DMEM Group Research Articles

Green tea leaf extract reduces viability and migration of cholesteatoma fibroblast of chronic suppurative otitis media cultured in vitro

Background: Chronic Suppurative Otitis Media (CSOM) is still a health problem, especially in developing countries. CSOM with cholesteatoma is a dangerous type. Cholesteatoma in CSOM sufferers can cause various complications. Therefore, alternative therapies are needed, such as green tea leaf extract. Green tea leaf extract can be an antioxidant and anti-inflammatory, but its effectiveness in treating CSOM has not been studied before, so research is needed.. Purpose: This research aimed to determine the effect of green tea leaf extract that could reduce viability and migration in cholesteatoma fibroblast of CSOM.Methods: This research was an in vitro experiment with a post-test-only control group design. The sample for this research was cholesteatoma fibroblast cells obtained from the isolation of patients with CSOM. The method used is Hoechst staining for viability and scratch techniques for cell migration with eight groups of cholesteatoma fibroblasts consisting of a negative control group (DMEM+FBS), DMEM group, two positive control groups dexamethasone (10 µM and 100 µM), and four green tea leaf extract group (10 µg/ml, 40 µg/ml, 80 µg/ml, and 160 µg/ml). Data analysis used One-Way ANOVA and Kruskal Wallis tests.Results: The results showed that the highest average cell viability was in the negative control group (165.33), and the lowest was in green tea leaf extract at a dose of 160 µg/ml (70.88). Cell migration at 72 hours showed that in the negative control group, there was faster closure of the scratch area (97,78%) and the slowest closure on green tea leaf extract at a dose of 160 µg/ml (13,81%).Conclusion: Green tea leaf extract can reduce the viability and migration in cholesteatoma fibroblast of CSOM. It shows the potential of green tea extract as an alternative to prevent cholesteatoma.

Limited Efficacy of Adipose Stromal Cell Secretome-Loaded Skin-Derived Hydrogels to Augment Skin Flap Regeneration in Rats.

Insufficient vascularization is a recurring cause of impaired pedicled skin flap healing. The administration of adipose tissue-derived stromal cells' (ASCs') secretome is a novel approach to augment vascularization. Yet, the secretome comprised of soluble factors that require a sustained-release vehicle to increase residence time. We hypothesized that administration of a hydrogel derived from decellularized extracellular matrix (ECM) of porcine skin with bound trophic factors from ASCs enhances skin flap viability and wound repair in a rat model. Porcine skin was decellularized and pepsin-digested to form a hydrogel at 37°C. Conditioned medium (CMe) of human ASC was collected, concentrated 20-fold, and mixed with the hydrogel. Sixty Wistar rats were included. A dorsal skin flap (caudal based) of 3 × 10 cm was elevated for topical application of DMEM (group I), a prehydrogel with or without ASC CMe (groups II and III), or ASC CMe (group IV). After 7, 14, and 28 days, perfusion was measured, and skin flaps were harvested for wound healing assessment and immunohistochemical analysis. Decellularized skin ECM hydrogel contained negligible amounts of DNA (11.6 ± 0.6 ng/mg), was noncytotoxic and well tolerated by rats. Irrespective of ASC secretome, ECM hydrogel application resulted macroscopically and microscopically in similar dermal wound healing in terms of proliferation, immune response, and matrix remodeling as the control group. However, ASC CMe alone increased vessel density after 7 days. Porcine skin-derived ECM hydrogels loaded with ASC secretome are noncytotoxic but demand optimization to significantly augment wound healing of skin flaps.

Establishment of Angora rabbits’ whisker hair follicle model and optimal culture conditions

To establish the model of whisker hair follicle culture in vitro and explore the best culture conditions, the whisker hair follicles of Angora rabbits were separated with stereomicroscope and cultured in William’s E, DMEM, MEM media. The surface of the cultured whisker hair follicles was not damaged due to manual operation, resulting in the hair shaft’s growth. This indicated the success of the in vitro whisker hair follicle model. The hair shaft grew at the fastest rate in the William’s E culture (p < 0.05), which was significantly higher than that in the DMEM and MEM media. The hematoxylin-eosin results showed that compared to the William’s E group, the atrophy of whisker hair follicles in the DMEM and MEM media was evident, especially in the MEM medium. PCNA immunofluorescence staining was employed to detect the expression of whisker hair follicles. The results showed that the PCNA positive expression of the William’s E group was significantly stronger than that of the DMEM and MEM groups. Furthermore, CCK-8 and Annexin V-FITC/PI methods were used to detect the proliferation and apoptosis of the dermal papilla cells (DPCs). The results of this study provide a model for studying the hair growth of fur animals.

