Carlos E. Astete

Synthesis and characterization of PLGA nanoparticles

Poly(lactide-co-glycolide) (PLGA) nanoparticles of different physical characteristics (size, size distribution, morphology, zeta potential) can be synthesized by controlling the parameters specific to the synthesis method employed. The aim of this review is to clearly, quantitatively and comprehensively describe the top–down synthesis techniques available for PLGA nanoparticle formation, as well as the techniques commonly used for nanoparticle characterization. Many examples are discussed in detail to provide the reader with an extensive knowledge base on the important parameters specific to the synthesis method described and ways in which these parameters can be manipulated to control the nanoparticle physical characteristics.

Nanoparticles with entrapped α-tocopherol: synthesis, characterization, and controlled release

An emulsion evaporation method was used to synthesize spherical poly(DL-lactide-co-glycolide) (PLGA) nanoparticles with entrapped α-tocopherol. Two different surfactants were used: sodium dodecyl sulfate (SDS) and poly(vinyl alcohol) (PVA). For SDS nanoparticles, the size of the nanoparticles decreased significantly with the entrapment of α-tocopherol in the PLGA matrix, while the size of PVA nanoparticles remained unchanged. The polydispersity index after synthesis was under 0.100 for PVA nanoparticles and around 0.150 for SDS nanoparticles. The zeta potential was negative for all PVA nanoparticles. The entrapment efficiency of α-tocopherol in the polymeric matrix was approximately 89% and 95% for nanoparticles with 8% and 16% α-tocopherol theoretical loading, respectively. The residual PVA associated with the nanoparticles after purification was approximately 6% ( w/w relative to the nanoparticles). The release profile showed an initial burst followed by a slower release of the α-tocopherol entrapped inside the PLGA matrix. The release for nanoparticles with 8% α-tocopherol theoretical loading (86% released in the first hour) was faster than the release for the nanoparticles with 16% α-tocopherol theoretical loading (34% released in the first hour).

Ca2+ Cross-Linked Alginic Acid Nanoparticles for Solubilization of Lipophilic Natural Colorants

The increased tendency toward healthy lifestyles has promoted natural food ingredients to the detriment of synthetic components of food products. The trend followed into the colorant arena, with consumers worried about potential health problems associated with synthetic colorants and demanding food products that use natural pigments. The goal of this study was to entrap a lipophilic natural pigment (beta-carotene) in a water-soluble matrix made of Ca(2+) cross-linked alginic acid, to allow its use as a colorant in water-based foods. The effects of different synthesis parameters such as type of solvent, alginic acid concentration, and calcium chloride concentration on nanoparticle characteristics (i.e., size, zeta potential, and morphology) were evaluated. The particle stability was assessed by measuring aggregation against pH, oxidation, and particle precipitation as a function of time. The particle synthesized measured 120-180 nm when formed with chloroform and 500-950 nm when synthesized with ethyl acetate. The particles were negatively charged (-70 to -80 mV zeta potential) and were stable at pH values ranging from 3 to 7. The presence of calcium was prevalent on the particles, indicating that the divalent ions were responsible for cross-linking lecithin with alginic acid and forming the matrix around the beta-carotene pockets. The addition of calcium increased nanoparticle density and improved beta-carotene protection against oxidation. It is concluded that the method proposed herein was capable of forming water-soluble nanoparticles with entrapped beta-carotene of controlled functionality, as a result of the type of solvent and the amounts of alginate and Ca(2+) used.

Chitosan/PLGA particles for controlled release of α-tocopherol in the GI tract via oral administration

AIM The physiochemical properties, controlled release characteristics, stability and cellular uptake of chitosan (Chi)/poly(D,L-lactide-co-glycolide) (PGLA) and PLGA particles with entrapped α-tocopherol were investigated to understand the behavior of these nanoparticles in the GI tract. MATERIALS & METHODS Chi/PLGA and PLGA particles stabilized by lecithin were synthesized and fully characterized for oral gastrointestinal delivery via transmission electron microscopy, dynamic light scattering, high-performance liquid chromatography and fluorescence microscopy. RESULTS Particle stability was pH- and system-dependent. In vitro release profiles showed a higher percentage of drug released in the intestinal domain by Chi/PLGA as opposed to the PLGA nanoparticles. Fluorescent counterparts of these particles were confirmed to associate with the surface of the intestinal villi, and penetrate deep in the endothelial lining of rabbit intestinal explants, indicating uptake. CONCLUSION In vitro and ex vivo results showed that PLGA and Chi/PLGA nanoparticles were efficiently taken up by the GI tract and could be optimized to deliver α-tocopherol to the intestine and improve its bioavailability.

