Cells
were maintained in Dulbecco’s minimum essential medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 50μg/mL gentamycine (Invitrogen Argentina), and 2mM L-glutamine (Invitrogen Argentina). Cells were cultured in 75cm2 culture flasks at 37°C in a humidified atmosphere of 5% CO2. 2.9. Cytotoxicity and In Vitro Performance of the Selected TMX-Loaded Inhibitors,research,lifescience,medical MEs For in vitro performance studies, cells were seeded in 96-well plates at a density of 5,000 cells/well. After 24 hours, medium was replaced by phenol-red-free media containing 2mM L-glutamine for 24 hours. To analyze effects of selected TMX-loaded formulations, cells were subsequently incubated with estradiol 10nM and the TMX-loaded MEs; in parallel, a TMX suspension containing 10mM of drug
in presence and in absence of estradiol was also evaluated. Cells were incubated Inhibitors,research,lifescience,medical further for 48 hours and then cell viability was assessed by the cell proliferation assay (MTS). 2.10. Statistical Analysis Statistical calculations were performed with the GraphPad InStat statistical package for Windows. Data shown in tables and figures Inhibitors,research,lifescience,medical of in vitro properties evaluation represent mean of three determinations ± standard deviation (SD). Statistical significance of the differences between the groups was calculated by the Tukey-Kramer multiple comparison test and probability value of P smaller than 0.05 indicated a statistically significant difference. 3. Results and Discussion 3.1. Preliminary Solubility Evaluation TMX resulted almost insoluble in IPM, Mygliol 840, Captex 355, Oleic acid, and Imwitor 408 and showed solubility near 20mg/g in the PC suspension and in Capmul MCM L (Figure 2(a)). Therefore, only Inhibitors,research,lifescience,medical PC and Capmul MCM L were selected for the forthcoming screening. The selection of the oily phase is very important because the drug solubility in the formulation depends mainly on it [23, 24].
So, this property results, fundamental in the search for high solubilizing Inhibitors,research,lifescience,medical capacity systems. Figure 2 (a) Solubility of Tamoxifen citrate in oil phases (expressed in mg/g). Each bar represents the mean of three samples ± SD. (b) Solubility of Tamoxifen Citrate in Polysorbate 80 and cosurfactants (expressed in mg/g). Each bar represents the mean … Lipid solubility values found in this work are in accordance with previous studies and significantly higher compared to other lipids Batimastat not considered in this study [25]. They also were significantly higher than TMX solubility in water (≈20mg/mL and ≈0.4μg/mL, resp.). Furthermore, the high solubility in PC is in accordance with previous works [26], which stated that active compounds with an intermediate lipophilicity (Log P of 4.0 and above, being 7.9 the value of the Tamoxifen) have a high tendency to be solubilized by phospholipids. Solubility of TMX in the five co-surfactants and in PS 80 is depicted in Figure 2(b).