Nanocrystal Suspensions Research Articles

In vitro - in vivo - in silico approach in the development of inhaled drug products: Nanocrystal-based formulations with budesonide as a model drug

This study aims to understand the absorption patterns of three different kinds of inhaled formulations via in silico modeling using budesonide (BUD) as a model drug. The formulations investigated in this study are: (i) commercially available micronized BUD mixed with lactose (BUD-PT), (ii) BUD nanocrystal suspension (BUD-NC), (iii) BUD nanocrystals embedded hyaluronic acid microparticles (BUD-NEM). The deposition patterns of the three inhaled formulations in the rats’ lungs were determined in vivo and in silico predicted, which were used as inputs in GastroPlus™ software to predict drug absorption following aerosolization of the tested formulations. BUD pharmacokinetics, estimated based on intravenous data in rats, was used to establish a drug-specific in silico absorption model. The BUD-specific in silico model revealed that drug pulmonary solubility and absorption rate constant were the key factors affecting pulmonary absorption of BUD-NC and BUD-NEM, respectively. In the case of BUD-PT, the in silico model revealed significant gastrointestinal absorption of BUD, which could be overlooked by traditional in vivo experimental observation. This study demonstrated that in vitro-in vivo-in silico approach was able to identify the key factors that influence the absorption of different inhaled formulations, which may facilitate the development of orally inhaled formulations with different drug release/absorption rates.

Hybrid chemoenzymatic heterogeneous catalyst prepared in one step from zeolite nanocrystals and enzyme-polyelectrolyte complexes.

The combination of inorganic heterogeneous catalysts and enzymes, in so-called hybrid chemoenzymatic heterogeneous catalysts (HCEHCs), is an attractive strategy to effectively run chemoenzymatic reactions. Yet, the preparation of such bifunctional materials remains challenging because both the inorganic and the biological moieties must be integrated in the same solid, while preserving their intrinsic activity. Combining an enzyme and a zeolite, for example, is complicated because the pores of the zeolite are too small to accommodate the enzyme and a covalent anchorage on the surface is often ineffective. Herein, we developed a new pathway to prepare a nanostructured hybrid catalyst built from glucose oxidase and TS-1 zeolite. Such hybrid material can catalyse the in situ biocatalytic formation of H2O2, which is subsequently used by the zeolite to trigger the epoxidation of allylic alcohol. Starting from an enzymatic solution and a suspension of zeolite nanocrystals, the hybrid catalyst is obtained in one step, using a continuous spray drying method. While enzymes are expectedly unable to resist the conditions used in spray drying (temperature, shear stress, etc.), we leverage on the preparation of “enzyme–polyelectrolyte complexes” (EPCs) to increase the enzyme stability. Interestingly, the use of EPCs also prevents enzyme leaching and appears to stabilize the enzyme against pH changes. We show that the one-pot preparation by spray drying gives access to hybrid chemoenzymatic heterogeneous catalysts with unprecedented performance in the targeted chemoenzymatic reaction. The bifunctional catalyst performs much better than the two catalysts operating as separate entities. We anticipate that this strategy could be used as an adaptable method to prepare other types of multifunctional materials starting from a library of functional nanobuilding blocks and biomolecules.

Progress in the development of stabilization strategies for nanocrystal preparations

In recent years, nanocrystal technology has been extensively investigated. Due to the submicron particle size and unique physicochemical properties of nanocrystals, they overcome the problems of low drug solubility and poor bioavailability. Although the structures of nanocrystals are simple, the further development of these materials is hindered by their stability. Drug nanocrystals with particle sizes of 1∼1000 nm usually require the addition of stabilizers such as polymers or surfactants to enhance their stability. The stability of nanocrystal suspensions and the redispersibility of solid nanocrystal drugs are the key factors for the large-scale production of nanocrystal preparations. In this paper, the factors that affect the stability of drug nanocrystal preparations are discussed, and related methods for solving the stability problem are put forward.

