Mucin Solution Research Articles

Study of Lactobacillus plantarum coated with Tremella polysaccharides to improve its intestinal adhesion.

The adhesion of probiotics to the intestine is crucial for their probiotic function. In previous studies, Tremella polysaccharides (TPS) (with sodium casein) have shown the potential to encapsulate probiotics and protect them in a simulated gastrointestinal tract. This study explored the effect of TPS (with sodium casein) on the adhesion of probiotics. Lactobacillus plantarum was coated with TPS and sodium casein in different proportions, and was freeze-dried. The rheological properties of the mixture of probiotics powder and mucin solution were determined by static and dynamic rheological analysis. Aqueous solutions of probiotic powder and mucin mixture exhibited pseudoplastic fluid rheological properties. The higher the proportion of TPS content, the higher the apparent viscosity and yield stress. The mixed bacterial powder and mucin fluid displayed thixotropy and was in accordance with the Herschel-Bulkley model. The TPS increased the bio-adhesive force of the probiotic powder and mucin. When using TPS as the only carbon source, the adhesion of L. plantarum to Caco-2 cells increased by 228% in comparison with glucose in vitro. Twelve adhesive proteins were also detected in the whole-cell proteome of L. plantarum. Among them, ten adhesive proteins occurred abundantly when grown with TPS as a carbon source. Tremella polysaccharides therefore possess probiotic properties and can promote the intestinal adhesion of L. plantarum. © 2024 Society of Chemical Industry.

A Novel Combined Dry Powder Inhaler Comprising Nanosized Ketoprofen-Embedded Mannitol-Coated Microparticles for Pulmonary Inflammations: Development, In Vitro-In Silico Characterization, and Cell Line Evaluation.

Pulmonary inflammations such as chronic obstructive pulmonary disease and cystic fibrosis are widespread and can be fatal, especially when they are characterized by abnormal mucus accumulation. Inhaled corticosteroids are commonly used for lung inflammations despite their considerable side effects. By utilizing particle engineering techniques, a combined dry powder inhaler (DPI) comprising nanosized ketoprofen-embedded mannitol-coated microparticles was developed. A nanoembedded microparticle system means a novel advance in pulmonary delivery by enhancing local pulmonary deposition while avoiding clearance mechanisms. Ketoprofen, a poorly water-soluble anti-inflammatory drug, was dispersed in the stabilizer solution and then homogenized by ultraturrax. Following this, a ketoprofen-containing nanosuspension was produced by wet-media milling. Furthermore, co-spray drying was conducted with L-leucine (dispersity enhancer) and mannitol (coating and mucuactive agent). Particle size, morphology, dissolution, permeation, viscosity, in vitro and in silico deposition, cytotoxicity, and anti-inflammatory effect were investigated. The particle size of the ketoprofen-containing nanosuspension was ~230 nm. SEM images of the spray-dried powder displayed wrinkled, coated, and nearly spherical particles with a final size of ~2 µm (nano-in-micro), which is optimal for pulmonary delivery. The mannitol-containing samples decreased the viscosity of 10% mucin solution. The results of the mass median aerodynamic diameter (2.4-4.5 µm), fine particle fraction (56-71%), permeation (five-fold enhancement), and dissolution (80% release in 5 min) confirmed that the system is ideal for local inhalation. All samples showed a significant anti-inflammatory effect and decreased IL-6 on the LPS-treated U937 cell line with low cytotoxicity. Hence, developing an innovative combined DPI comprising ketoprofen and mannitol by employing a nano-in-micro approach is a potential treatment for lung inflammations.

Preparation of Soluble Mucin Solutions from the Salivary Glands.

Studying salivary gland mucins is important for elucidating the pathogenesis of salivary gland diseases, including tumors and xerostomia, and developing diagnostic methods for them. Classic methods for isolating mucins from salivary glands require sacrificing several animals to obtain sufficient quantities of mucin and are time-consuming. Supported molecular matrix electrophoresis (SMME) was used to characterize mucins and their glycans. With this method, mucins can be analyzed within 2days using less than 100mg of tissue and without using expensive equipment, such as an ultracentrifuge. This chapter describes a method for preparing mucin solutions for SMME analysis of salivary gland mucins.

