Mechanical Characterization by Mesoscale Indentation: Advantages and Pitfalls for Tissue and Scaffolds.

Regenerative medicine and tissue engineering are hindered by the lack of consistent measurements and standards for the mechanical characterization of tissue and scaffolds. Indentation methods for soft matter are favored because of their compatibility with small, arbitrarily shaped samples, but contact mechanics models required to interpret data are often inappropriate for soft, viscous materials. In this study, we demonstrate indentation experiments on a variety of human biopsies, animal tissue, and engineered scaffolds, and we explore the complexities of fitting analytical models to these data. Although objections exist to using Hertz contact models for soft, viscoelastic biological materials since soft matter violates their original assumptions, we demonstrate the experimental conditions that enable consistency and comparability (regardless of arguable misappropriation). Appropriate experimental conditions involving sample hydration, the indentation depth, and the ratio of the probe size to sample thickness enable repeatable metrics that are valuable when comparing synthetic scaffolds and host tissue, and bounds on these parameters are carefully described and discussed. We have also identified a reliable quasistatic parameter that can be derived from indentation data to help researchers compare results across materials and experiments. Although Hertz contact mechanics and linear viscoelastic models may constitute oversimplification for biological materials, the reporting of such simple metrics alongside more complex models is expected to support researchers in tissue engineering and regenerative medicine by providing consistency across efforts to characterize soft matter. Impact Statement To engineer replacement tissue requires a deep understanding of its biomechanical properties. Mesoscale indentation (between micron and millimeter length scales) is well-suited to characterize tissue and engineered replacements as it accommodates small, oddly shaped samples. However, it is easy to run afoul of the assumptions for common contact models when working with biological materials. In this study, we describe experimental procedures and modeling approaches that allow researchers to take advantage of indentation for biomechanical characterization while minimizing its weaknesses.