High-throughput screening of respiratory hazards: Exploring lung surfactant inhibition with 20 benchmark chemicals.

As environmental air quality worsens and respiratory health injuries and diseases increase, it is essential to enhance our ability to develop better methods to identify potential hazards. One promising approach in emerging toxicology involves the utilization of lung surfactant as a model that addresses the limitations of conventional in vitro toxicology methods by incorporating the biophysical aspect of inhalation. This study employed a constrained drop surfactometer to assess 20 chemicals for potential surfactant inhibition. Of these, eight were identified as inhibiting lung surfactant function: 1-aminoethanol, bovine serum albumin, maleic anhydride, propylene glycol, sodium glycocholate, sodium taurocholate, sodium taurodeoxycholate, and Triton X-100. These results are consistent with previously reported chemical-induced acute lung dysfunction in vivo. The study provides information on each chemical's minimum and maximum surface tension conditions and corresponding relative area and contact angle values. Isotherms and box plots are reported for selected chemicals across doses, and vector plots are used to summarize and compare the results concisely. This lung surfactant bioassay is a promising non-animal model for hazard identification, with broader implications for developing predictive modeling and decision-making tools.

Molecular tension microscopy of the LINC complex in live cells.

We present a protocol to measure the effect of pharmacological treatments on the mechanical tension experienced by nesprins at the cytoplasmic surface of the nuclear envelope of mammalian cells in culture. We apply this protocol to MDCK epithelial cells exposed to the actin depolymerization agent cytochalasin D. To do so, we perform confocal spectral imaging of transiently expressed molecular tension sensors of mini-nesprin 2G and analyze the FRET signal from the sensors with a custom-made Fiji script. For complete details on the use and execution of this protocol, please refer to Déjardin et al. (2020).

Influence of the Medium Composition and the Culture Conditions on Surfactin Biosynthesis by a Native Bacillus subtilis natto BS19 Strain.

An effective microbial synthesis of surfactin depends on the composition of the culture medium, the culture conditions and the genetic potential of the producer strain. The aim of this study was to evaluate the suitability of various medium components for the surfactin producing strain and to determine the impact of the culture conditions on the biosynthesis of surfactin isoforms by the newly isolated native strain Bacillus subtilis natto BS19. The efficiency of surfactin biosynthesis was determined by measuring the surface tension of the medium before and after submerged culture (SmF) and by qualitative and quantitative analysis of the obtained compound by high performance liquid chromatography. The highest efficiency of surfactin biosynthesis was achieved using starch as the carbon source and yeast extract as the nitrogen source at pH 7.0 and 37 °C. Potato peelings were selected as an effective waste substrate. It was shown that the increase in the percentage of peel extract in the culture medium enhanced the biosynthesis of surfactin (mg/L) (2-30.9%; 4-46.0% and 6-58.2%), while reducing surface tension of the medium by about 50%. The obtained results constitute a promising basis for further research on biosynthesis of surfactin using potato peelings as a cheap alternative to synthetic medium components.

Ethanol as a Probe for the Mechanism of Bubble Nucleation in the Diet Coke and Mentos Experiment.

The Diet Coke and Mentos experiment involves dropping Mentos candies into carbonated beverages to produce a fountain. This simple experiment has enjoyed popularity with science teachers and the general public. Studies of the physicochemical processes involved in the generation of the fountain have been largely informed by the physics of bubble nucleation. Herein, we probe the effect of ethanol addition on the Diet Coke and Mentos experiment to explore the impact that beverage surface tension and viscosity have on the heights of fountains achieved. Our results indicate that current descriptions of the effects of surface tension and viscosity are not completely understood. We also extend and apply a previously reported, simplified version of Brunauer-Emmett-Teller theory to investigate kinetic and mechanistic aspects of bubble nucleation on the surface of Mentos candies in carbonated beverages. A combination of this new theory and experiment allows for the estimation that the nucleation sites on the Mentos candy that catalyze degassing are 1-3 µm in size, and that between 50,000 and 300,000 of these sites actively nucleate bubbles on a single Mentos candy. While the methods employed are not highly sophisticated, they have potential to stimulate fresh investigations and insights into bubble nucleation in carbonated beverages.