From a cholesteric non-aqueous cellulose nanocrystal suspension to a highly ordered film

AbstractA highly ordered cellulose nanocrystal (CNC) film was processed and characterized from a non-aqueous suspension. As a first step, by drawing upon the negative magnetic anisotropy of CNCs, a global order of the nanocrystals is achieved by magnetic field-assisted manipulation of a cholesteric suspension in n-methylformamide (NMF), and then the order is subsequently preserved into a solid-state film. We study the differences between the structures of the 4 T-dried film and the control film dried in the absence of magnetic field. Additionally, we compare the NMF-dried films to those dried from aqueous suspensions with and without magnetic field. Optical microscopy, cross-sectional imaging analysis, and sum frequency generation (SFG) spectroscopy show that the CNC-NMF film dried under magnetic field exhibited a highly ordered layered structure throughout the film, comparable to that observed when films were produced from aqueous suspensions. Extending the potential of the CNC alignment to non-aqueous systems will enable a broad spectrum of applications for CNC-based polymer composites.

Clinically established biodegradable long acting injectables: An industry perspective.

Long acting injectable formulations have been developed to sustain the action of drugs in the body over desired periods of time. These delivery platforms have been utilized for both systemic and local drug delivery applications. This review gives an overview of long acting injectable systems that are currently in clinical use. These products are categorized in three different groups: biodegradable polymeric systems, including microparticles and implants; micro and nanocrystal suspensions and oil-based formulations. Furthermore, the applications of these drug delivery platforms for the management of various chronic diseases are summarized. Finally, this review addresses industrial challenges regarding the development of long acting injectable formulations.

Influence of arabinoxylan on the drying of cellulose nanocrystals suspension: From coffee ring to Maltese cross pattern and application to enzymatic detection

Evaporation of sessile droplet containing suspension of cellulose nanocrystals (CNC) results on birefringent coffee ring pattern (CR), due to the concentration increase and self-assembly of CNC carried by the flow at the edge of evaporating droplet. In this work, we studied the apparition of Maltese cross pattern, (MC) after addition of an hydrosoluble biopolymer belonging to the hemicellulose family, i.e. arabinoxylan (AX). To investigate the mechanisms that control MC pattern apparition, distribution of the two components inside the dried droplet was investigated using FTIR. CNC and AX were found to be homogenously deposited and CNC self-assembly induces nanoparticles orientation in the CR deposit. We demonstrate that the increase of concentration during drying induces gelation of CNC/AX mixture leading to MC pattern apparition. We take advantage of the apparition of MC pattern to develop a novel catalytic activity detection assay based on the variation of viscosity. Indeed, addition of Endo-1,4-β-Xylanase (Xyl) addition to a suspension containing CNC/AX complex leads to hydrolysis of AX that decrease in droplet viscosity leading to MC disappearance. The enzymatic detection assay is thus simple, easy to handle, fast, sensitive and do not require complex analytical devices.

Preparation of parenteral nanocrystal suspensions of etoposide from the excipient free dry state of the drug to enhance in vivo antitumoral properties

Nanoparticle technology in cancer chemotherapy is a promising approach to enhance active ingredient pharmacology and pharmacodynamics. Indeed, drug nanoparticles display various assets such as extended blood lifespan, high drug loading and reduced cytotoxicity leading to better drug compliance. In this context, organic nanocrystal suspensions for pharmaceutical use have been developed in the past ten years. Nanocrystals offer new possibilities by combining the nanoformulation features with the properties of solid dispersed therapeutic ingredients including (i) high loading of the active ingredient, (ii) its bioavailability improvement, and (iii) reduced drug systemic cytotoxicity. However, surprisingly, no antitumoral drug has been marketed as a nanocrystal suspension until now. Etoposide, which is largely used as an anti-cancerous agent against testicular, ovarian, small cell lung, colon and breast cancer in its liquid dosage form, has been selected to develop injectable nanocrystal suspensions designed to be transferred to the clinic. The aim of the present work is to provide optimized formulations for nanostructured etoposide solutions and validate by means of in vitro and in vivo evaluations the efficiency of this multiphase system. Indeed, the etoposide formulated as a nanosuspension by a bottom-up approach showed higher blood life span, reduced tumor growth and higher tolerance in a murine carcinoma cancer model. The results obtained are promising for future clinical evaluation of these etoposide nanosuspensions.

Mechanistic study of preparation of drug/polymer/surfactant ternary hot extrudates to obtain small and stable drug nanocrystal suspensions.

