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Biofilms are soft multicomponent biological materials composed of microbial communities attached to surfaces. Despite the crucial relevance of biofilms to diverse industrial, medical, and environmental applications, the mechanical properties of biofilms are understudied. Moreover, most of the available techniques for the characterization of biofilm mechanical properties are destructive. Here, we detail a model-based approach developed to characterize the viscoelastic properties of soft materials and bacterial biofilms based on experimental data obtained using the nondestructive dynamic optical coherence elastography (OCE) technique. The model predicted the frequency- and geometry-dependent propagation velocities of elastic waves in a soft viscoelastic plate supported by a rigid substratum. Our numerical calculations suggest that the dispersion curves of guided waves recorded in thin soft plates by the dynamic OCE technique are dominated by guided waves, whose phase velocities depend on the viscoelastic properties and plate thickness. The numerical model was validated against experimental measurements in agarose phantom samples with different thicknesses and concentrations. The model was then used to interpret guided wave dispersion curves obtained by the OCE technique in bacterial biofilms developed in a rotating annular reactor, which allowed the quantitative characterization of biofilm shear modulus and viscosity. This study is the first to employ measurements of elastic wave propagation to characterize biofilms, and it provides a novel framework combining a theoretical model and an experimental approach for studying the relationship between the biofilm internal physical structure and mechanical properties.

Microbial granular biofilms are spherical, multi-layered aggregates composed of communities of bacterial cells encased in a complex matrix of hydrated extracellular polymeric substances (EPS). While granular aggregates are increasingly used for applications in industrial and municipal wastewater treatment, their underlying mechanical properties are poorly understood. The challenges of viscoelastic characterization for these structures are due to their spherical geometry, spatially heterogeneous properties, and their delicate nature. In this study, we report a model-based approach for nondestructive characterization of viscoelastic properties (shear modulus and shear viscosity) of alginate spheres with different concentrations, which was motivated by our measurements in granular biofilms. The characterization technique relies on experimental measurements of circumferential elastic wave speeds as a function of frequency in the samples using the Optical Coherence Elastography (OCE) technique. A theoretical model was developed to estimate the viscoelastic properties of the samples from OCE data through inverse analysis. This work represents the first attempt to explore elastic waves for mechanical characterization of granular biofilms. The combination of the OCE technique and the theoretical model presented in this paper provides a framework that can facilitate quantitative viscoelastic characterization of samples with curved geometries and the study of the relationships between morphology and mechanical properties in granular biofilms.