Micro-interaction of montmorillonite-loaded nanoparticles with mucin promotes retention of betaxolol hydrochloride on the ocular surface and the tear film microenvironment

This work developed a triple-encapsulated nano-delivery system for topical and long-term glaucoma treatment. We encapsulated betaxolol hydrochloride (BH), a glaucoma drug loaded in refined acid-treated montmorillonite (RAMt), in hyaluronic acid/chitosan (HA/CS) cross-linked nanoparticles with high encapsulation efficiency and further wrapped them with Eudragit (ED) films to achieve dual sustained release. RAMt-BH-HA/CS@ED NPs exhibited excellent physicochemical properties for the sustained release of betaxolol hydrochloride. In vitro release experiments were performed, and the results showed that the drug release lasted for up to 8 h. Then, particle surface hydrophobicity experiments showed that the nanoparticles had both hydrophilic and hydrophobic properties. The ocular safety of the nanoparticles was confirmed by the chick embryo allantoic membrane assay and the rabbit blink frequency test. In the mucin-nanoparticle interaction experiments, the interaction between the nanoparticles and the mucin solution with high pH and low concentration (simulating the aqueous layer microenvironment of tears) was stronger than that with low pH and high concentration (simulating the tear mucin layer microenvironment of tears). Therefore, the nanoparticles showed excellent retention properties in tears and cornea. In vivo experiments in a rabbit model of elevated intraocular pressure demonstrated that the IOP-lowering effect was sustained for more than 12 hours. In conclusion, nanoparticles effectively lower IOP with excellent tolerability, and offer promising applications for long-term treatment of glaucoma.

Tribology test design for friction measurements with application to oral medicines

In recent years tribology tests have been used to measure friction properties of oral consumables such as semi-solid foods and medicines. The tests aim to simulate thin-film mastication conditions and are intended to correlate with mouth feel or food texture properties. In this paper a new approach is proposed to better simulate shear conditions, fluid supply and friction data capture associated with mastication and swallowing. Two primary changes are suggested: these are the reduction of the inlet influence on lubricant film properties and the ability to measure transient and time-dependent friction. The new test was used to measure friction for a range of oral medicines including a viscous solution (cough syrup) and particulate suspensions (paediatric, calcium carbonate) in combination with an artificial saliva (mucin solution), The tongue-palate was replicated by a PCX glass lens loaded and reciprocating against a textured silicone surface. A short stroke length, comparable to the Hertzian diameter of the contact, was used so the contact operated in a partially replenished lubrication condition. This ensured the film in the contact region has the same composition as the bulk fluid. Friction was measured continuously during reciprocation for up to 5 cycles (comparable to mastication time) and data was sampled at 100 Hz to capture transient friction. Tests were run with and without a mucin layer present. The results showed that tests performed after 20 min adsorption of an artificial saliva solution reduced the friction coefficient from μ = 1 to μ = 0.2–0.3. Tests with the paracetamol suspensions, which contain hard particles, recorded transient friction spikes which were not recorded for the softer calcium carbonate suspensions. Key conclusions for the design of pertinent simulation tests are that the film properties in the oral cavity are not determined by the inlet as for classical lubrication. The (bulk) oral sample is captured in the tongue-palate contact and not continually replenished. When sheared it will exhibit time (transient, longer-term) and speed dependent friction responses which contribute to sensory and texture perception.

Size-Dependent Diffusion and Dispersion of Particles in Mucin.

