The effect of holding time on nanoindentation measurements of creep in bone.

Viscoelasticity may affect both the elastic and fracture characteristics of bone. Nanoindentation can be used to measure the creep behavior of bone by fitting the depth vs. time data at constant load to rheological models. However, the creep data may be influenced by latent effects arising during the loading phase of indentation. As such, the loading protocol, particularly the holding time, may affect the measured creep time constants. To characterize the effect of holding time on the measures, four cortical bone samples were prepared from four bovine femora and subjected to nanoindentation to measure the creep behavior. The creep time constants were found by fitting the indentation depth vs. time curve to three different rheological models: the standard linear solid, Burgers model, and a two-dashpot Kelvin model. All three models provided good fits to the data, which were relatively insensitive to the initial parameter estimates. The calculated creep time constants increased monotonically with increasing holding time for all three models. However, the relative differences between measurements within a single osteon, within a single sample, and between samples were maintained for creep holding times over 16s. Hence, while the creep time constants measured by nanoindentation with hold times up to 30s may not provide accurate property measurements, comparisons between samples are valid if all are assessed at the same holding time. Considering the long-term viscosity of bone tissue, Burgers model provided the best performance in terms of stability and goodness of fit, and is recommended for future studies.