scholarly journals Sitotoksisitas dan Induksi Apoptosis Ekstrak Etanol Teripang holothuria atra Jaeger, 1833 pada beberapa Sel Kanker

2018 ◽  
Vol 13 (2) ◽  
pp. 101 ◽  
Author(s):  
Ernie Halimatushadyah ◽  
Muhammad Da’i ◽  
Muhammad Nursid
Keyword(s):  
Double Staining ◽  
Tetrazolium Bromide ◽  
Diphenyl Tetrazolium Bromide ◽  
Holothuria Atra

Teripang Holothuria atra merupakan biota laut yang banyak ditemukan di perairan Indonesia yang termasuk dalam filum Echinodermata dan berpotensi sebagai antikanker. Penelitian ini bertujuan untuk mengetahui sitotoksisitas dan induksi apoptosis ekstrak etanol teripang H. atra secara in vitro terhadap beberapa sel lestari. Pengujian sitotoksisitas dilakukan dengan metode MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) menggunakan sel HeLa, T47D, WiDr dan sel normal Vero, sedangkan uji induksi apoptosis dilakukan terhadap sel dengan hasil uji sitotoksisitas terbaik menggunakan metode flowcytometry dan double staining. Hasil penelitian menunjukkan bahwa ekstrak etanol  teripang H. atra mampu menghambat pertumbuhan sel kanker HeLa, T47D dan WiDr. Nilai IC50 ekstrak H. atra terhadap ketiga sel tersebut masing-masing sebesar 41,06±4,21; 20,89±1,55; 26,50±4,43 µg/ml tetapi esktrak tersebut memiliki sitotoksisitas  yang lebih rendah terhadap sel Vero (IC50 sebesar 128,00). Analisis flowcytometry dan double staining pada sel T47D memperlihatkan bahwa ekstrak etanol teripang H.atra mampu menginduksi apoptosis pada sel tersebut.

scholarly journals Evaluation of 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyl tetrazolium bromide method for assessing biofilm formation in vitro by Trichosporon spp.

2018 ◽  
Vol 10 (04) ◽  
pp. 380-386 ◽  
Author(s):  
Thayanidhi Premamalini ◽  
Subramanian Anitha ◽  
Kanniappan Mohanapriya ◽  
Anupma Jyoti Kindo
Keyword(s):  
Mtt Assay ◽  
Biofilm Formation ◽  
Microtiter Plate ◽  
Test Tube ◽  
Tetrazolium Bromide ◽  
Biofilm Production ◽  
Invasive Infections ◽  
Polymerase Chain ◽  
Diphenyl Tetrazolium Bromide

ABSTRACT BACKGROUND: Invasive infections due to Trichosporon spp. have increased recently and are frequently associated with indwelling medical devices. Such infections which are associated with biofilm formation do not respond to the routinely used antifungal agents and are often persistent, associated with high mortality rate. Various methods have been described by researchers to evaluate and quantify the biofilm formation. AIM: This study was conducted to compare two methods of biofilm production by Trichosporon sp, i.e., test tube method with crystal violet (CV) staining and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. MATERIALS AND METHODS: Seventy-two clinical isolates of Trichosporon spp. collected from various sources were considered for the study. The identity of all the isolates was genotypically confirmed by Trichosporon-specific polymerase chain reaction (PCR). The isolates were further speciated phenotypically using biochemical profile and growth characteristics which identified the isolates as Trichosporon asahii (64/72), Trichosporon asteroides (5/72), Trichosporon cutaneum (2/72), and Trichosporon mucoides (1/72). Biofilm production was then evaluated and compared by test tube-CV method and MTT assay. RESULTS: All the Trichosporon isolates produced biofilm by MTT assay, whereas only 42 (53.6%) of the isolates were detected to be biofilm producers by CV method. Furthermore, MTT assay could differentiate better between weak and moderate biofilm producers as compared to CV method. CONCLUSION: Hence, MTT assay is a reliable method for quantification of biofilm produced by Trichosporon spp. using 96-well microtiter plate.

scholarly journals Nanocarriers of Miconazole or Fluconazole: Effects on Three-Species Candida Biofilms and Cytotoxic Effects In Vitro

2021 ◽  
Vol 7 (7) ◽  
pp. 500
Author(s):  
Anne Caroline Morais Caldeirão ◽  
Heitor Ceolin Araujo ◽  
Laís Salomão Arias ◽  
Wilmer Ramírez Carmona ◽  
Gustavo Porangaba Miranda ◽  
...  
Keyword(s):  
Fungal Infections ◽  
Artificial Saliva ◽  
Cytotoxic Effects ◽  
Promising Alternative ◽  
Colony Forming Units ◽  
Mtt Reduction ◽  
Diphenyl Tetrazolium Bromide ◽  
Negative Controls ◽  
Positive Controls

