Air Liquid Research Articles

Integrative genetic and genomic networks identify microRNA associated with COPD and ILD

Chronic obstructive pulmonary disease (COPD) and interstitial lung disease (ILD) are clinically and molecularly heterogeneous diseases. We utilized clustering and integrative network analyses to elucidate roles for microRNAs (miRNAs) and miRNA isoforms (isomiRs) in COPD and ILD pathogenesis. Short RNA sequencing was performed on 351 lung tissue samples of COPD (n = 145), ILD (n = 144) and controls (n = 64). Five distinct subclusters of samples were identified including 1 COPD-predominant cluster and 2 ILD-predominant clusters which associated with different clinical measurements of disease severity. Utilizing 262 samples with gene expression and SNP microarrays, we built disease-specific genetic and expression networks to predict key miRNA regulators of gene expression. Members of miR-449/34 family, known to promote airway differentiation by repressing the Notch pathway, were among the top connected miRNAs in both COPD and ILD networks. Genes associated with miR-449/34 members in the disease networks were enriched among genes that increase in expression with airway differentiation at an air–liquid interface. A highly expressed isomiR containing a novel seed sequence was identified at the miR-34c-5p locus. 47% of the anticorrelated predicted targets for this isomiR were distinct from the canonical seed sequence for miR-34c-5p. Overexpression of the canonical miR-34c-5p and the miR-34c-5p isomiR with an alternative seed sequence down-regulated NOTCH1 and NOTCH4. However, only overexpression of the isomiR down-regulated genes involved in Ras signaling such as CRKL and GRB2. Overall, these findings elucidate molecular heterogeneity inherent across COPD and ILD patients and further suggest roles for miR-34c in regulating disease-associated gene-expression.

Performance evaluation of a hydrostatic flow immersion cooling system for high-current discharge Li-ion batteries

A Li-ion battery can experience thermal runaway when its temperature rise and temperature distribution are uneven due to fast charging/discharging and extreme hot/cold ambient temperature conditions. In Electric vehicles, a battery thermal management system is used to address these challenges and assure superior battery pack performance. The conventional methods include air and indirect liquid (cold plate) cooling thermal management systems are influencing the high thermal gradients from the top to bottom of the cell and from the cell to cell in a battery pack. Also, when the battery is being discharged at a high current rate, it is difficult to achieve the expected heat dissipation requirements. In order to solve these issues, this research article provides a hydrostatic flow single-phase dielectric immersion cooling technique for an 18,650 cylindrical cell Li-ion battery. The three-dimensional electrochemical-thermal model of an 18,650-cell submerged in dielectric fluid is designed using a Multiscale, Multi-Dimensional (MSMD) approach with the Newman, Tiedemann, Gu, and Kim (NTGK) model. The accuracy of the numerical results is validated as being within 2 °C with experimental data at 3C discharge rate. The experimental results are exhibited a better cooling effect with a 33 % reduction in temperature rise at 3C battery discharge rate in 25 °C ambient temperature. With respect to both 25 °C and 60 °C ambient temperature conditions, this cooling system has a superior cooling effect of <10 °C temperature rise in a cell during a high current (3C) discharge rate. The study showed that at extremely high current discharge rates and various ambient temperature circumstances, the Li-ion battery submerged in a dielectric coolant provides enhanced cooling performance and uniform temperature distribution on the battery surface compared to natural convection. This article highlighted that the immersion cooling technique is an effective cooling phenomenon during electrical abuse conditions.

Patchy Bubble‐Propelled Colloids at Interfaces

AbstractLiquid–liquid or liquid–air interfaces provide interesting environments to study colloids and are ubiquitous in nature and industry, as well as relevant in applications involving emulsions and foams. They present a particularly intriguing environment for studying active particles which exhibit a host of phenomena not seen in passive systems. Active particles can also provide on‐demand controllability that greatly expands their use in future applications. However, research on active particles at interfaces is relatively rare compared to those at solid surfaces or in the bulk. Here, magnetically steerable active colloids at liquid–air interfaces that self‐propel by bubble production via the catalytic decomposition of chemical fuel in the liquid medium is presented. The bubble formation and dynamics of “patchy” colloids with a patch of catalytic coating on their surface is investigated and compared to more traditional Janus colloids with a hemispherical coating. The patchy colloids tend to produce smaller bubbles and undergo smoother motion which makes them beneficial for applications such as precise micro‐manipulation. This is demonstrated by manipulating and assembling patterns of passive spheres on a substrate as well as at an air–liquid interface. The propulsion and bubble formation of both the Janus and patchy colloids is characterized and it is found that previously proposed theories are insufficient to fully describe their motion and bubble bursting mechanism. Additionally, the colloids, which reside at the air–liquid interface, demonstrate novel interfacial positive gravitaxis towards the droplet edges which is attributed to a torque resulting from opposing downward and buoyant forces on the colloids.

