In vitro study of homeopathic medicines in macrophages co-cultured with Leishmania (L.) amazonensis

Author(s):  
Leoni Villano Bonamin ◽  
Fabiana Rodrigues De Santana ◽  
Fabiana Toshie De Camargo Konno ◽  
Luciane Costa Dalboni ◽  
Amarylis T. Cesar ◽  
...  

Background: In previous studies the effect of homeopathic medicines in murine cutaneous Leishmaniasis were described, indicating immunomodulation. The phagocytes migration and their activity increased after Thymulin 5cH and decreased after Antimonium crudum 30cH treatment. Methods: The morpho-functional features of RAW macrophages co-cultured with Leishmania (L.) amazonensis in RPMI medium were analyzed in vitro, after 24h treatment with 20% of Thymulin 5cH, 6cH and 7cH or Antimonium crudum 6cH, 30cH and 200cH. All assays were performed in blind and in three series of quadruplicate. The spreading was analyzed by the breadth and area of each macrophage, photographed in a NIKON Eclipse 200-Coolpix system and measured by Metamorph® Image analysis software. The phagocytosis was analyzed by the percentage of amastigotes incorporated into the phagocyte vacuoles. ANOVA and Tuckey Krammer were used as statistical methods. Results: A marked increase of macrophage spreading (considering breadth and area) was seen in Thymulin 5cH and Thymulin 7cH treated cells (p≤0.01), as in Antimonium crudum 30cH (p=0.05) and 200cH (p=0.001) treated cells. Only Antimonium crudum 200cH presented increase in phagocytosis index (p=0.001). Conclusions: The in vitro assay corroborates the findings obtained from in vivo studies and represents the first step to a deeper understanding of the mechanism of action.

2020 ◽  
Vol 11 (SPL4) ◽  
pp. 805-808
Author(s):  
Ravikumar Raju ◽  
Teja ◽  
Sravanathi P ◽  
Muthu Babu K

Breast cancer is the subsequent foremost reason of cancer death in a woman and ranks as the primary foremost reason of death in India. In its conduct, several measures and recommendation are considered. Homoeopathic medicines are one of the part of a corresponding, and another medicine is utilized for the treatment of cancer. The main purpose of the investigation is to evaluate the anticancer action of homoeopathic arrangements of Asterias rubens  on the basis of the similia principle. We directed an in vitro study using MTT assay to control the result of ultra diluted homoeopathic preparation in contradiction of two human breast glandular cancer cell lines(MCF-7 and MDA-MD- 231), frequently used for the breast cancer treatment, by testing the feasibility of breast cancer (MCF-7 and MDA-MD-231) cell line, with various attenuations of Asterias rubens  at 24 hrs. Multiple comparisons between tested reagents at different concentrations confirmed the significance of the said results. At a dilution of 1:25 6CH and 30CH potency shown superior activity on MCF-7 and no such significant changes on MDA-MD-231 at any dilutions As it fails to offer estrogen receptor(ER) Also progesterone receptor (PR) expression, and also HER2 (human epidermal development variable receptor2) so continuously a triple-negative breast cancer it will be a hostility manifestation for breast cancer with restricted medicine choices. However, further potency needs to be tested. These preliminary significant results warrant further in vitro and in vivo studies to estimate the possible of Asterias rubens  a medicine to treat breast cancer.


Marine Drugs ◽  
2019 ◽  
Vol 17 (5) ◽  
pp. 276 ◽  
Author(s):  
Marco Pelin ◽  
Jane Kilcoyne ◽  
Chiara Florio ◽  
Philipp Hess ◽  
Aurelia Tubaro ◽  
...  

