Tryphan Blue Assay Research Articles

An evaluative in vitro investigation of the delivery of cytarabine with RGD decorated solid lipid nanoparticles

Aim To synthesize cytarabine loaded SLNs modified with the RGD peptide as a ligand, suitable for effective cancer therapy. Methods SLNs were synthesized by the high shear, hot homogenization technique. A 2 level 3 factor analysis was used in optimization. Particle size, zeta potential, poly-dispersion index and surface morphology were measured. Drug encapsulation, drug release, release kinetics, nanoparticle stability and chemical structure were determined. LIVE/DEAD® Fluorescence Assay was used to qualify cytotoxicity and Tryphan Blue assay to quantify. Results Cyt-SLNs exhibited a size of 161 ± 2.25nm, a PDI of 0.49 ± 0.15 and zeta potential of -19.8mV. Entrapment fell at 88.87 ± 0.02% and release at 83.5 ± 0.95%. The in vitro release kinetics pointed towards a diffusion based drug release mechanism. SLNs remained stable for 60 days. Cytotoxicity studies revealed that conjugation of the ligand with the RDG peptide resulted in a significant decrease in cell viability in both cell lines. Conclusion Overall, the study suggests that RGD-SLN-cyt can be used for effective cancer therapy.

Green synthesis of silver, iron and gold nanoparticles of lycopene extracted from tomato: their characterization and cytotoxicity against COLO320DM, HT29 and Hella cell

Our study aimed at development of Silver, Iron and Gold nanoparticles of Lycopene isolated from tomato by using green synthesis technique and to evaluate its anticancer potential against colorectal and cervical cancer. Lycopene was extracted by benzene extraction method and the silver, iron and gold nanoparticles were developed by green synthesis method. 1% aqueous extract of isolated Lycopene was mixed with 1% solutions of AgNO3, FeCl3 and HAuCl4 solutions and incubated at ambient temperature for 3–4 h separately and observed for the color change which is an indicative of formation of the nanoparticles. The prepared nanoparticles were characterized by FTIR, SEM, XRD analysis and evaluated for their antimicrobial potential. The cytotoxicity studies were carried out by in vitro assay like MTT, SRB and Tryphan blue method against Colo 320 DM, HT 29, and Hella. SEM showed nanosized particles of 50–100 nm range, whereas no antimicrobial activity was exhibited by the prepared nanoparticles. In MTT assay the LyAgNP showed maximum 41.41 ± 0.4124% inhibition against COLO320DM, whereas LyGNP exhibited 41.47 ± 0.4469% inhibition against HT 29 and LyAgNP showed 40.9 ± 0.6908% inhibition against Hella cells. In SRB assay LyAgNP showed maximum 82.68 ± 1.1798% inhibition against COLO320DM, whereas LyGNP exhibited maximum 91.21 ± 0.2372% inhibition against HT29 and 87.98 ± 0.5878% inhibition against Hella cells. In tryphan blue assay against COLO320DM, HT29 and Hella cells, the maximum inhibition exhibited by the prepared nanoparticles were observed as LyGNP 83.45 ± 0.4694%, LyAgNP 88.05 ± 0.1870% and LyAgNP65.47 ± 0.4766%. We conclude that the developed nanoparticles of Lycopene exhibited potential anticancer activity against Colorectal and cervical cancer cell as compared with pure Lycopene.

GREEN SYNTHESIS OF GOLD NANOPARTICLES OF ISOLATED CITRUS BIOFLAVONOID FROM ORANGE: CHARACTERIZATION AND IN VITRO CYTOTOXICITY AGAINST COLON CANCER CELL LINES COLO 320DM AND HT29