Surfactant Impact on Interfacial Protein Aggregation and Utilization of Surface Tension to Predict Surfactant Requirements for Biological Formulations.

Biological drug products are formulated with excipients to maintain stability over the shelf life of the product. Surfactants are added to the drug product to stabilize air-water interfaces known to induce protein aggregation. Early formulation development is focused on maintaining protein conformation and colloidal stability over the course of the drug product shelf life but rarely considers stability through dose preparation and administration. Specifically, intravenous (IV) bag preparation exposes the therapeutic protein to a different solution environment concurrently diluting the stabilizing excipients that had been added to the drug product formulation. Mixing in IV bags can generate dynamic changes in the air-water interfacial area known to cause protein aggregation if not sufficiently protected. Therefore, understanding the surfactant requirements for drug product end-to-end stability in early formulation development provides critical information for a right-first-time approach to drug product formulation and robust clinical preparation. The goal of these studies was to understand if interfacial properties of proteins could predict surfactant formulation requirements for end-to-end stability. Specifically, the interfacial properties of five proteins were measured in 0.9% saline and 5% dextrose. Furthermore, shaking studies were conducted to identify the minimum surfactant concentration required to prevent subvisible and visible particle formulation in each diluent. The impact of surfactant type and concentration on particle generation and size was explored. A mathematical model was generated to predict the minimum surfactant concentration required to prevent interface-driven aggregation in each diluent based on the change in surface pressure upon exposure of the protein to the interface. The model was tested under typical IV-preparation conditions with experimental output closely matching the model prediction. By employing this model and better understanding the role of surfactants in interfacial stability, drug product development can generate robust end-to-end large molecule formulations across shelf life, dose preparation, and administration.

Excretory overexpression of hydrophobins as multifunctional biosurfactants in E. coli.

Hydrophobins are small amphipathic proteins excreted from filamentous fungi that self-assemble into the amphipathic film at hydrophobic/hydrophilic interfaces and can be used in a wide range of biotechnological application such as antimicrobial coatings, biosensors, and drug delivery. Here we describe a simple method for producing functionally active class I and class II hydrophobins in E. coli. The class I hydrophobin EAS (rodlet protein) from Neurospora crassa and class II hydrophobin HFBII from Trichoderma reesei were separately fused with fusion partner Ffu312 (ß-fructofuranosidase truncation with a native signal peptide) and successfully expressed in E. coli. Significantly, fused hydrophobins Ffu312-EAS and Ffu312-HFBII were excreted into the culture medium. The excretory expression of hydrophobins facilitated the correct disulfide-bond formation and simplified the purification. Both fusion hydrophobins reversed the glass surface hydrophilicity, reduced the water surface tension and improved emulsion stability. Ffu312 has little effect on surface coating, water surface tension and emulsion stabilization of hydrophobins. This study may provide an efficient approach for excretory and functional expression of class I and class II hydrophobins in E. coli.

Formulation of hyaluronan grafted with dodecanoic acid as a potential ophthalmic treatment.

This work concerns the chemical modification of medium molecular weight hyaluronan for ophthalmic applications. The synthesis of amphiphilic HA with dodecanoyl moities was carried out under mild aqueous conditions. Perfect control of the degree of substitution was obtained by varying the molar ratio of activated fatty acid used in the reaction feed. Moreover, the preparation of the derivatives was optimized to achieve the desired degree of substitution (DS = 9.0 ± 0.2 %). The prepared hyaluronan derivatives were water-soluble and exhibited self-associating properties (amphiphilicity). The structure of the prepared derivatives was elucidated by NMR spectroscopy, rheology, turbidity, SEC-MALLS, and gas chromatography (GC). The hydrophobic moieties increase the solution viscosity by physical crosslinking. Low concentration of HAC12 is needed to prepare highly viscous solutions with potential use for ophthalmic applications. Amphiphilic HA kept the biocompatibility of hyaluronan. The degree of substitution and Mw of the amphiphilic HA controls the sterilization by filtration. The protection against desiccation was tested using human keratinocytes (HaCaT) cells lines.

