Supported polymer/lipid hybrid bilayers formation resembles a lipid-like dynamic by reducing the molecular weight of the polymer.

Amphiphilic block copolymers form self-assembled bilayers even in combination with phospholipids. They represent an attractive alternative to native lipid-based membrane systems for supported bilayer formation with applications in biomedical research, sensoring and drug delivery. Their enhanced stability and excellent mechanical properties are linked to their higher molecular weight which generates thicker bilayers. Hypothesis: It is hypothesized that reducing the molecular weight of the polymer facilitates the formation of a thinner, more homogeneous polymer/lipid hybrid bilayer which would benefit the formation of supported bilayers on silicon oxide. Experiment: We investigated hybrid bilayers composed of mixtures of 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine and increasing amounts of a low molecular weight polybutadiene-b-polyethylene oxide copolymer (1050 g/mol). By assessing the bilayer thickness and the molecular packing behavior we sought to demonstrate how reducing the polymer molecular weight increases the tendency to form supported hybrid bilayers in a lipid-like manner. Findings: The formation of a supported hybrid bilayers occurs at polymer contents70 mol% in a lipid-like fashion and is proportional to the cohesive forces between the bilayer components and inversely related to the bilayer hydrophobic core thickness and the extended brush regime of the PEGylated polymeric headgroup

In vivo pharmacological activity and biodistribution of S-nitrosophytochelatins after intravenous and intranasal administration in mice

AbstractS-nitrosophytochelatins (SNOPCs) are novel analogues of S-nitrosoglutathione (GSNO) with the advantage of carrying varying ratios of S-nitrosothiol (SNO) moieties per molecule. Our aim was to investigate the in vivo pharmacological potency and biodistribution of these new GSNO analogues after intravenous (i.v.) and intranasal (i.n.) administration in mice. SNOPCs with either two or six SNO groups and GSNO were synthesized and characterized for purity. Compounds were administered i.v. or i.n. at 1 μmol NO/kg body weight to CD-1 mice. Blood pressure was measured and biodistribution studies of total nitrate and nitrite species (NOx) and phytochelatins were performed after i.v. administration. At equivalent doses of NO, it was observed that SNOPC-6 generated a rapid and significantly greater reduction in blood pressure (∼60% reduction compared to saline) whereas GSNO and SNOPC-2 only achieved a 30–35% decrease. The reduction in blood pressure was transient and recovered to baseline levels within ∼2 min for all compounds. NOx species were transiently elevated (over 5 min) in the plasma, lung, heart and liver. Interestingly, a size-dependent phytochelatin accumulation was observed in several tissues including the heart, lungs, kidney, brain and liver. Biodistribution profiles of NOx were also obtained after i.n. administration, showing significant lung retention of NOx over 15 min with minor systemic increases observed from 5 to 15 min. In summary, this study has revealed interesting in vivo pharmacological properties of SNOPCs, with regard to their dramatic hypotensive effects and differing biodistribution patterns following two different routes of administration

Comparison of Oral, Intranasal and Aerosol Administration of Amiodarone in Rats as a Model of Pulmonary Phospholipidosis.

‘Foamy’ alveolar macrophages (FAM) observed in nonclinical toxicology studies during inhaled drug development may indicate drug-induced phospholipidosis, but can also derive from adaptive non-adverse mechanisms. Orally administered amiodarone is currently used as a model of pulmonary phospholipidosis and it was hypothesized that aerosol administration would produce phospholipidosis-induced FAM that could be characterized and used in comparative inhalation toxicology. Han-Wistar rats were given amiodarone via (1) intranasal administration (6.25 mg/kg) on two days, (2) aerosol administration (3 mg/kg) on two days, (3) aerosol administration (10 mg/kg) followed by three days of 30 mg/kg or (4) oral administration (100 mg/kg) for 7 days. Alveolar macrophages in bronchoalveolar lavage were evaluated by di_erential cell counting and high content fluorescence imaging. Histopathology and mass-spectrometry imaging (MSI) were performed on lung slices. The higher dose aerosolised amiodarone caused transient pulmonary inflammation (p < 0.05), but only oral amiodarone resulted in FAM (p < 0.001). MSI of the lungs of orally treated rats revealed a homogenous distribution of amiodarone and a putative phospholipidosis marker, di-22:6 bis-monoacylglycerol, throughout lung tissue whereas aerosol administration resulted in localization of both compounds around the airway lumen. Thus, unlike oral administration, aerosolised amiodarone failed to produce the expected FAM responses.Peer reviewedFinal Published versio

