Experimental evidence in support of the biological effects and physical basis of homeopathic potencies

Background: Homeopathic potencies 12 cH and above cross the Avogadro number and, for this, do not contain any original drug molecules. Two major problems involved in the scientific study of potencies are (1) understanding the physical basis of potencies and (2) demonstrating the biological effects of potencies. The present study aims to address these questions. Methods and Results: In course of our experimental studies spanned over more than 30 years we have demonstrated significant effects of homeopathic potencies on man, animals and plants. We have also showed that potencies could be differentiated through their electronic spectra, and this difference in spectra can be attributed to the electron transfer interaction. In a molecular complex, electron of one molecule absorbs a quantum of visible radiation and is excited, not to a higher energy level of this molecule, but to one of the vacant high energy levels of the neighboring molecules. This process is known as electron or charge transfer interaction. This has been demonstrated in Iodine ө in two different solvents of CCl4 and aqueous ethanol (Sukul N C, Environ Ecol 17,866-872, 1999). We have further demonstrated that the effect of a homeopathic potency can be transmitted from one part of a plant to another, and also from one plant to another through water. I am presenting here a few selected cases of our experimental studies. Potentized Nux vomica significantly reduced ethanol consumption in rats by 73.7%and ethanol-induced sleep time in albino mice by 44.4%. Causticum 30 C and Rhus tox 30 C produced anti-inflamatory and anti-nocicptive effect on adjuvant arthritis in albino rats. Potentized homeopathic drugs reduced microfilaraemia by 28 to 100% and filariasis in two villages of West Bengal endemic for Bancroftian filaiasis. Potentized Cina and Thuja ameliorated trichinellosis in mice reducing larval population in muscles by 84% and 68%, respectively. Potencies of Agaricus and Nux vomica, produced excitatory effect on the isolated rat ileum. Potentized drugs altered firing rate of hypothalamic neurons in rats and cats. Nux vom 30 c and Merc cor 30c facilitated water permeability in erythrocytes of catfish in a test tube. Potencies of Merc cor and Merc iod enhanced α-amylase activity in vitro ,by 44 and 21%,respectively. Drugs ,that inhibit photosynthesis and plant growth in high doses, promote the same phenomena when applied on plants at ultra low doses. Potentized Cantharis, a homeopathic drug used for burn injuries, counter the effect of heat shock in Adhatoda vasica plants in terms of modulating the expression of heat-shock proteins in the plants. The effect of heat shock and of Cantharis treatment could be transmitted from one plant to another through water. The global network of surface water in a closed system is thought to be responsible for producing this effect. Several potentized homeopathic drugs show distinct variation from each other in their absorption spectra in ultra violet region of light. These drugs when mixed with sucrose solution, also show marked differences from each other at temperatures as low as 4 0 C and as high as 70 0 C. Electron transfer interaction may contribute to the characteristic spectral properties of a homeopathic potency. Conclusions: Homeopathic potencies could be detected and differentiated by their electronic spectra. Potencies show marked effect on animals, plants, ex vivo effect on isolated organs and in vitro effect on enzymes. Keywords: Homeopathic potencies, electronic spectra, ethanol intake, adjuvant arthritis, filariasis, Cantharis, Nux vomica

EVALUATION OF THE THERAPEUTIC EFFECT OF THE COMBINED USE OF CRYOPRESERVED PLACENTA EXTRACT AND DICLOFENAC SODIUM IN EXPERIMENTAL RHEUMATOID ARTHRITIS BY HEMATOLOGICAL PARAMETERS

Relevance. Cryopreserved placenta extract (CPE) increase the resistance of the mucous membrane of the gastrointestinal tract to the damaging effects of nonsteroidal anti-inflammatory drugs. Preventive administration of CPE can reduce the ulcerogenic effect of meloxicam, ibuprofen, diclofenac sodium (DS) and others. There is evidence of CPE's own anti-inflammatory activity, which can be successfully combined with the pharmacological properties of nonsteroidal anti-inflammatory drugs, while improving their safety profile. Objective: to characterize the therapeutic activity of the combined use of CPE and DS according to hematological parameters in the model of experimental rheumatoid arthritis (RA). Materials and methods. Studies were performed on 28 nonlinear laboratory rats. The rats were divided into 4 groups: I (n = 7) – intact rats; II (n = 7) – rats with experimental RA; ІІІ (n = 7) – rats with experimental RA, treated with DN; IV (n = 7) – rats with experimental RA, treated with DN and CPE. Adjuvant arthritis was modeled by subplantar administration of complete Freund's adjuvant. Treatment was performed from 14 to 28 days. CPE was administered on days 14, 17, 20, 23 and 26, and DS – daily. Blood tests were performed on day 28 of the experiment. Results. The combined use of CPE and DS is accompanied by a more pronounced leveling of inflammatory signs by hematological parameters – erythrocyte clotting rate decreased by 72.2% (p<0.001), and the number of leukocytes decreased by 54.81% (p<0.001) relative to rats with adjuvant arthritis without treatment. There was a leveling of signs of anemia of chronic inflammation – the level of hemoglobin and erythrocytes increased (p<0,001) by 17.6% and 36.8%, respectively, relative to rats with adjuvant arthritis without treatment. Conclusions. The combined use of CPE and DS is superior in therapeutic activity to monotherapy with this nonsteroidal anti-inflammatory drug of experimental rheumatoid arthritis.

