scholarly journals Evaluation of in vitro Cytotoxic Effects of Especifico Pessoa Phytotherapic Tincture on Ehrlich Tumor Cells and Mice Spleen Cells

2020 ◽  
Vol 9 (1) ◽  
pp. 458-472
Author(s):  
Lindsey Castoldi ◽  
Lucinéia Reuse Albiero ◽  
Eduardo Figueredo Nery ◽  
Taiany Oliveira Kelly ◽  
Jeniffer Charlene Silva Dalazen ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Spleen Cells ◽  
Ehrlich Tumor ◽  
Traditional Use ◽  
Trypan Blue Exclusion ◽  
Cytotoxic Effects ◽  
Trypan Blue Exclusion Method ◽  
Tumor Cell Suspension ◽  
Ehrlich Tumor Cells

Especifico Pessoa (EP) is traditionally used for the treatment of snakebite envenoming. The traditional use of EP and its properties have been reported. In this study, we evaluated the in vitro cytotoxic effects of EP on Ehrlich tumor and mice spleen cells. Cytotoxicity assay was carried out by using Trypan blue exclusion method. Spleen cell suspension was prepared (n=2) with RPMI medium and tumor cell suspension was prepared from ascitic fluid of Ehrlich tumor-bearing mice (n=1); both the suspensions contained 4 x 106 cells mL-1. Pure EP or EP diluted in RPMI (1:2; 1:4) was used. The results were expressed as percentage of cell viability and demonstrate that EP is toxic to Ehrlich cells at all concentrations (Control: 96.42 ± 3.40; Pure: 1.55 ± 2.91; 1:2: 4.85 ± 5.04; 1:4: 13.39 ± 5.08), but nontoxic to spleen cells in at the lowest dilution (Control: 72.86 ± 13.79; Pure: 13.52 ± 6,36; 1:2: 41.36 ± 13.51; 1:4: 56.59 ± 8.62). Therefore, the results demonstrate that EP has cytotoxic effects, depending on the dose and the cell line evaluated.

Alpha mangostin Inhibits Hepatic Stellate Cells Activation Through TGF-β/Smad and Akt Signaling Pathways: An in vitro Study in LX2

Drug Research ◽  
2017 ◽  
Vol 68 (03) ◽  
pp. 153-158 ◽  
Author(s):  
Rahmaniah Rahmaniah ◽  
Yuyuntia Yuyuntia ◽  
Vivian Soetikno ◽  
Wawaimuli Arozal ◽  
Radiana Antarianto ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Akt Signaling ◽  
Dependent Manner ◽  
Trypan Blue Exclusion ◽  
Ki 67 ◽  
Trypan Blue Exclusion Method ◽  
Dose Dependent

Abstract Background Alpha mangostin has been reported to have activity for the treatment of liver fibrosis in the rats. However, the mechanisms of action are poorly understood. This study was aimed to investigate the effect of alpha mangostin on hepatic stellate cells (HSC) activation and proliferation through TGF-β/Smad and Akt signaling pathways. Methods Immortalized HSC, LX2 cells, were incubated with TGF-β with or without alpha mangostin (5 or 10 μM). Sorafenib 10 µM was used as positive control. LX2 viability was counted using trypan blue exclusion method. The effect of alpha mangostin on TGF-β concentrations, and the expressions of proliferation and fibrogenic markers were evaluated. Results Alpha mangostin treatment resulted in a reduced proliferation of HSC, decreased Ki-67 and p-Akt expressions. These findings were followed with decreased concentrations of TGF-β in the medium of cells treated with alpha mangostin, decreased expressions of COL1A1, TIMP1, PAI1, α-SMA, and p-Smad3 as fibrogenic markers. These effects were shown to be dose-dependent. Conclusions Alpha mangostin inhibits hepatic stellate cells proliferation and activation through TGF-β/Smad and Akt signaling pathways in dose dependent manner.

scholarly journals INDUCTION OF A HEMOLYSIN RESPONSE IN VITRO

1970 ◽  
Vol 132 (6) ◽  
pp. 1267-1278 ◽  
Author(s):  
Klaus-Ulrich Hartmann
Keyword(s):  
Bone Marrow ◽  
Immune Response ◽  
T Cells ◽  
Cell Suspension ◽  
Cell Population ◽  
Cell Suspensions ◽  
Spleen Cells ◽  
Bone Marrow Derived Cells ◽  
Thymus Cells

