Bioactive Phenols-loaded chitosan nanoparticles (PL-CNps) were developed by ionic gelation from Persian lemon (Citrus latifolia) waste (PLW) and chitosan nanoparticles. Response Surface Methodology (RSM) was used to determine the optimal Ultrasound-Assisted Extraction (UAE) conditions for the total phenolic compounds (TPC) recovery from PLW (58.13 mg GAE/g dw), evaluating the ethanol concentration, extraction time, amplitude, and solid/liquid ratio. Eight compounds expressed as mg/g dry weight (dw) were identified by ultra-performance liquid chromatography coupled photo diode array (UPLC-PDA) analysis: eriocitrin (20.71 ± 0.09), diosmin (18.59 ± 0.13), hesperidin (7.30 ± 0.04), sinapic acid (3.67 ± 0.04), catechin (2.92 ± 0.05), coumaric acid (2.86 ± 0.01), neohesperidin (1.63 ± 0.00), and naringenin (0.44 ± 0.00). The PL-CNps presented size of 232.7 nm, polydispersity index of 0.182, Z potential of −3.8 mV, and encapsulation efficiency of 81.16%. The results indicated that a synergic effect between phenolic compounds from PLW and chitosan nanoparticles was observed in antioxidant and antibacterial activity, according to Limpel’s equation. Such results indicate that PLW in such bioprocesses shows excellent potential as substrates for the production of value-added compounds with a special application for the food industry.

The mucus gel covers the wet epithelia that forms the inner lining of the body. It constitutes our first line of defense protecting the body from infections and other deleterious molecules. Failure of the mucus barrier can lead to the inflammation of the mucosa such as in inflammatory bowel diseases. Unfortunately, there are no effective strategies that reinforce the mucus barrier properties to recover or enhance its ability to protect the epithelium. Herein, we describe a mucus engineering approach that addresses this issue where we physically crosslink the mucus gel with low molar mass chitosan variants to reinforce its barrier functions. We tested the effect of these chitosans on mucus using in-lab purified porcine gastric mucins, which mimic the native properties of mucus, and on mucus-secreting HT29-MTX epithelial cell cultures. We found that the lowest molar mass chitosan variant (degree of polymerisation of 8) diffuses deep into the mucus gels while physically crosslinking the mucin polymers, whereas the higher molar mass chitosan variants (degree of polymerisation of 52 and 100) interact only superficially. The complexation resulted in a tighter mucin polymer mesh that slowed the diffusion of dextran polymers and of the cholera toxin B subunit protein through the mucus gels. These results uncover a new use for low molar mass mucoadhesive polymers such a chitosan as non-cytotoxic mucosal barrier enhancers that could be valuable in the prevention and treatment of mucosal diseases.

In the context of gene delivery, chitosan has been widely used as a safe and effective polycation to complex DNA, RNA and more recently, siRNA. However, much less attention has been paid to chitosan oligosaccharides (COS) despite their biological properties. This study proposed to carry out a physicochemical study of COS varying in degree of polymerization (DP) from 5 to 50, both from the point of view of the solution properties and the complexing behavior with siRNA. The main parameters studied as a function of DP were the apparent pK_a, the solubility versus pH, the binding affinity with siRNA and the colloidal properties of complexes. Some parameters, like the pK_a or the binding enthalpy with siRNA, showed a marked transition from DP 5 to DP 13, suggesting that electrostatic properties of COS vary considerably in this range of DP. The colloidal properties of siRNA/COS complexes were affected in a different way by the COS chain length. In particular, COS of relatively high DP (≥50) were required to form small complex particles with good stability.

The objective of this work was the recovery of chitosan by ultrasound-assisted extraction (UAE) from white shrimp (Litopenaeus vannamei) chitin, and the physicochemical and structural characterization of the obtained biopolymer, as well as its antimicrobial effect on Pythium ultimum growth. A 2³ factorial design was used to evaluate chitosan extraction conditions. Instrumental analysis techniques for chitosan characterization and radial growth inhibition, as an antifungal activity test, were performed. The ultrasonically extracted chitosan (UC) reached a yield of 86.96% with 100% solubility, a degree of deacetylation (DDA) >78%, molecular weight (M_W) of 3.928 × 10⁵ g mol⁻¹, and a crystallinity index (Icr) of 87%, calculated through nuclear magnetic resonance (¹H NMR) and Fourier transform infrared spectroscopy (FTIR), size exclusion chromatography (SEC), and X-ray diffraction (XRD), respectively. The inhibitory activity of the chitosan was evaluated against the oomycete Pythium ultimum, observing a 93% radial inhibition over 24 h. UAE proved to be an excellent alternative to the conventional deacetylation, reducing reaction time and obtaining a UC with higher M_W and (Icr) than the commercial one, which could potentiate its applications.