We studied optimized conditions for preparing ternary hot extrudates (HEs) of glibenclamide (GLB)/polyvinylpyrrolidone (PVP)/sodium dodecyl sulfate to generate stable nanocrystal suspensions following aqueous dispersion. Raman and solid-state NMR measurements of ternary HEs prepared by altering HE conditions revealed that GLB crystallinity in HEs reduced with increased extrusion temperature and count and decreased screw speed. Aqueous dispersions of all HEs temporarily formed GLB nanoparticles with a diameter of 75-420nm. The suspension from the HEs with the low GLB crystallinity (<22%) precipitated after 4-h storage, while the HEs with the high GLB crystallinity (>22%) formed stable nanocrystal suspension. Interestingly, the number of GLB nanoparticles <150nm was different despite aqueous dispersion of HEs with similar GLB crystallinity, reflecting the different GLB crystalline size in those HEs. Although both the crushing by shear force and GLB dissolution into PVP reduced GLB crystalline size, the crushing GLB crystal by the shear force has a relatively high ability to decrease GLB crystalline size without excess amorphization of GLB. Performing the hot extrusion at a low temperature, a high screw speed, and maximizing extrusion count with GLB crystallinity >22% led to formation of small and stable nanocrystal suspensions.

Production of Itraconazole Nanocrystal-Based Polymeric Film Formulations for Immediate Drug Release.

In order to improve the solubility properties of BCS class II drug itraconazole, fast dissolving oral polymeric film formulations based on itraconazole nanocrystals were produced. Drug nanocrystals were manufactured by the wet pearl milling technique. In polymeric film formulations, hydroxypropyl methyl cellulose (HPMC) was used as a film forming polymer, and glycerin was used as a plasticizer. For nanocrystal suspensions and film formulations, thorough physicochemical characterization was performed, including particle sizing and size deviation, film appearance, weight variation, thickness, folding endurance, drug content uniformity, disintegration time, and dissolution profile. After milling, the nanoparticles were 369 nm in size with a PI value of 0.20. Nanoparticles were stable and after redispersion from film formulations, the particle size remained almost the same (330 nm and PI 0.16). The produced films were flexible, homogeneous, fast disintegrating, and drug release rate from both the nanosuspension and film formulations showed immediate release behavior. Based on the study, the film casting method for production of itraconazole nanocrystal based immediate release formulations is a good option for improved solubility.

Protein-crystal detection with a compact multimodal multiphoton microscope

There is an increasing demand for rapid, effective methods to identify and detect protein micro- and nano-crystal suspensions for serial diffraction data collection at X-ray free-electron lasers or high-intensity micro-focus synchrotron radiation sources. Here, we demonstrate a compact multimodal, multiphoton microscope, driven by a fiber-based ultrafast laser, enabling excitation wavelengths at 775 nm and 1300 nm for nonlinear optical imaging, which simultaneously records second-harmonic generation, third-harmonic generation and three-photon excited ultraviolet fluorescence to identify and detect protein crystals with high sensitivity. The instrument serves as a valuable and important tool supporting sample scoring and sample optimization in biomolecular crystallography, which we hope will increase the capabilities and productivity of serial diffraction data collection in the future.

Thermodynamics and kinetics of CO2 hydrate formation in the presence of cellulose nanocrystals with statistical treatment of data

Carbon dioxide hydrate was formed in aqueous suspensions of cellulose nanocrystals (CNC) and the induction time and rate of hydrate crystal growth was compared to data obtained when CO2 hydrate was formed in pure water (blank). CNC concentrations of 0.13, 0.5 and 0.75 wt% were employed to determine whether CNC could reduce the induction time and enhance hydrate growth rate as it has been reported for carbon nanotubes and other nanoparticle suspensions. Hydrate formation experiments were carried out in two different crystallizers. The first set-up was in semi-batch operation mode while the second set-up was in batch operation mode. The results show that the CNC did not have a statistically significant impact on CO2 hydrate formation. This study also raises questions about the validity of conclusions and claims made in the literature about the effect of carbon nanotubes and other nanoparticles on gas hydrate formation. Those claims were based on very small sample sizes and without statistical data analysis.

Interrogating helical nanorod self-assembly with fractionated cellulose nanocrystal suspensions

The helical self-assembly of cholesteric liquid crystals is a powerful motif in nature, enabling exceptional performance in many biological composites. Attempts to mimic these remarkable materials by drying cholesteric colloidal nanorod suspensions often yield films with a non-uniform mosaic-like character, severely degrading optical and mechanical properties. Here we show—using the example of cellulose nanocrystals—that these problems are due to rod length dispersity: uncontrolled phase separation results from a divergence in viscosity for short rods, and variations in pitch can be traced back to a twisting power that scales with rod length. We present a generic, robust and scalable method for fractionating nanorod suspensions, allowing us to interrogate key aspects of cholesteric self-assembly that were previously hidden by colloid dispersity. By controlled drying of fractionated suspensions, we can obtain mosaic-free films that are uniform in colour. Our findings unify conflicting observations and open routes to biomimetic artificial materials with performance that can compete with that of nature’s originals.

