Heterogeneous Diffusion of Polystyrene Nanoparticles through an Alginate Matrix: The Role of Cross-linking and Particle Size.

Most bacteria in natural and engineered environments grow and exist in biofilms. Recent investigations have shown that nanoparticles (NPs) interact with environmental biofilms, but these interactions are still not well characterized. Extracellular polymeric substances (EPS) are polymers secreted by bacteria to establish the functional and structural integrity of biofilms, and EPS porosity is a major contributor to NP access to and diffusion in biofilms. We used a synergistic combination of total internal reflection fluorescence microscopy and image correlation spectroscopy to monitor and map diffusion of fluorescent NPs in alginate yielding a detailed picture of the heterogeneous structure and connectivity of pores within a model EPS polymer. Using different sizes (20, 100, and 200 nm) of carboxylated polystyrene NPs, we examined how NP diffusive behaviors change as a result of calcium-induced cross-linking of the alginate matrix. This study reveals that cross-linking decreases NP diffusion coefficients and pore accessibility in an NP size-dependent manner and that NP movement through alginate matrices is anisotropic and heterogeneous. These results on heterogeneous and size-dependent movement within biofilms have important implications for future studies and simulations of NP-biofilm interactions.