An injectable, nanostructured implant for the delivery of adenosine triphosphate: towards long-acting formulations of small, hydrophilic drugs.

While long-acting injectable treatments are gaining increasing interest in managing chronic diseases, the available drug delivery systems almost exclusively rely on hydrophobic matrixes, limiting their application to either hydrophobic drugs or large and hydrophilic molecules such as peptides. To address the technological lock for long-acting delivery systems tailored to small, hydrophilic drugs such as anticancer and antiviral nucleoside/nucleotide analogues, we have synthesized and characterized an original approach with a multi-scale structure: (i) a nucleotide (adenosine triphosphate, ATP) is first incorporated in hydrophilic chitosan-Fe(III) nanogels; (ii) these nanogels are then transferred by freeze-drying and resuspension into a water-free, hydrophobic medium containing PLGA and an organic solvent, N-methyl-2-pyrrolidone. We show that this specific association allows an injectable and homogeneous dispersion, able to form in situ implants upon injection in physiological or aqueous environments. This system releases ATP in vitro without any burst effect in a two-step mechanism, first as nanogels acting as an intermediate reservoir over a week, then as free drug over several weeks. In vivo studies confirmed the potential of such nanostructured implants for sustained drug release following subcutaneous injection to mice hock, opening perspectives for sustained and targeted delivery through the lymphatic system.

Microwave-Assisted Extraction of Polyphenols from Blackcurrant By-Products and Possible Uses of the Extracts in Active Packaging.

The design of experiment (DoE) approach was used to optimize the extraction of polyphenols from blackcurrant by-products with microwave-assisted extraction and deionized water as a green solvent. Three factors (microwave power, extraction time, and solvent/matrix ratio) were evauated, and a central composite orthogonal design (CCO) was applied in order to reduce experimental runs. Empirical models relating the response and process parameters were developed. The validity of the models was tested using an analysis of variance (ANOVA). The optimal extraction conditions were found using the highest value of microwave power (780-800 W) and the lowest extraction time (60 min) and solvent/matrix ratio (10 m/g). Compared with conventional solvent extraction, the polyphenol yield increased by 25% after applying the optimized MAE process. The obtained extract was used to realize a sustainable active maltodextrin (Glucidex 2) pad using an electrospinning technique. The antimicrobial and antioxidant activities of the pads were tested on the post-harvest storage of raspberries. Two set of experiments were carried out. The recorded results showed that the pad had antimicrobial activity on the tested fruit samples and implied the possibility of using it to extend the shelf-life of the fruits.

The Postharvest Quality of Fresh Sweet Cherries and Strawberries with an Active Packaging System.

This study assessed the effect of the recently-introduced Life+ (ILIP, Valsamoggia, Italy) active packaging system on the postharvest quality of sweet cherries and strawberries. This system uses Equilibrium Modified Atmosphere Packaging (EMAP) to achieve specific intra-package conditions with three synergistic elements: an unvented and anti-mist heat sealable container, an active (naturally-antimicrobial) pad, and a heat-sealed, laser micro-perforated film of a specified gas permeability. Post-packaging quality parameters were monitored for 10 (strawberries) and 15 days (cherries): headspace gas concentration, weight loss, titratable acidity, total soluble solids, pH, disease incidence, and sensory quality. Results showed that use of the Life+ system delayed postharvest senescence by maintaining fruit color, acidity, and vitamin C content, and decreasing fruit weight loss and decay. The use of EMAP in sweet cherry resulted in enhanced sensory qualities compared to traditional perforated containers. The results suggest that the Life+ system leads to better sensory properties and improved shelf-life for strawberries and sweet cherries.

PLA-PEG Nanoparticles Improve the Anti-Inflammatory Effect of Rosiglitazone on Macrophages by Enhancing Drug Uptake Compared to Free Rosiglitazone.