The immune response to foreign erythrocytes was studied in vitro. Two subpopulations of cells were prepared. One was a population of bone marrow-derived spleen cells, taken from thymectomized, irradiated, and bone marrow-reconstituted mice; there was evidence that most of the precursors of the PFC had been present in this cell population, but few PFC developed in cultures of these cells alone in the presence of immunogenic erythrocytes. Another cell suspension was made from spleens of mice which had been irradiated and injected with thymus cells and erythrocytes; these cells were called educated T cells. The two cell suspensions together allow the formation of PFC in the presence of the erythrocytes which were used to educate the T cells, but not in the presence of noncross-reacting erythrocytes. If bone marrow-derived cells and T cells were kept in culture together with two different species of erythrocytes, and if one of the erythrocytes had been used to educate the T cells, then PFC against each of the erythrocytes could be detected.

scholarly journals Confocal Microscopy in Conjunction With Haemocytometry to Evaluate the Imperative Physical Characteristics of Multicellular Tumor Spheroids

2021 ◽  
Author(s):  
Aziz UR RAHMAN
Keyword(s):  
Confocal Microscopy ◽  
Drug Uptake ◽  
Live Cells ◽  
Multicellular Tumor Spheroids ◽  
Tumor Spheroids ◽  
Oriented Growth ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method

Abstract Background: Tumor tissues resist penetration of therapeutic molecules. Multicellular tumor spheroids (MCTSs) were used as an in vitro tumor model. The aim of this study was to determine the growth of MCTSs with the age of spheroids, which could be applied and compared with in vivo drug uptake and penetration. Method: Spheroids were generated by liquid overlay techniques, and their diameter was measured by confocal microscopy for up to two weeks. The trypan blue exclusion method was used to count dead and live cells separately via a hemocytometer. Results: The pentaphysical characteristics of spheroids, including diameter, cell number, volume per cell, viability status, and estimated shell of viable and core of dead cells, were determined. The growth of spheroids was linear over the first week but declined in the 2nd week, which may be due to an overconcentration of dead cells and degraded products inside the spheroids, hence lowering the ratio of live cells in spheroids. Compaction of spheroids occurs from day 3 to day 7, with the mature spheroids having a low amount of extracellular space compared to intracellular volume. Conclusion: Age-oriented growth of MCTSs provides a rationale to predict less rapid penetration as spheroids get older and could be correlated with in vivo tumors to predict pharmaceutical and therapeutic intervention.

scholarly journals Ethanolic extract of gabirobeira (Campomanesia adamantium) leaves reduces proliferation of osteosarcoma cells in vitro

2021 ◽  
Vol 43 ◽  
pp. e52783
Author(s):  
Nara Cristina Silva ◽  
Leandro Lopes Nepomuceno ◽  
Nayane Peixoto Soares ◽  
Vanessa de Souza Vieira ◽  
Vanessa de Sousa Cruz ◽  
...  
Keyword(s):  
Ethanolic Extract ◽  
Metastatic Potential ◽  
Chemotherapeutic Agents ◽  
In Vitro Cytotoxicity ◽  
Osteosarcoma Cells ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Ic50 Value ◽  
Ic50 Values

Osteosarcoma is the most commonly diagnosed malignant bone tumor in humans, with a higher incidence in children and young people. It is highly aggressive and has a high metastatic potential. Its treatment is based on both chemotherapy and surgical intervention. However, currently used chemotherapeutic agents, such as doxorubicin, have several adverse effects on the patient. Therefore, there is a growing demand for new chemotherapeutic agents that stimulate new researches, such as those involving compounds extracted from plants, such as the gabirobeira. In this study, we aimed to evaluate the cytotoxic effects of ethanolic extract, both crude and ethyl acetate, of gabirobeira leaves on osteosarcoma cells in vitro. Cytotoxicity was evaluated using the Trypan blue exclusion method and the IC50 values were calculated using the tetrazolium reduction method. The ethanolic extract of gabirobeira leaves showed a cytotoxic effect on osteosarcoma cells in vitro. The group treated with the crude extract at 1. 0μL mL-1 concentration for 48 hours showed higher cytotoxicity and the lowest IC50 value for this extract was found in the 24 to 48 hours interval. The ethanolic extract of gabirobeira leaves is cytotoxic for osteosarcoma cells.

scholarly journals Dental Pulp Fibroblasts Response after Stimulation with HEMA and Adhesive System

