Hair Growth Promoting Potential of Consciousness Energy Healing Treatment in Human Dermal Papilla Cells

Table of contents

1. Introduction

he hair follicle consists of mainly two types of components such as epithelial components that included the matrix and outer root sheath, and the dermal components, which included the dermal papilla and the connective tissue sheath [1] . There were three distinct stages of hair growth, which occurs during cellular proliferation like an active phase (anagen), an intermediate regressive (catagen), and a resting phase (telogen) [2,3] . Literature explored that generally loss of 50-100 hairs per day considered as hair fall disorders like alopecia. Modern trends of fast globalization and indiscriminate grow of industrialization led to the sacral therapy. Human Biofield Energy has subtle energy that can work effectively [12] . CAM therapies have been practised worldwide with reported clinical benefits in different health disease profiles [13] . This energy can be harnessed and transmitted by the experts into living and non-living things via the process of Biofield Energy II.

2. Materials and Methods

3. a) Chemicals and Reagents

Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were obtained from Gibco, India. Minoxidil sulphate (positive control) was purchased from Clearsynth Labs Ltd., Mumbai. Antibiotics solution (penicillin-streptomycin) was

4. b) BrdU Incorporation Cell Proliferation Assay in HFDPCs

The human follicular dermal papilla cells (HFDPCs) in DMEM supplemented with 10% FBS were counted using a hemocytometer and a single cell suspension was prepared. The single cell suspension was seeded at a density of 800 cells/well in a fresh DMEM supplemented with 10% FBS in 96-well plates. hours of incubation, the medium was replaced with a fresh DMEM supplemented with 0.1% FBS. Further, after 24 hours, cells were treated with the test items and positive control (minoxidil sulphate). After incubation for 48 hours, the effect of the test items on cell proliferation was assessed by bromodeoxyuridine (BrdU) incorporation using colorimetric ELISA kit. For that, 10 µL of BrdU solution was added per well and the cells were incubated for 90 minutes at 37ºC. After incubation, the medium was removed from each well by gentle pipetting. About 200 µL of a FixDenat solution was added to each well. After incubation, cells were incubated for 30 minutes at room temperature (RT) (15-25ºC). The FixDenat solution was removed by gentle pipetting. After incubation, 100 µL of anti-BrdU-POD (peroxidase) solution was added to each well. Then, the cells were incubated for 90 minutes at RT (15-25ºC). After incubation, the anti-BrdU-POD solution was removed by gentle pipetting. Each well was washed 3 times using 200 µL of washing solution. About 100 µL of substrate solution was added to each well. Cells were incubated for 30 minutes at RT (15-25ºC). After incubation, the absorbance of each well was measured at 370 nm.

Cellular proliferation was determined as following Equation ( 1):

% ???????????????? ?????????????????????????? = {(?? ? ??)/??} ?? 100(1)

Where, A = OD of Untreated DMEM wells B = OD of cells treated with the test item / positive control.

5. c) Experimental Design

The experimental groups composed of group 1 (G-I) with DMEM medium defined as the untreated DMEM group. Group 2 (G-II) contained positive control (minoxidil sulphate) at various concentrations. Further, group 3 (G-III) included the Biofield Energy Treated DMEM group.

6. d) Biofield Energy Healing Strategy

The test item, DMEM was divided into two parts. First part did not receive any sort of treatment and defined as the untreated DMEM group. The second part was treated with the Biofield Energy Treatment by a Alice Branton remotely for ~5 minutes under laboratory conditions and coded as the Biofield Energy Treated DMEM group. Healer in this study never visited the laboratory in person, nor had any contact with the test items (DMEM medium). Further, the untreated DMEM group was treated with a "sham" healer for comparative purposes. The "sham" healer did not have any knowledge about the Biofield Energy Treatment. After that, the Biofield Energy Treated and untreated test items were kept in similar sealed conditions for experimental study.

7. e) Statistical Analysis

All the values were represented as Mean ± SEM (standard error of mean) of three independent experiments. The statistical analysis was performed using SigmaPlot statistical software (v11.0). For two multiple group comparison, one-way analysis of variance (ANOVA) was used followed by post-hoc analysis by Dunnett's test. Statistically significant values were set at the level of p?0.05.

Healing. The Trivedi Effect ® -Consciousness Energy Healing Treatment has been reported with a significant revolution in the physicochemical properties of metals, chemicals, ceramics, and polymers [14][15][16] , improved agricultural crop yield, productivity, and quality [17,18] , transformed antimicrobial characteristics [19][20][21] , biotechnology [22,23] , improved bioavailability [24][25][26] , skin health [27,28] , nutraceuticals [29,30] , cancer research [31,32] , bone health [33][34][35] , human health and wellness. Considering the promising benefits of an alternative natural therapies-based literature information and importance of Biofield Energy Healing Treatment on various fields, the authors sought to evaluate the impact of the Biofield Energy Treatment (The Trivedi Effect ® ) on the test item (DMEM) for hair cells growth activity using standard assay in human follicular dermal papilla cells. procured from HiMedia, India. Other chemicals used in this study were analytical grade obtained from India.