Comparison of air freezing, liquid immersion freezing and pressure shift freezing on freezing time and quality of snakehead (Channa Argus) fillets

The objective of this study was to investigate the freezing time and quality differences in Snakehead fillets frozen by pressure shift freezing (PSF), conventional air freezing (AF) and liquid immersion freezing (LIF) at −20 °C, −40 °C and − 60 °C, respectively. The results showed that liquid immersion freezing at −60 °C maintained the quality best, with a freezing time of 3.62 min and the cross sectional area of 209.11 um2. Air freezing at −20 °C had the longest freezing time (184.58 min) and the largest cross sectional area (4470.79 um2), and lowest hardness and springiness of the fillets. Pressure shift freezing did not demonstrate the well established advantages of maintaining better product quality found in similar technique with some other foods. The samples of pressure shift freezing also had higher thawing loss and free water ratio after thawing. Therefore, the liquid immersion freezing at lower temperatures was demonstrated to better maintain the quality of frozen products and held significant potential for commercial application. Industrial relevanceFreezing is a widely used method for extending the shelf life of aquatic products, but some freezing methods, especially the slower ones, often lead to the decrease in the quality and commercial value of frozen products during storage. This paper explored the comparison of industrially used freezing techniques (air freezing and liquid immersion freezing) with the novel pressure shift freezing technique. Liquid immersion freezing at −60 °C was found to be the preferred freezing method for Snakehead fillets, which maintained better frozen product quality, with a simple freezing process and low cost.

Pseudomonas aeruginosa type IV pili actively induce mucus contraction to form biofilms in tissue-engineered human airways.

The opportunistic pathogen Pseudomonas aeruginosa causes antibiotic-recalcitrant pneumonia by forming biofilms in the respiratory tract. Despite extensive in vitro experimentation, how P. aeruginosa forms biofilms at the airway mucosa is unresolved. To investigate the process of biofilm formation in realistic conditions, we developed AirGels: 3D, optically accessible tissue-engineered human lung models that emulate the airway mucosal environment. AirGels recapitulate important factors that mediate host-pathogen interactions including mucus secretion, flow and air-liquid interface (ALI), while accommodating high-resolution live microscopy. With AirGels, we investigated the contributions of mucus to P. aeruginosa biofilm biogenesis in in vivo-like conditions. We found that P. aeruginosa forms mucus-associated biofilms within hours by contracting luminal mucus early during colonization. Mucus contractions facilitate aggregation, thereby nucleating biofilms. We show that P. aeruginosa actively contracts mucus using retractile filaments called type IV pili. Our results therefore suggest that, while protecting epithelia, mucus constitutes a breeding ground for biofilms.

Full thickness 3D in vitro conjunctiva model enables goblet cell differentiation

In vitro culture and generation of highly specialized goblet cells is still a major challenge in conjunctival 3D in vitro equivalents. A model comprising all physiological factors, including mucus-secreting goblet cells has the potential to act as a new platform for studies on conjunctival diseases. We isolated primary conjunctival epithelial cells and fibroblasts from human biopsies. 3D models were generated from either epithelial layers or a combination of those with a connective tissue equivalent. Epithelial models were investigated for marker expression and barrier function. Full-thickness models were analyzed for goblet cell morphology and marker expression via immunofluorescence and quantitative real-time PCR. Simple epithelial models cultured at the air–liquid interface showed stratified multi-layer epithelia with pathologic keratinization and without goblet cell formation. The combination with a connective tissue equivalent to generate a full-thickness model led to the formation of a non-keratinized stratified multi-layer epithelium and induced goblet cell differentiation. In our model, a high resemblance to natural conjunctiva was achieved by the combination of conjunctival epithelial cells with fibroblasts embedded in a collagen-hydrogel as connective tissue equivalent. In the future, our conjunctival in vitro equivalent enables the investigation of goblet cell differentiation, conjunctival pathologies as well as drug testing.

