Biofield Energy Healing Based Vitamin D3: An Improved Overall Bone Health Activity in MG-63 Cell Line- Juniper publishers
Journal of Trends in Technical and Scientific Research
Abstract
The potential of The Trivedi Effect®-Consciousness
Energy Healing vitamin D3 and DMEM medium was investigated in human
bone osteosarcoma cells (MG-63). The Test Items (TI) i.e. vitamin D3
and DMEM were separated into two parts. Bone health parameters were
tested such as Alkaline Phosphatase enzyme (ALP) activity, collagen
levels and bone mineralization. The Test Items (TI) i.e. vitamin D3 and
DMEM medium were divided into two parts. The test samples received
Consciousness Energy Healing Treatment by Karen Byrnes Allen and samples
were defined as the Biofield Energy Treated (BT) samples, while the
other parts of each sample were denoted as the Untreated test items
(UT). Cell viability using MTT assay exhibited increased cell viability
more than 71% with safe and nontoxic profile among test samples on MG-63
cell line. The level of ALP was significantly increased by116.4% at
50μmL in the UT-DMEM+BT-TI group, while 35.6%, 81.2%, and 47.9% at 10,
50 and 100 μmL, respectively in the BT-DMEM+UT-TI group. In addition,
ALP level was increased by 42.3% and 90.6% at 10 and 50 μ mL,
respectively in the BT-DMEM+BT-TI groups as compared with the untreated
group. The level of collagen was significantly increased by 10.5% and
110.1% at 0.1 and 1 μmL, respectively in the UT-DMEM+BT-TI group, while
22.6% and 190.6% at 0.1 and 1μmL, respectively in the BT-DMEM+UT-TI
group as compared with the untreated group. In addition, BT-DMEM+BT-TI
group showed a significant increased collagen level by 140.6% and 39.9%
at 0.1 and 1 μmL, respectively as compared with the untreated test item
and DMEM group. The percent of bone mineralization was significantly
increased by 128.7% at 50 μmL, in the UT-DMEM+BT-TI group, while 34.1%,
154.7%, and 18.7% at 1, 10, and 50 μmL, respectively in the
BT-DMEM+UT-TI group as compared with the untreated group. In addition,
BT-DMEM+BT-TI group showed a significant increased bone mineralization
by 105.7%, 78.1%, and 39.6% at 1, 10, and 50 μmL, respectively as
compared with the untreated group. Thus, the study results concluded
that The Trivedi Effect® Treatment would be the best alternative
treatment for the maintenance of strong and healthy bones and quality of
life. Further, it regulates the osteoblast function and improved the
level of collagen, ALP, and calcium absorption in wide range of bone
disorders along with wide range of adverse bone health conditions
Keywords: The Trivedi Effect®; Bone Health; Biofield Healing; Osteosarcoma Cells; Vitamin D; Bone mineralization
Abbreviations: CAM:
Complementary and Alternative Medicine, NCCAM: National Center for
Complementary and Alternative Medicine; MG- 63: Human Bone Osteosarcoma
Cells, ALP: Alkaline phosphatase, DMEM: Dulbecco's Modified Eagle's
Medium; FBS: Fetal bovine serum; EDTA: Ethylene Diamine Tetra Acetic
Acid, UT: Untreated, BT: Biofield Energy Treated, TI: Test Item.
Introduction
Vitamin D has multiple effects which regulate the
functions in different organs such as brain, lungs, liver, kidneys, and
heart, immune, skeletal, and reproductive systems. Moreover, it has
sig-nificant anti-inflammatory, anti-arthritic, anti-osteoporosis,
anti-stress, anti-aging, anti-apoptotic, wound healing, anti-cancer,
anti-psychotic, and anti-fibrotic roles. Vitamin D receptors (VDRs) are
widely present in most of the body organs like brain, heart, lungs,
kidney, liver, pancreas, large and small intestines, muscles,
reproductive, nervous system, etc. [1].
