Conventional methods used to image and quantify microdamage in bone tissue are limited to thin histological sections. Therefore recent studies have begun to investigate methods for non-destructive, three-dimensional (3-D) detection and imaging of microdamage in bone tissue. The objective of this study was to investigate gold nanoparticles (Au NPs) as a potential damage-specific X-ray contrast agent due to their relative biocompatibility, ease of surface functionalization, colloidal stability, and high X-ray attenuation. Au NPs were prepared using a citrate reduction reaction to ∼15 or 40 nm diameter, and functionalized with glutamic acid for targeting damaged bone tissue. As-synthesized and functionalized Au NPs were spherical, relatively monodispersed, and exhibited aqueous colloidal stability. Functionalized Au NPs were demonstrated to target damaged bovine cortical bone tissue as visually evidenced by surface scratches turning a characteristic red color after soaking in functionalized Au NP solutions. Individual Au NPs were observed on the surface of damaged tissue using backscattered electron imaging and atomic force microscopy. Therefore, functionalized Au NPs are a promising candidate for a targeted X-ray contrast agent for damaged bone tissue.