PAI-1 regulates AT2-mediated re-alveolarization and ion permeability

BackgroundAcute lung injury is characterized by overwhelmingly elevated PAI-1 in both lung edema fluid and the circulating system. The role of increased PAI-1, encoded by Serpine1 gene, in the regeneration of injured lung epithelium has not been understood completely. This study aimed to investigate the role of Serpine1 in the regulation of alveolar type 2 epithelial cell (AT2) fate in a humanized mouse line carrying diseased mutants (Serpine1Tg).MethodsWild-type (wt) and Serpine1Tg AT2 cells were either cultured as monolayers or 3D alveolospheres. Colony-forming assay and total surface area of organoids were analyzed. AT1 and AT2 cells in organoids were counted by immunohistochemistry and fluorescence-activated cell sorting (FACS). To test the potential effects of elevated PAI-1 on the permeability in the epithelial monolayers, we digitized the biophysical properties of polarized AT2 monolayers grown at the air–liquid interface.ResultsA significant reduction in total AT2 cells harvested in Serpine1Tg mice was observed compared with wt controls. AT2 cells harvested from Serpine1Tg mice reduced significantly over the wt controls. Spheroids formed by Serpine1Tg AT2 cells were lesser than wt control. Similarly, the corresponding surface area, a readout of re-alveolarization of injured epithelium, was markedly reduced in Serpine1Tg organoids. FACS analysis revealed a significant suppression in the number of AT2 cells, in particular, the CD44+ subpopulation, in Serpine1Tg organoids. A lesser ratio of AT1:AT2 cells in Serpine1Tg organoids was observed compared with wt cultures. There was a significant increase in transepithelial resistance but not amiloride inhibition.ConclusionsOur study suggests elevated PAI-1 in injured lungs downregulates alveolar epithelial regeneration by reducing the AT2 self-renewal, particularly in the CD44+ cells.

Marangoni Flow Investigation in Foam Fractionation Phenomenon

In this study, a numerical investigation of the Marangoni flow in foam fractionation was conducted, with a specific focus on the film of micro-foams in both the interior and exterior regions. A three-dimensional node–film–plateau border system was employed to model the system, utilizing time-dependent mass conservation equations. The study emphasized the influence of the surfactant concentration in the foam fractionation column and the mobility of the air–liquid interface on the Marangoni velocity within the film. The results indicated that higher surfactant concentration in the reflux column resulted in a significant increase in Marangoni velocities. Furthermore, a mobile interface enhanced the Marangoni flow, whereas a rigid interface reduced its intensity. The behaviour of the Marangoni flow was explored in both interior and exterior foams, revealing distinct characteristics. The presence of a wall in the exterior foam altered the flow dynamics, leading to a reduced Marangoni velocity compared to interior films.

Vitamin D promotes epithelial tissue repair and host defense responses against influenza H1N1 virus and Staphylococcus aureus infections

BackgroundEarly studies indicated that vitamin D (VD) exerted pleiotropic extra-skeletal effects in the airway, but the definite linkage between VD deficiency and airway host responses remains unclear.Methods142 cases of clinical data from Department of Otolaryngology, the Seventh Affiliated Hospital of Sun Yat-sen University, were collected to characterize the relationship between VD deficiency and chronic rhinosinusitis (CRS). Based on the clinical observations, 2.5-D airway epithelial organoids cultured at the air–liquid interface (ALI) were used to simulate the effects of VD treatment in the development of airway epithelium and the modulation of the host responses against influenza H1N1 virus (representing viral infections) and Staphylococcus aureus (representing bacterial infections) infections in the airway. The intrinsic mechanisms of VD deficiency underlying epithelial remodeling were mapped by transcriptomic as well as proteomic analyses.ResultsIn this study we observed prevailing VD deficiency among inpatients suffering from CRS, a common disease predominantly characterized by epithelial impairment and remodeling. Relative to control organoids cultured without VD, long-term incubation with VD accelerated basal cell proliferation during nasal epithelial development. Under infectious conditions, VD treatment protected the organoids against influenza H1N1 virus and Staphylococcus aureus invasions by reinforcing the respiratory host defenses, including upregulation of LL37, suppression (or inhibition) of proinflammatory cytokines, strengthening of epithelial integrity, and mucociliary clearance. In silico analysis of transcriptomics and proteomics suggested that VD modulated the epithelial development and remodeling, involving epithelial cell proliferation/differentiation, epithelial–mesenchymal transition (EMT), and cytokine signaling in the immune system, as well as responses to microbe, cell junction organization, and extracellular matrix organization via PTEN signaling, independent of TGF-β signaling.ConclusionsOur findings emphasize the importance of managing VD deficiency in clinical settings for the sake of alleviating pathological epithelial remodeling. Vitamin D promotes epithelial tissue repair and host defense responses against influenza H1N1 and Staphylococcus aureus infections.

