A Novel image processing technique for weighted particle size distribution assessment.

The objective of the study was to create a reliable method that could be used to evaluate the particle size distribution of samples and pre-mixes in real-world situations, particularly those consisting of typical formulation blends. The goal was to use this method to assess the uniformity of the samples and ensure that they met the required quality standards. The researchers aimed to create a method that could be easily incorporated into the manufacturing process, providing a practical and efficient solution. This study demonstrates the use of ImageJ software to analyze the particle size distribution (PSD) of powders. The technique produces qualitative data from microscopy images and quantitative data from analysis of parameters including average diameter, D10, D50, D90, and standard deviation. The method was tested with various treatments, showing differentiating outcomes in all cases. The alternate technique provides a rapid and cost-effective method for PSD analysis, surpassing the limitations of sieve analysis. Extensive testing of the method, using a variety of sample types, including typical formulation blends, was performed. The results suggest that the method can effectively assess the morphology of changing materials during batch manufacturing and characterize uniformity in blends. The methodology has the capability to identify attributes related to PSD that are typically required to be monitored during manufacturing. The technique allows for accurate and reliable quantification of the attributes through image capture technology. The technique has future potential and has important implications for material science, powder rheology, pharmaceutical formulation development, and continual process monitoring.

[Figure: see text]A Novel Image Processing Technique for Weighted Particle Size Distribution Assessment.

Deep Tissue Penetration of Bottle-Brush Polymers via Cell Capture Evasion and Fast Diffusion.

Drug nanocarriers (NCs) capable of crossing the vascular endothelium and deeply penetrating into dense tissues of the CNS could potentially transform the management of neurological diseases. In the present study, we investigated the interaction of bottle-brush (BB) polymers with different biological barriers in vitro and in vivo and compared it to nanospheres of similar composition. In vitro internalization and permeability assays revealed that BB polymers are not internalized by brain-associated cell lines and translocate much faster across a blood-brain barrier model compared to nanospheres of similar hydrodynamic diameter. These observations performed under static, no-flow conditions were complemented by dynamic assays performed in microvessel arrays on chip and confirmed that BB polymers can escape the vasculature compartment via a paracellular route. BB polymers injected in mice and zebrafish larvae exhibit higher penetration in brain tissues and faster extravasation of microvessels located in the brain compared to nanospheres of similar sizes. The superior diffusivity of BBs in extracellular matrix-like gels combined with their ability to efficiently cross endothelial barriers via a paracellular route position them as promising drug carriers to translocate across the blood-brain barrier and penetrate dense tissue such as the brain, two unmet challenges and ultimate frontiers in nanomedicine.

In Situ Characterization of the Protein Corona of Nanoparticles In Vitro and In Vivo.

A new theoretical framework that enables the use of differential dynamic microscopy (DDM) in fluorescence imaging mode to quantify in situ protein adsorption onto nanoparticles (NP) while simultaneously monitoring for NP aggregation is proposed. This methodology is used to elucidate the thermodynamic and kinetic properties of the protein corona (PC) in vitro and in vivo. The results show that protein adsorption triggers particle aggregation over a wide concentration range and that the formed aggregate structures can be quantified using the proposed methodology. Protein affinity for polystyrene (PS) NPs is observed to be dependent on particle concentration. For complex protein mixtures, this methodology identifies that the PC composition changes with the dilution of serum proteins, demonstrating a Vroman effect never quantitatively assessed in situ on NPs. Finally, DDM allows monitoring of the evolution of the PC in vivo. This results show that the PC composition evolves significantly over time in zebrafish larvae, confirming the inherently dynamic nature of the PC. The performance of the developed methodology allows to obtain quantitative insights into nano-bio interactions in a vast array of physiologically relevant conditions that will serve to further improve the design of nanomedicine.

Scratching the Surface of the Protein Corona: Challenging Measurements and Controversies.

This Review aims to provide a systematic analysis of the literature regarding ongoing debates in protein corona research. Our goal is to portray the current understanding of two fundamental and debated characteristics of the protein corona, namely, the formation of mono- or multilayers of proteins and their binding (ir)reversibility. The statistical analysis we perform reveals that these characterisitics are strongly correlated to some physicochemical factors of the NP-protein system (particle size, bulk material, protein type), whereas the technique of investigation or the type of measurement (in situ or ex situ) do not impact the results, unlike commonly assumed. Regarding the binding reversibility, the experimental design (either dilution or competition experiments) is also shown to be a key factor, probably due to nontrivial protein binding mechanisms, which could explain the paradoxical phenomena reported in the literature. Overall, we suggest that to truly predict and control the protein corona, future efforts should be directed toward the mechanistic aspects of protein adsorption.

Spontaneous shrinking of soft nanoparticles boosts their diffusion in confined media.

