Design and Optimization of Solid Lipid Nanoparticles Loaded with Triamcinolone Acetonide.

Principles of quality by design and design of experiments are acquiring more importance in the discovery and application of new drug carriers, such as solid lipid nanoparticles. In this work, an optimized synthesis of solid lipid nanoparticles loaded with Triamcinolone Acetonide is presented using an approach that involves Stearic Acid as a lipid, soy PC as an ionic surfactant, and Tween 80 as a nonionic surfactant. The constructed circumscribed Central Composite Design considers the lipid and nonionic surfactant quantities and the sonication amplitude in order to optimize particle size and Zeta potential, both measured by means of Dynamic Light Scattering, while the separation of unentrapped drug from the optimized Triamcinolone Acetonide-loaded solid lipid nanoparticles formulation is performed by Size Exclusion Chromatography and, subsequently, the encapsulation efficiency is determined by HPLC-DAD. The proposed optimized formulation-with the goal of maximizing Zeta potential and minimizing particle size-has shown good accordance with predicted values of Zeta potential and dimensions, as well as a high value of encapsulated Triamcinolone Acetonide. Experimental values obtained from the optimized synthesis reports a dimension of 683 ± 5 nm, which differs by 3% from the predicted value, and a Zeta potential of -38.0 ± 7.6 mV (12% difference from the predicted value).

Lipid-Stabilized Water-Oil Interfaces Studied by Microfocusing Small-Angle X-ray Scattering.

Water-in-oil (w/o) simple emulsions are dispersed microconfined systems that find applications in many areas of advanced materials and biotechnology, such as the food industry, drug delivery, and cosmetics, to name but a few. In these systems, the structural and chemical properties of the boundary layer at the w/o interface are of paramount importance in determining functionality and stability. Recently, microfluidic methods have emerged as a valuable tool for fabricating monodisperse emulsion droplets. Because of the intrinsic flexibility of microfluidics, different interfaces can be obtained, and general principles governing their stability are needed to guide the experimental approach. Herein, we investigate the structural characteristics of the region encompassing the liquid/liquid (L/L) interface of w/o emulsions generated by a microfluidic device in the presence of phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and other intercalating amphiphiles (dopants) using microfocused small-angle X-rays scattering (µ-SAXS). We show that phospholipids provide a stable and versatile boundary film of ∼100 µm whose basic units are swollen and uncorrelated DMPC bilayers. The internal arrangement of this interfacial film can be tuned by adding molecules with a different packing parameter, such as cholesterol, which is able to increase the stiffness of the lipid membranes and trigger interbilayer correlation.

Physiochemical Characterization of Lipidic Nanoformulations Encapsulating the Antifungal Drug Natamycin.

Natamycin is a tetraene polyene that exploits its antifungal properties by irreversibly binding components of fungal cell walls, blocking the growth of infections. However, topical ocular treatments with natamycin require frequent application due to the low ability of this molecule to permeate the ocular membrane. This limitation has limited the use of natamycin as an antimycotic drug, despite it being one of the most powerful known antimycotic agents. In this work, different lipidic nanoformulations consisting of transethosomes or lipid nanoparticles containing natamycin are proposed as carriers for optical topical administration. Size, stability and zeta potential were characterized via dynamic light scattering, the supramolecular structure was investigated via small- and wide-angle X-ray scattering and 1H-NMR, and the encapsulation efficiencies of the four proposed formulations were determined via HPLC-DAD.

Green Hydrogels Loaded with Extracts from Solanaceae for the Controlled Disinfection of Agricultural Soils.

The UN 2030 Agenda for Sustainable Development established the goal of cutting the use of pesticides in the EU by 50% by 2030. However, a ban on pesticides could seriously affect the productivity of agriculture, resulting in severe issues due to global hunger and food deficiency. Controlled release (CR) of bioactive chemicals could play a valid alternative in this context. To this aim, two biodegradable polymers, namely sodium alginate (AL) and sodium carboxymethylcellulose (CMC), were employed to obtain crosslinked hydrogel beads for the encapsulation and CR of glycoalkaloids extracted from tomato and potato leaves to be used as biocompatible disinfectants for agricultural soils. The physico-chemical characterization of the controlled-release systems was carried out by means of Attenuated Total Reflectance-Fourier Transform Infrared (ATR-FTIR) spectroscopy, Scanning Electron Microscopy (SEM), thermogravimetry (TGA), differential scanning calorimetry (DSC) (FWI > 80%) and drying kinetics. The plant extracts and the encapsulation efficiency (~84%) were, respectively, characterized and evaluated by High-performance Liquid Chromatography-Mass Spectrometry (HPLC-MS). Finally, preliminary microbiological tests were conducted to test the efficacy of the most promising systems as biocidal formulations both in the lab and on a model soil, and interesting results were obtained in the reduction of bacterial and fungal load, which could lead to sustainable perspectives in the field.

Structuring and De-Structuring of Nanovectors from Algal Lipids: Simulated Digestion, Preliminary Antioxidant Capacity and In Vitro Tests.

