Highly Cytocompatible Polylactic Acid Based Electrospun Microfibers Loaded with Silver Nanoparticles Generated onto Chestnut Shell Lignin for Targeted Antibacterial Activity and Antioxidant Action.

Electrospun (e-spun) fibers are generally regarded as powerful tools for cell growth in tissue regeneration applications, and the possibility of imparting functional properties to these materials represents an increasingly pursued goal. We report herein the preparation of hybrid materials in which an e-spun d,l-polylactic acid matrix, to which chitosan or crystalline nanocellulose was added to improve hydrophilicity, was loaded with different amounts of silver(0) nanoparticles (AgNP) generated onto chestnut shell lignin (CSL) (AgNP@CSL). A solvent-free mechanochemical method was used for efficient (85% of the theoretical value by XRD analysis) Ag(0) production from the reduction of AgNO3 by lignin. For comparison, e-spun fibers containing CSL alone were also prepared. SEM and TEM analyses confirmed the presence of AgNP@CSL (average size 30 nm) on the fibers. Different chemical assays indicated that the AgNP@CSL containing fibers exhibited marked antioxidant properties (EC50 1.6 ± 0.1 mg/mL, DPPH assay), although they were halved with respect to those of the CSL containing fibers, as expected because of the efficient silver ion reduction. All the fibers showed high cytocompatibility toward human mesenchymal stem cells (hMSCs) representative of the self-healing process, and their antibacterial properties were tested against the pathogens Escherichia coli (E. coli), Staphylococcus epidermidis, and Pseudomonas aeruginosa. Finally, competitive surface colonization as simulated by cocultures of hMSC and E. coli showed that AgNP@CSL loaded fibers offered the cells a targeted protection from infection, thus well balancing cytocompatibility and antibacterial properties.

All natural mussel-inspired bioadhesives from soy proteins and plant derived polyphenols with marked water-resistance and favourable antibacterial profile for wound treatment applications.

HYPOTHESIS: Implementation of tissue adhesives from natural sources endowed with good mechanical properties and underwater resistance still represents a challenging research goal. Inspired by the extraordinary wet adhesion properties of mussel byssus proteins resulting from interaction of catechol and amino residues, hydrogels from soy protein isolate (SPI) and selected polyphenols i.e. caffeic acid (CA), chlorogenic acid (CGA) and gallic acid (GA) under mild aerial oxidative conditions were prepared. EXPERIMENTS: The hydrogels were subjected to chemical assays, ATR FT-IR and EPR spectroscopy, rheological and morphological SEM analysis. Mechanical tests were carried out on hydrogels prepared by inclusion of agarose. Biological tests included evaluation of the antibacterial and wound healing activity, and hemocompatibility. FINDINGS: The decrease of free NH2 and SH groups of SPI, the EPR features, the good cohesive strength and excellent underwater resistance (15 days for SPI/GA) under conditions relevant to their use as surgical glues indicated an efficient interaction of the polyphenols with the protein in the hydrogels. The polyphenols greatly also improved the mechanical properties of the SPI/ agarose/polyphenols hydrogels. These latter proved biocompatible, hemocompatible, not harmful to skin, displayed durable adhesiveness and good water-vapour permeability. Excellent antibacterial properties and in some cases (SPI/CGA) a favourable wound healing activity on dermal fibroblasts was obtained.

Combining the Potent Reducing Properties of Pecan Nutshell with a Solvent-Free Mechanochemical Approach for Synthesizing High Ag0 Content-Silver Nanoparticles: An Eco-Friendly Route to an Efficient Multifunctional Photocatalytic, Antibacterial, and Antioxidant Material.

A straightforward, low-cost, and scalable solid-state mechanochemical protocol for the synthesis of silver nanoparticles (AgNP) based on the use of the highly reducing agri-food by-product pecan nutshell (PNS) is reported herein. Under optimized conditions (180 min, 800 rpm, PNS/AgNO3 ratio = 55/45 w/w), a complete reduction in silver ions was achieved, leading to a material containing ca. 36% w/w Ag0 (X-ray diffraction analysis). Dynamic light scattering and microscopic analysis showed a uniform size distribution (15-35 nm average diameter) of the spherical AgNP. The 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay revealed lower-although still absolutely high (EC50 = 5.8 ± 0.5 mg/mL)-antioxidant properties for PNS for the further incorporation of AgNP, supporting the efficient reduction of Ag+ ions by PNS phenolic compounds. Photocatalytic experiments indicated that AgNP-PNS (0.4 mg/mL) was able to induce the >90% degradation of methylene blue after 120 min visible light irradiation, with good recycling stability. Finally, AgNP-PNS demonstrated high biocompatibility and significantly light-enhanced growth inhibition properties against Pseudomonas aeruginosa and Streptococcus mutans at concentrations as low as 250 µg/mL, also eliciting an antibiofilm effect at 1000 µg/mL. Overall, the adopted approach allowed to reuse a cheap and abundant agri-food by-product and required no toxic or noxious chemicals, making AgNP-PNS a sustainable and easy-to-access multifunctional material.

A Model Eumelanin from 5,6-Dihydroxyindole-2-Carboxybutanamide Combining Remarkable Antioxidant and Photoprotective Properties with a Favourable Solubility Profile for Dermo-Cosmetic Applications.

