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Brewster angle microscopy (BAM) is a powerful technique that allows for real-time visualization of Langmuir monolayers. The lateral organization of these films can be investigated, including phase separation and the formation of domains, which may be of different sizes and shapes depending on the properties of the monolayer. Different molecules or small changes within a molecule such as the molecule's length or presence of a double bond can alter the monolayer's lateral organization that is usually undetected using surface pressure-area isotherms. The effect of such changes can be clearly observed using BAM in real-time, under full hydration, which is an experimental advantage in many cases. While previous BAM reviews focused more on selected compounds or compared the impact of structural variations on the lateral domain formation, this review provided a broader overview of BAM application using biological materials and systems including the visualization of amphiphilic molecules, proteins, drugs, extracts, DNA, and nanoparticles at the air-water interface.

There has been an increasing interest in the potential of nanomedicine, particularly in the use of nanoparticles between 10 nm and 1 μm in diameter as drug delivery vehicles. For pulmonary drug delivery, it is important to understand the effect of polymeric nanoparticles on the lung surfactant in order to optimize the carriers by reducing their potential toxicological effects. This work presents a biophysical study of the impact of gelatin nanoparticles on packing and lateral organization of simple and complex lipid layers containing the major components of lung surfactant. Zwitterionic phosphatidylcholines, negatively charged phosphatidylglycerols, and the sterol cholesterol were employed in the models. In addition, the impact of acyl chain length was investigated. Packing was determined by surface pressure-area isotherms, whereas direct imaging of the surfactant at the air-water interface was performed using Brewster angle microscopy. Our results indicate minor changes in the surface pressure-area isotherms but concomitantly significant effects on the lateral organization of the monolayers upon nanoparticle addition. The data also suggest differential interactions of nanoparticles with the major lipid classes. Gelatin nanoparticles interact stronger with negatively charged phosphatidyl-glycerols compared to zwitterionic phosphatidyl-cholines. Furthermore, charge distribution depending on the molar lipid ratio and acyl chain saturation is important as well. Even cholesterol, whose concentration is low compared to other components, plays an important role in nanoparticle interactions.

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Biopolymer microparticles have been developed for applications that require biocompatibility and biodegradability, such as drug delivery. In this study, we assessed the production of microparticles using carnauba wax, κ-carrageenan, alginate, and poly (lactic-co-glycolic acid) (PLGA) with the aim of developing a novel, DNA-tracer-loaded, biopolymer surrogate with a size, shape, surface charge, and relative hydrophobicity similar to stationary-phase Legionella pneumophila to mimic the bacteria’s mobility and persistence in engineered water systems. We found that the type and concentration of biopolymer, reaction conditions, and synthesis methods affected the morphology, surface charge, relative hydrophobicity, and DNA tracer loading efficiency of the biopolymer microparticles produced. Carnauba wax, κ-carrageenan, and alginate (Protanal®, and low and medium viscosity) produced highly polydisperse microspheres. In contrast, PLGA and alginate-CaCO3 produced uniform microspheres and rod-shaped microparticles, respectively, with high DNA tracer loading efficiencies (PLGA 70% and alginate-CaCO3 95.2 ± 5.7%) and high reproducibilities. Their synthesis reproducibility was relatively high. The relative hydrophobicity of PLGA microspheres closely matched the cell surface hydrophobicity of L. pneumophila but not the bacterial morphology, whereas the polyelectrolyte layer-by-layer assembly was required to enhance the relative hydrophobicity of alginate-CaCO3 microparticles. Following this surface modification, alginate-CaCO3 microparticles represented the best match to L. pneumophila in size, morphology, surface charge, and relative hydrophobicity. This new biopolymer surrogate has the potential to be used as a mimic to study the mobility and persistence of L. pneumophila in water systems where the use of the pathogen is impractical and unsafe.