Improving nanoparticles (NPs) transport across biological barriers is a significant challenge that could be addressed through understanding NPs diffusion in dense and confined media. Here, we report the ability of soft NPs to shrink in confined environments, therefore boosting their diffusion compared to hard, non-deformable particles. We demonstrate this behavior by embedding microgel NPs in agarose gels. The origin of the shrinking appears to be related to the overlap of the electrostatic double layers (EDL) surrounding the NPs and the agarose fibres. Indeed, it is shown that screening the EDL interactions, by increasing the ionic strength of the medium, prevents the soft particle shrinkage. The shrunken NPs diffuse up to 2 orders of magnitude faster in agarose gel than their hard NP counterparts. These findings provide valuable insights on the role of long range interactions on soft NPs dynamics in crowded environments, and help rationalize the design of more efficient NP-based transport systems.

Chitosan hydrogel micro-bio-devices with complex capillary patterns via reactive-diffusive self-assembly.

We present chitosan hydrogel microfluidic devices with self-assembled complex microcapillary patterns, conveniently formed by a diffusion-reaction process. These patterns in chitosan hydrogels are formed by a single-step procedure involving diffusion of a gelation agent into the polymer solution inside a microfluidic channel. By changing the channel geometry, it is demonstrated how to control capillary length, trajectory and branching. Diffusion of nanoparticles (NPs) in the capillary network is used as a model to effectively mimic the transport of nano-objects in vascularized tissues. Gold NPs diffusion is measured locally in the hydrogel chips, and during their two-step transport through the capillaries to the gel matrix and eventually to embedded cell clusters in the gel. In addition, the quantitative analyses reported in this study provide novel opportunities for theoretical investigation of capillary formation and propagation during diffusive gelation of biopolymers. STATEMENT OF SIGNIFICANCE: Hydrogel micropatterning is a challenging task, which is of interest in several biomedical applications. Creating the patterns through self assembly is highly beneficial, because of the accessible and practical preparation procedure. In this study, we introduced complex self-assembled capillary patterns in chitosan hydrogels using a microfluidic approach. To demonstrate the potential application of these capillary patterns, a vascularized hydrogel with microwells occupied by cells was produced, and the diffusion of gold nanoparticles travelling in the capillaries and diffusing in the gel were evaluated. This model mimics a simplified biological tissue, where nanomedicine has to travel through the vasculature, extravasate into and diffuse through the extracellular matrix and eventually reach targeted cells.

Interfacial Forces across Ionic Liquid Solutions: Effects of Ion Concentration and Water Domains †.

Using the surface force apparatus (SFA), the interaction forces between mica surfaces across ionic liquid (IL) solutions are studied. The IL solution, 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide in propylene carbonate solvent, is used at different concentrations to elucidate the ions' conformation at the interface from the analysis of short-range structural forces. A direct correlation between the ion layer thickness at the interface and the IL molar fraction in the solution is observed, suggesting conformational changes relative to the ion packing density. In addition, effects of large microscopic and macroscopic water domains at the interface are investigated. The microscopic water domains induced significant adhesion at contact because of the long-range capillary forces, which are found to depend on solvent concentration. The macroscopic water domains entirely cover the interaction area, ensuring that the long-range interfacial interactions occur entirely across the aqueous electrolyte solution with dissolved IL ions as the electrolyte. These results help elucidate the interfacial interactions in IL-charged solid interfaces with practical importance in green energy storage, catalysis, and lubrication.

Coiled Coil Affinity-Based Systems for the Controlled Release of Biofunctionalized Gold Nanoparticles from Alginate Hydrogels.

Affinity-based systems represent a promising solution to control the delivery of therapeutics using hydrogels. Here, we report a hybrid system that is based on the peptidic E/K coiled coil affinity pair to mediate the release of gold nanoparticles (NPs) from alginate scaffolds. On one hand, the gold NPs were functionalized with the Ecoil-tagged epidermal growth factor (EGF). The bioactivity of the grafted EGF and the bioavailability of the Ecoil moiety were confirmed by EGF receptor phosphorylation assays and by capturing the functionalized NPs on a Kcoil-derivatized surface. On the other hand, alginate chains were modified with azido-homoalanine Kcoil (Aha-Kcoil) by azide-alkyne click chemistry. The hybrid system was formed by dispersing NPs functionalized with the Ecoil-tagged EGF in alginate hydrogels containing either 0, 10, or 20% of Kcoil-modified alginate (Alg-Kcoil). With 20% of Alg-Kcoil, the release of Ecoil-functionalized NPs was reduced by half when compared to the release of NPs without Ecoil, whereas little to no differences were noticed with either 0 or 10% of Alg-Kcoil. Differential dynamic microscopy was used to determine the diffusion coefficient of the NPs, and the results showed a decrease in the diffusion coefficient of Ecoil-functionalized NPs, when compared to bare PEGylated NPs. Altogether, our data demonstrated that the E/K coiled coil system can control the release of NPs in a high Kcoil content alginate gel, opening diverse applications in drug delivery.

Effect of surface chemistry of polymeric nanoparticles on cutaneous penetration of cholecalciferol.

