Production of highly antioxidant lignin nanoparticles from a hardwood technical lignin.

Eucalypt kraft lignin isolated in a LignoBoost™ pilot plant was characterized by GC-MS, ICP-OES, DSC, HPSEC, 31P NMR, and HSQC 2D-NMR to be used without any further processing to produce lignin nanoparticles (LNPs) by nanoprecipitation. Tetrahydrofuran (THF) was used as a solvent, and water as a non-solvent. Microscopic analysis (TEM) showed that LNPs were regularly spherical with some hollow particles dispersed in-between, and sizes were tunable by changing the solvent dripping rate onto the non-solvent. LNP particle sizes had a bimodal distribution, with the largest population having an average apparent hydrodynamic diameter ranging from 105.6 to 75.6 nm. Colloidal dispersions of LNPs in water presented good stability in different dilutions without significant size changes upon storage at pH close to neutral for as long as 45 days. Zeta potentials around -40 mV were obtained for LNP suspensions at pH ranging from 7 to 9. The high carbohydrate content (circa 10 % on a dry basis, mostly xylans) of the lignin precursor did not interfere in LNP formation, whose antioxidant activity was expressive as demonstrated by the ABTS assay at pH 7.4, with an EC50 of 4.04 µg mL-1. Also, the Trolox® equivalent antioxidant capacity (TEAC) of LNPs reached 1.90 after 40 min reaction time.

Immobilization of the metagenomic lipase, LipG9, on porous pellets of poly-hydroxybutyrate produced by the double emulsion solvent evaporation technique.

This work aimed to produce porous poly-hydroxybutyrate (PHB) pellets in order to evaluate the pellets as a support for immobilization of the metagenomic lipase, LipG9. Four types of pelletized PHB particles with different morphological characteristics were obtained using the double emulsion and solvent evaporation technique (DESE). The micropores of these PHB pellets had similar average diameters (about 3 nm), but the pellets had different specific surface areas: 11.7 m2 g-1 for the PHB powder, 8.4 m2  g-1 for the control pellets (Ø < 0.5 mm, produced without the pore forming agent), 10.0 m2  g-1 for the small pellets (Ø < 0.5 mm), 9.5 m2  g-1 for the medium pellets (0.5 < Ø < 0.8 mm) and 8.4 m2  g-1 for the large pellets (Ø > 1.4 mm). Purified LipG9 was immobilized by adsorption on these pellets, and the results were compared with those obtained with PHB powder. The highest immobilization yield (83%) was obtained for the medium PHB pellets, followed by large (76%) and small (55%) PHB pellets. The activity of LipG9 immobilized on the pellets, for the synthesis of ethyl oleate in n-hexane, was highest for the medium pellets (22 U g-1 ). The immobilization yield was high for PHB powder (99%) but the esterification activity was slightly lower (20 U g-1 ). These results show that pelletized PHB beads can be used for the immobilization of lipases, with the advantage that pelletized PHB will perform better than PHB powder in large-scale enzyme bioreactors.

Spray-dried solid dispersions containing ferulic acid: comparative analysis of three carriers, in vitro dissolution, antioxidant potential and in vivo anti-platelet effect.

This article aimed to improve the relative solubility and dissolution rate of ferulic acid (FA) by the use of spray-dried solid dispersions (SDs) in order to ensure its in vitro antioxidant potential and to enhance its in vivo anti-platelet effect. These SDs were prepared by spray-drying at 10 and 20% of drug concentration using polyvinylpyrrolidone K30 (PVP-K30), polyethylene glycol 6000 (PEG 6000) and poloxamer-188 (PLX-188) as carriers. SDs and physical mixtures (PM) were characterized by SEM, XRPD, FTIR spectroscopy and TGA analysis. Spray-dried SDs containing FA were successfully obtained. Relative solubility of FA was improved with increasing carrier concentration. PVP-K30 and PEG 6000 formulations showed suitable drug content values close to 100%, whereas PLX-188 presented mean values between 70 and 90%. Agglomerates were observed depending on the carrier used. XRPD patterns and thermograms indicated that spray-drying led to drug amorphization and provided appropriate thermal stability, respectively. FTIR spectra demonstrated no remarkable interaction between carrier and drug for PEG 6000 and PLX-188 SDs. PVP-K30 formulations had changes in FTIR spectra, which denoted intermolecular O-H•••O = C bonds. Spray-dried SDs played an important role in enhancing dissolution rate of FA when compared to pure drug. The free radical-scavenging assay confirmed that the antioxidant activity of PEG 6000 10% SDs was kept. This formulation also provided a statistically increased in vivo anti-platelet effect compared to pure drug. In summary, these formulations enhanced relative solubility and dissolution rate of FA and chosen formulation demonstrated suitable in vitro antioxidant activity and improved in vivo anti-platelet effect.

Spray-dried Eudragit® L100 microparticles containing ferulic acid: Formulation, in vitro cytoprotection and in vivo anti-platelet effect.