Yielding of cellulose nanocrystal suspensions in the presence of electrolytes

The rheological properties and yielding behavior of 1 wt. % aqueous sulfonated cellulose nanocrystals (CNCs) in the presence of monovalent (Na+) ions have been investigated. The introduction of more than 20 mM NaCl to the system causes aggregation of neutralized CNCs and leads to the formation of self-similar clusters, which grow in size until they form a three-dimensional network. In the present work, we report a comprehensive study of nonlinear rheology and yielding behavior of CNC/salt gels in steady shear and oscillatory experiments. Two yield stresses have been determined. The first yield stress at low shear rates is attributed to the disconnected CNC clusters as a result of bond breakage. The second yield point occurs at higher shear rates, and it is related to the deformation of clusters, where individual nanorods are nearly separated and dispersed. The existence of these two yield stresses has been identified in both steady-shear scans (high to low and low to high) as well as oscillatory experiments, resulting in consistent results.

Quantitative Calorimetric Studies of the Chiral Nematic Mesophase in Aqueous Cellulose Nanocrystal Suspensions

Aqueous suspensions of cellulose nanocrystals (CNC) can spontaneously form a chiral nematic mesophase at a critical concentration (c*). Unfortunately, no current analytical technique permits rapid detection of c*. Herein, we introduce a facile and accurate approach to assess c* rapidly (< 2 h) from a small sample volume and compare our results to those obtained by conventional methods. Our strategy employs isothermal titration calorimetry (ITC) to measure the heat of interaction, which can identify the onset of mesophase formation since it is sensitive to suspension viscosity and thus capable of detecting small changes in the suspension environment. We measure c* for CNC samples differing in surface charge and aspect ratio and find that lower aspect ratios and higher surface charges both increase c*. Our ITC results reveal the role of CNC interactions prior to visual observation of mesophase formation and elucidate effects related to nanocrystals and their suspensions.

Brightly Luminescent CsPbBr3 Nanocrystals through Ultracentrifugation.

Using a combination of density-gradient and analytical ultracentrifugation, we studied the photophysical profile of CsPbBr3 nanocrystal (NC) suspensions by separating them into size-resolved fractions. Ultracentrifugation drastically alters the ligand profile of the NCs, which necessitates postprocessing to restore colloidal stability and enhance quantum yield (QY). Rejuvenated fractions show a 50% increase in QY compared to no treatment and a 30% increase with respect to the parent. Our results demonstrate how the NC environment can be manipulated to improve photophysical performance, even after there has been a measurable decline in the response. Size separation reveals blue-emitting fractions, a narrowing of photoluminescence spectra in comparison to the parent, and a crossover from single- to stretched-exponential relaxation dynamics with decreasing NC size. As a function of edge length, L, our results confirm that the photoluminescence peak energy scales a L-2, in agreement with the simplest picture of quantum confinement.

Nanoparticle-based 3D membrane for impedimetric biosensor applications

This paper reports on a comparison between nano-ZnO/CuO and nano-ZnO nitrocellulose membrane biosensors, both of which were fabricated using a simple and inexpensive sonication technique. To produce the nano-ZnO/CuO membranes, the technique involved sonication of 1% (w/v) ZnO and 1% (w/v) CuO nano-crystal colloidal suspensions, with a volume ratio of 1:2. The membranes were analysed by scanning electron microscopy and energy-dispersive spectroscopy, which showed the gradated distribution of nanoparticles in the membrane. Impedance spectroscopy demonstrated that the sonication resulted in a greater than two-fold enhancement of the output signal. Changes in impedance phase values, at a frequency of 100 Hz, were used to establish dose dependent responses for C-reactive protein (CRP). Limits of detection of 27 pg/mL for the 1% (w/v) nano-ZnO and 16 pg/mL for the 1% (w/v) nano-ZnO/CuO nitrocellulose membrane biosensors were demonstrated.