The aim of the present research was to perform green synthesis of gold nanoparticles of isolated citrus bioflavonoid from Citrus sinensis (orange) peel extract and to evaluate its anticancer potential. Methanolic extract of orange peel was obtained by Soxhlet extraction and citrus bioflavonoid was isolated by using column chromatography. Gold nanoparticles were developed by green synthesis method, wherein 1 % aqueous solution of isolated citrus bioflavonoid were mixed with 1% solution of HAuCl4 and incubated at ambient temperature for 4 to 5 hours and observed for the color change which indicated formation of nanoparticles. Obtained gold nanoparticles were evaluated by UV visible spectra, FTIR, SEM, XRD analysis and for antimicrobial potential against E coli, S.aureus and P. aeruginosa. Cytotoxicity study was carried out by using in vitro assays, namely MTT, SRB and Tryphan blue assay, against colon cancer cell line Colo 320 DM, and HT 29. results of SEM showed that nanosized particles in the range of 80-100nm were formed. Results of cytotoxicity studies revealed that CBFGNP exhibited 72.28% inhibition, against Colo320 DM whereas pure CBF showed 70.46% inhibition. Against HT 29, CBFGNP exhibited 69.79% inhibition, whereas for MTT assay and SRB assay, CBFGNP showed 80.15% and 58.29% inhibition, respectively. Moreover, CBFGNP exhibited 90.29% and 85% non viability against Colo320 DM and HT29. Based on the results, it can be concluded that gold nanoparticles of citrus bioflavonoid (CBFGNP) exhibits more cytotoxicity against Colo320 DM and HT29 as compared to pure citrus bioflavonoid when assessed by three different in vitro cytotoxicity assays.

Evaluation of Polyherbal Anticancer Tablets: A Review

Cancer is a malignant abnormal growth of cells, one of the most dreaded and complex diseases. It concerns with several tempo spatial changes in cell composition, which finally lead to neoplasia. Various types of cancers have been reported. Chemotherapy, radiation, and/or surgery may cure them. Herbal remedies are supposed to be harmless as they cause fewer complications and are less likely to habitual. Antioxidant compositions of therapeutic plants show the anticancer activity and therefore, use of different proportions of the active components to formulate various standardized preparation with single or multiple components for their synergistic effects play a crucial role in curing cancer. Evaluation parameters to assess the in vitro anticancer activity includes Caspase-3, Caspase-9, alamar blue, LDH assay, XTT assay, sulforhodamine-B assay, MTT assay, DNA fragmentation assay, neutral red uptake cytotoxic assay, tryphan blue assay. Evaluation of dried extract or granules includes bulk density, tapped density, Carr’s index, Hausner’s ratio, angle of repose while the tablets evaluated by drug-excipient compatibility study by FT-IR, stability studies, hardness, thickness, weight variation, friability, disintegration time and dissolution test.

Ca2+ mobilization induced by W-7 in MG63 human osteosarcoma cells

The effect of N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (W-7), a widely used calmodulin inhibitor, on intracellular free Ca2+levels ([Ca2+]i) in MG63 human osteosarcoma cells was explored using fura-2 as a Ca2+probe. W-7 (20–1000 μ m) induced an increase in [Ca2+]iin a dose-dependent manner, with an EC50of 100 μ m. The [Ca2+]isignal comprised an initial rise and a sustained plateau without significant decay within 5 min. External Ca2+removal decreased the Ca2+signals by reducing the peak and sustained phase, indicating W-7-activated intracellular Ca2+release and extracellular Ca2+influx. W-7 (500 μ m) failed to induce a [Ca2+]iincrease in a Ca2+-free medium after pre-treatment with thapsigargin (1 μ m), an endoplasmic reticulum Ca2+pump inhibitor. Conversely, W-7 pre-treatment abolished the Ca2+release induced by thapsigargin. This suggests that W-7 (500 μ m) released internal Ca2+mainly from the endoplasmic reticulum. The addition of 3 mm Ca2+increased [Ca2+]idose-dependently after preincubation with 20–1000 μ m W-7 in a Ca2+-free medium, implying that W-7 induced capacitative Ca2+entry. W-7-induced Ca2+release was not altered by inhibiting phospholipase C with 2 μ m 1-(6-((17β - 3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione) (U73122). Tryphan blue assay demonstrated that W-7 (200 μ m) caused gradual cell death within 30 min of the initial drug exposure. Together, it was found that W-7 induced [Ca2+]iincreases in human osteosarcoma cells by releasing internal Ca2+from the endoplasmic reticulum, and also by triggering Ca2+influx. W-7 may be cytotoxic to osteosarcoma cells.

봉독약침(蜂毒藥鍼)의 항암효과(抗癌效果)에 대한 분자생물학적(分子生物學的) 연구(硏究)

To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability and apoptosis were analyzed using MTT assay, tryphan blue assay, thymidine release assay, flow cytometric analysis, and activity of caspase-3 protease activity assay. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, thymidine release assay, and flow cytomet1 ric analysis of sub fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and Bcl- were down-regulated whereas Bax was up-regulated by bee venom treatment.