Mucus, composed significantly of glycosylated mucins, is a soft and rheologically complex material that lines respiratory, reproductive, and gastrointestinal tracts in mammals. Mucus may present as a gel, as a highly viscous fluid, or as a viscoelastic fluid. Mucus acts as a barrier to the transport of harmful microbes and inhaled atmospheric pollutants to underlying cellular tissue. Studies on mucin gels have provided critical insights into the chemistry of the gels, their swelling kinetics, and the diffusion and permeability of molecular constituents such as water. The transport and dispersion of micron and sub-micron particles in mucin gels and solutions, however, differs from the motion of small molecules since the much larger tracers may interact with microstructure of the mucin network. Here, using brightfield and fluorescence microscopy, high-speed particle tracking, and passive microrheology, we study the thermally driven stochastic movement of 0.5-5.0 μm tracer particles in 10% mucin solutions at neutral pH, and in 10% mucin mixed with industrially relevant dust; specifically, unmodified limestone rock dust, modified limestone, and crystalline silica. Particle trajectories are used to calculate mean square displacements and the displacement probability distributions; these are then used to assess tracer diffusion and transport. Complex moduli are concomitantly extracted using established microrheology techniques. We find that under the conditions analyzed, the reconstituted mucin behaves as a weak viscoelastic fluid rather than as a viscoelastic gel. For small- to moderately sized tracers with a diameter of lessthan 2 μm, we find that effective diffusion coefficients follow the classical Stokes-Einstein relationship. Tracer diffusivity in dust-laden mucin is surprisingly larger than in bare mucin. Probability distributions of mean squared displacements suggest that heterogeneity, transient trapping, and electrostatic interactions impact dispersion and overall transport, especially for larger tracers. Our results motivate further exploration of physiochemical and rheological mechanisms mediating particle transport in mucin solutions and gels.

A membrane-free microfluidic approach to mucus permeation for efficient differentiation of mucoadhesive and mucopermeating nanoparticulate systems.

The gastrointestinal mucus barrier is a widely overlooked yet essential component of the intestinal epithelium, responsible for the body's protection against harmful pathogens and particulates. This, coupled with the increasing utilisation of biological molecules as therapeutics (e.g. monoclonal antibodies, RNA vaccines and synthetic proteins) and nanoparticle formulations for drug delivery, necessitates that we consider the additional absorption barrier that the mucus layer may pose. It is imperative that in vitro permeability methods can accurately model this barrier in addition to standardised cellular testing. In this study, a mucus-on-a-chip (MOAC) microfluidic device was engineered and developed to quantify the permeation kinetics of nanoparticles through a biorelevant synthetic mucus layer. Three equivalently sized nanoparticle systems, formulated from chitosan (CSNP), mesoporous silica (MSNP) and poly (lactic-co-glycolic) acid (PLGA-NP) were prepared to encompass various surface chemistries and nanostructures and were assessed for their mucopermeation within the MOAC. Utilising this device, the mucoadhesive behaviour of chitosan nanoparticles was clearly visualised, a phenomenon not often observed via standard permeation models. In contrast, MSNP and PLGA-NP displayed mucopermeation, with significant differences in permeation pattern due to specific mucus-nanoparticle binding. Further optimisation of the MOAC to include a more biorelevant mucus mimic resulted in 5.5-fold hindered PLGA-NP permeation compared to a mucin solution. Furthermore, tracking of PLGA-NP at a single nanoparticle resolution revealed rank-order correlations between particle diffusivity and MOAC permeation. This device, including utilisation of biosimilar mucus, provides a unique ability to quantify both mucoadhesion and mucopenetration of nano-formulations and elucidate mucus binding interactions on a microscopic scale.

Bioadhesive behavior of Poloxamer/hyaluronic acid-based formulations analyzed by rheology method

<h2>Abstract</h2> Systems based on Poloxamer and hyaluronic acid have been developed for treatment of back pain as drug delivery formulations. Due that, it is important to study interactions among components and their ability to adhere to mucosa, since these formulations can release drugs on the intervertebral disc. Aqueous formulations with Poloxamer (PL) 407 15%, 30% (w/w), and binary system PL 407 15% + PL 338 15%, with hyaluronic acid (HA) at 0.5% (w/w) and the anesthetic bupivacaine (5 mg/mL) were used. To simulate mucosa, it was prepared mucin solutions (5%). The aim of this work is to compare the bioadhesive force (F) and relative rheological synergism (∆<i>G</i>′/<i>G</i>′) of all formulations when they interact with a simulated mucosa. It is pointed that binary systems show augmented ability to adhere to mucin, approximately 1000 times than other formulations. When it added hyaluronic acid to PL 407 15% and binary system, it is observed that F increases twice. It is seen increased interaction between PL 338-mucin and HA-mucin, demonstrating that more hydrophilic polymers have favored interactions with mucin. This work intends to show that the design of formulations of pharmacological interest can be favored with the use of simple models.