Among cardiovascular diseases, atherosclerosis remains the first cause of death in the United States of America and Europe, as it leads to myocardial infarction or stroke. The high prevalence of heart diseases is due to the difficulty in diagnosing atherosclerosis, since it can develop for decades before symptoms occur, and to the complexity of the treatment since targets are also important components of the host defenses. The antidiabetics thiazolidinediones, among which is rosiglitazone (RSG), have demonstrated anti-atherosclerotic effect in animal models, and are therefore promising candidates for the improvement of atherosclerosis management. Nevertheless, their administration is hindered by the insurgence of severe side effects. To overcome this limitation, rosiglitazone has been encapsulated into polymeric nanoparticles, which permit efficient delivery to its nuclear target, and selective delivery to the site of action, allowing the reduction of unwanted effects. In the present work, we describe nanoparticle formulation using polylactic acid (PLA) coupled to polyethylene glycol (PEG), their characterization, and their behavior on RAW264.7 macrophages, an important target in atherosclerosis treatment. RSG nanocarriers showed no toxicity on cells at all concentrations tested, an anti-inflammatory effect in a dose-dependent manner, up to 5 times more efficient than the free molecule, and an increased RSG uptake which is consistent with the effect shown. These biodegradable nanoparticles represent a valid tool to be further investigated for the treatment of atherosclerosis.

Liposome-supported peritoneal dialysis in the treatment of severe hyperammonemia: An investigation on potential interactions.

Peritoneal dialysis (PD) performed with transmembrane pH-gradient liposomes was reported to efficiently remove ammonia from the body, representing a promising alternative to current standard-of-care for patients with severe hepatic encephalopathy. In this study, we further characterized the properties of liposome-supported peritoneal dialysis (LSPD) by 1) assessing its in-use stability in the presence of ascitic fluids from liver-disease patients; 2) investigating its interactions with drugs that are commonly administered to acute-on-chronic liver failure patients; and 3) analyzing the in vivo extraction profile of LSPD. We found that LSPD fluid maintained its in vitro ammonia uptake capability when combined with ascitic fluids. The co-incubation of selected drugs (e.g., beta-blockers, antibiotics, diuretics) with LSPD fluids and ammonia resulted in limited interaction effects for most compounds except for two fluoroquinolones and propranolol. However, considering the experimental set-up, these results should be interpreted with caution and confirmatory drug-drug interaction studies in a clinical setting will be required. Finally, metabolite-mapping analysis on dialysates of LSPD-treated rats revealed that the liposomes did not remove important metabolites more than a conventional PD fluid. Overall, these findings confirm that LSPD is a potentially safe and effective approach for treating hyperammonemic crises in the context of acute-on-chronic liver failure.

Thyme and Savory Essential Oil Vapor Treatments Control Brown Rot and Improve the Storage Quality of Peaches and Nectarines, but Could Favor Gray Mold.

The effect of biofumigation, through slow-release diffusors, of thyme and savory essential oils (EO), was evaluated on the control of postharvest diseases and quality of peaches and nectarines. EO fumigation was effective in controlling postharvest rots. Naturally contaminated peaches and nectarines were exposed to EO vapors for 28 days at 0 °C in sealed storage cabinets and then exposed at 20 °C for five days during shelf-life in normal atmosphere, simulating retail conditions. Under low disease pressure, most treatments significantly reduced fruit rot incidence during shelf-life, while, under high disease pressure, only vapors of thyme essential oil at the highest concentration tested (10% v/v in the diffusor) significantly reduced the rots. The application of thyme or savory EO favored a reduction of brown rot incidence, caused by Monilinia fructicola, but increased gray mold, caused by Botrytis cinerea. In vitro tests confirmed that M. fructicola was more sensitive to EO vapors than B. cinerea. Essential oil volatile components were characterized in storage cabinets during postharvest. The antifungal components of the essential oils increased during storage, but they were a low fraction of the volatile organic compounds in storage chambers. EO vapors did not influence the overall quality of the fruit, but showed a positive effect in reducing weight loss and in maintaining ascorbic acid and carotenoid content. The application of thyme and savory essential oil vapors represents a promising tool for reducing postharvest losses and preserving the quality of peaches and nectarines.

