Synthesis of hesperetin-loaded PLGA nanoparticles by two different experimental design methods and biological evaluation of optimized nanoparticles

Abstract

Hesperetin was effectively encapsulated into poly (d,l-lactic-co-glycolic acid) nanoparticles by using experimental design methods. A seven-factor Plackett-Burman design was used in order to determine the major process parameters. A significant linear equation, which shows the effect of each process parameter on encapsulation efficiency was developed, and then the most effective factors were determined. Further investigation and optimization was carried out by applying the three-factor three-level Box-Behnken design. Significant second-order mathematical models were developed by regression analysis of the experimental data for both responses: encapsulation efficiency and nanoparticle size. The two step experimental design allowed the synthesis of the desired nanoparticle formulations with maximum encapsulation efficiency (80.5 ± 4.9%) and minimum particle size (260.2 ± 16.5 nm) at optimum process conditions: 0.5% polyvinyl alcohol (PVA) concentration, 5.13 water:organic phase ratio, and 3.59 ml min flow rate of the emulsified solution into 0.1% PVA. Furthermore, the biological activity of these optimized nanoparticles were determined with antimicrobial activity and cytotoxicity studies; results were then compared to the free hesperetin. The cytotoxicity result revealed that hesperetin and hesperetin-loaded nanoparticles were biocompatible with normal cell line L929 fibroblast cells up to 184.83 and 190.88 μg ml for 24 h, and up to 133.24 and 134.80 μg ml for 48 h, respectively. In the antimicrobial study, the optimized nanoparticle showed inhibition activity (minimal inhibitory concentration (MIC) values were 125 μg ml for Escherichia coli, and 200 μg ml for Staphylococcus aureus), while the free hesperetin did not demonstrate activity in both strains (MIC value >200 μg ml). These in vitro results may provide useful information for the investigation of hesperetin-loaded nanoparticles in diagnostic and therapeutic applications.

Comparative evaluation of hesperetin loaded nanoparticles for anticancer activity against C6 glioma cancer cells

“…On the other hand, incubation with HspNPs for 24 and 48 h resulted in dose and time dependent decrease in cell viability, with IC 50 of 28 and 21 lg/mL, respectively. In contrast, it has been recently reported that Hsp and HspNPs have showed very low inhibition effect on cell viability of L929 fibroblast cells, which indicates they did not show any significant cytotoxicity to normal cells [23]. At the same concentration, HspNPs drastically decreased cell viability compared to free Hsp.…”

“…HspNPs were prepared by a modified oil in water (o/w) single emulsion solvent evaporation method described as in our previous study [23]. According to this method, the organic phases consisted of PLGA and hesperetin were dissolved into DCM and acetone, respectively.…”

Potentials of Encapsulated Flavonoids in Biologics: A Review

“…The result revealed that the hesperetin and hesperetin-loaded nanoparticles were biocompatible with normal cell line L929 fibroblast cells up to 184.83 and190.88 µg ml -1 for 24 h and up to 133.24 and 134.80 µg ml -1 for 48 h. The antimicrobial activity studies were carried using two different methods against Staphylococcus aureus and Escherichia coli. The MIC (minimal inhibitory concentration) values were 125 µg ml -1 for Escherichia coli and 200 µg ml -1 for Staphylococcus aureus, while the free hesperetin did not demonstrate activity in both strains (MIC value >200 µg ml-1) [111].…”

2,4-Dichlorophenoxyacetic Acid Loaded Polymeric Nanoparticle Synthesis and Its Effect on Biomass in Medicago sativa Cell Suspension Cultures

“…advantages, it has been reported that many hydrophobic molecules are loaded into nanoparticular systems, thus increasing their biological activities [15,[27][28][29]. There are many studies on nanoparticular systems in mammalian cell cultures and nanoparticles have been used quite successfully until now in mammalian cell cultures and drug targeting systems [30,31].…”

“…The Poly-(D,L-lactic-co-glycolic acid) (PLGA) is the most commonly used polymer in the production of polymeric nanoparticles. It has been determined that the systems created by loading the active substances into the PLGA nanoparticle have increased the pharmacokinetic properties, therapeutic indices and biocompatibilities of the active substances compared to the free form [12][13][14][15].…”