Thermo‐responsive fluorine‐free foam stabilized by PEO–PPO–PEO triblock copolymer (EO)100(PO)65(EO)100 for pool fire suppression

AbstractResearch and development of novel fluorine‐free materials to replace fluorinated aqueous film‐forming foam (AFFF) are crucial for improving pool fire suppression performance and protecting the environment. In this study, we report the thermo‐responsive fluorine‐free foam stabilized by triblock PEO–PPO–PEO copolymers (EO)100(PO)65(EO)100 for pool fire suppression. Small‐angle X‐ray scattering (SAXS) and reflected light interferometric techniques are conducted to study the molecular self‐assembly in bulk and film thinning behavior, and the foaming kinetics of copolymer solution and thermophysical properties of the liquid foam are studied by dynamic surface tension and oscillatory rheology analysis. At room temperature, the amphipathic structure of PEO–PPO–PEO makes it possible to absorb at the air–liquid interface forming large‐scale liquid foams containing the mobile films with a detergent state. Upon heating to the surface cooling temperature of burning oil, the mobile films can be actively switched into mechanically strong films with rigid surfaces. The in situ switching of the two interfacial states leads to the significant enhancement of the foam stability, especially under the dual defoaming effects of heat and oil. What's more, it is observed that the confinement of organized copolymer micelles in the Plateau borders and micellar self‐layering in film confinement induce drainage delay of foam and film's stepwise thinning phenomenon, further increasing film thickness and enhancing the thermal stability of the foam. In standard fire‐fighting tests, it is proved that the burnback performance exhibited by thermo‐responsive copolymer foams is three times better than that for classical fluorine‐free foams and almost 1.5 times higher than that for commercial AFFF.

An effective air–liquid contactor for CO2 direct air capture using aqueous solvents

The development of a cost-effective, corrosion resistant, high-flux direct air capture (HiDAC) contactor for a solvent-based direct air capture (DAC) process is reported. Literature technoeconomic analyses suggest that the air–liquid contactor can cost over 20 % of the overall DAC plant’s annualized capital costs. To bring down the overall cost of DAC, it is imperative that an effective contactor is developed. A hybrid contactor consisting of a commercial polyvinylchloride structured packing enhanced with stainless-steel 410 random packing has been developed to provide a high surface area for air–liquid contact. The contactor geometry, wettability, corrosion resistance, pressure drop, along with its CO₂ uptake efficiency, CO₂ uptake rates, and extended loading potential using potassium sarcosinate solutions are investigated. Results show that the HiDAC contactor has a relatively high specific surface area (885 m2/m3), which allows for CO₂ uptake efficiencies of up to 75 % and capture rates of up to 550 g of CO₂ per day for a 0.3 × 0.25 × 0.3 m contactor. The contactor also exhibits high levels of wettability and corrosion resistance with amino acid-based DAC solvents. A CO₂ uptake model was developed, and modeling results are compared to experimental data to simulate and predict the performance of the contactor in a DAC process using potassium sarcosinate solvent. The system’s overall mass transfer coefficient and theoretical pressure drop were also calculated, and these results were compared to DAC data found in literature. The results presented indicate that the HiDAC contactor is well suited for DAC due to its high specific surface area, resistance to corrosion, and high degree of wettability.

