Chronic respiratory diseases, whose one of the hallmarks is oxidative stress, are still incurable and need novel therapeutic tools and pharmaceutical agents. The phenolic compounds contained in grape are endowed with well-recognized anti-oxidant, anti-inflammatory, anti-cancer, and anti-aging activities. Considering that natural anti-oxidants, such as proanthocyanidins, have poor water solubility and oral bioavailability, we have developed a drug delivery system based on solid lipid nanoparticles (SLN), apt to encapsulate grape seed extract (GSE), containing proanthocyanidins. Plain, 6-coumarin (6-Coum), DiR- and GSE-loaded SLN were produced with the melt-emulsion method. Physicochemical characterization of all prepared SLN was determined by photon correlation spectroscopy and laser Doppler anemometry. MTT assay (spectrophotometry) and propidium iodide (PI) assay (cytofluorimetry) were used to assess cell viability. Flow cytometry coupled with cell imaging was performed for assessing apoptosis and necrosis by Annexin V/7-AAD staining (plain SLE), cell internalization (6-Coum-SLN) and reactive oxygen species (ROS) production (SLN-GSE). NF-κB nuclear translocation was studied by immunofluorescence. In vivo bio-imaging was used to assess lung deposition and persistence of aerosolized DiR-loaded SLN. Plain SLN were not cytotoxic when incubated with H441 airway epithelial cells, as judged by both PI and MTT assays as well as by apoptosis/necrosis evaluation. 6-Coum-loaded SLN were taken up by H441 cells in a dose-dependent fashion and persisted into cells at detectable levels up to 16 days. SLN were detected in mice lungs up to 6 days. SLN-GSE possessed 243 nm as mean diameter, were negatively charged, and stable in size at 37 °C in Simulated Lung Fluid up to 48 h and at 4 °C in double distilled water up to 2 months. The content of SLN in proanthocyanidins remained unvaried up to 2 months. GSE-loaded SLN determined a significant reduction in ROS production when added 24–72 h before the stimulation with hydrogen peroxide. Interestingly, while at 24 h free GSE determined a higher decrease of ROS production than SLN-GSE, the contrary was seen at 48 and 72 h. Similar results were observed for NF-κB nuclear translocation. SLN are a biocompatible drug delivery system for natural anti-oxidants obtained from grape seed in a model of oxidative stress in airway epithelial cells. They feature stability and long-term persistence inside cells where they release proanthocyanidins. These results could pave the way to novel anti-oxidant and anti-inflammatory therapies for chronic respiratory diseases.

A widely investigated approach to bypass the blood brain barrier is represented by the intranasal delivery of therapeutic agents exploiting the olfactory or trigeminal connections nose-brain. As for Parkinson’s disease (PD), characterized by dopaminergic midbrain neurons degeneration, currently there is no disease modifying therapy. Although several bio-nanomaterials have been evaluated for encapsulation of neurotransmitter dopamine (DA) or dopaminergic drugs in order to restore the DA content in parkinsonian patients, the premature leakage of the therapeutic agent limits this approach. To tackle this drawback, we undertook a study where the active was linked to the polymeric backbone by a covalent bond. Thus, novel nanoparticles (NPs) based on N,O-Carboxymethylchitosan-DA amide conjugate (N,O-CMCS-DA) were prepared by the nanoprecipitation method and characterized from a technological view point, cytotoxicity and uptake by Olfactory Ensheating Cells (OECs). Thermogravimetric analysis showed high chemical stability of N,O-CMCS-DA NPs and X-ray photoelectron spectroscopy evidenced the presence of amide linkages on the NPs surface. MTT test indicated their cytocompatibility with OECs, while cytofluorimetry and fluorescent microscopy revealed the internalization of labelled N,O-CMCS-DA NPs by OECs, that was increased by the presence of mucin. Altogether, these findings seem promising for further development of N,O-CMCS-DA NPs for nose-to-brain delivery application in PD.