Genome-wide DNA (hydroxy) methylation reveals the individual epigenetic landscape importance on osteogenic phenotype acquisition in periodontal ligament cells.

Mesenchymal cells' biology has been an important investigative tool to maximize bone regeneration through tissue engineering. Here we used mesenchymal cells from periodontal ligament (PDLCs) with high (h-) and low (l-) osteogenic potential, isolated from different donors, to investigate the impact of the individual epigenetic and transcriptional profiles on the osteogenic potential. Genome-wide and gene-specific DNA (hydroxy) methylation, mRNA expression and immunofluorescence analysis were carried out in h- and l-PDLCs at DMEM (non-induced to osteogenesis) and OM (induced-3rd and 10th days of osteogenic differentiation) groups in vitro. Genome-wide results showed distinct epigenetic profile among PDLCs with most of the differences on 10th day of OM; DMEMs showed higher concentrations (xOM) of differentially methylated probes in gene body, intronic and open sea (3rd day), increasing this concentration in TSS200 and island regions, at 10 days. At basal levels, h- and l-PDLCs showed different transcriptional profiles; l-PDLCs demonstrated higher levels of NANOG/OCT4/SOX2, BAPX1, DNMT3A, TET1/3, and lower levels of RUNX2 transcripts, confirmed by NANOG/OCT4 and RUNX2 immunofluorescence. After osteogenic induction, the distinct transcriptional profile of multipotentiality genes was maintained among PDLCs. In l-PDLCs, the anti-correlation between DNA methylation and gene expression in RUNX2 and NANOG indicates methylation could play a role in modulating both transcripts. Epigenetic and transcriptional distinct profiles detected at basal levels among PDLCs were maintained after osteogenic induction. We cannot discard the existence of a complex that represses osteogenesis, suggesting the individual donors' characteristics have significant impact on the osteogenic phenotype acquisition.

A rabies virus vectored severe fever with thrombocytopenia syndrome (SFTS) bivalent candidate vaccine confers protective immune responses in mice

The Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne hemorrhagic zoonotic disease, which is potentially fatal in human with mortality rates ranging from 16.2%–32%. The rabies virus (RABV) LBNSE vector expressing foreign antigens have shown considerable promise as vaccines against viral diseases, which is effective and safe. In the present study, we generated a recombinant RABV rLBNSE-Gn expressing a SFTSV glycoprotein Gn by reverse genetic technology to control rabies and SFTS in animals. An extra insertion of Gn gene did not impact replication of the recombinant virus rLBNSE-Gn in NA and BHK-21 cells compared to the parent rLBNSE strain. The SFTSV Gn gene together with RABV N and G genes were efficiently expressed in rLBNSE-infected Vero cells by immunostaining and immune blots. A single dose of 107 FFU of the rLBNSE-Gn intramuscularly inoculated in BALB/c mice induced rapid and robust humoral responses against both RABV and SFTSV without any signs of disease or weight loss. Compared to the rLBNSE and DMEM groups, the extra Gn expression contributed to the recruitments and/or activations of the dendritic cells and B cells from inguinal lymph nodes of BALB/c mice vaccinated with rLBNSE-Gn. The protective efficacy of rLBNSE-Gn against SFTSV in C57BL/6 mice was evaluated, and the virus loading in the spleens reduced to 10 TCID50/mg at 7 days post SFTSV infections, which indicated that the rLBNSE-Gn conferred efficacious protective immune responses from SFTSV in C57BL/6 mice. All the mice immunization with rLBNSE-Gn and rLBNSE survived after a lethal RABV challenge, suggesting a 100 % protection from RABV. Therefore, the rLBNSE-Gn would be a promising bivalent candidate vaccine against SFTS and rabies in animals.

Downregulation of Stemness Genes and Induction of Necrosis in Rat LA7 Cancer Stem Cells Induced Tumors Treated with Starved Fibroblasts Culture Supernatant.