Optimisation of a High-Throughput Model for Mucus Permeation and Nanoparticle Discrimination Using Biosimilar Mucus.

High-throughput permeation models are essential in drug development for timely screening of new drug and formulation candidates. Nevertheless, many current permeability assays fail to account for the presence of the gastrointestinal mucus layer. In this study, an optimised high-throughput mucus permeation model was developed employing a highly biorelevant mucus mimic. While mucus permeation is primarily conducted in a simple mucin solution, the complex chemistry, nanostructure and rheology of mucus is more accurately modelled by a synthetic biosimilar mucus (BSM) employing additional protein, lipid and rheology-modifying polymer components. Utilising BSM, equivalent permeation of various molecular weight fluorescein isothiocyanate-dextrans were observed, compared with native porcine jejunal mucus, confirming replication of the natural mucus permeation barrier. Furthermore, utilising synthetic BSM facilitated the analysis of free protein permeation which could not be quantified in native mucus due to concurrent proteolytic degradation. Additionally, BSM could differentiate between the permeation of poly (lactic-co-glycolic) acid nanoparticles (PLGA-NP) with varying surface chemistries (cationic, anionic and PEGylated), PEG coating density and size, which could not be achieved by a 5% mucin solution. This work confirms the importance of utilising highly biorelevant mucus mimics in permeation studies, and further development will provide an optimal method for high-throughput mucus permeation analysis.

Synthetic Mucin Gels with Self-Healing Properties Augment Lubricity and Inhibit HIV-1 and HSV-2 Transmission.

Mucus is a self-healing gel that lubricates the moist epithelium and provides protection against viruses by binding to viruses smaller than the gel's mesh size and removing them from the mucosal surface by active mucus turnover. As the primary nonaqueous components of mucus (≈0.2%-5%, wt/v), mucins are critical to this function because the dense arrangement of mucin glycans allows multivalence of binding. Following nature's example, bovine submaxillary mucins (BSMs) are assembled into "mucus-like" gels (5%, wt/v) by dynamic covalent crosslinking reactions. The gels exhibit transient liquefaction under high shear strain and immediate self-healing behavior. This study shows that these material properties are essential to provide lubricity. The gels efficiently reduce human immunodeficiency virus type 1 (HIV-1) and genital herpes virus type 2 (HSV-2) infectivity for various types of cells. In contrast, simple mucin solutions, which lack the structural makeup, inhibit HIV-1 significantly less and do not inhibit HSV-2. Mechanistically, the prophylaxis of HIV-1 infection by BSM gels is found to be that the gels trap HIV-1 by binding to the envelope glycoprotein gp120 and suppress cytokine production during viral exposure. Therefore, the authors believe the gels are promising for further development as personal lubricants that can limit viral transmission.

Mucus-penetrating nanoparticles based on chitosan grafted with various non-ionic polymers: Synthesis, structural characterisation and diffusion studies

Transmucosal administration offers numerous advantages for drug delivery as it usually helps to avoid first pass metabolism, provides rapid onset of action, and is a non-invasive route. Mucosal surfaces are covered by a viscoelastic mucus gel layer which acts as a protective barrier preventing the entrance of harmful substances into the human tissues. This function of mucus also inhibits the diffusion of drugs and nano-formulations and can result in a significant reduction of their efficacy. The design of mucus-penetrating nanoparticles can overcome the barrier function of mucus which may lead to better therapeutic outcomes. In this study, chitosan was chemically modified by grafting short chains of poly(ethylene glycol), poly(2-hydroxyethyl acrylate), poly(2-ethyl-2-oxazoline), or poly(N-vinyl pyrrolidone) and the resulting chitosan derivatives were used to prepare nanoparticles using an ionic gelation method with sodium tripolyphosphate. These nanoparticles were characterised using dynamic light scattering, transmission electron microscopy, small-angle neutron scattering and nanoparticle tracking analysis. Small-angle neutron scattering data revealed the presence of a large amount of water inside these nanoparticles and lack of a heterogeneous internal structure. The nanogel model with low crosslinking density is suggested as the most feasible model to describe the structure of these nanoparticles. The studies of the behaviour of these nanoparticles in bovine submaxillary mucin solutions and their penetration into sheep nasal mucosa indicated greater diffusivity of modified chitosan nanoparticles compared to unmodified chitosan nanoparticles with the best results achieved for the chitosan grafted with poly(N-vinyl pyrrolidone).