2018 ◽  
Vol 29 (5) ◽  
pp. 419-426 ◽  
Author(s):  
Karin Cristina da Silva Modena ◽  
Adriana Maria Calvo ◽  
Carla Renata Sipert ◽  
Thiago José Dionísio ◽  
Maria Fidela de Lima Navarro ◽  
...  
Keyword(s):  
Gene Expression ◽  
Nitric Oxide ◽  
Dental Pulp ◽  
Close Contact ◽  
Adhesive System ◽  
Single Bond ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Nitric Oxide Release

Abstract This study evaluated in vitro cell viability and metabolism, nitric oxide release and production of chemokines by cultured human dental pulp fibroblasts (DPF) under contact with HEMA and Single Bond. Cultures of DPF were established by means of an explant technique. Once plated, cells were kept under contact with increasing concentrations of HEMA (10, 100 and 1000 nM) or Single Bond (SB) [10-fold serially diluted in culture medium (10-4, 10-3 and 10-2 v/v)] and also with polymerized SB components. Cytotoxicity was assessed by Trypan Blue exclusion method and MTT [3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Nitric oxide release on cell supernatant was detected by Griess Method whereas chemokines (CXCL12 and CXCL8) were detected by ELISA. RT-qPCR was employed for chemokines gene expression analysis. Cytotoxic tests showed significant differences for SB 10-2. None of the tested materials significantly altered NO levels. Protein levels of CXCL12 were significantly decreased only by HEMA. On the other hand, while CXCL12 mRNA remained unaltered, gene expression of CXCL8 had significant decrease with all materials, except for polymerized SB. In conclusion, Single Bond and HEMA at various concentrations, decreased expression and production of molecules involved in inflammatory processes and, therefore, the use of adhesive systems such as pulp capping materials must be viewed with caution due to its large cytotoxic effect when in close contact with the pulp.

scholarly journals ANTIOXIDATIVE ACTIVITY OF ALPHA-MANGOSTIN IN ACETALDEHYDE-INDUCED HEPATIC STELLATE CELLS: AN IN VITRO STUDY

2019 ◽  
pp. 164-167
Author(s):  
NOVRIANTIKA LESTARI ◽  
SAMUEL PRATAMA ◽  
KELVIN THEANDRO GOTAMA ◽  
VIVIAN SOETIKNO ◽  
MELVA LOUISA
Keyword(s):  
Liver Fibrosis ◽  
Cell Viability ◽  
Hepatic Stellate Cells ◽  
In Vitro Study ◽  
Stellate Cells ◽  
Trypan Blue Exclusion ◽  
Trypan Blue Exclusion Method ◽  
Antioxidative Properties ◽  
Causes Of Mortality

Objective: Alcohol accumulation in the liver can cause pathological disorders such as liver fibrosis that can develop into hepatocellular carcinoma,one of the main causes of mortality associated with liver disease. The previous studies have shown that a plant compound, alpha-mangostin, has anantioxidant effect in the inhibition of pancreatic tumor growth in vitro. This study aimed to analyze the antioxidative properties of alpha-mangostin inacetaldehyde-induced liver fibrosis in vitro.Methods: Immortalized hepatic stellate cells (HSCs), of the LX-2 cell line, were incubated with acetaldehyde in the presence or absence of alphamangostin(10 and 20 μM). The cells were then counted and lysed, and LX-2 cell viability was determined with the trypan blue exclusion method. Themalondialdehyde levels, superoxide dismutase activity, and glutathione (GSH) levels were also determined using the cell lysates.Results: Acetaldehyde treatment resulted in a significant increase in HSC cell viability and decreased the production of GSH. Alpha-mangostintreatment resulted in reduced cell viability of the HSCs and prevention of the loss of intracellular GSH.Conclusion: Alpha-mangostin reduced acetaldehyde-induced cell proliferation, and this was affected at least in part by its antioxidative properties

scholarly journals Glucosinolate-Degradation Products as Co-Adjuvant Therapy on Prostate Cancer in Vitro

2019 ◽  
Vol 20 (20) ◽  
pp. 4977 ◽  
Author(s):  
María Jesús Núñez-Iglesias ◽  
Silvia Novío ◽  
Carlota García ◽  
Elena Pérez-Muñuzuri ◽  
Pilar Soengas ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Adjuvant Therapy ◽  
Degradation Products ◽  
Allyl Isothiocyanate ◽  
Chemotherapeutic Drug ◽  
Trypan Blue Exclusion ◽  
Cytotoxic Effects ◽  
Cycle Arrest ◽  
Anticancer Effects