Acute Inhalation Toxicity After Inhalation of ZnO Nanoparticles: Lung Surfactant Function Inhibition In Vitro Correlates With Reduced Tidal Volume in Mice.

People can be exposed to zinc oxide (ZnO) by inhalation of consumer products or during industrial processes. Zinc oxide nanoparticle (NP) exposure can induce acute inhalation toxicity. The toxicological mechanisms underlying the acute effects on the lungs have long focused on the phagolysosomal dissolution of ZnO NPs in macrophages followed by the release of free Zn2+ ions. However, we postulate an alternative mechanism based on the direct interaction of ZnO NPs with the lung surfactant (LS) layer covering the inside of the alveoli. Therefore, we tested the effect of ZnO NPs and Zn2+ ions on the function of LS in vitro using the constrained drop surfactometer. We found that the ZnO NPs inhibited the LS function, whereas Zn2+ ions did not. To examine the role of lung macrophages in the acute toxicity of inhaled ZnO NPs, mice were treated with Clodrosome, a drug that depletes alveolar macrophages, or Encapsome, the empty carrier of the drug. After macrophage depletion, the mice were exposed to an aerosol of ZnO NPs in whole body plethysmographs recording breathing patterns continuously. Mice in both groups developed shallow breathing (reduced tidal volume) shortly after the onset of exposure to ZnO NPs. This suggests a macrophage-independent mechanism of induction. This study shows that acute inhalation toxicity is caused by ZnO NP interaction with LS, independently of NP dissolution in macrophages.

Comparison of mono-rhamnolipids and di-rhamnolipids on microbial enhanced oil recovery (MEOR) applications.

Rhamnolipids (RMLs) have more effectiveness for specific uses according to their homologue proportions. Thus, the novelty of this work was to compare mono-RMLs and di-RMLs physicochemical properties on microbial enhanced oil recovery (MEOR) applications. For this, RML produced by three strains of Pseudomonas aeruginosa containing different homologues proportion were used: a mainly mono-RMLs producer (mono-RMLs); a mainly di-RMLs producer (di-RMLs), and the other one that produces relatively balanced amounts of mono-RML and di-RML homologues (mono/di-RML). For mono-RML, the most abundant molecules were Rha-C10 C10 (m/z 503.3), for di-RML were RhaRha-C10 C10 (m/z 649.4) and for Mono/di-RML were Rha-C10 C10 (m/z 503.3) and RhaRha-C10 C10 (m/z 649.4). All RMLs types presented robustness under high temperature and variation of salinity and pH, and high ability for oil displacement, foam stability, wettability reversal and were classified as safe for environment according to the European Union Directive No. 67/548/EEC. For all these properties, it was observed a highlight for mono-RML. Mono-RML presented the lowest surface tension (26.40 mN/m), interfacial tension (1.14 mN/m), and critical micellar concentration (CMC 27.04 mg/L), the highest emulsification index (EI24 100%) and the best wettability reversal (100% with 25 ppm). In addition, mono-RML showed the best acute toxicity value (454 mg/L), making its application potential even more attractive. Based on the results, it was concluded that all RMLs homologues studied have potential for MEOR applications. However, results showed that mono-RML stood out and have the best mechanism of oil incorporation in micelles due their most effective surface-active physicochemical features.

Evaluation of Thickened Fluids Used in Dysphagia Management Using Extensional Rheology.