Stem cell differentiation therapy is a promising strategy in cancer treatment. we show that protein cocktail prepared from serum starved fibroblasts has therapeutic potential based on this strategy. The condition medium was prepared from foreskin isolated fibroblasts and analyzed by Liquid chromatography electrospray ionization mass spectrometry-mass spectrometry (LC-ESI-MS/MS). LA7 mammary gland cancer stem cells originated tumors were induced in Sprague Dawley rats. The rats treated subcutaneously with DMEM (group A), condition medium (group B), or normal saline (group C) once daily for 7 days. Then the tumors were removed and divided into the two parts, one part was used to quantify gene expression by stem-loop RT-qPCR assay and the other part was used for Hematoxylin & Eosin (H & E), Giemsa, and immunohistochemistry (IHC) staining. All induced tumors appeared as sarcomatoid carcinoma (SC). Immunohistochemistry staining confirmed this conclusion by recognizing the tumor as Ki67+, cytokeratin+, vimentine+, and estrogen receptor negative SC. RT-qPCR analysis revealed that Oct4-, Sox-2, Nanog- gene expression was much reduced in the condition medium treated tumors versus proper controls (p< 0.05). Tissue necrosis was more prevalent in this group while tumors volume was diminished almost by 40%. The LC-ESI-MS/MS analysis unrevealed the stemness reducing and the cell death inducing proteins such as, pigment epithelium-derived factor (PEDF), insulin like growth factor binding protein-5 (IGFBP-5) and -7 (IGFBP-7) in the condition medium. This study showed that the substances released from starved human fibroblasts were able to down-regulate the stemness-related genes and induce necrosis in LA7 derived tumors.

The adipose-derived mesenchymal stem cell secretome promotes hepatic regeneration in miniature pigs after liver ischaemia-reperfusion combined with partial resection

BackgroundHepatic ischaemia-reperfusion injury (HIRI) is inevitable in complicated liver surgery and is a major factor leading to postoperative complications and liver dysfunction. Studies have shown that the paracrine mechanisms of stem cell may be essential to tissue repair and functional improvement after transplantation. However, the role of the adipose-derived mesenchymal stem cell secretome (ASC-secretome) in liver regeneration in large animals remains to be determined.MethodsTwenty-four miniature pigs were subjected to laparoscopic liver ischaemia-reperfusion combined with partial hepatectomy and divided into the following four groups: the saline group, the DMEM group, the ASC group and the ASC-secretome group. Serum and liver tissue samples were collected before the operation and at 1, 3 and 7 days after the operation, and changes in tissue pathology, serum inflammation, liver function, angiogenesis-related factors and liver tissue regeneration-related genes and proteins were evaluated.ResultsDetailed histological analysis showed that ASCs and the ASC-secretome changed pathological damage to liver tissue after liver ischaemia-reperfusion combined with partial hepatectomy (1 and 3 days: p < 0.01). Compared with the saline and DMEM control groups, the ASC-secretome group had significantly reduced expression levels of ALP (1 and 3 days: p < 0.05), ALT (1 day: p < 0.01; 3 days: p < 0.05) and AST (1 and 3 days: p < 0.01), which promoted the recovery of liver function. Moreover, detection of the expression levels of TNF-α and IL-1β (1 day: p < 0.01; 3 days: p < 0.05), IL-6 (1 and 3 days: p < 0.05) and IL-10 (1 and 3 days: p < 0.01) in serum confirmed that the ASC-secretome had obvious anti-inflammatory effects. In addition, the ASC-secretome increased the expression levels of ANG-1 (3 days: p < 0.01), ANG-2 (3 and 7 days: p < 0.01) and VEGF (1 and 7 days: p < 0.05; 3 days: p < 0.01) and promoted angiogenesis during liver regeneration. Moreover, it promoted the mRNA expression of HGF and Cyclin D1 (1 and 3 days: p < 0.01); increased the levels of p-STAT3 (1 and 3 days: p < 0.01), PCNA and Ki67 (1 and 3 days: p < 0.01; 7 days: p < 0.05); inhibited the negative feedback of SOCS3 (1 and 3 days: p < 0.01); and decreased the mRNA expression of TGF-β (3 days: p < 0.01). The cytokines and growth factors detected in the ASC-secretome included TNF-α, IL-6, IL-1β, ANG-1, ANG-2, VEGF and b-FGF.ConclusionThe ASC-secretome alleviates the inflammatory response induced by ischaemia-reperfusion combined with partial hepatectomy in miniature pigs and promotes liver regeneration.

Construction and immunogenicity of a gE/gI/TK-deleted PRV based on porcine pseudorabies virus variant