Biocompatible nanocarriers can be obtained by lipid extraction from natural sources such as algal biomasses, which accumulate different lipid classes depending on the employed culture media. Lipid aggregates can be distinguished according to supramolecular architecture into lamellar and nonlamellar structures. This distinction is mainly influenced by the lipid class and molecular packing parameter, which determine the possible values of interfacial curvature and thus the supramolecular symmetries that can be obtained. The nanosystems prepared from bio-sources are able to self-assemble into different compartmentalized structures due to their complex composition. They also present the advantage of increased carrier-target biocompatibility and are suitable to encapsulate and vehiculate poorly water-soluble compounds, e.g., natural antioxidants. Their functional properties stem from the interplay of several parameters. Following previous work, here the functionality of two series of structurally distinct lipid nanocarriers, namely liposomes and cubosomes deriving from algal biomasses with different lipid composition, is characterized. In the view of their possible use as pharmaceutical or nutraceutical formulations, both types of nanovectors were loaded with three well-known antioxidants, i.e., curcumin, α-tocopherol and piperine, and their carrier efficacy was compared considering their different structures. Firstly, carrier stability in biorelevant conditions was assessed by simulating a gastrointestinal tract model. Then, by using an integrated chemical and pharmacological approach, the functionality in terms of encapsulation efficiency, cargo bioaccessibility and kinetics of antioxidant capacity by UV-Visible spectroscopy was evaluated. Subsequently, in vitro cytotoxicity and viability tests after administration to model cell lines were performed. As a consequence of this investigation, it is possible to conclude that nanovectors from algal lipids, i.e., cubosomes and liposomes, can be efficient delivery agents for lipophilic antioxidants, being able to preserve and enhance their activity toward different targets while promoting sustained release.

Lipids from algal biomass provide new (nonlamellar) nanovectors with high carrier potentiality for natural antioxidants.

Lipid mesophases are lyotropic liquid crystalline systems which differ from liposomes and other globular aggregates in dilute regimes due to their inner ordering. It is known that natural lipids enable to obtain a rich variety of nanosystems and many of them have been proposed as delivery agents for bioactive compounds. Due to their packing parameters, several classes of lipids found in natural sources are able to self-assemble into nonlamellar structures. Among lipids occurring in plants and algae, triglycerides display this tendency. In the present study we examine new nanosystems built with lipids extracted from the marine microalga Nannochloropsis sp and their use as carriers for lipophilic antioxidants. The antioxidants studied, curcumin and tocopherol were encapsulated with high rate in the carriers. The physico-chemical characterization of plain and loaded vectors showed their structure and localization site, as well as the structure-functionality relationship related to potential drug delivery. The results show that the cargo molecules play an active role in driving the interactions which characterize the overall structure of the aggregates. The systems studied showed several coexisting mesophases, the most predominant structure being of cubic symmetry.

A new method for the direct tracking of in vivo lignin nanocapsules in Eragrostis tef (Poaceae) tissues.

Environmental concerns have driven scientists to research new eco-friendly approaches for the preparation of nanosystems. For this purpose, novel bio-polymers have been selected. Among these, one of the most promising is lignin, which is biodegradable and biocompatible. Additionally, lignin is one of the main by-products of the paper industry and can be re-used in nanosystems building. Lignin-based nanosystems could be used in agriculture, to improve the uptake of bioactive compounds, thus avoiding soil pollution. However, the mechanism of penetration in the plant and the route of transportation within the internal plant tissues are unknown and need to be clearly elucidated. Here we present a method of lignin nanocapsules staining and tracking by fluorochrome: Fluoral Yellow 088, which is a well-suited dye for the tracking of lipids and other oil phases. Two different applications were applied: in the first one fourteen-day plants were soaked with fluorescent nanocapsules (fNCs) pure solution and in the second one, Eragrostis tef plants were laid down on blotting paper and soaked with diluted fNCs solution. Wetting the roots of Teff plantlets with the pure fNCs solution resulted in the most efficient way of nanocapsule entrance. The dyeing of lignin nanocapsules allowed us to track them in Eragrostis tef plant tissues through microscopic observations. In particular, fNCs were proven to be able to permeate roots, reaching xylem vessels where, through water pressure, they reached the leaf.

Exploring the water/oil/water interface of phospholipid stabilized double emulsions by micro-focusing synchrotron SAXS.

Surfactant stabilized water/oil/water (w/o/w) double emulsions have received much attention in the last years motivated by their wide applications. Among double emulsions, those stabilized by phospholipids present special interest for their imitation of artificial cells, allowing the study of the effect of confining chemical reactions in biomimetic environments. Upon evaporation of the oil shell, phospholipid stabilized double emulsions can also serve as templates for giant vesicles. In this context, general assumptions have been made on the self-assembly and structural organization/arrangement of amphiphilic molecules, at the aqueous/oil liquid interface. However, to the best of our knowledge, no detailed evidence of the interfacial structuring have been reported. In this paper, w/o/w double emulsions formulated using the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and a mixture of chloroform and cyclohexane as the oil phase were produced using a microfluidic device. To obtain information on the phospholipid arrangement, the w/o/w interface was investigated by spatially resolved micro-focusing SAXS. We observed that (i) the basic units forming both the w/o and o/w interfaces were oil-swollen DMPC bilayers, arranged into a substantially disordered shell of ∼45 µm thickness surrounding the internal oil phase; (ii) the evaporation process was slow, i.e. in the order of one hour at 50 °C and (iii) oil evaporation led to a shrinkage of the interfacial shell, but not to an increase of the ordering of the lipid bilayers. Interestingly, no stacked DMPC bilayers were observed during the evaporation process, as shown by the absence of Bragg's peaks in the SAXS intensity profiles.