Probing nanoliposomes using single particle analytical techniques: effect of excipients, solvents, phase transition and zeta potential.

There has been a steady increase in the interest towards employing nanoliposomes as colloidal drug delivery systems, particularly in the last few years. Their biocompatibility nature along with the possibility of encapsulation of lipid-soluble, water-soluble and amphipathic molecules and compounds are among the advantages of employing these lipidic nanocarriers. A challenge in the successful formulation of nanoliposomal systems is to control the critical physicochemical properties, which impact their in vivo performance, and validating analytical techniques that can adequately characterize these nanostructures. Of particular interest are the chemical composition of nanoliposomes, their phase transition temperature, state of the encapsulated material, encapsulation efficiency, particle size distribution, morphology, internal structure, lamellarity, surface charge, and drug release pattern. These attributes are highly important in revealing the supramolecular arrangement of nanoliposomes and incorporated drugs and ensuring the stability of the formulation as well as consistent drug delivery to target tissues. In this article, we present characterization of nanoliposomal formulations as an example to illustrate identification of key in vitro characteristics of a typical nanotherapeutic agent. Corresponding analytical techniques are discussed within the context of nanoliposome assessment, single particle analysis and ensuring uniform manufacture of therapeutic formulations with batch-to-batch consistency.

Optimization on preparation condition of polyunsaturated fatty acids nanoliposome prepared by Mozafari method.

This study presents the application of the response surface methodology (design) to develop an optimal preparation condition (independent variables) namely shear rate (600-1000 rpm), mixing time (30-60 min), and sonication time (10-20 min) for polyunsaturated fatty acids (docosahexaenoic acid and eicosapentaenoic acid) nanoliposomes. Fifteen lipid mixtures were generated by the Box-Behnken design and nanoliposomes were prepared by the Mozafari (direct hydration and without using organic solvents) method. Nanoliposomes were characterized with respect to entrapment efficiency (EE) and vesicle size as Y1 and Y2 dependent variables, respectively. The results were then applied to estimate the coefficients of response surface model and to find the optimal preparation conditions with maximum EE and minimum vesicle size. The response surface analysis exhibited that the significant (p < 0.05) second-order polynomial regression equations were successfully fitted for all dependent variables with no significant (p > 0.05) lack of fit for the reduced models. The response optimization of experiments was the shear rate: 795 rpm; mixing time: 60 min; and sonication time: 10 min. The optimal nanoliposome had an average diameter of 81.4 nm and EE of 100%. The experimental results of optimal nanoliposomes characterization confirmed an accurate fitness of the predicted values by reduced response surface models.

Comparative study of the oxidative and physical stability of liposomal and nanoliposomal polyunsaturated fatty acids prepared with conventional and Mozafari methods.

The relative oxidative stability of freshly prepared and stored liposomal and nanoliposomal systems of docosahexaenoic acid (DHA, 22:6 n-3) and eicosapentaenoic acid (EPA, 20:5 n-3) were investigated. The effects of organic solvents on the oxidative stability of liposomal polyunsaturated fatty acids (PUFAs) produced by two methods, the Bangham thin-film hydration (conventional rotary evaporation method and using organic solvents) and Mozafari (direct hydration and without using organic solvents) methods, were compared. The highest physicochemical stability was observed in PUFA liposomes prepared by the Mozafari method, followed by conventional liposomes and bulk PUFAs. There was no significant change in physicochemical stability during 10 months of cold storage (4°C) in the dark. Moreover, the comparison between liposomes (>200 nm) and nanoliposomes (50-200 nm) revealed that the surface charge, physical stability and oxidative stability of liposomal PUFAs increased as the size of the liposomes decreased. The differences in the oxidative stability of PUFAs may be due to the protective effects of aqueous systems, which indicate the advantage of using non-organic solvent (water and CO(2)) techniques in liposome manufacturing.

Gastrointestinal tract morphological alteration by unpleasant physical treatment and modulating role of Lactobacillus in broilers.

1. An experiment was conducted to determine the effects of supposedly unpleasant physical treatment on broiler performance, small intestinal development and ameliorating role of probiotics. 2. The following treatments were applied from day one: (1) chicks exposed to normal human contact fed basal diet (control); (2) chicks were exposed to unpleasant physical treatment and fed basal diet (UPT-BD); and (3) chicks were exposed to unpleasant physical treatment and fed basal diet supplemented with Lactobacillus (UPT-BDL). Chicks were exposed to UPT from days 1 to 21. Different segments of gastrointestinal tract were sampled at 14, 28, 35 and 42 d of age. 3. Broilers of UPT-BD had lower feed consumption compared with control group at 7 d of age. Overall, UPT-BDL birds showed higher body weight gain (BWG) and better feed conversion ratio (FCR) over the course of the experiment. 4. Birds of UPT-BD had lower concentrations of lactic, propionic and butyric acids in the caecum as compared with other groups at 14 d of age. Acetic acid concentration was profoundly decreased in both UPT groups compared to the control. 5. Duodenal villus height of UPT-BD broilers showed a slight reduction compared to the control and UPT-BDL birds at 14 d of age. Afterwards until day 42, UPT-BDL birds showed the highest villus height among treatments in different parts of the small intestine. 6. The results suggested that, even though UPT did not have significant inhibitory effects on the development of the small intestine and broiler performance, it negatively affected bacterial metabolic end products in the caecum, which could be ameliorated by the addition of Lactobacillus.

In vitro fermentation of broiler cecal content: the role of lactobacilli and pH value on the composition of microbiota and end products fermentation.

AIM: To assess the probiotic effects of Lactobacillus agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230 and the pH on the cecal microflora of chicken and metabolic end products. METHODS AND RESULTS: An in vitro system, operated with batch bioreactor, was used for this assessment. Selected bacterial species were monitored at two pH values, over 24 h of batch culture incubation. The concentration of short chain fatty acids (SCFA) and lactate in the fermented material was also determined. The addition of L. agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230 into vessel 2 (Cc + P) increased the total anaerobes, lactobacilli and bifidobacteria after 24 h incubation. Moreover, lactobacilli supplementation decreased the total aerobes and streptococci, but it did not have any effects on coliforms. The supplementation of lactobacilli in vessel 2 (Cc + P) was found to significantly increase the production of lactate, propionate and butyrate. Furthermore, pH did not alter the formation of butyrate, whereas the production of acetate and propionate was significantly decreased at pH = 5.8. CONCLUSIONS: L. agilis JCM 1048 and L. salivarius ssp. salicinius JCM 1230, as probiotic bacteria, have the ability to re-establish proper microbial balance by the formation of lactate as well as propionate, and stimulate butyrate-producing bacteria to produce butyrate in the chicken cecum. SIGNIFICANCE AND IMPACT OF THE STUDY: This study was the first to report this under in vitro conditions, highlighting the probiotic roles of the two Lactobacillus strains in broiler cecal fermentation at different initial pH. These useful data can be helpful in improving the fermentation process in chicken cecum.