The contribution of different Candida species in oral fungal infections has stimulated the search for more effective therapies. This study assessed the antibiofilm effects of nanocarriers of miconazole (MCZ) or fluconazole (FLZ) on Candida biofilms, and their cytotoxic effects on murine fibroblasts. Three-species biofilms (Candida albicans/Candida glabrata/Candida tropicalis) were formed on 96-well plates, and they were treated with nanocarriers (iron oxide nanoparticles coated with chitosan—“IONPs-CS”) of MCZ or FLZ at 39/78/156 µg/mL; antifungals alone at 156 µg/mL and artificial saliva were tested as positive and negative controls, respectively. Biofilms were analyzed by colony forming units (CFU), biomass, metabolic activity, and structure/viability. The cytotoxicity (L929 cells) of all treatments was determined via 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) reduction assay. Data were submitted to one- or two-way ANOVA, followed by Tukey’s or Fisher LSD’s tests (p < 0.05). IONPs-CS-MCZ at 78 µg/mL promoted similar antibiofilm and cytotoxic effects compared with MCZ at 156 µg/mL. In turn, IONPs-CS-FLZ at 156 µg/mL was overall the most effective FLZ antibiofilm treatment, surpassing the effects of FLZ alone; this nanocarrier was also less cytotoxic compared with FLZ alone. It can be concluded that both nanocarriers are more effective alternatives to fight Candida biofilms compared with their respective positive controls in vitro, being a promising alternative for the treatment of oral fungal infections.

scholarly journals Osteogenic Properties of 3D-Printed Silica-Carbon-Calcite Composite Scaffolds: Novel Approach for Personalized Bone Tissue Regeneration

2021 ◽  
Vol 22 (2) ◽  
pp. 475
Author(s):  
Parastoo Memarian ◽  
Francesco Sartor ◽  
Enrico Bernardo ◽  
Hamada Elsayed ◽  
Batur Ercan ◽  
...  
Keyword(s):  
Osteogenic Differentiation ◽  
Bone Tissue ◽  
Cell Attachment ◽  
Three Dimensional ◽  
Bone Tissue Regeneration ◽  
Hemolysis Assay ◽  
Novel Approach ◽  
Related Transcription Factor ◽  
Diphenyl Tetrazolium Bromide

Carbon enriched bioceramic (C-Bio) scaffolds have recently shown exceptional results in terms of their biological and mechanical properties. The present study aims at assessing the ability of the C-Bio scaffolds to affect the commitment of canine adipose-derived mesenchymal stem cells (cAD-MSCs) and investigating the influence of carbon on cell proliferation and osteogenic differentiation of cAD-MSCs in vitro. The commitment of cAD-MSCs to an osteoblastic phenotype has been evaluated by expression of several osteogenic markers using real-time PCR. Biocompatibility analyses through 3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) activity, hemolysis assay, and Ames test demonstrated excellent biocompatibility of both materials. A significant increase in the extracellular alkaline phosphatase (ALP) activity and expression of runt-related transcription factor (RUNX), ALP, osterix (OSX), and receptor activator of nuclear factor kappa-Β ligand (RANKL) genes was observed in C-Bio scaffolds compared to those without carbon (Bio). Scanning electron microscopy (SEM) demonstrated excellent cell attachment on both material surfaces; however, the cellular layer on C-Bio fibers exhibited an apparent secretome activity. Based on our findings, graphene can improve cell adhesion, growth, and osteogenic differentiation of cAD-MSCs in vitro. This study proposed carbon as an additive for a novel three-dimensional (3D)-printable biocompatible scaffold which could become the key structural material for bone tissue reconstruction.