Development of a novel air–liquid interface airway tissue equivalent model for in vitro respiratory modeling studies

The human airways are complex structures with important interactions between cells, extracellular matrix (ECM) proteins and the biomechanical microenvironment. A robust, well-differentiated in vitro culture system that accurately models these interactions would provide a useful tool for studying normal and pathological airway biology. Here, we report the development and characterization of a physiologically relevant air–liquid interface (ALI) 3D airway ‘organ tissue equivalent’ (OTE) model with three novel features: native pulmonary fibroblasts, solubilized lung ECM, and hydrogel substrate with tunable stiffness and porosity. We demonstrate the versatility of the OTE model by evaluating the impact of these features on human bronchial epithelial (HBE) cell phenotype. Variations of this model were analyzed during 28 days of ALI culture by evaluating epithelial confluence, trans-epithelial electrical resistance, and epithelial phenotype via multispectral immuno-histochemistry and next-generation sequencing. Cultures that included both solubilized lung ECM and native pulmonary fibroblasts within the hydrogel substrate formed well-differentiated ALI cultures that maintained a barrier function and expressed mature epithelial markers relating to goblet, club, and ciliated cells. Modulation of hydrogel stiffness did not negatively impact HBE differentiation and could be a valuable variable to alter epithelial phenotype. This study highlights the feasibility and versatility of a 3D airway OTE model to model the multiple components of the human airway 3D microenvironment.

P170 Trikafta® modulates release of extracellular vesicles in cystic fibrosis

Dysregulation of CFTR leads to chronic inflammation in Cystic Fibrosis (CF) airways. Extracellular vesicles (EVs) have emerged as an important biomarkers of disease. We previously reported elevated levels of EVs in CF with distinct age-related protein profiles. The levels of EVs and its associated proteins in CF in the presence of CF modulators Trikafta (elexacaftor, tezacaftor, ivacaftor), however remains unexplored. The study aims to characterise and study the effect of Trikafta on EVs and its associated protein targets in CF airway models and serum samples from patients with cystic fibrosis (PWCF). Bronchial brushings from CF patients were passaged and differentiated by air liquid interface (ALI) cultures. After 21 days, TEER was recorded and differentiation of mucociliary layer was confirmed with MUC5AC staining. Further the cultures were treated with Trikafta. EVs released into the apical and basal compartments before and after treatment were extracted by differential centrifugation and characterised by NTA and mass spectrometry (MS). The therapeutic restoration of CFTR was examined using immunoblotting. Serum samples from PWCF on Trikafta were also examined for EV release and content before and after 12 months treatment. After 21 days, of ALI culture. EVs were isolated before and after treatment with Trikafta showed a significant 1.5-fold decrease in EVs in both apical and basal compartments respectively (PWCF (Apical) Control: 4.2E+07 ± 3E+06; Treated 3.2E+07 ± 2E+06; (Basal) Control: 1.3E +08 ± 1.6E+07; Treated 8.1E+07 ± 5E+06) demonstrating that Trikafta treatment diminishes epithelial EV releases. MS based proteomics revealed unique EV protein fingerprints post Trikafta treatment for both epithelial EVs and serum EVs with changes in expression of immune proteins were observed. The study demonstrates that Trikafta can modulate EV release and content in PWCF demonstrating novel mechanisms by which CFTR modulators can regulate the inflammatory landscape in CF.

Q1291H-CFTR molecular dynamics simulations and ex vivo theratyping in nasal epithelial models and clinical response to elexacaftor/tezacaftor/ivacaftor in a Q1291H/F508del patient.