A Bio-synthetic Hybrid Hydrogel Formed under Physiological Conditions Consisting of Mucin and a Synthetic Polymer Carrying Boronic Acid.

Mucin-containing bio-synthetic hybrid hydrogel is successfully formed under physiological conditions upon mixing aqueous solutions of native mucin and synthetic polymers carrying boronic acids. The mechanical properties and stability of the hydrogel in physiological solutions, e.g., cell culture media, are tunable depending on the boronic acid content of polymers. The hydrogel dissolved in the physiological solutions releases native mucin and boronic acid-containing polymer, which can control the adhesion of mammalian cells to the surface.

Complexation between porcine gastric mucin (PGM) and lysozyme: Influence of heat treatment of lysozyme on the tribological properties

The influence of complexation between porcine gastric mucin (PGM) and lysozyme (LYZ) solutions (pH⁓7.0) on their lubricating properties was studied at a hydrophobic self-mated polydimethylsiloxane (PDMS) tribopair. To this end, LYZ solutions with varying heating time, namely 1hr, 3hr-, and 6hr at 90 °C, as well as unheated LYZ solution, were prepared. The lubricating capability of PGM and LYZ solutions and also their mixtures was characterized using pin-on-disk tribometry. In parallel, to precisely investigate the interaction between PGM and LYZ solutions, an array of the well-known experiments including electrophoretic-dynamic light scattering, circular dichroism spectroscopy and optical waveguide light-mode spectroscopy were employed. These experiments were utilized to elucidate the key features e.g. zeta potential, hydrodynamic diameter, conformational structure and mass adsorption. The tribometry results indicated that both PGM and unheated LYZ solutions had poor lubricating properties in the boundary lubrication regime (sliding speed lower than 10 mm/s). Mixing PGM with unheated LYZ led to a slight decrease in the friction coefficient, but no desirable lubricity was observed. An optimum slippery characteristic was achieved by incorporating 1hr heated LYZ solution into PGM one. Excellent lubricity of PGM/1hr heated LYZ may stem from surface charge compensation, tenaciously compact aggregation, unique conformational structure and considerable mass adsorption onto PDMS. This finding revealed that a strong interaction between PGM and LYZ molecules and as a result, the promising lubricating capability of PGM/LYZ mixtures, can be administered by varying heat-treatment duration of LYZ proteins.

SEDDS facilitate cinnamaldehyde crossing the mucus barrier: The perspective of mucus and Caco-2/HT29 co-culture models

Self-emulsifying drug delivery systems (SEDDS) have potential applications in the delivery of hydrophobic components. Oral drugs are readily captured and cleared by intestinal mucus, a natural barrier that covers the mucosal epithelium and prevents the entry of foreign substances. In this study, we investigated for the first time the ability of SEDDS to deliver the lipophilic aldehyde cinnamaldehyde (CA-SEDDS) in rat mucus, mucin solution, Caco-2 and Caco-2/HT29 co-culture monolayer systems. CA-SEDDS was characterized by particle size, Zeta potential and the logDSEDDS/release medium. The capacity of CA-SEDDS to enhance mucus permeability was investigated in rat intestinal mucus gel and mucin solution with the period of in 12 h by Transwell® diffusion. We evaluated the potential of CA-SEDDS delivery of CA in a co-culture system of absorptive Caco-2 and mucus-secreting HT29 cells. CA-SEDDS exhibited excellent mucus permeability in mucus and mucin solutions, 5.1- and 2.8-fold higher than the free CA group, respectively. CA-SEDDS penetration increased by 2.5-fold compared with free CA when using the mucus-secreting co-culture cell model as a barrier. The relative oral bioavailability of CA-SEDDS was 242% compared to CA without formulation. These findings suggest that SEDDS exhibited good release and superior mucus permeability, displaying great potential for the future of hydrophobic oral applications.