Stealth doxorubicin conjugated bimetallic selenium/silver nanoparticles for targeted cervical cancer therapy

2021 ◽  
Vol 12 (4) ◽  
pp. 045006
Author(s):  
Thoko Malinga ◽  
Tukayi Kudanga ◽  
Londiwe Simphiwe Mbatha
Keyword(s):  
Cervical Cancer ◽  
Folic Acid ◽  
Colloidal Stability ◽  
Controlled Drug Release ◽  
Delivery Systems ◽  
Drug Binding ◽  
Cervical Cancer Cells ◽  
Diphenyl Tetrazolium Bromide

Abstract Bimetallic nanosized delivery systems are attracting a lot of research interest as alternatives to monometallic delivery systems. This study evaluated the ability of bimetallic selenium silver chitosan pegylated folic acid targeted nanoparticles (SeAgChPEGFA NPs) to deliver doxorubicin (DOX) in cervical cancer cells. Comparison studies using monometallic selenium chitosan pegylated folic acid (SeChPEGFA NPs) targeted NPs and free DOX were also conducted. The prepared NPs and their drug nanocomplexes were characterised morphologically and physico-chemically. Drug binding and releasing studies were conducted under a simulated environment in vitro. The cytotoxicity and apoptosis studies were studied using the 3-[(4, 5-dimethylthiazol-2-yl)−2, 5-diphenyl tetrazolium bromide] (MTT) assay and the dual dye staining. The findings revealed that the bimetallic SeAgChPEGFA NPs displayed better colloidal stability, superior physico-chemical qualities, and higher binding abilities in comparison with monometallic SeChPEGFA NPs. In addition, the SeAgChPEGFA NPs showed the pH-triggered controlled drug release and cell-specific cytotoxicity. These findings suggest that the bimetallic NPs are superior delivery systems when compared to their monometallic NPs and free drug counterparts, thus, setting a platform for further in vivo examination.

In Vitro Cytotoxicity and Mutagenicity Evaluation of Dental Leucite Glass-Ceramics from Local High Grade Silica Sand

2015 ◽  
Vol 754-755 ◽  
pp. 979-984
Author(s):  
Siti Mazatul Azwa Bt Saiyed Mohd Nurddin ◽  
Malek B. Selamat
Keyword(s):  
Mtt Assay ◽  
Glass Ceramics ◽  
Mutagenic Effect ◽  
Negative Control ◽  
Silica Sand ◽  
High Grade ◽  
Reverse Mutation ◽  
Diphenyl Tetrazolium Bromide ◽  
Mutation Assay

The objective of the study was to determine the degree of biocompatibility of leucite glass-ceramics that have been produced from local high grade silica sand in terms of cytotoxicity and mutagenicity assays. In the present study, the cyctotoxicity and mutagenicity were studied using the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay (MTT) and Ames Reverse Mutation. In the MTT assay, a dose response cytotoxicity of leucite sample was evaluated in L929 cells. The cells were treated with the concentrations of 6.25, 12.5, 25.0, 50.00, 100.00 and 200.00 mg/ml of the leucite sample for 24 hours. The cytotoxicity was determined by assessing the cell viability through the reduction of tetrazolium salts (MTT). The mutagenenicity of leucite sample was evaluated inS. typhiriumTA98. TA100, TA1535, TA1537 andE. coliWP2 in the Ames Reverse Mutation assay. Mutagenic effects were evaluated by comparing the mean number of revertant colonies of each extract concentraction with mean number of revertant colonies of the negative control. In results of MTT assay evaluated that the leucite did not show a cytotoxic effect at all concentrations under the condition of the study. Ames Reverse Mutation assay result proven that the leucite sample did not demonstrate a mutagenic effect under the condition of this study withSalmonella typhimuriumandEscherichia coli.

scholarly journals Interleukin-1β Augments Angiogenic Responses of Murine Endothelial Progenitor Cells in Vitro

2009 ◽  
Vol 29 (5) ◽  
pp. 933-943 ◽  
Author(s):  
Anna Rosell ◽  
Ken Arai ◽  
Josephine Lok ◽  
Tongrong He ◽  
Shuzhen Guo ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Endothelial Progenitor Cells ◽  
Mononuclear Cells ◽  
Therapeutic Angiogenesis ◽  
Interleukin 1Β ◽  
Endothelial Progenitor ◽  
Angiogenic Potential ◽  
Diphenyl Tetrazolium Bromide ◽  
In Vitro Angiogenesis

