Technical note: Effects of chemical, radiational, and thermal treatment of bone tissue on material stiffness and anisotropy.

Abstract

Objectives: We evaluated the effects of four treatments for pathogen inactivation in bone tissue (ethanol storage, formalin fixation, gamma irradiation, and heat treatment via autoclave) on stiffness and anisotropy values in bone samples.

Materials and methods: Cortical bone samples from the humerus of 14 bovine specimens were subjected to Knoop microindentation analysis in longitudinal and transverse planes of section and four indenter orientations within each section. From each specimen, individual samples were assigned to one of five treatment conditions: 50% ethanol-saline solution, formalin immersion, gamma irradiation, autoclave, and buffered saline (controls). Each sample was microindented 100 times and the resultant stiffness data were analyzed by a resampled factorial analysis of variance.

Results: First- and second-order interactions as well as main effects of indenter orientation, treatment, and specimen were significant for both transverse and longitudinal sections, with the sole exception that indenter orientation-treatment interaction was nonsignificant for longitudinal sections. Interaction plots reveal that thermal (autoclave) and irradiation treatments depress stiffness values the most, while patterns of indentation stiffness at different orientations are relatively unaffected.

Discussion: Patterns of anisotropy are relatively unaffected by preservative treatments, but elastic modulus changes are consistent and unambiguous. Formalin and ethanol treatments are most comparable to controls and represent the best preservative media for mechanical testing. These options, however, are likely to be the least effective for ensuring the inactivation or sterilization of potentially contaminated samples.