A Methodological Approach for Interpreting and Comparing the Viscoelastic Behaviors of Soft Biological Tissues and Hydrogels at the Cell-Length Scale

The behavior of a cell is strongly influenced by the physical properties and stimuli in its microenvironment. Furthermore, the activation and modulation of mechanotransduction pathways are involved in tissue development and homeostasis and even pathological processes. Thus, when developing materials aimed at mimicking the extracellular matrixes of healthy or pathological tissues, their mechanical features should be closely considered. In this context, nanoindentation represents a powerful technique for mechanically characterizing biological tissues and hydrogels at the cell-length scale. However, standardized experimental protocols and data analysis techniques are lacking. Here, we proposed a methodological approach based on the nanoindentation technique for quantitatively analyzing and comparing the time-dependent load relaxation responses of soft biological tissues and hydrogels. As this was an explanatory study, stress-relaxation nanoindentation tests were performed on samples of pig and human lung tissues and of a specific gelatin-methacryloyl (GelMA) hydrogel to quantify and compare their viscoelastic properties. The proposed method allowed for identifying the characteristic parameters needed for describing the behavior of each sample, permitting us to quantitatively compare their mechanical behaviors. All samples showed load relaxation at a defined indentation depth because of their intrinsic viscoelastic behaviors, and the GelMA samples showed the highest relaxation capabilities. The distribution of the characterization parameters showed that the biological samples presented similar time-dependent responses, while differences were observed in the GelMA samples. Overall, the proposed methodological approach allows for providing key insights into the time-dependent behaviors of soft biological tissues and hydrogels at the cell-length scale in view of supporting tissue engineering and pathophysiological investigations.