Abstract
Fracture repair is a complex process that involves the interaction of numerous molecular factors, cell lineages and tissue types. These biological processes allow for an impressive feat of engineering: an elastic soft callus is progressively replaced by a more rigid and mineralized callus. During this reparative phase, the healing bone is exposed to a risk of re-fracture. Bone volume and bone quality are the two major factors determining the strength of the callus. Although both factors are important, often only bone volume is analyzed and reported in preclinical studies. Recent developments in techniques for examining bone quality in the callus will enable the rapid and detailed analysis of its material properties and its microstructure. This review aims to give an overview of the methods available for quantitatively phenotyping the bone callus in preclinical studies such as Raman spectroscopy, nanoindentation, scanning acoustic microscopy, in vivo micro-computed tomography (micro-CT) and high-resolution micro-CT. Consolidated and emerging experimental methods are described with a focus on their applicability, and with examples of their utilization.