VDRs influence cell-to- cell communication, normal cell growth, cell
differentiation, cell cycling and proliferation, hormonal balance,
neurotransmission, skin health, immune and cardiovascular functions.
Bone-related health issues become a major problem among the population
from village to the cities. Vitamin D plays a vital role in preserving a
healthy mineralized skeleton of most of the vertebrates including
humans. Cod liver oil, irradiation of other foods including plants,
sunlight, etc. are found to be effective against bone related disorders,
which lead to discovering the active principle- vitamin D [1].
The role of vitamin D has been well defined not only for improving the
bone mineralization but also with increased bone resorption, aging,
inflammation and overall quality of life. Vitamin D3 is
synthesized in the skin by sunlight and once formed it sequentially
metabolized in the liver and kidney to 1,25-dihydroxyvitamin D
(calcitriol, the vitamin D hormone) [2].
Calcitriol play an important role in maintaining the normal level of
calcium and phospho-rus, promotes bone mineralization, induce or repress
the genes responsible for conserving the mineral homeostasis and
skeletal integrity, and inhibit hypertension, kidney damage,
cardiovascular and immune disorders (such as Lupus, Addison Disease,
Graves' Disease, Hashimoto Thyroiditis, Multiple Sclerosis, Myasthenia
Gravis, Anemia, Sjogren Syndrome, Systemic Lupus Erythemato-sus,
Diabetes, Alopecia Areata, Fibromyalgia, Vitiligo, Psoriasis,
Scleroderma, Chronic Fatigue Syndrome and Vasculitis), and the secondary
hyperparathyroidism [3].
Vitamin D insufficiency and deficiency is the major health problem,
which causes metabolic bone disease in the young and elderly populations
[4].
Fortified foods have a variable amount of vitamin D and most of the
foods do not contain vitamin D, which can be fulfilled using some
supplements. In order to avoid the bone related disorders such as
osteomalacia, exacerbate osteoporosis, hyperparathyroidism, immune
disorders, etc. calcium 1000-1500mg/day along with vitamin D supplement
around 400 IU/day is very important for maintaining the good bone health
[5].
Various in vitro studies have readily
established the role of bone health using cell lines and its resorbing
effects using three important key biomarkers, such as alkaline
phosphatase (ALP), collagen and calcium. MG-63 cell line derived from
juxta cortical osteosarcoma, which represents an immature osteoblast
phenotype and undergoes temporal development in long term culture. The
response of MG-63 cells to 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) administration has been studied to be similar to normal human osteoblast cells [6]. Hence, MG-63 cell line is widely used for studying the potential of any test compounds to improve the bone health [7].
The formation of new bone involves a complex series of events including
the proliferation and differentiation of osteoblasts, and eventually
the formation of a mineralized extracellular matrix. ALP is a phenotypic
marker for the early differentiation and maturation of osteoblasts. ALP
increases the local concentration of inorganic phosphate for bone
mineralization and hence is an important marker for osteogenic activity [8].
Similarly, active osteoblasts synthesize and extrude collagen, which
plays an important role in the formation of bone extracellular matrix by
providing strength and flexibility. Collagen fibrils formed an arrays
of an organic matrix known as Osteoid [9].
Likewise, calcium phosphate is deposited in the Osteoid and gets
mineralized (combination of calcium phosphate and hydroxyapatite) and
provides rigidity to the bone [10]. Thus, these parameters are very essential in order to study the bone health in cell lines. Authors evaluated the in vitro effect of the Biofield Energy Treated vitamin D3 as a test item, a Complementary and Alternative Medicine (CAM) on bone health using MG-63 cell line for major biomarkers.
Within the burgeoning ground of CAM therapies,
Biofield En-ergy Treatment or energy medicine, is emerging with
significant benefits in various scientific fields. The effects of the
CAM ther-apies have great potential, which include external qigong,
Johrei, Reiki, therapeutic touch, yoga, Qi Gong, polarity therapy, Tai
Chi, pranic healing, deep breathing, chiropractic/osteopathic
manipu-lation, guided imagery, meditation, massage, homeopathy,
hypno-therapy, progressive relaxation, acupressure, acupuncture, special
diets, relaxation techniques, Rolfing structural integration, healing
touch, movement therapy, pilates, mindfulness, Ayurvedic medi-cine,
traditional Chinese herbs and medicines in biological systems both in vitro and in vivo[11].