Copper-chitosan (Cu-CS) nanoantimicrobials are a novel class of bioactive agents, providing enhanced and synergistic efficiency in the prevention of biocontamination in several application fields, from food packaging to biomedical. Femtosecond laser pulses were here exploited to disrupt a Cu solid target immersed into aqueous acidic solutions containing different CS concentrations. After preparation, Cu-CS colloids were obtained by tuning both Cu/CS molar ratios and laser operating conditions. As prepared Cu-CS colloids were characterized by Fourier transform infrared spectroscopy (FTIR), to study copper complexation with the biopolymer. X-ray photoelectron spectroscopy (XPS) was used to elucidate the nanomaterials’ surface chemical composition and chemical speciation of the most representative elements. Transmission electron microscopy was used to characterize nanocolloids morphology. For all samples, ξ-potential measurements showed highly positive potentials, which could be correlated with the XPS information. The spectroscopic and morphological characterization herein presented outlines the characteristics of a technologically-relevant nanomaterial and provides evidence about the optimal synthesis parameters to produce almost monodisperse and properly-capped Cu nanophases, which combine in the same core-shell structure two renowned antibacterial agents.

(1) Backgrond: Considering the positive effects of citicoline (CIT) in the management of some neurodegenerative diseases, the aim of this work was to develop CIT-Loaded Solid Lipid Nanoparticles (CIT-SLNs) for enhancing the therapeutic use of CIT in parkinsonian syndrome; (2) Methods: CIT-SLNs were prepared by the melt homogenization method using the self-emulsifying lipid Gelucire^® 50/13 as lipid matrix. Solid-state features on CIT-SLNs were obtained with FT-IR, thermal analysis (DSC) and X-ray powder diffraction (XRPD) studies. (3) Results: CIT-SLNs showed a mean diameter of 201 nm, −2.20 mV as zeta potential and a high percentage of entrapped CIT. DSC and XRPD analyses evidenced a greater amorphous state of CIT in CIT-SLNs. On confocal microscopy, fluorescent SLNs replacing unlabeled CIT-SLNs released the dye selectively in the cytoplasm. Biological evaluation showed that pre-treatment of SH-SY5Y dopaminergic cells with CIT-SLNs (50 µM) before the addition of 40 µM 6-hydroxydopamine (6-OHDA) to mimic Parkinson’s disease’s degenerative pathways counteracts the cytotoxic effects induced by the neurotoxin, increasing cell viability with the consistent maintenance of both nuclear and cell morphology. In contrast, pre-treatment with CIT 50 and 60 µM or plain SLNs for 2 h followed by 6-OHDA (40 µM) did not significantly influence cell viability. (4) Conclusions: These data suggest an enhanced protection exerted by CIT-SLNs with respect to free CIT and prompt further investigation of possible molecular mechanisms that underlie this difference.

Introduction. Larvae of the insect Hermetia illucens can represent an alternative source for low-molecular-weight chitosan (CS) production compared with CS from crustaceans (CS_crustac), making it appealing in terms of pharmaceutical applications. Hence, the performances of CS_larvae and CS_crustac were compared herein by investigating the in vitro features of nanoparticles (NPs) made from each polysaccharide and administered with the antioxidant quercetin (QUE). Methods. X-ray diffraction and FT-IR spectroscopy enabled the identification of each type of CS. Following the ionic gelation technique and using sulfobutylether-β-cyclodextrin as a cross-linking agent, NPs were easily obtained. Results. Physicochemical data, release studies in PBS, and the evaluation of antioxidant effects via the 1,1-diphenyl-2-picrylhydrazyl (DPPH) test were studied for both CS_larvae and CS_crustac. QUE-loaded NP sizes ranged from 180 to 547 nm, and zeta potential values were between +7.5 and +39.3 mV. In vitro QUE release in PBS was faster from QUE-CS_larvae NPs than from CS_crustac, and high antioxidant activity—according to the DPPH test—was observed for all tested NP formulations. Discussion. The agar diffusion assay, referring to Escherichia coli and Micrococcus flavus, as well as the microdilution assay, showed the best performance as antimicrobial formulations in the case of QUE-CS_larvae NPs. QUE-CS_larvae NPs can represent a promising vehicle for QUE, releasing it in a sustained manner, and, relevantly, the synergism noticed between QUE and CS_larvae resulted in a final antimicrobial product. Conclusions. New perspectives for low-molecular-weight CS are disclosed by adopting renewable sources from insects instead of the commercial CS_crustac.