Bone tissue ultrastructural response to elastic deformation probed by Raman spectroscopy

Abstract

Raman spectroscopy is used as a probe of ultrastructural (molecular) changes in both the mineral and matrix (protein and glycoprotein, predominantly type I collagen) components in real time of murine cortical bone as it responds to elastic deformation. Because bone is a composite material, its mechanical properties are dependent on the structure and composition at a variety of dimensional scales. At the ultrastructural level, crystal structure and protein secondary structure distort as the tissue is loaded. These structural changes are followed as perturbations to tissue spectra. We load murine femora in a custom-made mechanical tester that fits on the stage of a Raman microprobe and can accept hydrated tissue specimens. As the specimen is loaded in tension, the shifts in mineral P–O₄ν₁ are followed with the microprobe. Average load and strain are measured using a load cell. These devices ensure that specimens are not loaded to or beyond the yield point. Changes occur in the mineral component of bone as a response to loading in the elastic regime. We propose that the mineral apatitic crystal lattice is deformed by movement of calcium and other ions. Raman microspectroscopy shows that bone mineral is not a passive contributor to tissue strength. The mineral active response to loading may function as a local energy storage and dissipation mechanism, thus helping to protect tissue from catastrophic damage.