This paper aimed to obtain new spray-dried microparticles containing ferulic acid (FA) prepared by using a methacrylic polymer (Eudragit® L100). Microparticles were intended for oral use in order to provide a controlled release, and improved in vitro and in vivo biological effects. FA-loaded Eudragit® L100 microparticles were obtained by spray-drying. Physicochemical properties, in vitro cell-based effects, and in vivo platelet aggregation were investigated. FA-loaded Eudragit® L100 microparticles were successfully prepared by spray-drying. Formulations showed suitable encapsulation efficiency, i.e. close to 100%. Microparticles were of spherical and almost-spherical shape with a smooth surface and a mean diameter between 2 and 3µm. Fourier-transformed infrared spectra demonstrated no chemical bond between FA and polymer. X-ray diffraction and differential scanning calorimetry analyses indicated that microencapsulation led to drug amorphization. FA-loaded microparticles showed a slower dissolution rate than pure drug. The chosen formulation demonstrated higher in vitro cytoprotection, anti-inflammatory and immunomodulatory potential and also improved in vivo anti-platelet effect. These results support an experimental basis for the use of FA spray-dried microparticles as a feasible oral drug delivery carrier for the controlled release of FA and improved cytoprotective and anti-platelet effects.

PCL/PHBV microparticles as innovative carriers for oral controlled release of manidipine dihydrochloride.

Microparticles of poly(ε-caprolactone) (PCL) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) containing manidipine dihydrochloride (MAN) were successfully prepared by the simple emulsion/solvent evaporation method. All formulations showed loading efficiency rates greater than 80% and average particle size less than 8 µm. Formulations had spherical shape with smooth and porous surface for PCL and PHBV, respectively. According to Fourier-transform infrared spectroscopy, initial components were not chemically modified during microencapsulation. X-ray diffraction patterns and differential scanning calorimetry demonstrated that this process led to drug amorphization. In vitro dissolution studies showed that all microparticles prolonged MAN release, mainly which one obtained using PCL that contained 5% of drug loaded (PCL-M5). Animal studies demonstrated that formulation PCL-M5 was able to keep the variation of mean arterial pressure after phenylephrine administration up to 24 hours. These data confirmed the sustained antihypertensive effect of the investigated microparticles. Results provided an experimental basis for using formulation PCL-M5 as a feasible carrier for oral controlled release of MAN intended for treating high blood pressure.

An innovative quaternary ammonium methacrylate polymer can provide improved antimicrobial properties for a dental adhesive system.

A quaternary ammonium methacrylate polymer (QAMP) with antimicrobial potential was synthesized. The resulting product (QAMP) was characterized by FTIR spectroscopy, NMR spectroscopy, visible spectrophotometry, XRPD and TGA. The in vitro susceptibility tests against Streptococcus mutans of QAMP were investigated prior and after incorporation into a commercial adhesive system (Clearfil™ SE Bond). The release of quaternary ammonium compounds from the experimental adhesive system (Clearfil™ SE Bond + 5% QAMP) was performed during 1, 7, 14, 21 and 30 days. Spectroscopic data confirmed that QAMP was successfully obtained. Thermogravimetric analysis indicated that QAMP was heat stable. Prior incorporation into the adhesive system, QAMP revealed an inhibition halo of 18.33 ± 0.6 mm. By agar disk diffusion test, Clearfil™ SE Bond containing 5% QAMP presented an inhibition halo (16.67 ± 1.5 mm) similar to Clearfil™ Protect Bond (positive control, 17.00 ± 1.7, p = 0.815) and significantly higher than Clearfil™ SE Bond (negative control, 11.00 ± 1.0, p = 0.006). The minimum inhibitory/bactericidal concentrations for Clearfil™ SE Bond containing 5% QAMP were 20 µL mL(-1). The release of quaternary ammonium compounds from the experimental adhesive containing QAMP was very low (5.1%) when compared to Clearfil™ Protect Bond that released 47.2% of its quaternary ammonium monomer (MDPB) after 30 days. The QAMP can offer enhanced antimicrobial properties for self-etching adhesive systems.

PHBV/PCL microparticles for controlled release of resveratrol: physicochemical characterization, antioxidant potential, and effect on hemolysis of human erythrocytes.

Microparticles of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly(ε-caprolactone) (PCL) containing resveratrol were successfully prepared by simple emulsion/solvent evaporation. All formulations showed suitable encapsulation efficiency values higher than 80%. PHBV microparticles revealed spherical shape with rough surface and presence of pores. PCL microparticles were spherically shaped with smooth surface. Fourier-transformed infrared spectra demonstrated no chemical bond between resveratrol and polymers. X-ray powder diffraction patterns and differential scanning calorimetry analyses indicated that microencapsulation led to drug amorphization. These PHBV/PCL microparticles delayed the dissolution profile of resveratrol. Release profiles were better fitted to biexponential equation. The hypochlorous-acid-scavenging activity and 2,2-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical cation discoloration assay confirmed that the antioxidant activity of PHBV/PCL microparticles was kept, but was dependent on the microparticle morphology and dissolution profile. Resveratrol-loaded PHBV/PCL microparticles showed no cytotoxic effect on red blood cells.