Recent studies show that understanding the rheological properties of thickened fluids, such as viscosity and yield stress, is advantageous in designing optimal thickened fluids for the treatment of dysphagia. To date, these studies have focused on the rheological behavior of thickened fluids in shear deformation, while limited information is available on the surface tension of thickened fluids or their rheological behavior in extensional deformation. Knowledge of the extensional properties of thickened fluids (extensional viscosity and cohesiveness) is important to fully understand the behavior of such fluids while swallowing. Our aim in this work, therefore, was to characterize water and skim milk thickened with a commercial thickener (xanthan gum based) to determine extensional deformation and surface tension properties. It was observed that the surface tension decreases as the thickener concentration increases due to the accumulation of the biopolymer at the surface of the fluid when it dissolves in water. In addition, the extensional viscosity of the fluid increased over time as the filament thinned (i.e., as the Hencky strain increased) until it reached a plateau. It was observed that the maximum extensional viscosity, which is related to the cohesiveness of the fluid, increases with the higher concentrations of thickener. However, the cohesiveness of thickened skim milk was lower than that of the thickened water at a given thickener concentration due to lower surface tension. This study confirms that by increasing the concentration of thickener, it will not only increase the shear viscosity (i.e., bolus thickness) of the fluid, but also the extensional viscosity (i.e., bolus cohesiveness).

Wetting and adhesion behavior on apple tree leaf surface by adding different surfactants.

OBJECTIVE: This study was conducted to investigate the wetting behavior of different surfactant solutions on the leaf surfaces of apple during the fruit formation stage. METHODS: Five surfactants, including C12E5, Tween-20, Triton X-100, DTAB, and SDS were evaluated in this study. The contact angle, surface tension, adhesion tension, work of adhesion, and solid-liquid interface tension of droplets on the leaf surface were determined by the drop method. RESULTS: The results showed that the nonionic surfactants C12E5 and Triton X-100 had better wetting effects than other surfactants. Moreover, when the concentration of C12E5 and Triton X-100 was 1 × 10-3 mol/L, the leaves reached a completely wet state. Toxicity measurement showed that the incubation rate of Carposina niponensis eggs decreased gradually with increasing content of C12E5 or Triton X-100. Additionally, field efficacy analysis showed that adding C12E5 or Triton X-100 significantly improved the beta-cyfluthrin 3% water emulsion (EW) against C. niponensis. CONCLUSIONS: These results indicate that the surfactants C12E5 and Triton X-100 can significantly improve pesticide application, which will be helpful for reducing pesticide use and developing new pesticides.

How a lipid bilayer membrane responds to an oscillating nanoparticle: Promoted membrane undulation and directional wave propagation.

Mechanical forces acting on a plasma membrane are of essential importance to cellular functioning via inducing delicate change of the membrane shape with the underlying mechanism yet to be elucidated. Here, we introduce an oscillating nanoparticle (NP) interaction with a lipid bilayer membrane, using the coarse-grained simulation to investigate the dynamic membrane response to constrained mechanical stimulation, which is ubiquitous in biology. Our results demonstrate that, the membrane responds to an oscillating NP by generating nanoscale undulation waves, which immediately propagate through the membrane. In dynamics, propagation of the generated membrane undulation waves always starts from flattening of the region where the NP locates, thus producing a lateral force to propel the waves away from the point of stimulation. The speed of membrane undulation wave propagation is proportional to that of NP oscillation and accelerated by increasing the integral membrane surface tension, suggesting that both the membrane bending and stretching contribute to the energy driving the unique response of membrane undulation wave propagation.

Determination of rheology and surface tension of airway surface liquid: a review of clinical relevance and measurement techniques.