Background: Cystic fibrosis (CF) is caused by a wide spectrum of mutations in the CF transmembrane conductance regulator (CFTR) gene, with some leading to non-classical clinical presentations. We present an integrated in vivo, in silico and in vitro investigation of an individual with CF carrying the rare Q1291H-CFTR allele and the common F508del allele. At age 56years, the participant had obstructive lung disease and bronchiectasis, qualifying for Elexacaftor/Tezacaftor/Ivacaftor (ETI) CFTR modulator treatment due to their F508del allele. Q1291H CFTR incurs a splicing defect, producing both a normally spliced but mutant mRNA isoform and a misspliced isoform with a premature termination codon, causing nonsense mediated decay. The effectiveness of ETI in restoring Q1291H-CFTR is largely unknown. Methods: We collected clinical endpoint measurements, including forced expiratory volume in 1s percent predicted (FEV1pp) and body mass index (BMI), and examined medical history. In silico simulations of the Q1291H-CFTR were compared to Q1291R, G551D, and wild-type (WT)-CFTR. We quantified relative Q1291H CFTR mRNA isoform abundance in patient-derived nasal epithelial cells. Differentiated pseudostratified airway epithelial cell models at air liquid interface were created and ETI treatment impact on CFTR was assessed by electrophysiology assays and Western blot. Results: The participant ceased ETI treatment after 3months due to adverse events and no improvement in FEV1pp or BMI. In silico simulations of Q1291H-CFTR identified impairment of ATP binding similar to known gating mutants Q1291R and G551D-CFTR. Q1291H and F508del mRNA transcripts composed 32.91% and 67.09% of total mRNA respectively, indicating 50.94% of Q1291H mRNA was misspliced and degraded. Mature Q1291H-CFTR protein expression was reduced (3.18% ± 0.60% of WT/WT) and remained unchanged with ETI. Baseline CFTR activity was minimal (3.45 ± 0.25μA/cm2) and not enhanced with ETI (5.73 ± 0.48μA/cm2), aligning with the individual's clinical evaluation as a non-responder to ETI. Conclusion: The combination of in silico simulations and in vitro theratyping in patient-derived cell models can effectively assess CFTR modulator efficacy for individuals with non-classical CF manifestations or rare CFTR mutations, guiding personalized treatment strategies and optimizing clinical outcomes.

WS17.06 F/HN pseudotyped lentiviral vector-mediated transduction of non-human primates

Objectives: We have developed a lentiviral vector platform pseudotyped with the Sendai virus F and HN envelope proteins (rSIV.F/HN), including the clinical candidate BI 3720931 for cystic fibrosis (CF) gene therapy. We have previously demonstrated restoration of CFTR function in CF-patient bronchial epithelial cell air–liquid interface cultures and intestinal organoids, as well as efficient and persistent in vivo transduction of murine airways. We have now assessed transduction efficiency and acute toxicology in non-human primates (NHPs).

Cover Feature: Enabling Ultrahigh Surface Area of Covalently‐linked Organic Framework for Boosted CO2 Capture: An Air Liquid Interfacial Plasma as Post‐furnishing Protocol (Chem. Eur. J. 38/2023)

The underlying features of the presented work is a newly devised strategic economical post-furnishing protocol, that is, air–liquid interfacial plasma (ALIP), on a covalently linked organic framework (COF) material for explicitly fine-tuning the surface properties and subsequently developing a new pathway towards boosted CO2 capture. More information can be found in the Research Article by A. Kalita and co-workers (DOI: 10.1002/chem.202300756).

Genetic characterization of a novel organoid from human malignant giant-cell tumor

Malignant giant-cell tumors are extremely rare bone sarcomas that transform from conventional giant-cell tumors during long periods of treatment. Owing to their rarity, no further analysis of their molecular pathogenesis exists, and thus, no standard treatment has been established. Recently, organoid culture methods have been highlighted for recapturing the tumor microenvironment, and we have applied the culture methods and succeeded in establishing patient-derived organoids (PDO) of rare sarcomas. This study aimed to investigate the genomic characteristics of our established novel organoids from human malignant giant-cell tumors. At our institute, we treated a patient with malignant giant-cell tumor. The remaining sarcoma specimens after surgical resection were cultured according to the air–liquid interface organoid-culture method. Organoids were xenografted into NOD-scid IL2Rgnull mice. The developed tumors were histologically and genomically analyzed to compare their characteristics with those of the original tumors. Genetic changes over time throughout treatment were analyzed, and the genomic status of the established organoid was confirmed. Organoids from malignant giant-cell tumors could be serially maintained using air–liquid interface organoid-culture methods. The tumors developed in xenografted NOD-scid IL2Rgnull mice. After several repetitions of the process, a patient-derived organoid line from the malignant giant-cell tumor was established. Immunohistochemical analyses and next-generation sequencing revealed that the established organoids lacked the H3-3A G34W mutation. The xenografted organoids of the malignant giant-cell tumor had phenotypes histologically and genetically similar to those of the original tumor. The established organoids were confirmed to be derived from human malignant giant-cell tumors. In summary, the present study demonstrated a novel organoid model of a malignant giant-cell tumor that was genetically confirmed to be a malignant transformed tumor. Our organoid model could be used to elucidate the molecular pathogenesis of a malignant giant-cell tumor and develop novel treatment modalities.