Pseudorabies (PR) caused by re-emerging pseudorabies virus (PRV) variant has outbroken among PRV vaccine-immunized swine herds on many Chinese pig farms, with severe socioeconomic consequences since late 2011. Here, a gE/gI/TK-deleted recombinant virus (rPRV NY-gE−/gI−/TK−) was constructed based on PRV NY strain from 2012 through homologous DNA recombination and gene-editing technology termed clustered regularly interspaced palindromic repeats (CRISPR)/associated (Cas9) system. The rPRV NY-gE−/gI−/TK− strain showed similar growth kinetics to the parental PRV NY strain in vitro, and was safe for mice. Sixty mice were injected subcutaneously (s.c.) twice with 106.0 TCID50 of rPRV NY-gE−/gI−/TK− and DMEM, respectively, with two-week interval. The levels of PRV gB antibodies and neutralizing antibodies against PRV NY in mice immunized with rPRV NY-gE−/gI−/TK− were higher than those in the DMEM control group. The number of T lymphocyte subclasses CD3+, CD4+ and CD8+ in rPRV NY-gE−/gI−/TK−-immunized mice was higher than that in DMEM-injected mice. After challenge with 106.0 TCID50 PRV NY at 42 dpi, all rPRV NY-gE−/gI−/TK−-immunized mice survived without exhibiting any pathological lesions in different tissues and intranuclear eosinophilic inclusions of the brain, and the viral genomic copy numbers in various organs of mice were obviously lower than DMEM group. These results showed the rPRV NY-gE−/gI−/TK− could be a promising next-generation vaccine to control now epidemic PR in China.

The Combination of Adipose-derived Schwann-like Cells and Acellular Nerve Allografts Promotes Sciatic Nerve Regeneration and Repair through the JAK2/STAT3 Signaling Pathway in Rats

Schwann cells (SCs) combined with acellular nerve allografts (ANAs) effectively promote the regeneration and repair of peripheral nerves, but the exact mechanism has not been fully elucidated. However, the disadvantages of SCs include their limited source and slow rate of expansion in vitro. Previous studies have found that adipose-derived stem cells have the ability to differentiate into Schwann-like cells. Therefore, we speculated that Schwann-like cells combined with ANAs could profoundly facilitate nerve regeneration and repair. The aim of the present study was to investigate the cellular and molecular mechanisms of regeneration and repair. In this study, tissue-engineered nerves were first constructed by adipose-derived Schwann-like cells and ANAs to bridge missing sciatic nerves. Then, the rats were randomly divided into five groups (n = 12 per group): a Control group; a Model group; an ADSC group; an SC-L group; and a DMEM group. Twelve weeks postsurgery, behavioral function tests and molecular biological techniques were used to evaluate the function of regenerated nerves and the relevant molecular mechanisms after sciatic nerve injury (SNI). The results showed that adipose-derived Schwann-like cells combined with ANAs markedly promoted sciatic nerve regeneration and repair. These findings also demonstrated that the expression of neurotrophic factors (NFs) was increased, and the expression of Janus activated kinase2 (JAK2)/P-JAK2, signal transducer and activator of transcription-3 (STAT3)/P-STAT3 was decreased in the spinal cord after SNI. Therefore, these results suggested that highly expressed NFs in the spinal cord could promote nerve regeneration and repair by inhibiting activation of the JAK2/STAT3 signaling pathway.

The growth inhibitory effect of human gingiva-derived mesenchymal stromal cells expressing interferon-\u03b2 on tongue squamous cell carcinoma cells and xenograft model

BackgroundInterferon-β (IFN-β) is a cytokine with pleiotropic cellular functions, including antiviral, antiproliferative, and immunomodulatory activities. IFN-β inhibits multiple tumor cell growth in vitro. However, the contradiction between the therapeutic dose of IFN-β and its maximally tolerated dose is still inextricable in vivo. Human gingiva-derived mesenchymal stromal cells (GMSCs) represent promising vehicles for cancer gene therapy. This study evaluated the potential of GMSCs genetically engineered to produce IFN-β as a targeted gene delivery system to treat tongue squamous cell carcinoma (TSCC) in vitro and in vivo.MethodsA lentiviral vector encoding IFN-β was constructed and transfected into GMSCs to obtain IFN-β gene-modified GMSCs (GMSCs/IFN-β). Enzyme-linked immunosorbent assay (ELISA) was used to measure the IFN-β concentration in conditioned medium (CM) from GMSCs/IFN-β. The Cell Counting Kit-8 (CCK8), colony formation assay, and flow cytometry were used to detect the effects of GMSCs/IFN-β on TSCC cell line CAL27 cell growth and apoptosis in vitro. TSCC xenograft model was developed by subcutaneous injection of CAL27 cells into BALB/c nude mouse, and the role of intravenously injected GMSCs/IFN-β in engrafting in TSCC and controlling tumor progression was measured in vivo.ResultsGMSCs/IFN-β expressed a high level of IFN-β. Both CCK8 and colony forming assay showed that GMSCs/IFN-β significantly inhibited the proliferation of CAL27 cells compared with the GMSCs, GMSCs/vector, or DMEM group. Flow cytometry analysis demonstrated that the CAL27 cell apoptosis rate was higher in the GMSCs/IFN-β group than in the other three groups. The in vivo experiment revealed that GMSCs/IFN-β engrafted selectively in TSCC xenograft and expressed a high level of IFN-β. There were smaller tumor volume and lower number of Ki67-positive cells in the GMSCs/IFN-β group than in the GMSCs, GMSCs/vector, or phosphate-buffered saline (PBS) group. Interestingly, GMSCs and GMSCs/vector also presented the potential of CAL27 cell growth inhibition in vitro and in vivo, although such an effect was weaker than GMSCs/IFN-β.ConclusionsGMSCs/IFN-β inhibits the proliferation of TSCC cells in vitro and in vivo. These results provide evidence that delivery of IFN-β by GMSCs may be a promising approach to develop an effective treatment option for TSCC therapy.