By airway surface liquid, we mean a thin fluid continuum consisting of the airway lining layer and the alveolar lining layer, which not only serves as a protective barrier against foreign particles but also contributes to maintaining normal respiratory mechanics. In recent years, measurements of the rheological properties of airway surface liquid have attracted considerable clinical attention due to new advances in microrheology instruments and methods. This article reviews the clinical relevance of measurements of airway surface liquid viscoelasticity and surface tension from four main aspects: maintaining the stability of the airways and alveoli, preventing ventilator-induced lung injury, optimizing surfactant replacement therapy for respiratory syndrome distress, and characterizing the barrier properties of airway mucus to improve drug and gene delivery. Primary measuring techniques and methods suitable for determining the viscoelasticity and surface tension of airway surface liquid are then introduced with respect to principles, advantages and limitations. Cone and plate viscometers and particle tracking microrheometers are the most commonly used instruments for measuring the bulk viscosity and microviscosity of airway surface liquid, respectively, and pendant drop methods are particularly suitable for the measurement of airway surface liquid surface tension in vitro. Currently, in vivo and in situ measurements of the viscoelasticity and surface tension of the airway surface liquid in humans still presents many challenges.

Evaluation of Yield and Surface Tension-lowering Activity of Iturin A Produced by Bacillus subtilis RB14.

Bacillus subtilis RB14 produces the lipopeptide antibiotic iturin A by submerged and biofilm fermentation. In this study, we optimized the conditions for iturin A production in a jar fermentor. The maximum yield of iturin A was 932 mg L-1 after 120 h. The surface tension of water decreased from 72.0 to 39.0 mN m-1 as the concentrations of C14 iturin A increased, indicating that C14 iturin A behaves as a surfactant in water. The critical micellar concentration obtained from the intersection of two fitted lines was 1.2 × 10-4 M. Moreover, the surface tension of water decreased as the length of the alkyl chain of iturin A increased.

Biophysical and chemical stability of surfactant/budesonide and the pulmonary distribution following intra-tracheal administration.

Intra-tracheal instillation of budesonide using surfactant as a vehicle significantly decreased the incidence of bronchopulmonary dysplasia or death in preterm infants. The formularity of surfactant supplemented with budesonide and biophysical and chemical stability of the suspension has not been well reported. The aims are to investigate the biophysical and chemical stability of two surfactant preparations, Survanta and Curosurf, supplemented with budesonide. Biophysical property of the surface tension of Survanta and Survanta/budesonide suspension and of Curosurf and Curosurf/budesonide suspension was conducted by a pulsating bubble surfactometer and by a drop shape tensiometer. Chemical stability of Survanta/budesonide and of Curosurf/budesonide suspensions was tested by high-performance liquid chromatography analysis (HPLC). Pulmonary distribution of Survanta/18F-budesonide suspension was examined by a Nano/PET digital scan in rats. The Marangoni effect of Survanta, Curosurf, and budesonide was tested by digital high speed photography. For Survanta supplemented with budesonide, with a concentration ratio of ≥50, the surface tension-lowering activity was minimally affected. Similarly, the surface tension-lowering activity of Curosurf was not significantly affected by addition of budesonide, if the concentration ratio was ≥160. With these concentration ratios of both suspensions, HPLC analysis revealed no new compounds identified. Curosurf as compared to Survanta exhibited a significantly higher Marangoni effect. We conclude that with current dosage recommended for Survanta and Curosurf, both surfactant/budesonide suspensions are biophysically and chemically stable. Both surfactants can act as an effective vehicle for budesonide delivery.

Effects of Polymer/Surfactant as Carriers on the Solubility and Dissolution of Fenofibrate Solid Dispersion.