Improving bioavailability and biodistribution of anti-HIV chemotherapy.

In the context of the treatment of HIV/AIDS, many improvements have been achieved since the introduction of the combination therapy (HAART). Nevertheless, no cure for this disease has been so far possible, because of some particular features of the therapies. Among them, two important ones have been selected and will be the subject of this review. The first main concern in the treatments is the poor drug bioavailability, resulting in repeated administrations and therefore a demanding compliance (drug regimens consist of multiple drugs daily intake, and non-adherence to therapy is among the important reasons for treatment failure). A second important challenge is the need to target the drugs into the so-called reservoirs and sanctuaries, i.e. cells or body compartments where drugs cannot penetrate or are distributed in sub-active concentrations. The lack of antiviral action in these regions allows the virus to lie latent and start to replicate at any moment after therapy suspension. Recent drug delivery strategies addressing these two limitations will be presented in this review. In the first part, strategies to improve the bioavailability are proposed in order to overcome the absorption or the target cell barrier, or to extend the efficacy time of drugs. In the second section, the biodistribution issues are considered in order to target the drugs into the reservoirs and the sanctuaries, in particular the mononuclear phagocyte system and the brain.

Anthocyanins, phenolics and antioxidant capacity after fresh storage of blueberry treated with edible coatings.

The influence of different edible coatings on total phenolic content, total anthocyanin and antioxidant capacity in highbush blueberry (Vaccinium corymbosum L. cv Berkeley and O'Neal) was investigated, mainly for industrial applications. Also titratable acidity, soluble solids content, firmness and weight loss of berries were determined at harvest and at 15-day intervals during 45 storage days at 0 °C, in order to optimize coating composition. Application of chitosan coating delayed the decrease in anthocyanin content, phenolic content and antioxidant capacity. Coating samples showed no significant reduction in the weight loss during storage period. In cv Berkeley, the use of alginate coating showed a positive effect on firmness, titratable acidity and maintained surface lightness of treated berries. In cv O'Neal, no significant differences in total soluble solids content were found, and the chitosan-coated berries showed the minimum firmness losses. In both cultivars, the addition of chitosan to coatings decreases the microbial growth rate.

Stabilization and cellular delivery of chitosan-polyphosphate nanoparticles by incorporation of iron.

Chitosan (CS) nanoparticles are typically obtained by complexation with tripolyphosphate (TPP) ions, or more recently using triphosphate group-containing drugs such as adenosine triphosphate (ATP). ATP is an active molecule we aim to deliver in order to restore its depletion in macrophages, when associated with their death leading to plaque rupture in atherosclerotic lesions. Despite high interest in CS nanoparticles for drug delivery, due to the biodegradability of CS and to the ease of the preparation process, these systems tend to readily disintegrate when diluted in physiological media. Some stabilization strategies have been proposed so far but they typically involve the addition of a coating agent or chemical cross-linkers. In this study, we propose the complexation of CS with iron ions prior to nanoparticle formation as a strategy to improve the carrier stability. This can be achieved thanks to the ability of iron to strongly bind both chitosan and phosphate groups. Nanoparticles were obtained from either TPP or ATP and chitosan-iron (CS-Fe) complexes containing 3 to 12% w/w iron. Isothermal titration calorimetry showed that the binding affinity of TPP and ATP to CS-Fe increased with the iron content of CS-Fe complexes. The stability of these nanoparticles in physiological conditions was evaluated by turbidity and by fluorescence fluctuation in real time upon dilution by electrolytes, and revealed an important stabilization effect of CS-Fe compared to CS, increasing with the iron content. Furthermore, in vitro studies on two macrophage cell lines (J774A.1 and THP-1) revealed that ATP uptake is improved consistently with the iron content of CS-Fe/ATP nanoparticles, and correlated to their lower dissociation in biological medium, allowing interesting perspectives for the intracellular delivery of ATP.