Biofield Energy Healing Treatment (The Trivedi Effect®) contain a
putative bioenergy, which is channeled by a renowned practitioners from a
distance. Biofield Energy Healing as a CAM showed a significant results
in biological studies [12].
However, the National Center for Complementary and Alternative Medicine
(NCCAM), well-defined Biofield therapies in the subcategory of Energy
Therapies [13].
The Trivedi Effect®- Consciousness Energy Healing Treatment has been
reported with significant revolution in the physicochemical properties
of metals, chemicals, ceramics and polymers [14-16], improved agricultural crop yield, productivity, and quality [17,18], transformed antimicrobial characteristics [19,20], bone health [21,22], biotechnology [23], improved bioavailability [24-26], skin health [27,28], nutra- ceuticals [29,30], cancer research [31,32], and human health and wellness.
Based on the significant outcomes of Biofield Energy Treatment and vital role of vitamin D3 on bone health, authors sought to evaluate the impact of the Biofield Energy Treatment (The Trivedi Effect®) on vitamin D3
as test sample for bone health activity with respect to the assessment
of different bone health parameters like ALP, collagen content, and bone
mineralization using standard in vitro assays in MG-63 cells.
Material and Methods
Chemicals and reagents
Rutin hydrate was purchased from TCI, Japan, while vitamin D3
(denoted as test item) and L-ascorbic acid were obtained from
Sigma-Aldrich, USA. Fetal Bovine Serum (FBS) and Dulbec- co's Modified
Eagle's Medium (DMEM) were purchased from Life Technology, USA.
Antibiotics solution (penicillin-streptomycin) was procured from
HiMedia, India, while 3-(4, 5-diamethyl-2-thi- azolyl)-2,
5-diphenyl-2H-tetrazolium) (MTT), Direct Red 80, and ethylene diamine
tetra acetic acid (EDTA) were purchased from Sigma, USA. All the other
chemicals used in this experiment were analytical grade procured from
India.
Cell culture
Human bone osteosarcoma cell line -MG-63 was used as
test system in the present study. The MG-63 cell line was maintained in
DMEM growth medium for routine culture supplemented with 10% FBS. Growth
conditions were maintained as 37 °C, 5% CO2 and 95% humidity
and sub cultured by trypsinisation followed by splitting the cell
suspension into fresh flasks and supplementing with fresh cell growth
medium. Three days before the start of the experiment, the growth medium
of near-confluent cells was replaced with fresh phenol-free DMEM,
supplemented with 10% charcoal dextran stripped FBS (CD-FBS) and 1%
penicillin-strep-tomycin [33].
Experimental design
The experimental groups consisted of cells in
baseline control, vehicle control groups (0.05% DMSO with Biofield
Energy Treated and untreated DMEM), positive control group (rutin
hydrate) and experimental test groups. The experimental groups included
the combination of the Biofield Energy Treated and untreated vitamin D3/DMEM.
It consisted of four major treatment groups on specified cells with
Untreated-DMEM + Untreated-Test item (UT-TI), UT-DMEM + Biofield Energy
Treated test item (BT-TI), BT-DMEM + UT-TI, and BT-DMEM + BT-TI.
Consciousness energy healing treatment strategies
The test item and DMEM were divided into two parts.
One part each of the test item and DMEM was treated with the Biofield
Energy by a renowned Biofield Energy Healer (also known as The Trivedi
Effect®) and coded as the Biofield Energy Treated item, while the second
part did not receive any sort of treatment. This Biofield Energy
Healing Treatment was provided by Karen Byrnes Allen remotely for ~5
minutes. Biofield Energy Healer was remotely located in the USA, while
the test samples were located in the research laboratory of Dabur
Research Foundation, New Delhi, India. This Biofield Energy Treatment
was administered for 5 minutes through the Healer's unique Energy
Transmission process remotely to the test samples under laboratory
conditions. Karen Byrnes Allen in this study never visited the
laboratory in person, nor had any contact with the test item and medium.