Background: Azaspiracids (AZAs) are marine toxins that are produced by Azadinium and Amphidoma dinoflagellates that can contaminate edible shellfish inducing a foodborne poisoning in humans, which is characterized by gastrointestinal symptoms. Among these, AZA1, -2, and -3 are regulated in the European Union, being the most important in terms of occurrence and toxicity. In vivo studies in mice showed that, in addition to gastrointestinal effects, AZA1 induces liver alterations that are visible as a swollen organ, with the presence of hepatocellular fat droplets and vacuoles. Hence, an in vitro study was carried out to investigate the effects of AZA1, -2, and -3 on liver cells, using human non-tumor IHH hepatocytes. Results: The exposure of IHH cells to AZA1, -2, or -3 (5 × 10−12–1 × 10−7 M) for 24 h did not affect the cell viability and proliferation (Sulforhodamine B assay and 3H-Thymidine incorporation assay), but they induced a significant concentration-dependent increase of mitochondrial dehydrogenases activity (MTT reduction assay). This effect depends on the activity of mitochondrial electron transport chain complex I and II, being counteracted by rotenone and tenoyl trifluoroacetone, respectively. Furthermore, AZAs-increased mitochondrial dehydrogenase activity was almost totally suppressed in the K+-, Cl−-, and Na+-free media and sensitive to the specific inhibitors of KATP and hERG potassium channels, Na+/K+, ATPase, and cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels. Conclusions: These results suggest that AZA mitochondrial effects in hepatocytes derive from an imbalance of intracellular levels of K+ and, in particular, Cl− ions, as demonstrated by the selective reduction of toxin effects by CFTR chloride channel inhibition.


2004 ◽  
Vol 43 (5) ◽  
pp. A328
Author(s):  
Xiaokui Li ◽  
Hui Jiang ◽  
Diane Paine ◽  
Zuhua Mao ◽  
Aarti Hejmadi Bhat ◽  
...  

Author(s):  
Matthew B. Fisher ◽  
Nicole Söegaard ◽  
David R. Steinberg ◽  
Robert L. Mauck

Given the limitations of current surgical approaches to treat articular cartilage injuries, tissue engineering (TE) approaches have been aggressively pursued over the past two decades. Although biochemical and biomechanical properties on the order of the native tissue have been achieved (1–5), several in-vitro and in-vivo studies indicate that increased tissue maturity may limit the ability of engineered constructs to remodel and integrate with surrounding cartilage, although results are highly variable (2, 6–8). Thus, “static” measures of construct maturity (e.g. compressive modulus) upon implantation may not be the best indicators of in-vivo success, which likely requires implanted TE constructs to mature, remodel, and integrate with the host over time to achieve optimal results. We recently introduced the concept of “trajectory-based” tissue engineering (TB-TE), which is based on the general hypothesis that time-dependent increases in construct maturation in-vitro prior to implantation (i.e. positive rates) may provide a better predictor of in-vivo success (9). As a first step in evaluating this concept, in the current study we hypothesized that time-dependent increases in equilibrium modulus (a metric of growth) would be correlated to ability of constructs to integrate to cartilage using an in-vitro assay. To test this hypothesis, the current objective was to determine and model the time course of maturation of TE constructs during in-vitro culture and to assess the ability of these constructs to integrate to cartilage at various points during their maturation.


2011 ◽  
Vol 22 (6) ◽  
pp. 479-485 ◽  
Author(s):  
Lucas Amaral Fontanari ◽  
Shelon Cristina Souza Pinto ◽  
Rodrigo Cavassim ◽  
Rubens Spin-Neto ◽  
Eduardo de Paula Ishi ◽  
...  

Although in vitro studies have shown encouraging results for root surface conditioning with demineralizing agents, in vivo studies have failed to show its benefits in periodontal healing. This can be attributed to several factors, among which, the hypermineralization of dental surface. Therefore, this in vitro study compared, using scanning electron microscopy (SEM), the effect of root surface conditioning with different conditioners (1% and 25% citric acid, 24% EDTA and 50 mg/mL tetracycline hydrochloride) in impacted teeth and in teeth that had their roots exposed to the oral environment. One trained examiner assessed the SEM micrographs using a root surface modification index. There was a tendency of more root surface modification in the group of impacted teeth, suggesting that the degree of root mineralization influences its chemical demineralization.


2021 ◽  
Vol 14 (12) ◽  
pp. 1337
Author(s):  
Akhtar Ali ◽  
YoungJoon Park ◽  
Jeonghoon Lee ◽  
Hyo-Jin An ◽  
Jong-Sik Jin ◽  
...  