Human Bone Marrow Mesenchymal Stem Cell Conditioned Medium Promotes Wound Healing in Deep Second-Degree Burns in Male Rats

Burn wound treatment is difficult and one of the most challenging problems in the clinic. Researchers have examined the applications of mesenchymal stem cells as a cell-based therapy for skin regeneration. But the role of human bone marrow mesenchymal stem cell conditioned medium (hBM-MSC-CM) in the treatment of burn injury remains unclear. This research aims at detecting whether hBM-MSC-CM can increase the wound healing of deep second-degree burns in male rats. In this study, 32 adult male rats per each time point were randomly divided into four groups: (1) control group, (2) sham group (DMEM), (3) common treatment group (CT), and (4) conditioned media group (CM). A 3 × 3 cm circular burn was created on the back of the rats. On postsurgical days 7, 15, and 28, the wound closure area of each wound was measured and then the skin samples were removed and analyzed using stereological methods. Wound closure area was significantly increased in the CM and CT groups on the 15th and the 28th day after burn injury compared to the control and DMEM groups. The stereological parameters and immunohistochemistry analysis of the wounds revealed significantly improved healing in the CM group compared to the control and other groups. It is concluded that these findings indicate that hBM-MSC-CM promotes skin wound healing by increasing cell proliferation, regulating collagen synthesis and collagen composition, and inducing angiogenesis at the injury site.

In Vitro Effect of Biofield Energy Treated DMEM On Mitochondrial Biogenesis Using Myoblasts Cell Line, C2C12

Mitochondria, a signaling organelles are responsible for energy metabolism within the cell, while its dysfunction in any form could results in the cell death due to an increased oxidative stress and that leads to various metabolic diseases. The aim of the present study was to examine the effect of Consciousness Energy Healing based DMEM medium on murine myoblasts (C2C12) cells to evaluate the mitochondrial mass content using 10-N-nonyl acridine orange (NAO) dye assay. The test item, DMEM was divided into three parts. First part did not receive any sort of treatment and defined as the untreated DMEM group. The second and third parts were treated with the one-time and two-times Biofield Energy Treatment by a renowned Biofield Energy Healer, Alice Branton and coded as the one-time Biofield Energy Treated DMEM (BT-I) and two-times Biofield Energy Treated DMEM (BT-II) groups, respectively. Cell viability of the test items using MTT assay showed 72.32%, 97.89%, and 104.96% viable cells in the untreated DMEM, BT-I, and BT-II groups, respectively suggested that the test items were nontoxic and safe in murine myoblasts (C2C12) cells. Further, the mitochondrial mass content in terms of Fluorescence Unit (FU) was significantly (p≤0.05) increased by 43.11% and 64.52% in the BT-I and BT-II groups, respectively in C2C12 cells compared to the untreated DMEM group. Thus, overall experimental data suggested that two-times blessed DMEM showed a significant improvement of mitochondrial mass content and results in better thermogenesis. In the present study, results demonstrated that an increased mitochondrial mass content in the cells, when treated with The Trivedi Effect®. This indicates that the Biofield Energy Treated DMEM has the great potential to improve thermogenesis, which can be used against various metabolic diseases, such as insulin resistance, type-2 diabetes, and cardiovascular diseases.