Further, the control 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 samples were kept in similar sealed conditions for
experimental study.
Determination of non-cytotoxic concentration
The cell viability was performed by MTT assay in
human bone osteosarcoma cell line (MG-63). The cells were counted and
plated in 96 well plates at the density corresponding to 5 X 103 to 10 X 103
cells/well/180 μL of cell growth medium. The above cells were incubated
overnight under growth conditions and allowed the cell recovery and
exponential growth, which were subjected to serum stripping or
starvation. The cells were treated with the test item, DMEM, and
positive control. The untreated cells were served as baseline control.
The cells in the above plate(s) were incubated for a time point ranging
from 24 to 72 hours in CO2 incubator at 37 °C, 5% CO2,
and 95% humidity. Following incubation, the plates were taken out and
20μL of 5mg/mL of MTT solution were added to all the wells followed by
additional incubation for 3 hours at 37 °C. The supernatant was
aspirated and 150μL of DMSO was added to each well to dissolve formazan
crystals. The absorbance of each well was read at 540 nm using Synergy
HT micro plate reader, BioTek, USA [34]. The percentage cytotoxicity at each tested concentrations of the test substance were calculated using the
following equation (1):
% Cytotoxicity = (1 - X / R) *100 (1)
Where, X = Absorbance of treated cells; R = Absorbance of untreated cells
The percentage cell viability corresponding to each treatment was obtained using the following equation (2):
% Cell Viability = 100 - % Cytotoxicity (2)
The concentrations exhibiting >70% Cell viability was considered as non-cytotoxic.
Assessment of alkaline phosphatase (ALP) activity
The cells were counted using an hemocytometer and plated in a 24-well plate at the density corresponding 1 x 104
cells/well in phenol free DMEM supplemented with 10% CD-FBS. Following
respective treatments, the cells in the above plate were incubated for
48 hours in CO2 incubator at 37 °C, 5% CO2, and
95% humidity. After 48hours of incubation, the plate was taken out and
processed for the measurement of ALP enzyme activity. The cells were
washed with 1X PBS and lysed by freeze thaw method i.e,
incubation at -80 °C for 20 minutes followed by incubation at 37 °C for
10 minutes. To the lysed cells, 50μL of substrate solution i.e, 5mM of p-nitrophenyl phosphate (pNPP) in 1M diethanolamine and 0.24 mM magnesium chloride (MgCl2)
solution (pH 10.4) was added to all the wells followed by incubation
for 1 hour at 37 °C. The absorbance of the above solution was read at
405 nm using Synergy HT micro plate reader (Biotek, USA). The absorbance
val-ues obtained were normalized with substrate blank (pNPP solution
alone) absorbance values [33].
The percentage increase in ALP enzyme activity with respect to the
untreated cells (baseline group) was calculated using equation (3):
% Increase =[(X -R) /R)]*100 (3)
Where, X = Absorbance of cells corresponding to positive control and test groups
R = Absorbance of cells corresponding to baseline group (untreated cells).
Assessment of collagen synthesis
The MG-63 cells were counted using a hemocytometer and plated in 24-well plate at the density corresponding to 10 x 103
cells/well in phenol free DMEM supplemented with 10% CD-FBS. Following
respective treatments, the cells in the above plate were incubated for
48 hours in CO2 incubator at 37°C, 5% CO2, and 95%
humidity. After 48 hours of incubation, the plate was taken out and the
amount of collagen accumulated in MG-63 cells corresponding to each
treatment was measured by Direct Sirius red dye binding assay. In brief,
the cell layers were washed with PBS and fixed in Bouin's solution (5%
acetic acid, 9% formaldehyde and 0.9% picric acid) for 1 hours at room
temperature (RT). After 1 hour of incubation, the above wells were
washed with milliQ water and air dried. The cells were then stained with
Sirius red dye solution for 1 hour at RT followed by washing in 0.01 N
HCl to remove un-bound dye. The collagen dye complex obtained in the
above step was dissolved in 0.1 N NaOH and absorbance was read at 540 nm
using Biotek Synergy HT micro plate reader. The level of collagen was
extrapolated using standard curve obtained from purified Calf Collagen
Bornstein and Traub Type I (Sigma Type III) [33].