Antioxidant poly(lactic-co-glycolic) acid nanoparticles made with α-tocopherol-ascorbic acid surfactant.

The goal of the study was to synthesize a surfactant made of α-tocopherol (vitamin E) and ascorbic acid (vitamin C) of antioxidant properties dubbed as EC, and to use this surfactant to make poly(lactic-co-glycolic) acid (PLGA) nanoparticles. Self-assembled EC nanostructures and PLGA-EC nanoparticles were made by nanoprecipitation, and their physical properties (size, size distribution, morphology) were studied at different salt concentrations, surfactant concentrations, and polymer/surfactant ratios. EC surfactant was shown to form self-assembled nanostructures in water with a size of 22 to 138 nm in the presence of sodium chloride, or 12 to 31 nm when synthesis was carried out in sodium bicarbonate. Polymeric PLGA-EC nanoparticles presented a size of 90 to 126 nm for 40% to 120% mass ratio PLGA to surfactant. For the same mass ratios, the PLGA-Span80 formed particles measured 155 to 216 nm. Span80 formed bilayers, whereas EC formed monolayers at the interfaces. PLGA-EC nanoparticles and EC showed antioxidant activity based on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay measurements using UV and EPR techniques, antioxidant activity which is not characteristic to commercially available Span80. The thiobarbituric acid reactive substances (TBARS) assay for lipid peroxidation showed that PLGA nanoparticles with EC performed better as antioxidants than the EC nanoassembly or the free vitamin C. Nanoparticles were readily internalized by HepG2 cells and were localized in the cytoplasm. The newly synthesized EC surfactant was therefore found successful in forming uniform, small size polymeric nanoparticles of intrinsic antioxidant properties.

Cellular uptake, antioxidant and antiproliferative activity of entrapped α-tocopherol and γ-tocotrienol in poly (lactic-co-glycolic) acid (PLGA) and chitosan covered PLGA nanoparticles (PLGA-Chi).

The aim of this study was to formulate and characterize α-tocopherol (α-T) and tocotrienol-rich fraction (TRF) entrapped in poly (lactide-co-glycolide) (PLGA) and chitosan covered PLGA (PLGA-Chi) based nanoparticles. The resultant nanoparticles were characterized and the effect of nanoparticles entrapment on the cellular uptake, antioxidant, and antiproliferative activity of α-T and TRF were tested. In vitro uptake studies in Caco2 cells showed that PLGA and PLGA-Chi nanoparticles displayed a greater enhancement in the cellular uptake of α-T and TRF when compared with the control without causing toxicity to the cells (p<0.0001). Furthermore, the cellular internalization of both PLGA and PLGA-Chi nanoparticles labeled with FITC was investigated by fluorescence microscopy; both types of nanoparticles were able to get internalized into the cells with reasonable amounts. However, PLGA-Chi nanoparticles showed significantly higher (3.5-fold) cellular uptake compared to PLGA nanoparticles. The antioxidant activity studies demonstrated that entrapment of α-T and TRF in PLGA and PLGA-Chi nanoparticles exhibited greater ability in inhibiting cholesterol oxidation at 48 h compared to the control. In vitro antiproliferative studies confirmed marked cytotoxicity of TRF on MCF-7 and MDA-MB-231 cell lines when delivered by PLGA and PLGA-Chi nanoparticles after 48 h incubation compared to control. In summary, PLGA and PLGA-Chi nanoparticles may be considered as an attractive and promising approach to enhance the bioavailability and activity of poorly water soluble compounds such as α-tocopherol and tocotrienols.

Bioavailability and biodistribution of nanodelivered lutein.