Chitin Pickering Emulsion for Oil Inclusion in Composite Films

A film containing a stable and well-dispersed hydrophobic phase in a surfactant-free bio-based hydrophilic matrix is proposed. In this study, an aqueous suspension of rod-like chitin nanocrystals (ChiNCs), mixed with paraffin oil, form an oil-in-water Pickering emulsion with a droplet diameter of 3 μm. These emulsions mixed with a 5 wt% starch solution formed homogeneous composite films by solvent casting. Various amounts of emulsion were incorporated, leading to self-supported films with a volume of oil as high as 45 vol%, with less than 1% of ChiNCs. This model inclusion system leads to droplets homogeneously dispersed throughout the composite films, as revealed by microscopy (SEM and CLSM) with mechanical properties controlled by the matrix. Finally, the droplets were easily released from the matrix by enzymatic hydrolysis. This easy-to-implement transparent film proved to be a good candidate when it is desirable to disperse a poor water-soluble component in a hydrophilic edible matrix.

Nonlinear viscoelastic characterization of charged cellulose nanocrystal network structure in the presence of salt in aqueous media

The change in ionic strength of cellulose nanocrystal (CNC) suspensions is shown to contribute to a respective change in colloidal behavior, such as stiffness and fractal gelation. In this study, dynamic colloidal behavior and stability of aqueous CNC suspensions and their correlation with nonlinear viscoelastic properties of the CNC gel structures in the presence of different concentrations of sodium chloride (NaCl) salt were investigated. The microstructure of CNC/salt suspensions/gels were investigated with a wide range of characterization technique. To obtain further insight into the network structure of CNC/salt systems, for the first time, nonlinear rheology of the suspensions/gels was analyzed to correlate macro-mechanical viscoelastic response of the CNC/salt aqueous systems to structural changes as a response to strain. The intra-cycle viscoelasticity, explained utilizing qualitative Lissajous–Bowditch plots and quantitative nonlinear parameters, demonstrates a strong dependence of the nonlinear response of the samples to salt concentration, CNC concentration, and frequency of deformation. Higher intra-cycle nonlinearity was observed upon increasing the salt loading.

Self-assembly behaviors of colloidal cellulose nanocrystals: A tale of stabilization mechanisms

HypothesisIn solvent casting, colloidal nanocrystal self-assembly patterns are controlled by a mix of cohesive and repulsive interactions that promote destabilization-induced self-assembly (DISA) or evaporation-induced self-assembly (EISA). Tuning the strength and nature of the stabilization mechanisms may allow repulsive interactions to govern self-assembly during the casting of colloidal cellulose nanocrystal (CNC) suspensions. ExperimentsWe propose a tool to classify the level of electrostatic and solvation-induced stabilizations based on two solvent parameters only: dielectric constant, ε, and chemical affinity for CNCs, in terms of Hansen Solubility Parameters, Ra. These criteria are applied to study CNC self-assembly in solvent casting experiments in various media and binary mixtures. FindingsIn solvent casting of suspensions stabilized through a combination of electrostatic and solvation effects, the primarily governing mechanism is EISA, which leads to the formation of chiral nematic domains and optically active thin films. In electrostatically-stabilized suspensions, EISA and DISA are in competition and casting may yield anything from a continuous film to a powder. In other suspensions, DISA prevails and evaporation yields a powder of CNC agglomerates. By classifying media according to their stabilization mechanisms, this work establishes that the behavior of CNC suspensions in solvent casting may be predicted from solvent parameters only.

Retrieving the Coassembly Pathway of Composite Cellulose Nanocrystal Photonic Films from their Angular Optical Response.

Aqueous suspensions of cellulose nanocrystals (CNCs) are known to self-assemble into a chiral nematic liquid crystalline phase, leading to solid-state nanostructured colored films upon solvent evaporation, even in the presence of templating agents. The angular optical response of these structures, and therefore their visual appearance, are completely determined by the spatial arrangement of the CNCs when the drying suspension undergoes a transition from a flowing and liquid crystalline state to a kinetically arrested state. Here, it is demonstrated how the angular response of the final film allows for retrieval of key physical properties and the chemical composition of the suspension at the onset of the kinetic arrest, thus capturing a snapshot of the past. To illustrate this methodology, a dynamically evolving sol-gel coassembly process is investigated by adding various amounts of organosilica precursor, namely, 1,2-bis(trimethoxysilyl)ethane. The influence of organosilica condensation on the kinetic arrest can be tracked and thus explains the angular response of the resulting films. The a posteriori and in situ approach is general; it can be applied to a variety of additives in CNC-based films and it allows access to key rheological information of the suspension without using any dedicated rheological technique.