The percentage increase in collagen level with respect to the untreated
cells (baseline group) was calculated using equation (4):
% Increase = [(X - R) / R] *100 (4)
Where, X = Collagen levels in cells corresponding to positive control and test groups
R = Collagen levels in cells corresponding to baseline group (untreated cells).
Assessment of bone mineralization by alizarin red S staining
The MG-63 cells were counted using an hemocytometer and plated in 24-well plate at the density corresponding to 10 x103cells/well
in phenol free DMEM supplemented with 10% CD- FBS. Following respective
treatments, the cells in the above plate were incubated for 48 hours in
CO2 incubator at 37°C, 5% CO2, and 95% humidity
to allow cell recovery and exponential growth. Following overnight
incubation, the above cells will be subjected to serum stripping for 24
hours. The cells will be then be treated with non-cytotoxic
concentrations of the test samples and positive control. After 3-7days
of incubation with the test samples and positive control, the plates
were taken out cell layers and processed further for staining with
Alizarin Red S dye. The cells were fixed in 70% ethanol for 1 hour,
after which Alizarin Red solution (40 μm; pH 4.2) was added to the
samples for 20 minutes with shaking. The cells were washed with
distilled water to remove unbound dye. For quantitative analysis by
absorbance evaluation, nodules were solubilized with 10% cetylpyridinium
chloride for 15 minutes with shaking. Absorbance was measured at 562 nm
using Biotek Synergy HT micro plate reader [33].
The percentage increase in bone mineralization with respect to the
untreated cells (baseline group) was calculated using the following
equation (5):
% Increase = [(X - R) / R] *100 (5)
Where, X = Absorbance in cells corresponding to
positive control or test groups; R = Absorbance in cells corresponding
to baseline (untreated) group.
Statistical analysis
All the values were represented as percentage of
respective parameters. For multiple group comparison, one-way analysis
of variance (ANOVA) was used followed by post-hoc analysis by Dun- netts
test. Statistically significant values were set at the level of p≤0.05.
Results and Discussion
Cell viability using MTT assay
Cell viability data showed that cell viability was significant im-proved percentage among the Biofield Energy Treated vitamin D3 and DMEM in MG-63 cells are shown in Figure 1.
The data showed that the test samples found as nontoxic and safe (as
evidence of cell viability approximately more than 71%) across all the
tested concentrations up to 100μg/mL. Hence, the same concentrations
were used for the evaluation of other bone health parameters such as
alkaline phosphatase (ALP) activity, collagen synthesis, and bone
mineralization in MG-63 cells.
Alkaline phosphatase (ALP) enzyme activity
The Trivedi Effect®-Energy of Consciousness Healing based test items displayed an increased ALP level in various groups (Figure 2)
as compared with the untreated group. The percentage change in ALP data
at various concentrations in different groups were presented in Figure 2
in terms of percentage values. The positive control, rutin showed a
significant increased value by 43.44%, 53.55%, and 83.33% at 0.01, 0.1,
and 1 μg/mL, respectively with respect to the untreated cells. The
experimental test group's untreated medium and Biofield Treated Test
item (UT-DMEM+BT- TI) showed a significant increased level of ALP by
116.4% and 2.3% at 50 and 100 μg/mL, respectively while Biofield Treated
medium and untreated Test item (BT-DMEM+UT-TI) showed a significant
increased ALP level by 35.6%, 81.2%, and 47.9% at 10, 50 and 100 μg/mL,
respectively as compared with the untreated test item and DMEM group.