Implications of Polystyrene Microplastics on the Gastric Digestion of Bovine Milk

The prevalence of microplastics (MP) pollution in different zones of the environment has been established by several studies [1]. Due to its widespread presence, MP have found its way into food items. Fish, shellfish, water, milk, salt, and sugar are just some examples of the food we commonly consume that are contaminated with MP [2]. Human ingestion of MP is already well-established but there is limited data regarding how MP affect human gastric digestion of food components, especially proteins. In this study, we investigated the effects of polystyrene (PS) MP on pepsin, the major protease in human gastric digestion. Pepsin activity was tested during exposure to two different sizes -10 μm (PS10) and 100 μm (PS100), and three different quantities- low count (142 particles), moderate count (1420 particles), and high count (14200 particles), of PS using haemoglobin as substrate. Results showed that exposure to PS100 has no effect on enzyme activity. However, exposure to high count PS10 considerably reduced pepsin activity from 2957 ± 310 U/mg to 1674 ± 270 U/mg. To test the effect on food digestion, high count PS10 was added to a sample of commercially available liquid bovine milk (defatted). In this case, the static in vitro simulation of gastric digestion was followed to mimic human digestion of food [3]. Milk digesta at different time points (5, 10, 15, 20, 30, 60, 90, 120 minutes) were obtained to monitor the progress of protein degradation. SDS-PAGE showed no difference in the peptide bands from 30-120 minutes. However, bands corresponding to caseins were not observed at 5 minutes when PS10 was present. Additionally, 14 kDa fragments were not observed at 10-20 minutes. Washing of the PS particles followed by SDSPAGE revealed a faint pepsin band from all time points. At 5 and 10 minutes, faint peptide bands >10kDa were also observed. These suggest that pepsin and some milk peptides were adsorbed on the surface of PS10. Zeta potential analysis of PS revealed a slightly negative surface charge which could explain the adsorption and disappearance of peptide bands. This adsorption of pepsin on PS did not seem to affect its overall protease activity. However, the interaction of milk peptides with PS may reduce the nutrients human could acquire from milk. Acknowledgements This study was supported by Ghent University Global Campus; Special Research Fund (BOF) of Ghent University (grant number 01N01718) and IMPTOX European Union’s Horizon 2020 research and innovation program (grant number 965173). References [1] S. Sharma, S. Basu, N. P. Shetti, M. N. Nadagouda, T. M. Aminabhavi (2021) Chem. Eng. J., 408, 127317. [2] K. D. Cox, G. A. Covernton, H. L. Davies, J. F. Dower, F. Juanes, S. E. Dudas (2019) Environ. Sci. Technol., 53(12), 7068–7074. [3] A. Brodkorb et al. (2019) Nat. Protoc., 14(4), 991– 1014

Imaging drugs, metabolites and biomarkers in rodent lung: a DESI MS strategy for the evaluation of drug-induced lipidosis

© The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.Within drug development and pre-clinical trials, a common, significant and poorly understood event is the development of drug-induced lipidosis in tissues and cells. In this manuscript, we describe a mass spectrometry imaging strategy, involving repeated analysis of tissue sections by DESI MS, in positive and negative polarities, using MS and MS/MS modes. We present results of the detected distributions of the administered drug, drug metabolites, lipid molecules and a putative marker of lipidosis, di-docosahexaenoyl (22:6)-bis(monoacylglycerol) phosphate (di-22:6-BMP). A range of strategies have previously been reported for detection, isolation and identification of this compound, which is an isomer of di-docosahexaenoic (22:6 n-3) phosphatidylglycerol (di-22:6 PG), a commonly found lipid that acts as a surfactant in lung tissues. We show that MS imaging using MS/MS can be used to differentiate these compounds of identical mass, based upon the different distributions of abundant fragment ions. Registration of images of these fragments, and detected drugs and metabolites, is presented as a new method for studying drug-induced lipidosis in tissues. Graphical abstract.Peer reviewe

Dynamics of aerosol size during inhalation : Hygroscopic growth of commercial nebulizer formulations