The effect of complex herbal extract and methotrexate on suppressing adjuvant arthritis in rats

The present study evaluated the therapeutic benefits of complex herbal preparation named CBMDS, consisting of turmeric (Curcuma longa), Boswellia (Boswellia serrata), Methylsulphonylmethane, Devil’s Claw (Harpagophytum procumbens) and Silymarin, using it in combination with methotrexate, in order to suppress adjuvant arthritis in rats, and to attenuate methotrexate-induced liver damage. Adjuvant arthritis was induced in 28 rats by a single subplantar injection of complete Freund’s adjuvant (0.1 mL) into the left hind paw. The animals were divided into four groups (with seven animals in each). Group I received CBMDS, Group II - CBMDS in combination with methotrexate, and Group III just methotrexate. The treatment lasted from day 0 to day 17 (CBMDS was given daily except weekends in a dose of 160 mg/kg, methotrexate - 2 mg/kg once a week). Group IV was the control group. Clinical (body weight, hind paw volume, erythrocyte sedimentation rate, leukocyte count), biochemical (serum pro-/antioxidant activity markers), immunological (serum interleukin levels) and histological changes in joint and liver tissues were evaluated. CBMDS significantly alleviated arthritis and reduced hepatic damage, which was more evident in the methotrexate group. The combined treatment also markedly reduced arthritic symptoms and levels of malondialdehyde. Antioxidant activity was significantly higher in treated Groups I and II. CBMDS and its combination with methotrexate promoted anti-arthritic action, reduced histological changes in the joint tissues, and minimized methotrexate-induced liver toxicity.

Course correction of adjuvant arthritis with cryopreserved multipotent mesenchymal stromal cells

Rheumatoid arthritis is an inflammatory autoimmune disease that occurs as a result of impaired immune tolerance, leading to an aberrant immune response to autologous antigens. Multipotent mesenchymal stromal cells (MMSCs) and the biologically active substances they produce can promote the activation of regenerative processes in the organism not only by direct cell differentiation, but also due to their inherent trophic and immunosuppressive potentials. The aim of the study was to experimentally evaluate changes in the course of the acute phase of adjuvant arthritis upon local and generalized administration of cryopreserved MMSCs from adipose and cartilage tissues. The results of histological, imunohistochemical and biochemical studies showed that the animals of the control group throughout the observation period developed an inflammatory process, which manifested in joint swelling (increased arthritis index), leukocytosis, spread of chondrocyte-free zones, weakening of staining, loss of clarity of cartilage tissue contours, increased content of cyclooxygenase-2, reduced glycosaminoglycan content and total antioxidant defense system activity. At the same time, the local administration of cryopreserved MMSCs from adipose and cartilage tissues contributed to the normalization of the structural and functional organization, content of glycosaminoglycans and cyclooxygenase-2 with complete recovery of blood parameters. Less pronounced regeneration processes in articular cartilage occurred under generalized administration of cryopreserved MMSCs from adipose and cartilage tissues in comparison with the local method. However, the difference between the control and experimental groups indicates the ability of cryopreserved MMSCs to influence the intensity of regenerative processes in damaged cartilage tissue with both methods of administration. Comparative evaluation of the use of cryopreserved MMSCs from adipose and cartilage tissues showed the absence of significant changes in the studied indicators. These data can be used to substantiate and develop methods of arthritis treatment in clinical practice.

Evaluation of the anti-inflammatory activity of melatonin in a model of experimental arthritis

The epiphyseal hormone melatonin (1 and 5 mg/kg) significantly limited locomotor and psychoemotional disorders, indicators of inflammation and immunological reactivity in rats with experimental adjuvant arthritis.