However, the Biofield Energy Treated medium and Biofield Energy Treated
Test item (BT-DMEM+BT-TI) showed a significant increased ALP level by
42.3% and 90.6% at 10 and 50 μg/mL, respectively as compared with the
untreated test item and DMEM group. ALP (zinc metalloprotein enzymes) is
considered as one of the important bone biomarker proteins for
osteoblast differentiation. Reduced level of bone ALP leads to various
disorders such as obstructive liver disease, osteoblastic activity,
metabolic bone disease, reduced bone growth, acromegaly, osteogenic
sarcoma, or bone metastases, healing fracture, myelo-fibrosis, leukemia,
and rarely myeloma [35-37].
Thus, the overall data suggested a significant improved ALP level after
Biofield Energy Treatment. Biofield Energy Treated test samples could
be useful against patients suffering from bone disorders to improve the
skeletal structure and overall bone-related disorders.
Estimation of collagen synthesis
The Trivedi Effect®-Biofield Energy Treated test
samples showed a significant enhanced level of collagen synthesis as
compared with the untreated test samples. All the results are presented
in percentage collagen values in Figure 3.
The rutin hydrate (positive control group) showed a significant
increased value of collagen by 46.59%, 51.97%, and 65.41% at 0.01, 0.1,
and 1 μg/ mL, respectively. In addition, the experimental test groups
such as UT-DMEM+BT-TI showed a significant increased collagen level by
10.5% and 110.1% at 0.1 and 1 μg/mL, respectively while BT- DMEM+UT-TI
group showed a significant increased collagen level by 22.6%, 190.6%,
and 1.6% at 0.1, 1, and 10 μg/mL, respectively as compared with the
untreated test item and DMEM group. On the other hand, BT-DMEM+BT-TI
group showed a significant increased collagen level by 140.6% and 39.9%
at 0.1 and 1 μg/mL, respectively as compared with the untreated test
item and DMEM group. Collagen synthesis leads to build stronger bone and
muscles, which provide strength to skeletal structure. Collagen fibers
provide strong mechanical force and strength to life. The most abundant
matrix protein, collagen type I play a significant role in overall bone
health [38,39]. Overall, the Consciousness Energy treated vitamin D3
had significantly improved the synthesis of collagen fibers in the
human osteosarcoma cells with respect to all the treatment groups.
Hence, it is assumed that The Trivedi Effect® has the significant
potential to improve the bone health in various skeletal disorders
against weaken joints, tendons, and ligaments.
Bone mineralization
The effect of Biofield Energy Treatment (Trivedi
Effect®-Bio- field Energy Healing) on bone mineralization in MG-63 cells
is shown in Figure 4,
which showed a significant increased bone mineralization percentage in
the test samples. Supplementation with calcium and vitamin D3
increased the chance of degree of bone mineralization. As, bioactive
vitamin D or calcitriol is a steroid hormone, which has an important
long term role in regulating the body levels of calcium and phosphorus,
and in bone mineralization process. However, lack of vitamin D and
calcium is directly linked to an inability of any person to fight
against infections effectively, depression, muscle weakness, multiple
sclerosis, fatigue and the development of diabetes, bone cancers, heart
disease, high blood pressure, and stroke [40,41]. The results in term of percentage bone mineralization was presented in Figure 4.
The positive control, rutin group showed a significant increased value
of bone mineralization by 47.98%, 59.73%, and 139.02% at 5, 10, and 25
μg/mL, respectively. The experimental data among test group's
UT-DMEM+BT-TI showed a significant increased bone mineralization by
128.7% at 50 μg/mL, while BT-DMEM+UT- TI group showed a significantly
increased bone mineralization by 34.1%, 154.7%, and 18.7% at 1, 10, and
50 μg/mL, respectively as compared with the untreated test item and DMEM
group. However, BT-DMEM+BT-TI group showed a significant increased bone
mineralization by 105.7%, 78.1%, and 39.6% at 1, 10, and 50 μg/ mL,
respectively as compared with the untreated test item and DMEM group.