The purpose of this work is to investigate the effects of polymer/surfactant as carriers on the solubility and dissolution of fenofibrate solid dispersions (FF SDs) with the aid of systematic research on the physicochemical properties of the polymer/surfactant system and further highlight the importance of studying polymer/surfactant interaction in the preformulation. The critical micelle concentration (CMC) of sodium lauryl sulfate (SLS) and critical aggregation concentration (CAC) of polymer/SLS solutions were obtained through conductivity measurement. Meanwhile, surface tension, viscosity, morphology, and wettability of polymer/SLS with different weight ratios of SLS were analyzed to screen out the suitable content of SLS (weight%, 5% in carriers) incorporated in SDs. Polymer/SLS coprecipitate and FF SDs were prepared by the solvent evaporation method. The results from differential scanning calorimetry (DSC) and X-ray diffraction (XRD) analysis showed that FF was molecularly dispersed in SDs. Compared to the solubility of FF in povidone/SLS (PVP/SLS) solutions, the increment of FF solubility in copovidone/SLS (VA64/SLS) solutions was due to the formation of free SLS micelles, which have been confirmed by transmission electron microscopy (TEM). Particularly, the wettability of FF SDs and physical mixtures (PMs) was also determined by the sessile drop technique. A linear relationship between the wettability of carriers and that of FF SDs was found, which revealed the significant role of carriers on the surface composition of FF SDs. As the molecular weight of PVP increased, the wettability of carriers decreased, thus leading to the reduction of the dissolution rate of SDs. Although the presence of SLS did not enhance the dissolution of FF SDs, it increased the amount of drug released at the initial stage. All these results indicated that the polymer/SLS interaction would affect the performance of SDs; hence, it was necessary to study their properties in the preformulation.

Purification and characterization of a biosurfactant produced by Bacillus subtilis in cashew apple juice and its application in the remediation of oil-contaminated soil.

The ability of some microorganisms to use clarified cashew apple juice as carbon and energy source for biosurfactant production was assessed under strict controlled conditions. Twelve strains of Bacillus were isolated and evaluated regarding their biosurfactant production capabilities. The biosurfactant obtained with these selected strains showed the capacity of decreasing the surface tension of water from 72.0 to 31.8 mN.m-1 and the interfacial tension of n-hexadecane to 27.2 mN.m-1, with a critical micelle concentration of 12.5 mg.L-1. Not only did the biosurfactant present excellent stability to pH, temperature and salinity, it also showed emulsifying properties in different hydrocarbons. The behavior of the phase diagrams showed the potential of the produced biosurfactant to obtain relatively-stable emulsions for up to 96 h, which allows for its application in several areas. The semi-purified biosurfactant did not show toxicity against Lactuca sativa (lettuce) or Artemia salina (microcrustacean), presenting an LC50 of 612.27 µ mL-1. The surfactant was characterized as being a cyclic lipopeptide with molecular structure similar to that of surfactin. Furthermore, through the employment of the surfactant produced, the remediation effect in oil-contaminated soil could be significantly improved.

Qualitative analysis of biosurfactants from Bacillus species exhibiting antifungal activity.

Bacillus spp. produce a broad spectrum of lipopeptide biosurfactants, among which surfactin, iturin and fengycin are widely studied families. The goals of this study were to characterize the biosurfactant activity of Bacillus spp. and to investigate their motility and biofilm formation capabilities. In addition, we extracted lipopeptides from these bacteria to assess their antifungal activities and analyzed these products by mass spectrometry (MS). B. amyloliquefaciens FZB42, Bacillus sp. NH 217 and B. subtilis NH-100 exhibited excellent biosurfactant and surface spreading activities, whereas B. atrophaeus 176s and Paenibacillus polymyxa C1225 showed moderate activity, and B. subtilis 168 showed no activity. Strains FZB42, NH-100, NH-217, 176s and CC125 exhibited excellent biofilm formation capabilities. Lipopeptide extracts displayed good antifungal activity against various phytopathogens and their associated diseases, such as Fusarium moniliforme (rice bakanae disease), Fusarium oxysporum (root rot), Fusarium solani (root rot) and Trichoderma atroviride (ear rot and root rot). Lipopeptide extracts of these strains also showed hemolytic activity, demonstrating their strong potential to produce surfactants. LCMS-ESI analyses identified the presence of surfactin, iturin and fengycin in the extracts of Bacillus strains. Thus, the strains assayed in this study show potential as biocontrol agents against various Fusarium and Trichoderma species.