Osteoarthritis (OA) is a common degenerative joint disorder that affects joint function, mobility, and pain. The release of proinflammatory cytokines stimulates matrix metalloproteinases (MMPs) and aggrecanase production which further induces articular cartilage degradation. Hypertrophy-like changes in chondrocytes are considered to be an important feature of OA pathogenesis. A Glycyrrhiza new variety, Wongam (WG), was developed by the Korea Rural Development Administration to enhance the cultivation and quality of Glycyrrhizae Radix et Rhizoma (licorice). This study examined the regulatory effect of WG against hypertrophy-like changes such as RUNX2, Collagen X, VEGFA, MMP-13 induction, and Collagen II reduction induced by IL-1β in SW1353 human chondrocytes. Additionally, in silico methods were performed to identify active compounds in licorice to target chondrocyte hypertrophy-related proteins. WG showed inhibitory effects against IL-1β-induced chondrocyte hypertrophy by regulating both HDAC4 activation via the PTH1R/PKA/PP2A pathway and the SOX9/β-catenin signaling pathway. In silico analysis demonstrated that 21 active compounds from licorice have binding potential with 11 targets related to chondrocyte hypertrophy. Further molecular docking analysis and in vivo studies elicited four compounds. Based on HPLC, isoliquiritigenin and its precursors were identified and quantified. Taken together, WG is a potential therapeutic agent for chondrocyte hypertrophy-like changes in OA.


2018 ◽  
Vol 97 (10) ◽  
pp. 1137-1143 ◽  
Author(s):  
Y. Itoh ◽  
J.I. Sasaki ◽  
M. Hashimoto ◽  
C. Katata ◽  
M. Hayashi ◽  
...  

Dental pulp regeneration therapy for the pulpless tooth has attracted recent attention, and clinical trial studies are underway with the tissue engineering approach. However, there remain many concerns, including the extended period for regenerating the dental pulp. In addition, the use of scaffolds increases the risk of inflammation and infection. To establish a basic technology for novel dental pulp regenerative therapy that allows transplant of pulp-like tissue, we attempted to fabricate scaffold-free 3-dimensional (3D) cell constructs composed of dental pulp stem cells (DPSCs). Furthermore, we assessed viability of these 3D DPSC constructs for dental pulp regeneration through in vitro and in vivo studies. For the in vitro study, we obtained 3D DPSC constructs by shaping sheet-like aggregates of DPSCs with a thermoresponsive hydrogel. DPSCs within constructs remained viable even after prolonged culture; furthermore, 3D DPSC constructs possessed a self-organization ability necessary to serve as a transplant tissue. For the in vivo study, we filled the human tooth root canal with DPSC constructs and implanted it subcutaneously into immunodeficient mice. We found that pulp-like tissues with rich blood vessels were formed within the human root canal 6 wk after implantation. Histologic analyses revealed that transplanted DPSCs differentiated into odontoblast-like mineralizing cells at sites in contact with dentin; furthermore, human CD31–positive endothelial cells were found at the center of regenerated tissue. Thus, the self-organizing ability of 3D DPSC constructs was active within the pulpless root canal in vivo. In addition, blood vessel–rich pulp-like tissues can be formed with DPSCs without requiring scaffolds or growth factors. The technology established in this study allows us to prepare DPSC constructs with variable sizes and shapes; therefore, transplantation of DPSC constructs shows promise for regeneration of pulpal tissue in the pulpless tooth.


2020 ◽  
Vol 21 (18) ◽  
pp. 6892
Author(s):  
Hiroaki Hirata ◽  
Shusuke Ueda ◽  
Toru Ichiseki ◽  
Miyako Shimasaki ◽  
Yoshimichi Ueda ◽  
...  

Mitochondrial injury has recently been implicated in the pathogenesis of glucocorticoid-induced osteonecrosis. Using cultured osteocytes and a rabbit model, we investigated the possibility that taurine (TAU), which is known to play a role in the preservation of mitochondrial function, might also prevent the development of osteonecrosis. To reduplicate the intraosseous environment seen in glucocorticoid-induced osteonecrosis, dexamethasone (Dex) was added to MLO-Y4 cultured in 1% hypoxia (H-D stress environment). An in vitro study was conducted in which changes in mitochondrial transcription factor A (TFAM), a marker of mitochondrial function, and ATP5A produced by mitochondria, induced by the presence/absence of taurine addition were measured. To confirm the effect of taurine in vivo, 15 Japanese White rabbits were administered methylprednisolone (MP) 20 mg/kg as a single injection into the gluteus muscle (MP+/TAU− group), while for 5 consecutive days from the day of MP administration, taurine 100 mg/kg was administered to 15 animals (MP+/TAU+ group). As a control 15 untreated rabbits were also studied. The rabbits in each of the groups were sacrificed on the 14th day after glucocorticoid administration, and the bilateral femora were harvested. Histopathologically, the incidence of osteonecrosis was quantified immunohistochemically by quantifying TFAM and ATP5A expression. In the rabbits exposed to an H-D stress environment and in MP+/TAU− group, TFAM and ATP5A expression markedly decreased. With addition of taurine in the in vitro and in vivo studies, the expression of TFAM and ATP5A was somewhat decreased as compared with Dex−/hypoxia− or MP−/TAU− group, while improvement was noted as compared with Dex+/hypoxia+ or MP+/TAU− group. In rabbits, the incidence of osteonecrosis was 80% in MP+/TAU− group, in contrast to 20% in the taurine administered group (MP+/TAU+), representing a significant decrease. Since taurine was documented to exert a protective effect on mitochondrial function by inhibiting the mitochondrial dysfunction associated with glucocorticoid administration, we speculated that it might also indirectly help to prevent the development of osteonecrosis in this context. Since taurine is already being used clinically, we considered that its clinical application would also likely be smooth.


