpose. The sham healer did not have any knowledge about the Biofield Energy Healing Treatment. The other portion of the test item was received one-time Biofield Energy Treatment and referred as the BT-I and was also given two-times Biofield Energy Treatment and defined as the BT-II. Both the test items (BT-I and BT-II) were received Biofield Energy Healing Treatment (known as The Trivedi Effect®) under laboratory conditions for ~3 minutes through Mahendra Kumar Trivedi’s unique Biofield Energy Transmission process. Biofield Energy Healer was located in the USA, however the test items were located in the research laboratory of Dabur Research Foundation, New Delhi, India. Biofield Energy Healer in this experiment did not visit the laboratory, nor had any contact with the test samples. After that, the Biofield Energy Treated and untreated test items were kept in similar sealed conditions and used for the study as per the study plan.
Assessment of Cell Viability Using MTT Assay
The cell viability was performed by MTT assay in H9c2 cells (ATCC® CRL-1446™). The cells were counted and plated in a 96-well plate at the density corresponding to 10 X 103 cells/well/180 µL in DMEM + 10% FBS. The cells in the above plate(s) were incubated for 24 hours in a CO2incubator at 37°C, 5% CO2, and 95% humidity. Following incubation, the medium was removed and the following treatments were given. In the test item group, 200 µL of the test item was added to wells. Besides, in the positive control group, added 180 µL of SFM with 20 µL of positive controls were added from the respective 10X stock solutions. After incubation for 48 hours, the effect of test item on cell viability was assessed by MTT assay. 20 µL of 5 mg/mL of MTT was added to all the wells and incubated at 37°C for 3 hours. The supernatant was aspirated and 150 µL of DMSO was added to all wells to dissolve formazan crystals. The optical density (OD) of each well was read at 540 nm using Biotek Reader.
Effect of the test items on viability of H9c2 cells was determined using Equation (1):