Micro-computed tomography (μCT) is the gold standard for nondestructive 3D imaging of biomedical samples in the
centimeter scale, but it has limited effectiveness in revealing intricate soft tissue details due to the limited attenuation contrast.
Radiopaque contrast agents that accumulate in the structures of interest are employed to enhance their visibility. However, the
increased attenuation provided by the contrast agents does not guarantee discrimination among tissues. This issue can be solved by
spectral μCT (SμCT) systems employing small-pixel chromatic photon-counting detectors. These detectors, combined with material
decomposition algorithms, allow the generation of high-resolution material-specific 3D maps. This work aims to demonstrate the
potential of photon-counting X-ray SμCT on osteochondral samples loaded with a cationic iodinated contrast agent (CA4+) at a
spatial resolution below 50 μm, and to compare the results against a conventional μCT system. An osteochondral sample extracted
from a bovine stifle joint was loaded with CA4+ and imaged with a novel multimodal X-ray imaging system, featuring a 62 μm pixel
CdTe spectral detector (Pixirad1-PixieIII). After material decomposition, quantitative 3D density maps of iodine and hydroxyapatite
were reconstructed. The same sample was also scanned with a commercial μCT scanner with matched spectrum and exposure time.
SCTμ images at a (measured) spatial resolution comparable with the commercial scanner (∼45 μm) were obtained. Spectral images
allowed for a fully automatic segmentation of cartilage and subchondral bone. The unambiguous discrimination between iodine and
hydroxyapatite revealed a more realistic representation of proteoglycan distribution compared to conventional imaging.