APPLICATION OF MULTIPOTENT MESENCHYMAL STEM CELL SECRETOME IN THE TREATMENT OF ADJUVANT ARTHRITIS AND CONTACT-ALLERGIC DERMATITIS IN ANIMAL MODELS

The therapeutic effect of multipotent mesenchymal stem cells has been proven on various disease models. One of the mechanisms is the paracrine effect of the cells on the surrounding tissues.The aim. To investigate the secretome effectiveness of the multipotent mesenchymal stem cells in the treatment of adjuvant arthritis and contact-allergic dermatitis in Wistar rats.Materials and methods. Adjuvant arthritis was simulated in 26 female rats by the administration of Freund's complete adjuvant and then treated with the administration of 100 µl of multipotent mesenchymal stem cell secretome or saline. Contact-allergic dermatitis was modeled on 30 female rats by applying 200 μl of an oil solution of dinitrofluorobenzene to the skin on days 1, 5 and 6. Then the rats were treated with fluocinolone ointment (a positive control), baby cream (a negative control), baby cream with a secretome of native multipotent mesenchymal stem cells or from the cells processed with dexamethasone.Results. Judging by the indicators of the longitudinal and transverse dimensions of the paws in rats and a histological examination, the secretome did not have any anti-inflammatory effect on adjuvant arthritis. A cream with a secret from multipotent mesenchymal stem cells processed with dexamethasone, was the most effective on the model of contact-allergic dermatitis: the clinical improvement occurred on the 2nd day. The secretome from native multipotent mesenchymal stem cells and fluocinolone had a therapeutic effect on the 3rd day of application, the negative control - on the 4th day. The lymphocytic infiltration coefficient was significantly lower (p <0.05) in all the cases compared to the negative control (2.8 ± 0.1). However, the lowest infiltration was observed when the cream with secretome from native (1.75 ± 0,1) and dexamethasone-stimulated (1.76 ± 0.1) multipotent mesenchymal stem cells was being used.Conclusion. The cream with the secretome of multipotent mesenchymal stem cells suppresses lymphocytic infiltration more strongly than the highly active topical glucocorticosteroid - fluocinolone - on the model of contact-allergic dermatitis, which is a classic local delayed-type hypersensitivity reaction. However, a further study of the therapeutic effect of the secretome on models of systemic inflammatory diseases is required after its preliminary purification from large-molecular proteins.

Blockade of TRPM7 Alleviates Chondrocyte Apoptosis and Articular Cartilage Damage in the Adjuvant Arthritis Rat Model Through Regulation of the Indian Hedgehog Signaling Pathway

Articular cartilage damage with subsequent impairment of joint function is a common feature of articular diseases, in particular, rheumatoid arthritis and osteoarthritis. While articular cartilage injury mediated by chondrocyte apoptosis is a known major pathological feature of arthritis, the specific mechanisms remain unclear at present. Transient receptor potential melastatin-like seven channel (TRPM7) is reported to play an important regulatory role in apoptosis. This study focused on the effects of TRPM7 on arthritic chondrocyte injury and its underlying mechanisms of action. Sodium nitroprusside (SNP)-induced rat primary chondrocyte apoptosis and rat adjuvant arthritis (AA) were used as in vitro and in vivo models, respectively. Blockage of TRPM7 with 2-APB or specific siRNA resulted in increased chondrocyte viability and reduced toxicity of SNP. Moreover, treatment with 2-APB enhanced the Bcl-2/Bax ratio and reduced cleaved PARP and IL-6, MMP-13 and ADAMTS-5 expression in SNP-treated chondrocytes. Activation of Indian Hedgehog with purmorphamine reversed the protective effects of 2-APB on SNP-induced chondrocyte apoptosis. Blockage of TRPM7 with 2-APB relieved the clinical signs of AA in the rat model and reduced the arthritis score and paw swelling. Similar to findings in SNP-treated chondrocytes, 2-APB treatment increased the Bcl-2/Bax ratio and suppressed cleaved PARP, IL-6, MMP-13, ADAMTS-5, TRPM7, and Indian hedgehog expression in articular cartilage of AA rats. Our collective findings suggest that blockade of TRPM7 could effectively reduce chondrocyte apoptosis and articular cartilage damage in rats with adjuvant arthritis through regulation of the Indian Hedgehog signaling pathway.

Bioflavonoid Robinin from Astragalus falcatus Lam. Mildly Improves the Effect of Metothrexate in Rats with Adjuvant Arthritis

Anti-inflammatory potential of orally administrated bioflavonoid-robinin, active sub-stance of original drug Flaroninum™ (FL), was investigated in the combination with methotrexate (MTX) and in monotherapy in rats suffering from adjuvant-induced arthritis (AA). Robinin (kaempferol-3-O-robinoside-7-O-rhamnoside) was isolated from the aerial parts of Astragalus falcatus Lam. The monotherapy with robinin was not efficient in alleviating symptoms of AA. The combination of MTX with robinin was similarly active as MTX alone in reducing the hind paw volume and change of body weight during the whole experiment. The combination, however, reduced plasma levels of Interleukin-17Aand activity of gamma-glutamyl transferase in joint more efficiently then MTX alone. Our results demonstrate that the novel combination of robinin and MTX mildly improved the reduction of inflammation in experimental arthritis.