Drug-induced nanocarrier assembly as a strategy for the cellular delivery of nucleotides and nucleotide analogues.

The natural nucleotide adenosine triphosphate (ATP) and nucleotide analogues such as azidothymidine triphosphate (AZT-TP) display important pharmacological activities for the treatment of ischemia and HIV infections, respectively. Their clinical use is, however, limited mostly due to their hydrophilicity, which highly restricts their diffusion into the target cells. Few nanocarriers have been proposed to address the challenge of ATP/AZT-TP cellular delivery, but the loading efficiency, preparation complexity, and efficient cellular delivery remain important barriers to their development. In this study, we propose an original, straightforward and versatile design of nucleotide and nucleotide analogue nanocarriers based on the natural polysaccharide chitosan (CS). We show that the drugs ATP and AZT-TP can induce ionotropic gelation of CS, leading to CS/ATP and CS/AZT-TP nanoparticles with high drug entrapment efficiency and loading rate-up to 44%. Such nanocarriers release ATP and AZT-TP in physiological media and allow an efficient in vitro cellular delivery of these molecules down to the cell cytoplasm.

Contrast-enhanced magnetic resonance imaging of fibrovascular tissue ingrowth within synthetic hydroxyapatite orbital implants in children.

PURPOSE: To assess the usefulness of magnetic resonance imaging (MRI) in assessing fibrovascularization progression into synthetic hydroxyapatite (HA) implants inserted in anophthalmic sockets of children submitted to enucleation. METHODS: We studied 23 HA orbital implants in 23 children who underwent enucleation for retinoblastoma. Each patient was examined by MRI within to 9 to 69 weeks after implant insertion (mean 34 weeks, median 30 weeks). No patient had received chemotherapy or radiotherapy at MRI examination. From each T1-weighted, fat-suppressed enhanced axial examination, the image depicting the center of the implant was identified. Enhancement was evaluated using a 5-point scale. Additionally, possible associated orbital and intracranial pathologies and implant migration or extrusion were evaluated. RESULTS: All patients showed areas of enhancement of the implant consistent with the presence of fibrovascular ingrowth. There was no grade 1 enhancement in our series. Grade 2 was observed in 1 patient (4.34%), grade 3 in 7 patients (30.43%), grade 4 in 11 patients (47.82%), and grade 5 in 4 patients (17.39%). During follow-up there were no cases of clinically evident orbital infection, implant migration, or implant extrusion. No second tumor, optic nerve invasion, orbital extension, tumor relapse, or leptomeningeal brain seeding were noted. CONCLUSIONS: In this series, enhanced MRI showed satisfactory fibrovascular ingrowth of orbital implants since the 13th week after HA spheres insertion, with a trend towards progressive enhancement during the following weeks. The data also confirm the advice to wait at least 5-6 months after enucleation to perform drilling and peg placement.

Shelf-life extension of highbush blueberry using 1-methylcyclopropene stored under air and controlled atmosphere.

The potential of 1-methylcyclopropene for controlling ripening in 'Lateblue' blueberry fruit was explored. After harvest, blueberry fruits were exposed to 1-MCP (0.3 and 0.6µll(-1)). After treatment, samples were stored in air at 0°C for 35days and in a controlled atmosphere (3kPa O2+11kPa CO2) for 60days. Quality parameters were monitored (weight loss, total soluble solids content, titratable acidity, firmness, anthocyanin content, phenolic content, total antioxidant capacity). Blueberries treated with 1-MCP showed a reduced weight loss during storage and a lower total soluble solid content compared to untreated fruit. High titratable acidity values were observed after controlled atmosphere storage, but no significant effect of 1-MCP on this parameter was observed. 1-MCP had no significant effects on anthocyanins, phenolics or antioxidant activities.