2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Anna Lorenzin ◽  
Llaria Godi ◽  
Massimo De Cal ◽  
Claudio Ronco

Abstract Background and Aims The rationale for blood purification as adjunctive therapy during sepsis involved the capacity in removing endogenous and exogenous toxins, but currently no recommendations exists [1]. A critical point may be the potential interaction with antimicrobial therapy, which remains the mainstay of sepsis treatment. HA330 cartridge (Jafron, Zhuhai City, China), widely used in China and actually available in Europe, is used in hemoperfusion for blood purification in septic patients.The aim of this in vitro study was to investigate the adsorptive capacity of HA330 related to vancomycin (VAN). Reference: 1. Rhodes A et al. Crit Care Med 45:486-552. 2017 Method This is an experimental study, simulating an hemoperfusion treatment with HA330. We circulated (250 ml/min) in a closed loop 500 ml of normal saline solution enriched with VAN, stirred and maintained at 37°C. We spiked 100 mg of VAN every 15 minutes to increase the antibiotic load until reaching 1500 mg, the last injection was 500 mg to get a total amount of 2 g. Samples were collected from the inlet line at each VAN adjunct, after system stabilization. Measured VAN concentrations were used to get the adsorption isotherm: for each VAN load, the adsorbed amount of VAN was obtained by multiplying VAN reduction ratio and the quantity of VAN injected. Results Figure 1 shows VAN adsorption isotherm obtained with HA330 cartridge. We observed that, at each injection, after 15 minutes VAN was almost entirely adsorbed by the cartridge, with an average reduction ratio of 0.96. Interestingly, this was confirmed even adding 500 mg at once. Even increasing the VAN load to 500 mg at once, the reduction ratio was maintained. Conclusion In our study, simulating hemoperfusion using HA330, a rapid and clinically relevant removal of VAN has been shown. A significant amount of VAN (2g) was adsorbed without reaching membrane saturation nor reducing its adsorptive capacity. In a clinical setting, we recommend a therapeutic drug monitoring to optimize VAN levels during blood purification with HA330 and a VAN loading dose may be considered. Further in vivo studies are warranted to confirm these findings.


Electronics ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 1629
Author(s):  
Mauricio Guerra-Hernández ◽  
Gabriela Josefina Vidaña-Martínez ◽  
José S. Camacho-Juárez ◽  
Hugo Barragán-Villegas ◽  
José Enrique Calacuayo-Rojas ◽  
...  

A low-cost video laryngoscope (VDL) called Hybrid 1.0 was developed using smart devices for visualization. To test its performance, we compared it with a high-end VDL device, using both in vitro and in vivo studies. During the in vitro study, medical students without experience in airway intubation were randomly asked to intubate a mannequin with different degrees of difficulty (Cormack–Lehane scales) by using either the Hybrid 1.0 VDL (GI) or a conventional laryngoscope (GII). During the in vivo study, N = 60 endotracheal intubations were performed by resident and base physicians, divided into two groups; the first group intubated with the Hybrid 1.0 VDL (GI) while the second group used a VDL C-Mac shovel (GII). As performance indexes, both studies reported the number of successful intubations (correct capnography signal) and intubation time. For the in vitro testing, no statistically significant differences were found regarding the number of successful intubations, while statistically significant differences were found regarding the intubation times. During the in vivo tests, procedures were performed by residents and by base physicians, and no statistically significant differences were found. The provided results point out that the VDL proposed can be clinically useful and offers technical characteristics similar to other VDLs that currently exist on the market.


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