We investigated the influence of nanoparticle (NP) surface composition on different aspects of skin delivery of a lipophilic drug: chemical stability, release and skin penetration. Cholecalciferol was chosen as a labile model drug. Poly(lactic acid) (PLA)-based NPs without surface coating, with a non-ionic poly(ethylene glycol) (PEG) coating, or with a zwitterionic poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC) coating were prepared using flash nanoprecipitation. Process was optimized to obtain similar hydrodynamic diameters. Polymeric NPs were compared to non-polymeric cholecalciferol formulations. Cholecalciferol stability in aqueous medium was improved by polymeric encapsulation with a valuable effect of a hydrophilic coating. However, the in vitro release of the drug was found independent of the presence of any polymer, as for the drug penetration in an intact skin model. Such tendency was not observed in impaired skin since, when stratum corneum was removed, we found that a neutral hydrophilic coating around NPs reduced drug penetration compared to pure drug NPs and bare PLA NPs. The nature of the hydrophilic block (PEG or PMPC) had however no impact. We hypothesized that NPs surface influenced drug penetration in impaired skin due to different electrostatic interactions between NPs and charged skin components of viable skin layers.

Mechanistic Insights into the Directed Assembly of Hydrogel Blocks Mediated by Polyelectrolytes or Microgels.

In this study, we report the directed assembly of hydrogel blocks mediated by electrostatic interactions. We compared two different assembly mechanisms, one mediated by microgel particles and another mediated by direct interaction between oppositely charged blocks. The system consisted of hydrogel blocks made of an interpenetrated network of (hydroxyethyl)methacrylate-poly(ethylene glycol)dimethacrylate (HEMA-PEGDMA) and either positively charged polyethylenimine (PEI) or negatively charged hyaluronic acid (HA). Positively charged hydrogel blocks were pretreated with negatively charged microgel particles (MG) made of N-isopropylacrylamide-methacrylic acid. Both systems (PEI/HA and PEI/MG) demonstrated spontaneous directed assembly, meaning that positive blocks were systematically found in contact with oppositely charged blocks. Directed assembly in water of PEI/HA blocks resulted in large and open aggregates, while PEI/MG blocks exhibited more compact aggregates. Effects of salt and pH were also assessed for both systems. Inhibition of blocks aggregation was found to appear above a critical salt concentration (CSalt*) which was significantly higher for the PEI/HA system (80 mM) compared to the PEI/MG system (5-20 mM). The observed difference was interpreted in terms of the nanostructure of the contact area between blocks. Blocks aggregation was also found to be controlled by the content of negatively charged groups in the microgels as well as the concentration of MG in the suspension (CMG) used to treat the hydrogel block surfaces. Our results shine light on the subtle differences underlying the adhesion mechanisms between hydrogel blocks and suggest new routes toward the design of innovative complex soft materials.

A simple method for the subnanomolar quantitation of seven ophthalmic drugs in the rabbit eye.

This study describes the development and validation of a new liquid chromatography-tandem mass spectrometry (MS/MS) method capable of simultaneous quantitation of seven ophthalmic drugs-pilocarpine, lidocaine, atropine, proparacaine, timolol, prednisolone, and triamcinolone acetonide-within regions of the rabbit eye. The complete validation of the method was performed using an Agilent 1100 series high-performance liquid chromatography system coupled to a 4000 QTRAP MS/MS detector in positive TurboIonSpray mode with pooled drug solutions. The method sensitivity, evaluated by the lower limit of quantitation in two simulated matrices, yielded lower limits of quantitation of 0.25 nmol L(-1) for most of the drugs. The precision in the low, medium, and high ranges of the calibration curves, the freeze-thaw stability over 1 month, the intraday precision, and the interday precision were all within a 15% limit. The method was used to quantitate the different drugs in the cornea, aqueous humor, vitreous humor, and remaining eye tissues of the rabbit eye. It was validated to a concentration of up to 1.36 ng/g in humors and 5.43 ng/g in tissues. The unprecedented low detection limit of the present method and its ease of implementation allow easy, robust, and reliable quantitation of multiple drugs for rapid in vitro and in vivo evaluation of the local pharmacokinetics of these compounds.

2D, 3D and 4D active compound delivery in tissue engineering and regenerative medicine.

Recent advances in tissue engineering and regenerative medicine have shown that controlling cells microenvironment during growth is a key element to the development of successful therapeutic system. To achieve such control, researchers have first proposed the use of polymeric scaffolds that were able to support cellular growth and, to a certain extent, favor cell organization and tissue structure. With nowadays availability of a large pool of stem cell lines, such approach has appeared to be rather limited since it does not offer the fine control of the cell micro-environment in space and time (4D). Therefore, researchers are currently focusing their efforts on developing strategies that include active compound delivery systems in order to add a fourth dimension to the design of 3D scaffolds. This review will focus on recent concepts and applications of 2D and 3D techniques that have been used to control the load and release of active compounds used to promote cell differentiation and proliferation in or out of a scaffold. We will first present recent advances in the design of 2D polymeric scaffolds and the different techniques that have been used to deposit molecular cues and cells in a controlled fashion. We will continue presenting the recent advances made in the design of 3D scaffolds based on hydrogels as well as polymeric fibers and we will finish by presenting some of the research avenues that are still to be explored.