Endothelial progenitor cells (EPCs) may provide novel opportunities for therapeutic angiogenesis after ischemic diseases. However, it is unclear how the angiogenic potential of EPCs might be affected by an inflammatory environment. We examine how the potent cytokine interleukin-1β (IL-1β) affects angiovasculogenic responses in EPCs in culture. Mononuclear cells isolated from mouse spleen were plated on fibronectin-coated wells and grown in EGM-2 MV media. Endothelial progenitor cells were phenotyped using multiple markers (UEA-Lectin, ac-LDL, CD133, CD34, vWillebrand Factor, Flk-1) and to identify the IL-1 Receptor-I. We quantified cell and colony counts and performed MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide) and Matrigel assays, in vitro, under control and IL-1β (10 ng/mL) conditions. Endothelial progenitor cells exposed to IL-1β increased in the number of cells and colonies compared with untreated cells, without any effect on cell metabolic integrity. Furthermore, IL-1β treatment augmented EPC angiogenic function, significantly increasing the number of vessel-like structures in the Matrigel assay. An early phosphorylation of ERK1/2 occurred after IL-1β stimulation, and this pathway was inhibited if IL-1 Receptor-I was blocked. Our results suggest that IL-1β is a potent stimulator of in vitro angiogenesis through ERK signaling in mouse EPCs. Further studies are warranted to assess how interactions between proinflammatory environments and EPC responses may be leveraged to enhance therapeutic angiogenesis.

scholarly journals The activity of extracts chara vulgaris against promastigotes of leishmania tropica.

2018 ◽  
Vol 12 (2) ◽  
pp. 66-72
Author(s):  
Ban Hussein Ali ◽  
Thaer A. Saleh ◽  
Mohammed M. Al-Halbosiy
Keyword(s):  
Major Health Problem ◽  
Organic Extract ◽  
Major Health ◽  
Treatment Groups ◽  
Chara Vulgaris ◽  
Incubation Periods ◽  
Potential Activity ◽  
Diphenyl Tetrazolium Bromide ◽  
Better Than

            Leishmaniasis is a widespread parasitic disease caused by Leishmania parasite, this disease considers as a major health problem worldwide. The available therapy is unsatisfactory expensive with a cytotoxic side effects. Studies of marine algae as a source of pharmacological active compounds have increased worldwide.  This study was aimed to investigate the effect of a type of green algae (Chara vulgaris) on promastigotes of L. tropica, by using. various concentrations (500, 250, 125, 62.5, 31.25, 15.6 µg/mL) in vitro by MTT assay [3-(4.5-dimethylthiazol-2-yl)- 2.5-diphenyl tetrazolium bromide)], to investigate its effect on the proliferation of promastigotes, by three incubation periods (24, 48, 72 hr.) The results showed a significant (p< 0.05)   decrease in survived of promastigotes in treatment groups with concentrations that ranged between 15 to 500 μg/ ml.  This study revealed a major growth inhibition effect of the organic extract of C. vulgaris against L. tropica promastigotes, and the extract of ethyl acetate showed potential activity is better than the aqueous extract.

scholarly journals EVALUATION AND COMPARISON OF CYTOTOXIC EFFECT OF VILAZODONE HYDROCHLORIDE WITH 5-FLUOROURACIL IN HT-29 BOWEL CANCER CELL LINE

2019 ◽  
pp. 124-127
Author(s):  
LATHA PRIYA A ◽  
ANUSHA D ◽  
DARLING CHELLATHAI K ◽  
HEMALATHA A ◽  
JEGAN MOHAN Y
Keyword(s):  
Cell Line ◽  
Cytotoxic Effect ◽  
Depressive Disorders ◽  
Cancer Cell Line ◽  
Test Sample ◽  
Standard Drug ◽  
Drug Sample ◽  
Diphenyl Tetrazolium Bromide ◽  
Ht 29

Objectives: Vilazodone hydrochloride is a novel selective serotonin reuptake inhibitor (SSRI) used to treat major depressive disorders. There are only sparse data available to know about the SSRI’s and its association with colon cancer. This study aims to evaluate and compare the in vitro cytotoxic effect of vilazodone with 5-fluorouracil (5-FU) in HT-29 cell line. Methods: Cell viability was tested by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay (Mosmann, 1983). Test sample and standard drug in variable concentrations were added to the HT-29 cell lines for incubation over 24 h under ideal conditions. After washing the test and standard drug sample from the well with saline, MTT was added and incubated for 4 h. Dimethyl sulfoxide of 1 ml was added in all wells after incubation with MTT. The absorbance at 570 nm was measured with an ultraviolet - spectrophotometer. Results: The values were tabulated, and the graph was plotted to find the IC-50 value (inhibitory concentration at 50%) which was struck at 28.5 μg/ml and12. 8 μg/ml for vilazodone hydrochloride and 5-FU, respectively. Conclusion: The results show that vilazodone hydrochloride has good anticancer property comparable with 5-FU, which would probably play a role as a cytotoxic agent in tumor cells. The proposed mechanism of action could be by activation of caspase-3 enzyme, thereby increasing apoptosis and indicates its use in coexisting depression and colon carcinoma. Other mechanism includes suppression of oncogene p53, which can be confirmed by future studies.

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