Mimicking the Gastrointestinal Mucus Barrier: Laboratory-Based Approaches to Facilitate an Enhanced Understanding of Mucus Permeation.

The gastrointestinal mucus layer plays a significant role in maintaining gut homeostasis and health, offering protective capacities against the absorption of harmful pathogens as well as commensal gut bacteria and buffering stomach acid to protect the underlying epithelium. Despite this, the mucus barrier is often overlooked during preclinical pharmaceutical development and may pose a significant absorption barrier to high molecular weight or lipophilic drug species. The complex chemical and physical nature of the dynamic mucus layer has proven problematic to reliably replicate in a laboratory setting, leading to the development of multiple mucus models with varying complexity and predictive capacity. This, coupled with the wide range of analysis methods available, has led to a plethora of possible approaches to quantifying mucus permeation; however, the field remains significantly under-represented in biomedical research. For this reason, the development of a concise collation of the available approaches to mucus permeation is essential. In this review, we explore widely utilized mucus mimics ranging in complexity from simple mucin solutions to native mucus preparations for their predictive capacity in mucus permeation analysis. Furthermore, we highlight the diverse range of laboratory-based models available for the analysis of mucus interaction and permeability with a specific focus on in vitro, ex vivo, and in situ models. Finally, we highlight the predictive capacity of these models in correlation with in vivo pharmacokinetic data. This review provides a comprehensive and critical overview of the available technologies to analyze mucus permeation, facilitating the efficient selection of appropriate tools for further advancement in oral drug delivery.

374: Rheological comparison of solubility and gel permanence between airway mucin solutions and mucus systems

374: Rheological comparison of solubility and gel permanence between airway mucin solutions and mucus systems

Biofilm Eradication Four-Step Strategy: Study of Using Self-Assembled Azithromycin/Rhamnolipid Nanoparticles for Removing Pseudomonas aeruginosa Biofilm

To investigate the in vitro eradicative effect of self-assembled azithromycin/rhamnolipid nanoparticles (AZI-RHL NPs) on P seudomonas aeruginosa ( P. aeruginosa) biofilm. AZI-RHL NPs were prepared and characterized. The minimum inhibitory concentration (MIC) of AZI-RHL NPs on planktonic P. aeruginosa was measured by the broth microdilution method. The eradicative effect of AZI-RHL NPs on P. aeruginosa biofilm was evaluated via crystal violet staining and SYTO 9/PI live/dead staining. Fluorescence labeling was used to measure the eradicative effect of NPs on extracellular polymeric substances (EPS). In addition, crystal violet staining was performed to evaluate the inhibitory effect of AZI-RHL NPs on the adhesion of P. aeruginosa on human bronchial epithelial BEAS-2B cells. To investigate the ability of AZI-RHL NPs to penetrate mucus, the interaction between NPs and mucin was measured via particle size changes after co-incubation with mucin solution. The AZI-RHL NPs had a particle size of about 121 nm and were negatively charged on the surface, displaying a high encapsulation efficiency and a high drug loading capacity of 96.72% and 45.08% for AZI, respectively and 99.38% and 53.07% for RHL, respectively. The MIC of AZI-RHL NPs on planktonic P. aeruginosa was half of that of using AZI alone. AZI-RHL NPs displayed the capacity to effectively destroy the biofilm structure and remove the proteins and polysaccharides in EPS, eradicating biofilms in addition to reducing the survival rate of bacteria in the biofilm. AZI-RHL NPs were shown to have inhibited P. aeruginosa adhesion on BEAS-2B cells and prevented the residual bacteria from forming a new biofilm. There was no significant change in the particle size of NPs after co-incubation with mucin solution, indicating a weak interaction between NPs and mucin, and suggesting that NPs could penetrate the mucus and reach the P. aeruginosa infection sites. AZI-RHL NPs were able to effectively enhance the removal of P. aeruginosa biofilm through a four-step strategy of biofilm eradication, including penetrating the mucus, disintegrating the biofilm structure, killing the bacteria dispersed from biofilm, and preventing the adhesion of residual bacteria. We hope that this study will provide a replicable common strategy for the treatment of refractory infections caused by P. aeruginosa and other types of biofilms.