Virtual High‐Throughput Screening of Vapor‐Deposited Amphiphilic Polymers for Inhibiting Biofilm Formation

AbstractAmphiphilic copolymers (AP) represent a class of novel antibiofouling materials whose chemistry and composition can be tuned to optimize their performance. However, the enormous chemistry‐composition design space associated with AP makes their performance optimization laborious; it is not experimentally feasible to assess and validate all possible AP compositions even with the use of rapid screening methodologies. To address this constraint, a robust model development paradigm is reported, yielding a versatile machine learning approach that accurately predicts biofilm formation by Pseudomonas aeruginosa on a library of AP. The model excels in extracting underlying patterns in a “pooled” dataset from various experimental sources, thereby expanding the design space accessible to the model to a much larger selection of AP chemistries and compositions. The model is used to screen virtual libraries of AP for identification of best‐performing candidates for experimental validation. Initiated chemical vapor deposition is used for the precision synthesis of the model‐selected AP chemistries and compositions for validation at solid–liquid interface (often used in conventional antifouling studies) as well as the air–liquid–solid triple interface. Despite the vastly different growth conditions, the model successfully identifies the best‐performing AP for biofilm inhibition at the triple interface.

Three-Dimensional Oral Mucosal Equivalents as Models for Transmucosal Drug Permeation Studies.

Oral transmucosal administration, where drugs are absorbed directly through the non-keratinized, lining mucosa of the mouth, represents a solution to drug delivery with several advantages. Oral mucosal equivalents (OME) developed as 3D in vitro models are of great interest since they express the correct cell differentiation and tissue architecture, simulating the in vivo conditions better than monolayer cultures or animal tissues. The aim of this work was to develop OME to be used as a membrane for drug permeation studies. We developed both full-thickness (i.e., connective plus epithelial tissue) and split-thickness (i.e., only epithelial tissue) OME using non-tumor-derived human keratinocytes OKF6 TERT-2 obtained from the floor of the mouth. All the OME developed here presented similar transepithelial electrical resistance (TEER) values, comparable to the commercial EpiOral™. Using eletriptan hydrobromide as a model drug, we found that the full-thickness OME had similar drug flux to EpiOral™ (28.8 vs. 29.6 µg/cm2/h), suggesting that the model had the same permeation barrier properties. Furthermore, full-thickness OME showed an increase in ceramide content together with a decrease in phospholipids in comparison to the monolayer culture, indicating that lipid differentiation occurred due to the tissue-engineering protocols. The split-thickness mucosal model resulted in 4-5 cell layers with basal cells still undergoing mitosis. The optimum period at the air-liquid interface for this model was twenty-one days; after longer times, signs of apoptosis appeared. Following the 3R principles, we found that the addition of Ca2+, retinoic acid, linoleic acid, epidermal growth factor and bovine pituitary extract was important but not sufficient to fully replace the fetal bovine serum. Finally, the OME models presented here offer a longer shelf-life than the pre-existing models, which paves the way for the further investigation of broader pharmaceutical applications (i.e., long-term drug exposure, effect on the keratinocytes' differentiation and inflammatory conditions, etc.).

Liquid Film Thickness for Gas-Liquid Annular Flow Across Grid Spacer with Vanes in Simple 3 × 3 Rod Bundle Channel

This study reports the effects of the grid spacer with mixing vane (MV) on gas velocity for air single-phase flow and liquid film thickness for air-water two-phase annular flow in a 3 × 3 rod bundle channel. To investigate the effects of the shape of the spacers, three kinds of spacers were installed into the channel: spacer without MV (without MV), spacer with four MVs (4-MV30), and spacer with two MVs (2-MV30). The 4-MV30 and 2-MV30 had vanes that were inclined 30 deg from the vertical axis. Gas velocity was measured with a hot-wire anemometer and liquid film thickness was measured with the constant electric current method. From the gas velocity measurement, it was found that the irregularities in the velocity distribution became smaller toward downstream of the spacer. For the 2-MV30, the flow distribution was asymmetric and the flow was biased. From the liquid film thickness measurement, it was found that the liquid film thickness became thicker downstream of the spacer.