Morphological Estimation of DMPC/DHPC Self-Assemblies in Diluted Condition: Based on Physicochemical Membrane Properties

Self-assembly membranes, composed of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC), were characterized at the total lipid concentration below 20 mM. The sizes of the assemblies varied depending on the molar ratio of DMPC and DHPC (q = [DMPC]/[DHPC]). The small assemblies with diameter of ca. 10 nm were formed at q ≤ 2.0 at 20 ºC (below phase transition temperature of DMPC). The physicochemical membrane properties were then studied using fluorescence probes, 1,6-diphenyl-1,3,5-hexatriene and 6-dodecanoyl-N,N-dimethyl-2-naphthylamine, upon the dilution. DHPC micelle showed a higher membrane fluidity, while the DMPC/DHPC membranes at q ≥ 0.5 showed lower membrane fluidities as well as DMPC vesicle in gel (ordered) phase. Upon dilution, the ordered membrane properties were maintained while the solution turbidities increased, implying the morphological change of the self-assembly, bicelle to the vesicle in gel phase. Based on the obtained results, a phase diagram of DMPC/DHPC binary system (at 20 ºC) is described: (i) the bicelle suspension is transparent and the membrane is in ordered state, (ii) the micelle suspension is transparent and the membrane is in disordered state, (iii) the vesicle suspension is turbid and the membrane is in ordered state.

STUDY ON CAUSE–EFFECT RELATIONS AND OPTIMIZATION OF EXEMESTANE-LOADED NANOSTRUCTURED LIPID CARRIERS

Objective: Exemestane is an anti-breast cancer drug, possesses low water solubility and low permeability. This work aimed at the cause-effect relations and optimization of exemestane-loaded nanostructured lipid carriers (EXE-NLCs) for oral delivery.Methods: Excipient screening was based on exemestane solubilities and the emulsification efficiency of surfactants. A D-optimal design based on three independent variables was applied to evaluate the cause-effect relations and optimise EXE-NLCs formulation. The particle size (PS), polydispersity index (PDI), entrapment efficiency (EE) and drug loading (DL) were investigated with respect to three independent variables including liquid lipid to total lipid ratio (X1), surfactant concentration (X2), total lipid concentration (X3).Results: EXE-NLCs were prepared by a hot sonication method employing Labrafac CC and Compritol 888ATO as liquid and solid lipids, respectively, and Cremophor RH40 as a surfactant and Lutrol E-400 as a co-surfactant. All investigated factors: liquid lipid to total lipid ratio, surfactant concentration and total lipid concentration showed significant influences on physicochemical characteristics of EXE-NLCs. The optimal EXE-NLC formulation was composed of liquid lipid to total lipid ratio (X1) of 24 % (w/w), surfactant concentration (X2) of 4 % (w/v) and total lipid concentration (X3) of 4 % (w/v). The PS, PDI, EE and DL of the optimized EXE-NLCs were found to be 41.787 nm; 0.11; 97.605 % and 1.935 %, respectively. The optimized formulation was experimentally examined which demonstrated a good agreement between experimental and predicted values.Conclusion: The cause-effect relations and optimization of EXE-NLCs were investigated and reported for the first time. EXE-NLCs formulation was successfully optimized using D-optimal design and merits further study.

Preparation, Optimization and Characterization of Ketoprofen Proniosomes for Transdermal Delivery

The aim of the present study was to prepare, optimize and characterize ketoprofen proniosomes. The niosomes were prepared using a slurry method followed by in vitro evaluation after embedding the proniosomes-derived niosomes into a carbopol matrix. A central, composite Box-Wilson design was used for the optimization with the total lipid concentration (X1), surfactant loading (X2) and amount of drug (X3) as the independent variables. Prepared proniosomes were characterized for percentage drug entrapment (PDE) and mean volume diameter (MVD). Multiple regression analysis and contour plots were used to relate the dependent and independent variables. Checkpoint batches were also prepared to prove the validity of the evolved mathematical model and contour plots. The optimization model predicted the levels of X1, X2 and X3 (-1, -0.3 and 0.92, respectively), for a maximized response of PDE with constraints of £ 5 mm on MVD. Optimized batch was used to prepare a niosomal gel, which showed significantly higher cumulative amount of drug permeated and steady state transdermal flux compared to plain gel. This work has demonstrated the use of the central composite Box-Wilson design, regression analysis, and contour plots in optimizing ketoprofen proniosomes. Developed niosomal gel formulation has also demonstrated permeation enhancement of ketoprofen compared to plain gel.

Transfer of n–3 and n–6 polyunsaturated fatty acids from yolk to embryo during development of the king penguin

This study examines the transfer of lipids from the yolk to the embryo of the king penguin, a seabird with a high dietary intake of n–3 fatty acids. The concentrations of total lipid, triacylglycerol (TAG), and phospholipid (PL) in the yolk decreased by ∼80% between days 33 and 55 of development, indicating intensive lipid transfer, whereas the concentration of cholesteryl ester (CE) increased threefold, possibly due to recycling. Total lipid concentration in plasma and liver of the embryo increased by twofold from day 40 to hatching due to the accumulation of CE. Yolk lipids contained high amounts of C20–22 n–3 fatty acids with 22:6(n–3) forming 4 and 10% of the fatty acid mass in TAG and PL, respectively. Both TAG and PL of plasma and liver contained high proportions of 22:6(n–3) (∼15% in plasma and >20% in liver at day 33); liver PL also contained a high proportion of 20:4(n–6) (14%). Thus both 22:6(n–3) and 20:4(n–6), which are, respectively, abundant and deficient in the yolk, undergo biomagnification during transfer to the embryo.

Quantification of lipid in cultured 3T3-L1 adipocytes

A preliminary study was conducted to quantify the lipid produced by differentiated 3T3-L1 cells after incubation in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% foetal bovine serum (FBS), supplemented with or without dimethyl-sulphoxide (DMSO; 9•6 g/l) and acetone (1•2 g/l). The two media treatments were applied to 3T3-L1 cells, plated at either 15K or 30K cells per well in 24-well plates. Cells were grown to confluence (96 h) and then treated with dexamethasone, methyl-isobutylxanthine and insulin for 48 h and later maintained in their respective media treatments for another 144 h. Cells from each treatment were recovered after two, 5-min incubations with trypsin, washed and resuspended in DMEM and counted on a haemocytometer. The lipid in the cells was extracted with hexane derivatized with tetramethyl-guanidine and analysed by gas chromatography. Final mean cell density was 6•8 (s.e. 0•18) ✕ 105 and 4•6 (s.e. 0•19) ✕ 105 when initially plated at 30K and 15K cells per well, respectively. Inclusion of DMSO and acetone in the medium did not affect final cell numbers. Plating density did not affect concentration of lipid (0•55 (s.e. 0•08) mg per 1 ✕ 105 cells) but inclusion of DMSO and acetone led to overall decreases in total lipid concentration. Results indicate that initial plating density influenced final cell number in treatment cultures, but that DMSO and acetone treatments only had an effect on final lipid concentration. Collectively, these data suggest that the application of treatments to cell cultures may be influenced by the carrier vehicle that the treatment is contained in and this should be considered when developing an in vitro system to evaluate growth and development of adipocytes.