Modulation of Gastrointestinal Mucus Properties with Ionic Liquids for Drug Delivery.

The mucus barrier lining the gastrointestinal tract poses a significant barrier to the oral delivery of macromolecular drugs. Successful approaches to overcoming this barrier have primarily focused on reducing drug and carrier interactions with mucus or disrupting the mucus layer directly. Choline-based ionic liquids (ILs) such as choline geranate and choline glycolate (CGLY) have recently been shown to be effective in enhancing the intestinal absorption of macromolecules such as insulin and immunoglobulin (IgG), respectively. Herein, the use of choline-based ILs as mucus-modulating agents for safely improving drug penetration through mucus is described. Choline-based ILs significantly increase the diffusion rates of cationic dextrans through mucin solution. Choline-maleic acid (CMLC 2:1) enhances the diffusion of 4 kDa cationic dextran in mucin solution by more than fourfold when compared to phosphate-buffered saline control. Choline-based ILs also reduce mucus viscosity without significantly impacting the native mucus gel structure. In vitro studies in a mucus-secreting coculture model with Caco-2 and HT29MTX-E12 cells further demonstrate the effectiveness of ILs in improving transport of cationic molecules in the presence of secreted mucus. This work demonstrates the potential for choline-based ionic liquids to be used as nondestructive mucus-modulating agents for enabling enhanced oral delivery of macromolecular drugs.

Effects of Initially Adsorbed Proteins on Substrate Surfaces during Multilayer Heterogeneous Protein Adsorption

When multilayer heterogeneous proteins are adsorbed on substrate surfaces, the effects of the adsorption state of the initially adsorbed proteins may affect subsequent adsorption. In this study, the relationships between the adsorption state of the initially adsorbed proteins and the amount of secondary adsorbed proteins were examined. A carboxylate-terminated self-assembled monolayer was applied to bovine serum albumin (BSA) solutions of different concentrations for 180 min and subsequently applied to phosphate-buffered saline (PBS) for an additional 180 min to remove weakly adsorbed proteins. The amount of adsorbed proteins was measured using a quartz crystal microbalance with dissipation. The obtained BSA adsorption layer was then applied to mucin solution for 60 min. When a 1.7 mg/mL BSA solution was applied to the surface, the amount of adsorbed BSA after 10 min of adsorption and after washing with PBS for 167 min was >5 × 102 ng/cm2, representing the saturation amount of monolayer-adsorbed BSA in a side-on orientation. In contrast, the amount of adsorbed BSA after 10 min adsorption was <5 × 102 ng/cm2 when a BSA solution with a concentration <0.43 mg/mL was used. The velocity of BSA adsorption plateaued at approximately 0.43 mg/mL, suggesting that the orientation of the adsorbed protein was determined by protein treatment concentration immediately after the proteins were adsorbed. Furthermore, the amount of adsorbed mucin on the BSA adsorption layer decreased as initial BSA treatment concentration increased up to 0.43 mg/mL and plateaued at concentrations above 0.43 mg/mL. These results indicated that the orientation of the initially adsorbed protein was preserved for several hours and affected the subsequent adsorption of mucin.

Viscoelastic particle focusing in human biofluids.

Saliva and blood plasma are non-Newtonian viscoelastic fluids that play essential roles in the transport of particulate matters (e.g., food and blood cells). However, whether the viscoelasticity of such biofluids alters the dynamics of suspended particles is still unknown. In this study, we report that under pressure-driven microflows of both human saliva and blood plasma, spherical particles laterally migrate and form a focused stream along the channel centerline by their viscoelastic properties. We observed that the particle focusing varied among samples on the basis of sampling times/donors, thereby demonstrating that the viscoelasticity of the human biofluids can be affected by their compositions. We showed that the particle focusing, observed in bovine submaxillary mucin solutions, intensified with the increase in mucin concentration. We expect that the findings from this study will contribute to the understanding of the physiological roles of viscoelasticity of human biofluids.