Thus, The Trivedi Effect®-Biofield Energy Treated vit D3
could be significantly useful to maintain a healthy skeletal structure
for the patients suffering from various bone-related disorders.
Conclusion
The experimental results of cell viability data using
MTT assay showed more than 71% cells were viable and Consciousness
Energy Healing based vitamin D3 further improved the cell
viability. Thus, MTT data indicated that the test samples were safe and
nontoxic in all the tested concentrations. However, various bone health
parameters were significantly improved such as ALP was increased by
116.4% at 50 μg/mL in the UT-DMEM+BT-TI, while 35.6%, 81.2%, and 47.9%
at 10, 50 and 100 μg/mL, respectively in the BT-DMEM+UT-TI group as
compared with the untreated test item and DMEM group. In addition,
BT-DMEM+BT-TI group showed an increased ALP level by 42.3% and 90.6% at
10 and 50 μg/mL, respectively The level of collagen was significantly
increased by 10.5% and 110.1% at 0.1 and 1 μg/mL, respectively in the
UT-DMEM+BT-TI, while 22.6%, 190.6%, and 1.6% at 0.1, 1, and 10 μg/mL,
respectively in the BT-DMEM+UT-TI group. The level of collagen was
increased by 140.6% and 39.9% at 0.1 and 1μg/mL, respectively in
BT-DMEM+BT-TI group as compared with the untreated test item and DMEM
group. Similarly, the bone mineralization percent was significantly
increased by 128.7% at 550 μg/mL in the UT-DMEM+BT-TI group, while
34.1%, 154.7%, and 18.7% at 1, 10, and 50 μg/mL, respectively in the
BT-DMEM+UT-TI group as compared with the untreated group. In addition,
BT-DMEM+BT- TI group showed a significant increased bone mineralization
by 105.7%, 78.1%, and 39.6% at 1, 10, and 50 μg/mL, respectively as
compared with the untreated group. The Bone health parameters were
significantly improved among the Biofield Energy Treated vitamin D3
test samples in MG-63 cells. Overall, the Biofield Energy Treated (The
Trivedi Effect®) test samples were found to have a significant impact on
tested bone health parameters collagen, bone mineralization, and ALP,
which are very vital to combat the bone disorders. Therefore, the
Consciousness Energy Healing based vitamin D3 might be a
suitable alternative nutritional supplement, which could be useful for
the management of various bone related disorders osteoporosis, Paget's
disease of bone, rickets, deformed bones, osteomalacia, bone and/or
joint pain, increased frequency of fractures, osteoma, hormonal
imbalance, stress, aging, bone loss and fractures, and other bone
diseases that are caused by poor nutrition, genetics, or problems with
the rate of bone growth or rebuilding. Biofield Energy Treated Vitamin D3can
be useful as anti-inflammatory, anti-aging, anti-stress,
anti-arthritic, anti-osteoporosis, anti-cancer, anti-apoptotic, wound
healing, anti-psychotic and anti-fibrotic roles. It also influence
cell-to-cell communication, normal cell growth, cell differentiation,
neurotransmission, cell cycling and proliferation, hormonal balance,
skin health, immune and cardiovascular functions. Besides, it can also
be utilized in hormonal imbalance, aging, and various immune related
disease conditions such as Multiple Sclerosis, Alzheimer's Disease,
Asthma, Atherosclerosis, Pernicious Anemia, Aplastic Anemia,
Diverticulitis, Graves' Disease, Dermatomyositis, Dermatitis, stress,
Irritable Bowel Syndrome, Systemic Lupus Ery-thematosus, Hepatitis,
Hashimoto Thyroiditis,Diabetes, Myasthe-nia Gravis, Ulcerative Colitis,
Sjogren Syndrome, Parkinson's Dis-ease, etc. with a safe therapeutic
index to improve overall health, and quality of life.
Acknowledgement
Authors are grateful to Dabur Research Foundation, Trivedi
Global, Inc., Trivedi Science, Trivedi Testimonials, and Trivedi Mas-ter Wellness for their support throughout the work.
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