Nanoindentation of Bone in a Physiological Environment

Nanoindentation has been established as an effective method to measure the mechanical properties of bone tissue at the micron and sub-micron length scale. Although it is well-documented that the mechanical properties of macroscopic bone specimens vary depending on whether the samples are tested dry or wet, nanoindentation is generally conducted on dehydrated bone tissue at room temperature, primarily because nanoindentation systems are extremely sensitive to changes in environmental conditions such as humidity and temperature. In this study, these problems were overcome by using a specially constructed liquid cell with an extension piece that allowed the indenter tip to be submerged under 5 mm of liquid. The custom setup was used to test cortical bovine bone and cancellous human bone specimens in three distinct conditions – dehydrated, rehydrated in simulated body fluid (SBF) at 20°C, and rehydrated in SBF at 37.5°C. A heating element with a temperature control unit was used to test at 37.5°C. The hardness and elastic modulus of the bone samples were found to decrease when dry specimens were rehydrated and tested in physiological conditions. It is suggested that nanoindentation in physiological conditions gives a better estimate of the mechanical properties of the microstructural components of bone in vivo rather than nanoindentation under conventional conditions.