Assessment of Biofield Energy Treatment on Lung Health Using Lung Adenocarcinoma Cell Line (A549)

Oxidative stress is one of the major molecular mechanism responsible for human lung diseases. The objective of the study was to examine the effect of a Consciousness Energy Healing (The Trivedi Effect®) Treated DMEM medium for its anti-oxidative potential using various parameters such as the protection against oxidative stress and Superoxide Dismutase (SOD), antioxidant enzyme activity in A549 cells. The test item, DMEM was divided into two parts. One part received the Consciousness Energy Healing Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and was labeled as the Biofield Energy Treated test item, while the other part did not receive any sort of treatment and is defined as the untreated DMEM group. The cell viability of the test sample using MTT assay showed 122.67% viable cells, indicating a safe and nontoxic profile of the test item. Protection against oxidative stress was significantly increased by 64.70% in the Biofield Energy Treated DMEM group compared to the untreated DMEM group. In addition, Antioxidant Enzyme (SOD) activity was significantly increased by 45.1% in the Biofield Energy Treated DMEM group compared to the untreated DMEM group. Thus, these data suggest that Biofield Energy Healing Treatment showed a significant improvement of the Antioxidant Enzyme Activity (SOD level) along with an improved protection against oxidative damage, which can be used in various human lung disorders such as asthma, Chronic Obstructive Pulmonary Disease (COPD), lung malignancies and parenchymal lung diseases like idiopathic pulmonary fibrosis and lung granulomatous diseases.

Impact of Biofield Energy Treated DMEM on Mitochondrial Biogenesis Using Myoblast Cell Line, C2C12: Implications for Metabolic Disease

Author(s): Trivedi, Mahendra Kumar; Jana, Snehasis | Abstract: Mitochondria play an important role in the body, and its dysfunction lead to the defective cellular energy production results to heterogeneous group of disorders such as early aging, type-2 diabetes, Alzheimer’s disease, and many more metabolic diseases. The study was aimed to examine the effect of Consciousness Energy Healing based DMEM on myoblasts cells (C2C12) for mitochondrial mass content. The test item, DMEM was divided into three parts. First part did not receive any sort of treatment and defined as the untreated DMEM group. The second and third portions were treated with the one-time and two-times Biofield Energy Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and coded as the one-time Biofield Energy Treated DMEM (BT-I) and two-times Biofield Energy Treated DMEM (BT-II) groups, respectively. Cell viability of the test items by MTT assay showed more than 72% viable cells, which suggested that the test items were nontoxic and safe in nature. Moreover, the mitochondrial mass content in terms of fluorescence unit (FU) was significantly (p≤0.05) increased by 43.11% and 64.16% in the BT-I and BT-II groups, respectively in C2C12 cells compared to the untreated DMEM group. Overall, the experimental data suggested that two-times treated DMEM (BT-II) was found more significant improvement in mitochondrial mass content compared with the one-time treated DMEM (BT-I) group and results in better respiratory capacity. Thus, data envisaged that the Biofield Energy Treated DMEM has a potential to improve respiratory activity, which could be used against various metabolic diseases, such as insulin resistance, type-2 diabetes, and cardiovascular diseases, etc.

Impact of the Trivedi Effect ® - Energy of Consciousness Healing on the Proliferation of Plant, Mouse and Human Stem Cells

The objective of the present study was to evaluate the effect of Biofield Energy Healing on the plant callus, mouse and human derived Mesenchymal Stem Cells (MSCs) for their cell count and cellular proliferation. All the three type stem cells and DMEM media for the stem cell culture were divided into two parts, one received Biofield Energy Treatment (known as The Trivedi Effect®-Energy of Consciousness Healing Treatment) by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi through the Healer’s unique Energy Transmission process remotely and defined as Biofield Energy Treated (BT) group. On day 9 after treatment, the plant callus flasks of mandukparni, amla, and Centella were observed and visualized which showed an increased percentage in weight of the callus by 11.4%, 24.9%, and 51.7%, respectively as compared with the untreated groups. In addition, stem cell assay in murine bone marrow derived Mesenchymal Stem Cells (MSCs) showed an increased cellular proliferation by 115.2%, 127.6%, 161.9%, and 123.6%, in Biofield Energy Treated DMEM in the passage 1, 2, 3, and 4, respectively. Similarly, human derived Mesenchymal Stem Cells, Adipose-Derived (AD-MSC) cells showed a significant increased cell count by 139%, 119%, and 182% in the Biofield Energy Treated DMEM group with the passage 1, 2, and 3, respectively. Similarly, bone marrow (BM-MSC) cells showed a significant increased cell count by 102%, 170%, and 141% in the Biofield Energy Treated DMEM group with the passage 1, 2, and 3, respectively. However, BrdU assay for % cellular proliferation in the AD-MSC cells showed a significant increased cellular proliferation by 122%, 120%, and 184% in the passage 1, 2, and 3, respectively. On the other hand, BM-MSC cells showed a significant increased cellular proliferation by 125%, 146%, and 157% in the passage 1, 2, and 3, respectively. Hence, the results suggest that there was a significant growth in plant callus weight, mouse and human stem cell growth and proliferation after The Trivedi Effect®-Energy of Consciousness Treatment. It can be concluded that The Trivedi Effect® can be used as a complementary and alternate therapy for the stem cells regeneration, improvement of tissue injury that might be every effective against various autoimmune human diseases such as arthritis, osteoarthritis, and different neurodegenerative disorders.