The aim of the study was to evaluate the ability of poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NP) to enhance lutein bioavailability. The bioavailability of free lutein and PLGA-NP lutein in rats was assessed by determining plasma pharmacokinetics and deposition in selected tissues. Lutein uptake and secretion was also assessed in Caco-2 cells. Compared to free lutein, PLGA-NP increased the maximal plasma concentration (Cmax) and area under the time-concentration curve in rats by 54.5- and 77.6-fold, respectively, while promoting tissue accumulation in the mesenteric fat and spleen. In comparison with micellized lutein, PLGA-NP lutein improved the Cmax in rat plasma by 15.6-fold and in selected tissues by ⩾ 3.8-fold. In contrast, PLGA-NP lutein had a lower uptake and secretion of lutein in Caco-2 cells by 10.0- and 50.5-fold, respectively, compared to micellized lutein. In conclusion, delivery of lutein with polymeric NP may be an approach to improve the bioavailability of lutein in vivo.

Synthesis of Poly(DL-Lactide-Co-Glycolide) Nanoparticles with Entrapped Magnetite by Emulsion Evaporation Method

The goal of this study was to synthesize Poly(DL-lactide-Co-glycolide) nanoparticles with entrapped magnetite, of under 100 nm in diameter, for future drug delivery applications. The emulsion evaporation method was selected to form poly(lactide-co-glycolide) (PLGA) nanoparticles with entrapped magnetite (Fe3O4) in the polymeric matrix, in the presence of sodium dodecyl sulfate (SDS) as a surfactant. Magnetite, a water-soluble compound, was surface functionalized with oleic acid to ensure its efficient entrapment in the PLGA matrix. The inclusion of magnetite with oleic acid (MOA) into the PLGA nanoparticles was accomplished in the organic phase. Synthesis was followed by dialysis, performed to eliminate the excess SDS, and lyophilization. The synthesized nanoparticles ranged in size from 38.6 to 67.1 nm for naked PLGA nanospheres and from 78.8 to 87.2 nm for MOA-entrapped PLGA nanospheres. The entrapment efficiency ranged from 57.36% to 77.3%.

Cytotoxicity and intracellular fate of PLGA and chitosan-coated PLGA nanoparticles in Madin–Darby bovine kidney (MDBK) and human colorectal adenocarcinoma (Colo 205) cells

Polymeric nanoparticles (NPs) are known to facilitate intracellular uptake of drugs to improve their efficacy, with minimum bioreactivity. The goal of this study was to assess cellular uptake and trafficking of PLGA NPs and chitosan (Chi)-covered PLGA NPs in Madin-Darby bovine kidney (MDBK) and human colorectal adenocarcinoma (Colo 205) cells. Both PLGA and Chi-PLGA NPs were not cytotoxic to the studied cells at concentrations up to 2500 μg/mL. The positive charge conferred by the chitosan deposition on the PLGA NPs improved NPs uptake by MDBK cells. In this cell line, Chi-PLGA NPs colocalized partially with early endosomes compartment and showed a more consistent perinuclear localization than PLGA NPs. Kinetic uptake of PLGA NPs by Colo 205 was slower than that by MDBK cells, detected only at 24 h, exceeding that of Chi-PLGA NPs. This study offers new insights on NP interaction with target cells supporting the use of NPs as novel nutraceuticals/drug delivery systems in metabolic disorders or cancer therapy.

Synthesis of Vitamin E-Carnosine (VECAR): New Antioxidant Molecule with Potential Application in Atherosclerosis

Abstract Natural antioxidants such as carnosine and α-tocopherol (vitamin E) provide protection against several oxidative stress–related diseases such as atherosclerosis and Alzheimers. The synthetic combination of α-tocopherol and carnosine can take advantage of the cellular transport mechanism of α-tocopherol by α-tocopherol transfer protein (α-TTP) to colocate α-tocopherol and carnosine at the interface between a lipophilic and a hydrophilic domain and protect both from oxidation. Successful synthesis of a novel heterodimer of α-tocopherol (vitamin E) and carnosine, VECAR was carried out in a total of nine steps. The VECAR design uses a 13-carbon phytyl-chain mimic to link carnosine to Trolox at the C2 carbon position. This design feature is anticipated to maintain binding to α-TTP, while maintaining the antioxidant activity of the two heterodimer components. Our results confirmed that there was no loss in antioxidant activity in VECAR using an in-vitro DPPH assay, versus α-tocopherol and Trolox. Supplemental materials are available for this article. Go to the publishers online edition of Synthetic Communications® to view the free supplemental. GRAPHICAL ABSTRACT