The search for new synthetic melanin-related pigments that maintain the antioxidant and photoprotective properties of naturally occurring dark eumelanins, while overcoming their unfavorable solubility, and molecular heterogeneity is presently a very active issue for dermo-cosmetic purposes. In this work, we explored the potential of a melanin obtained from the carboxybutanamide of a major eumelanin biosynthetic precursor, 5,6-dihydroxyindole-2-carboxylic acid (DHICA), by aerobic oxidation under slightly alkaline conditions. Analysis of the pigment by EPR, ATR-FTIR and MALDI MS indicated a substantial structural similarity to DHICA melanin, while investigation of the early intermediates confirmed unchanged regiochemistry of the oxidative coupling. The pigment exhibited a UVA-visible absorption even more intense than that of DHICA melanin, and a noticeable solubility in polar solvents of dermo-cosmetic relevance. The hydrogen- and/or electron-donor ability, and the iron (III) reducing power as determined by conventional assays provided evidence for marked antioxidant properties not merely ascribable to the more favorable solubility profile, while the inhibitory action of the radical- or photosensitized solar light-induced lipid peroxidation was more marked compared to that of DHICA melanin. Overall, these results hint at this melanin, which remarkable properties are, in part, due to the electronic effects of the carboxyamide functionality as a promising functional ingredient for dermo-cosmetic formulations.

Recovery of Lignins with Potent Antioxidant Properties from Shells of Edible Nuts by a Green Ball Milling/Deep Eutectic Solvent (DES)-Based Protocol.

Lignins are phenolic polymers endowed with potent antioxidant properties that are finding increasing applications in a variety of fields. Consequently, there is a growing need for easily available and sustainable sources, as well as for green extraction methodologies of these compounds. Herein, a ball milling/deep eutectic solvent (DES)-based treatment is reported as an efficient strategy for the recovery of antioxidant lignins from the shells of edible nuts, namely chestnuts, hazelnuts, peanuts, pecan nuts, and pistachios. In particular, preliminarily ball-milled shells were treated with 1:2 mol/mol choline chloride:lactic acid at 120 °C for 24 h, and the extracted material was recovered in 19-27% w/w yields after precipitation by the addition of 0.01 M HCl. Extensive spectroscopic and chromatographic analysis allowed for confirmation that the main phenolic constituents present in the shell extracts were lignins, accompanied by small amounts (0.9% w/w) of ellagic acid, in the case of chestnut shells. The recovered samples exhibited very promising antioxidant properties, particularly in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay (EC50 values ranging from 0.03 to 0.19 mg/mL). These results open new perspectives for the valorization of nut shells as green sources of lignins for applications as antioxidants, e.g., in the biomedical, food, and/or cosmetic sector.

A Straightforward Access to New Amides of the Melanin Precursor 5,6-Dihydroxyindole-2-carboxylic Acid and Characterization of the Properties of the Pigments Thereof.

We report herein an optimized procedure for preparation of carboxamides of 5,6-dihydroxyindole-2-carboxylic acid (DHICA), the main biosynthetic precursor of the skin photoprotective agents melanins, to get access to pigments with more favorable solubility properties with respect to the natural ones. The developed procedure was based on the use of a coupling agent (HATU/DIPEA) and required protection of the catechol function by easily removable acetyl groups. The O-acetylated compounds could be safely stored and taken to the reactive o-diphenol form just before use. Satisfactorily high yields (>85%) were obtained for all amides. The oxidative polymerization of the synthesized amides carried out in air in aqueous buffer at pH 9 afforded melanin-like pigmented materials that showed chromophores resembling those of DHICA-derived pigments, with a good covering of the UVA and the visible region, and additionally exhibited a good solubility in alcoholic solvents, a feature of great interest for the exploitation of these materials as ingredients of dermocosmetic formulations.

Non-covalent small molecule partnership for redox-active films: Beyond polydopamine technology.

HYPOTHESIS: The possibility to use hexamethylenediamine (HMDA) to impart film forming ability to natural polymers including eumelanins and plant polyphenols endowed with biological activity and functional properties has been recently explored with the aim to broaden the potential of polydopamine (PDA)-based films overcoming their inherent limitations. 5,6-dihydroxyindole-2-carboxylic acid, its methyl ester (MeDHICA) and eumelanins thereof were shown to exhibit potent reducing activity. EXPERIMENTS: MeDHICA and HMDA were reacted in aqueous buffer, pH 9.0 in the presence of different substrates to assess the film forming ability. The effect of different reaction parameters (pH, diamine chain length) on film formation was investigated. Voltammetric and AFM /SEM methods were applied for analysis of the film redox activity and morphology. HPLC, MALDI-MS and 1HNMR were used for chemical characterization. The film reducing activity was evaluated in comparison with PDA by chemical assays and using UV stressed human immortalized keratinocytes (HaCat) cells model. FINDINGS: Regular and homogeneous yellowish films were obtained with moderately hydrophobic properties. Film deposition was optimal at pH 9, and specifically induced by HMDA. The film consisted of HMDA and monomeric MeDHICA accompanied by dimers/small oligomers, but no detectable MeDHICA/HMDA covalent conjugation products. Spontaneous assembly of self-organized networks held together mainly by electrostatic interactions of MeDHICA in the anion form and HMDA as the dication is proposed as film deposition mechanism. The film displayed potent reducing properties and exerted significant protective effects from oxidative stress on HaCaT.