We thank the Elizabeth Blackwell Institute (EBI) for financial support through the EBI Early Career Research Fellowship awarded to AEH, and the EPSRC for financial support through a Leadership Fellowship awarded to JPR (grant reference EP/G007713/1). This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are creditedThe size of aerosol particles prior to, and during, inhalation influences the site of deposition within the lung. As such, a detailed understanding of the hygroscopic growth of an aerosol during inhalation is necessary to accurately model the deposited dose. In the first part of this study, it is demonstrated that the aerosol produced by a nebulizer, depending on the airflows rates, may experience a (predictable) wide range of relative humidity prior to inhalation and undergo dramatic changes in both size and solute concentration. A series of sensitive single aerosol analysis techniques are then used to make measurements of the relative humidity dependent thermodynamic equilibrium properties of aerosol generated from four common nebulizer formulations. Measurements are also reported of the kinetics of mass transport during the evaporation or condensation of water from the aerosol. Combined, these measurements allow accurate prediction of the temporal response of the aerosol size prior to and during inhalation. Specifically, we compare aerosol composed of pure saline (150 mM sodium chloride solution in ultrapure water) with two commercially available nebulizer products containing relatively low compound doses: Breath, consisting of a simple salbutamol sulfate solution (5 mg/2.5 mL; 1.7 mM) in saline, and Flixotide Nebules, consisting of a more complex stabilized fluticasone propionate suspension (0.25 mg/mL; 0.5 mM in saline. A mimic of the commercial product Tobi (60 mg/mL tobramycin and 2.25 mg/mL NaC1, pH 5.5-6.5) is also studied, which was prepared in house. In all cases, the presence of the pharmaceutical was shown to have a profound effect on the magnitude, and in some cases the rate, of the mass flux of water to and from the aerosol as compared to saline. These findings provide physical chemical evidence supporting observations from human inhalation studies, and suggest that using the growth dynamics of a pure saline aerosol in a lung inhalation model to represent nebulizer formulations may not be representative of the actual behavior of the aerosolized drug solutions. (C) 2014 Published by Elsevier B.V.Peer reviewe

Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion

Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion

Lung inflammation does not affect the clearance kinetics of lipid nanocapsules following pulmonary administration

Lipid nanocapsules (LNCs) are semi-rigid spherical capsules with a triglyceride core that present a promising formulation option for the pulmonary delivery of drugs with poor aqueous solubility. Whilst the biodistribution of LNCs of different size has been studied following intravenous administration, the fate of LNCs following pulmonary delivery has not been reported. We investigated quantitatively whether lung inflammation affects the clearance of 50nm lipid nanocapsules, or is exacerbated by their pulmonary administration. Studies were conducted in mice with lipopolysaccharide-induced lung inflammation compared to healthy controls. Particle deposition and nanocapsule clearance kinetics were measured by single photon emission computed tomography/computed tomography (SPECT/CT) imaging over 48 h. A significantly lower lung dose of (111)In-LNC50 was achieved in the lipopolysaccharide (LPS)-treated animals compared with healthy controls (p<0.001). When normalised to the delivered lung dose, the clearance kinetics of (111)In-LNC50 from the lungs fit a first order model with an elimination half-life of 10.5±0.9h (R(2)=0.995) and 10.6±0.3h (R(2)=1.000) for healthy and inflamed lungs respectively (n=3). In contrast, (111)In-diethylene triamine pentaacetic acid (DTPA), a small hydrophilic molecule, was cleared rapidly from the lungs with the majority of the dose absorbed within 20min of administration. Biodistribution to lungs, stomach-intestine, liver, trachea-throat and blood at the end of the imaging period was unaltered by lung inflammation. This study demonstrated that lung clearance and whole body distribution of lipid nanocapsules were unaffected by the presence of acute lung inflammation

Influence of Surfactant Structure on Photoluminescent ?-Conjugated Polymer Nanoparticles: Interfacial Properties and Protein Binding

π-Conjugated polymer nanoparticles (CPNs) are under investigation as photoluminescent agents for diagnostics and bioimaging. To determine whether the choice of surfactant can improve CPN properties and prevent protein adsorption, five nonionic polyethylene glycol alkyl ether surfactants were used to produce CPNs from three representative π-conjugated polymers. The surfactant structure did not influence size or yield, which was dependent on the nature of the conjugated polymer. Hydrophobic interaction chromatography, contact angle, quartz crystal microbalance, and neutron reflectivity studies were used to assess the affinity of the surfactant to the conjugated polymer surface and indicated that all surfactants were displaced by the addition of a model serum protein. In summary, CPN preparation methods which rely on surface coating of a conjugated polymer core with amphiphilic surfactants may produce systems with good yields and colloidal stability in vitro, but may be susceptible to significant surface alterations in physiological fluids