Glucosinolate-degradation products (GS-degradation products) are believed to be responsible for the anticancer effects of cruciferous vegetables. Furthermore, they could improve the efficacy and reduce side-effects of chemotherapy. The aim of the present study was to determine the cytotoxic effects of GS-degradation products on androgen-insensitive human prostate cancer (AIPC) PC-3 and DU 145 cells and investigate their ability to sensitize such cells to chemotherapeutic drug Docetaxel (DOCE). Cells were cultured under growing concentrations of allyl-isothiocyanate (AITC), sulforaphane (SFN), 4-pentenyl-isothiocyanate (4PI), iberin (IB), indole-3-carbinol (I3C), or phenethyl-isothiocyanate (PEITC) in absence or presence of DOCE. The anti-tumor effects of these compounds were analyzed using the trypan blue exclusion, apoptosis, invasion and RT-qPCR assays and confocal microscopy. We observed that AITC, SFN, IB, and/or PEITC induced a dose- and time-dependent cytotoxic effect on PC-3 and DU 145 cells, which was mediated, at least, by apoptosis and cell cycle arrest. Likewise, we showed that these GS-degradation products sensitized both cell lines to DOCE by synergic mechanisms. Taken together, our results indicate that GS-degradation products can be promising compounds as co-adjuvant therapy in prostate cancer.

scholarly journals Alpha Mangostin Inhibits the Proliferation and Activation of Acetaldehyde Induced Hepatic Stellate Cells through TGF-β and ERK 1/2 Pathways

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Novriantika Lestari ◽  
Melva Louisa ◽  
Vivian Soetikno ◽  
Averina Geffanie Suwana ◽  
Putra Andito Ramadhan ◽  
...  
Keyword(s):  
Liver Fibrosis ◽  
Mechanism Of Action ◽  
Hepatic Stellate Cells ◽  
Stellate Cells ◽  
Trypan Blue Exclusion ◽  
Ki 67 ◽  
Activation Markers ◽  
Trypan Blue Exclusion Method ◽  
And Migration

Liver fibrosis is characterized by excessive accumulation of extracellular matrix in chronic liver injury. Alcohol-induced fibrosis may develop into cirrhosis, one of the major causes of liver disease mortality. Previous studies have shown that alpha mangostin can decrease ratio of pSmad/Smad and pAkt/Akt in TGF-β-induced liver fibrosis model in vitro. Further investigation of the mechanism of action of alpha mangostin in liver fibrosis model still needs to be done. The present study aimed to analyze the mechanism of action of alpha mangostin on acetaldehyde induced liver fibrosis model on TGF-β and ERK 1/2 pathways. Immortalized HSCs, LX-2 cells, were incubated with acetaldehyde, acetaldehyde with alpha mangostin (10 and 20 μM), or alpha mangostin only (10 μM). Sorafenib 10 μM was used as positive control. LX-2 viability was counted using trypan blue exclusion method. The effect of alpha mangostin on hepatic stellate cells proliferation and activation markers and its possible mechanism of action via TGF-β and ERK1/2 were studied. Acetaldehyde was shown to increase proliferation and expression of profibrogenic and migration markers on HSC, while alpha mangostin treatment resulted in a reduced proliferation and migration of HSC and decreased Ki-67 and pERK 1/2 expressions. These findings were followed with decreased expressions and concentrations of TGF-β; decreased expression of Col1A1, TIMP1, and TIMP3; increased expression of MnSOD and GPx; and reduction in intracellular reactive oxygen species. These effects were shown to be dose dependent. Therefore, we conclude that alpha mangostin inhibits hepatic stellate cells proliferation and activation through TGF-β and ERK 1/2 pathways.

scholarly journals REGULATION OF LYMPHOCYTE RESPONSES IN VITRO

1972 ◽  
Vol 136 (4) ◽  
pp. 956-961 ◽  
Author(s):  
Masaru Yoshinaga ◽  
Aiko Yoshinaga ◽  
Byron H. Waksman
Keyword(s):  
T Cell ◽  
Dna Synthesis ◽  
Cell Suspension ◽  
Direct Effect ◽  
Adherent Cells ◽  
Spleen Cells ◽  
Large Numbers ◽  
Normal Rat ◽  
Lymphocyte Responses

DNA synthesis of normal rat spleen cells in response to endotoxin increases markedly if adherent cells are first removed from the cell suspension. Addition of small numbers of purified macrophages to the cultures restores the response to a low level. T-deprived cells show these effects in very much lesser degree. Large numbers of macrophages completely suppress the response of both normal and T-deprived spleen. We conclude that two mechanisms of suppression are at work: a direct effect of macrophages and a macrophage-dependent "suppressor T cell" effect.

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