Then, the cells were incubated in a CO 2 incubator for 24 hours at 37°C, 5%CO 2 , and 95% humidity. After 24 renowned Biofield Energy Healer (The Trivedi Effect ® ), groups comparison student's t-test was used. For III.

8. Results and Discussion

9. a) BrdU Incorporated Cell Proliferation of Dermal Papilla Cells

The effect of the test item son Biofield Energy Treatment and the percent of cellular proliferation of DPCs is presented in Figure 1. The immortalized human follicular dermal papilla cells suspension were treated with the positive control and test item (DMEM). Topical application of minoxidil is a well-established therapeutic for various types of hair growth-related disorders like alopecia [36] . The untreated DMEM group exhibited 100% cells proliferation of DPCs. Additionally, the positive control, minoxidil showed 68.57%, 187.14% (p?0.001), and 230.95% (p?0.001) increase the cellular proliferation of DPCs at 0.001, 0.01, and 0.1 µM, respectively in a concentration-dependent manner compared to the untreated DMEM group. Moreover, the percent proliferation of DPCs was significantly (p?0.001) increased by 219.30% in the Biofield Energy Treated DMEM group with respect to the untreated DMEM (Figure 1). For the study of hair follicle biology, human hair growth in vitro model is used as a prototype [37] . Hair follicles undergo different cycles of growth (anagen), regression (catagen), quiescence (telogen), and regeneration [38] . The current experimental results demonstrated that Biofield Energy Treatment significantly increased the proliferation of dermal papilla cells (DPCs). Further, DPCs are responsible for the regulation of hair follicles development and periodic regeneration [5,36,39,40] . Also, the DPCs can generate a signal that regulates the behaviour of keratinocytes in the follicle during the hair cycle [41,42] . Apart from this, Wnt/?-catenin signaling pathway plays a critical role to initiate generation of hair follicle via stimulation of keratinocytes [43,44] . Based on the literature and findings of this study, it is assumed that the increment of DPCs The representative photo images signified the intensity of proliferative DPCs after treatment with the Biofield Energy Treated test item (DMEM) in HFDPCs (Figure 2). Overall, data suggested that the Biofield Energy Treatment significantly improved the growth and proliferation of human dermal papilla cells, which is due to the Biofield Energy Healing (The Trivedi Effect ® ). Based on that it is concluded that the Consciousness Energy Therapy could be beneficial to maintain a steady-state proliferation of hair follicles. The study findings was observed that the Biofield Energy Treated test item (DMEM) group showed a significant (p?0.001)increase the percent of dermal papilla cells (DPCs) by 219.30% in human follicular dermal papilla cells (HFDPCs) in vitro. In conclusion, The Trivedi Effect ® -Consciousness Energy Healing Treatment might act as a hair growth promoter, and it can be used as a complementary and alternative treatment for the prevention of various types of skin and hair-related disorders viz. necrotizing fasciitis, actinic keratosis, sebaceous cysts, diaper rash, decubitus ulcer, androgenetic alopecia, telogen effluvium, trichodystrophy, alopecia areata, etc. Besides, it could be useful to improve cell-to-cell communication, normal cell growth, cell differentiation, neurotransmission, cell cycling and proliferation, hormonal balance, skin health, immune and cardiovascular functions. Moreover, it can also be utilized in organ transplants (i.e., kidney transplants, liver transplants and heart transplants), hormonal imbalance, aging, and various immune-related disease conditions such as Ulcerative Colitis (UC), Alzheimer's Disease (AD), Dermatitis, Irritable Bowel Syndrome (IBS), Asthma, Hashimoto Thyroiditis, Pernicious Anemia, Sjogren Syndrome, Multiple Sclerosis, Aplastic Anemia, Hepatitis, Diverticulitis, Graves' Disease, Dermatomyositis, Diabetes, Myasthenia Gravis, Parkinson's Disease, Atherosclerosis, Systemic Lupus Erythematosus (SLE), stress, etc. with a safe therapeutic index to improve overall health and Quality of Life.

Figure 1. Fig. 1 :
1Fig. 1: Effect of the test samples on hair growth regarding dermal papilla cells (DPCs) proliferation after 48 hours of treatment in immortalized human follicular dermal papilla cell line (HFDPC). All the values are represented as mean
Figure 2. ±
SEM of three independent experiments. ***p?0.001 vs. untreated DMEM group.

Appendix A

Appendix A.1 Acknowledgements

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

Appendix B

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Date: 2018 2018-01-15