AFM analysis of the lacunar-canalicular network in demineralized compact bone

Atomic force microscopy has been successfully used to examine a wide range of cellular and biomolecular structures and interactions. The application of atomic force microscopy in the analysis of organs and tissues, however, has been limited. In this study, we present a new method for high-resolution atomic force microscopy imaging of compact bone tissue. We performed atomic force microscopy imaging on demineralized compact bone from bovine tibia to obtain structural information about the bone matrix and the lacunar-canalicular network. Knowledge of the dimensions and distributions of the network allows quantitative analysis of the microfluidics of bone tissue. Results from our study show that (1) the canalicular distribution and dimensions are homogenous in transverse, radial and longitudinal orientations; (2) the lamellae of an osteon consist of alternating high and low bands; (3) the canaliculi follow the contour of lamellar bands and (4) globular structures cover much of the bone matrix, including canalicular walls. Our work demonstrates that atomic force microscopy studies of thin-section tissue samples can provide structural details at nanometre resolution.