Tracheal acid or surfactant instillation raises alveolar surface tension.

Whether alveolar liquid surface tension, T, is elevated in the acute respiratory distress syndrome (ARDS) has not been demonstrated in situ in the lungs. Neither is it known how exogenous surfactant, which has failed to treat ARDS, affects in situ T. We aim to determine T in an acid-aspiration ARDS model before and after exogenous surfactant administration. In isolated rat lungs, we combine servo-nulling pressure measurement and confocal microscopy to determine alveolar liquid T according to the Laplace relation. Administering 0.01 N (pH 1.9) HCl solution by alveolar injection or tracheal instillation, to model gastric liquid aspiration, raises T. Subsequent surfactant administration fails to normalize T. Furthermore, in normal lungs, tracheal instillation of control saline or exogenous surfactant raises T. Lavaging the trachea with saline and injecting the lavage solution into the alveolus raises T, suggesting that tracheal instillation may wash T-raising airway contents to the alveolus. Adding 0.01 N HCl or 5 mM CaCl2-either of which aggregates mucins-to tracheal lavage solution reduces or eliminates the effect of lavage solution on alveolar T. Following tracheal saline instillation, liquid suctioned directly out of alveoli through a micropipette contains mucins. Additionally, alveolar injection of gastric mucin solution raises T. We conclude that 1) tracheal liquid instillation likely washes T-raising mucins to the alveolus and 2) even exogenous surfactant that could be delivered mucin-free to the alveolus might not normalize T in acid-aspiration ARDS. NEW & NOTEWORTHY We demonstrate in situ in isolated lungs that surface tension is elevated in an acid-aspiration acute respiratory distress syndrome (ARDS) model. Following tracheal liquid instillation, also in isolated lungs, we directly sample alveolar liquid. We find that liquid instillation into normal lungs washes mucins to the alveolus, thereby raising alveolar surface tension. Furthermore, even if exogenous surfactant could be delivered mucin-free to the alveolus, exogenous surfactant might fail to normalize alveolar surface tension in acid-aspiration ARDS.

Enhancing the Spreading Behavior on Pulmonary Mucus Mimicking Subphase via Catanionic Surfactant Solutions: Toward Effective Drug Delivery through the Lungs.

Effective and efficient spreading of drug formulations on the pulmonary mucosal layer is key to successful delivery of therapeutics through the lungs. The pulmonary mucus layer, which covers the airway surface, acts as a barrier against therapeutic agents, especially in the case of chronic lung diseases due to increased thickness and viscosity of the mucus. Therefore, spreading of the drug formulations on the airways gets harder. Although spreading experiments have been conducted with different types of formulations on mucus-mimicking subphases, a highly effective formulation is yet to be discovered. Adding surfactant to such formulations decreases the surface tension and triggers the Marangoni forces to enhance the spreading behavior. In this study, catanionic (cationic + anionic) surfactant mixtures composed of dodecyltrimethylammonium bromide (DTAB) and dioctyl sulfosuccinate sodium salt (AOT) mixed at various mole ratios are prepared and their spreading behavior on both mucin and cystic fibrosis (CF) mucus models is investigated for the first time in the literature. Synergistic interaction is obtained between the components of the DTAB/AOT mixtures, and this interaction has enhanced the spreading of the formulation drop on both the mucin and CF mucus models when compared with the spreading performances of selected conventional surfactants. It is proposed that the catanionic surfactant mixtures, especially when mixed at the molar ratios of 8/2 and 7/3 (DTAB/AOT), improve the spreading even on the cystic fibrosis sputum model. As it is vital to transport a sufficient amount of drug to the targeted region for the treatment of diseases, this study presents an important application of the fundamentals of colloidal science to pharmaceutical nanotechnology.