Improved Metabolic Cardiac Biomarkers Activity Using Rat Cardiomyocytes Cell Line (H9c2) against Biofield Energy Treated Test Sample

Cardiac biomarkers are important targets for any test sample to discover ideal cardiac health supplement. The study was aimed to evaluate the cardioprotective activity of Biofield Energy Treated test sample (DMEM) using rat cardiomyocytes (H9c2) cells. The test item (DMEM medium) was divided into three parts, first part received one-time Biofield Treatment by Alice Branton, a renowned Biofield Energy Healer, and defined as the one-time Biofield Energy Treated (BT-I) DMEM, while second part received two-times Biofield Energy Treatment and denoted as the BT-II DMEM. The third part did not give treatment and denoted as the untreated DMEM group. The cell viability of the test sample by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay showed an increase level of viable cells by 86.58% and 91.37% in the BT-I and BT-II groups, respectively. Besides, BT-I and BT-II groups showed 10.56% and 23.53% cytoprotective action compared to the tert-Butyl hydroperoxide (t-BHP) induced group. Further, lactate dehydrogenase (LDH) level was significantly (p≤0.001) reduced by 54.43% and 53.52% in the BT-I and BT-II groups, respectively compared to t-BHP induced group. Moreover, the level of creatine kinase-myocardial band (CK-MB) was reduced by 63.48% and 60.68% in the BT-I and BT-II groups, respectively compared to the t-BHP induced group. Additionally, reactive oxygen species (ROS) level was significantly inhibited by 55.6% and 112.8% in the BT-I and BT-II groups, respectively compared to the t-BHP induced group. Besides, apoptotic activity was significantly inhibited by 16% and 10% in the BT-I and BT-II groups, respectively compared to the t-BHP induced group. Overall, the experimental data demonstrated that The Trivedi Effect® has significant capacity to improve the overall cardiac parameters in H9c2 cells, which can be used against various cardiac disorders such as congestive heart failure (CHF), cardiomyopathy, heart valve disorders, arrhythmias, myocardial infarction, etc.

Bone marrow-derived mesenchymal stem cells attenuate tubulointerstitial injury through multiple mechanisms in UUO model.

Current evidence supports the use of bone marrow-derived mesenchymal stem cells (MSCs) for a diverse range of clinical applications, and many studies have shown that MSCs have renal-protective effects, but the mechanism is not well understood. Therefore, in this study, we aim to further identify whether MSCs can attenuate renal fibrosis by decreasing tubulointerstitial injury in a unilateral ureteral obstruction (UUO) model. In this study, we cultured MSCs and then transplanted them into a UUO model through the tail vein. Histology, cell proliferation, peritubular capillary (PTC) loss and myofibroblast markers were examined on days 3, 7 and 14 after surgery. We demonstrated that renal interstitial fibrosis in the MSC group was significantly attenuated compared with the UUO and DMEM groups. Moreover, MSC treatment inhibited the loss of PTCs and increased parenchymal cell proliferation. In addition, UUO-induced activation and proliferation of myofibroblasts were suppressed by MSC infusion. Furthermore, MSCs attenuated tubulointerstitial infiltration of macrophages in UUO mice. Tubulointerstitial damage plays a very important role in the progression of chronic kidney disease (CKD). PTC loss, macrophage recruitment, and myofibroblast activation are directly correlated with the development of renal tubulointerstitial fibrosis. Our results suggest that MSC infusion in the UUO model is a promising therapeutic strategy for promoting kidney repair.

In Vitro Assessment of Estrogenic Potential of Biofield Energy Treatment using Human Endometrial Adenocarcinoma Cell Line

