Each experiment was repeated broadcast in triplicate and the mean values and standard deviations were calculated. 2.6. Stability tsstudien Of the physical and chemical stability t of mixed micelles over a period of 45 days. Stability tsstudien MPC-3100 Were carried out again dispersed w Bombers loaded mixed micelles DSPE PEG2000 / Vitamin E TPGS with both paclitaxel and parthenolide, at each drug loading of 2.3%, to a final concentration of 0, 1% for each drug in the dispersion . Stability Tsstudien of mixed micelles from the visual observation for signs of growth or Kristallausf Precipitation, particle Enanalyse and HPLC analysis for the amount of drug that are enclosed in micelles. Micellar dispersions were more for the 1H NMR signals for inspection free of drugs including the UN.
The absence of free drug was determined by analysis of samples of the filtrate centrifuged with Amicon Ultra centrifugal best units with a 30,000 MWCO CONFIRMS. The chemical stability of t was determined by HPLC analysis by examining the development of Peakfl Surface, shape, residence time and the presence of degradation peaks in the chromatograms of the mixed micellar dispersions containing paclitaxel shops protected and parthenolide. The results were the averages of three replicates. 2.7. The ability Lebensf The cell studies in vitro studies of Lebensf Ability of the cells were performed on A549 taxol taxol sensitive and resistant lines T24 A549 human lung adenocarcinoma cells. A549 cells were obtained from American Type Culture Collection. Taxol-resistant A549-T24 cell clone was a big generous donation from Dr.
Susan Horwitz at the Albert Einstein College of Medicine. The cells were f in medium with 12 FK heat-inactivated 10% Fetal calf serum K And 1% penicillin-streptomycin erg Held complements. For the Lebensf Ability of the cells studies the cells were incubated in monolayers in tissue culture flasks at C in an atmosphere of 5% CO 2 Re and cultured 90% relative humidity. For the Lebensf Ability test were added, the cells on a 96-well plate at a cell density of 5000 cells / well. The number of cells / well was calculated against a calibration curve by plating various concentrations of cells, prepared as determined by hemocytometry at the beginning of each experiment.
The growth medium was removed after 24 h incubation and liberated by 100 LL fresh medium containing various concentrations of free paclitaxel, parthenolide, combination or their corresponding micellar formulations. L Solutions of paclitaxel and parthenolide-free or a combination thereof produced by first Highest the drug, alone or in combination, in a mixture of 1:01 Cremophor EL and ethanol and then diluting the mixture in a culture medium. The plates were then 48 h, after which the Lebensf Ability of the cells using the test-3 2,5 diphenyl tetrazolium color was incubated. The optical density of each sample was measured at 570 nm using a microplate Leseger t. Rst Were the dose-response of the free paclitaxel or parthenolide in L Generated solution at concentrations in the range of 37.5 to 1000 nm for paclitaxel and 5 to 20 lm parthenolide. Then, the cells with a combination of parthenolide and paclitaxel at concentrations in the range of 37.5 to 1000 nM. Once the optimum concentration of active substance was identified in the combination, the cells were treated