The objective of the study was to investigate the effect of Consciousness Energy Healing based DMEM medium on the level of alkaline phosphatase enzyme (ALP) activity in Ishikawa cells. The test item, DMEM medium was divided into two parts. One part of the test item received Consciousness Energy Healing Treatment by a renowned Biofield Energy Healer, Mahendra Kumar Trivedi and was labeled as the Biofield Energy Treated DMEM, while the other part did not receive any treatment, and defined as the untreated DMEM group. The cell viability using MTT assay of the Biofield Energy Treated DMEM group was observed as 108%, which indicated that the test item was safe and non-toxic. The estrogenic potential using ALP level showed a significantly increase by 73.21% in the Biofield Energy Treated DMEM group as compared to the untreated DMEM group. Overall, the experimental data suggested that the Biofield Energy Treated DMEM has significantly improved ALP level, which play a vital role for the promotion and maintenance of estrogen level. Based on the study outcomes, it is concluded that Biofield Energy Healing Treatment showed a significant improved ALP level, which can be used in various estrogenic disorders such as hypophosphatasia, osteoporosis, severe anemia, malnutrition, hypothyroidism, magnesium deficiency, heart surgery, aplastic anemia, chronic myelogenous leukemia, enteritis in children, Wilson’s disease, pernicious anemia, bacterial infection and intrauterine infection is a leading cause of pelvic inflammatory disease, subfertility, infertility, endometritis, early pregnancy loss, fetal defects, and preterm birth.

Evaluation of Phosphodiesterase-5 Inhibitory Potential of Biofield Energy Treated DMEM by Determining cGMP Level in Human Endothelial Cell Line

Erectile dysfunction is a common disorder found in men, which occurs due to multiple factors such as psychogenic, hormonal imbalance, and neurovascular disturbances. The present investigation was undertaken to examine the effect of a Consciousness Energy Healing based DMEM medium on the Human Endothelial Hybrid Cell Line (EA. hy926) to evaluate the level of cyclic guanosine monophosphate (cGMP). The test item (DMEM medium) was divided into three parts, first part received a one-time Consciousness Energy Healing Treatment by a renowned Biofield Energy Healer, Alice Branton and was labeled as the one-time Biofield Energy Treated (BT-I) DMEM, while second part received the two-times the 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. The level of cGMP for the inhibition of PDE-5 enzyme was assessed using cGMP ELISA assay kit (colorimetric). Sildenafil citrate, used as positive control, which showed a significant increase of cGMP levels in Ea. hy926 cells. The one-time Biofield Energy Healing Treated DMEM (BT-I) showed 29.92% alteration, while the two-times Biofield Energy Healing Treated DMEM (BT-II) showed a significantly (p≤0.001) increased level of intracellular cGMP by 307.09% in Ea. hy926 cells compared to the untreated DMEM group. Overall, experimental data suggested that the two-times Biofield Treated DMEM showed a significantly improved level of cGMP compared with the one-time Biofield Treated DMEM group. Therefore, data indicated that the Biofield Energy Healing Treatment can be used to treat erectile dysfunction along with other sexual disorders such as orgasmic disorders, female sexual arousal disorder, fetishistic disorder, frotteuristic disorder, hypoactive sexual desire disorder, sex addiction, sexual masochism and sadism, vaginismus, voyeuristic disorder, premature or delayed ejaculation.

In vitro Assessment of Biofield Energy Treated DMEM on Thermogenesis Using Myoblasts Cell Line (C2C12)

Author(s): Trivedi, Dahryn; Jana, Snehasis | Abstract: Mitochondrial dysfunction lead to various serious disorders, which are considered as one of the important components related with the aging, such as type-2 diabetes and Alzheimer’s disease. The aim of the present study was to examine the effect of Consciousness Energy Healing based DMEM medium on murine myoblasts (C2C12) cells to evaluate the mitochondrial mass content using 10-N-nonyl acridine orange (NAO) dye assay. The test item (DMEM medium) was divided into two parts, one part received Consciousness Energy Healing Treatment by a renowned Biofield Energy Healer, Dahryn Trivedi and was labeled as the Biofield Energy Treated DMEM group, while the other part did not receive any kind of Treatment and denoted as the untreated DMEM group. The level of mitochondrial mass content was assessed using 10-N-nonyl acridine orange (NAO) dye method. Cell viability of the test items using MTT assay showed 72.32% and 125.32% viable cells in the untreated DMEM and Biofield Energy Treated DMEM groups, respectively suggested a safe and nontoxic profile of the test items. Besides, the mitochondrial mass content in terms of Fluorescence Unit (FU) was significantly (p≤0.05) increased by 81.78% in the Biofield Energy Treated DMEM group compared to the untreated DMEM group. Overall, the experimental data suggested that the Consciousness Energy Healing Based DMEM showed a significant improvement of mitochondrial mass content and results in better thermogenesis with respect to naive DMEM. Thus, an increased level of NAO dye accumulation in muscle cells indicated increased mitochondrial mass content and hence, better thermogenesis. In the present study, results demonstrated that an increased mitochondrial mass content in the cells when treated with The Trivedi Effect®. This indicates that the test sample has the potential to improve thermogenesis, which can be used against various metabolic diseases, such as insulin resistance, type-2 diabetes, and cardiovascular diseases, etc.