Optimization of High-Frequency Ultrasound Imaging to Detect Incremental Changes in Mineral Content at the Cartilage–Bone Interface Ex Vivo

(1) Background: Osteoarthritis is a degenerative disease of the whole joint marked by cartilage-bone interface (CBI) remodeling, but methods to monitor subtle changes in mineralization are lacking. We optimized a non-destructive ultrasound imaging method to monitor incremental shifts in mineralization, using brief decalcification as a mimetic of CBI remodeling. (2) Methods: We used a 35-MHz transducer to scan 3 mm diameter bovine osteochondral explants wrapped with parafilm to produce surface-directed decalcification and dedicated 3D-printed holders to maintain sample orientation. Customized MATLAB codes and a matched pair design were used for quantitative hypothesis testing. (3) Results: Optimal scan precision was obtained when the High-Frequency Ultrasound (HFUS) focal distance was trained at the CBI. HFUS cartilage thickness increased by 53 ± 21 µm or 97 ± 28 µm after three or seven hours of ethylene diamine tetra-acetic acid (EDTA) (but not PBS), respectively, and was highly correlated with histological cartilage thickness (R = 0.98). The en face CBI backscatter pattern was irregular and shifted after the EDTA-displacement of the mineral front. Collective data suggested that the -10 dB echogenic CBI signal originated from the mineral front and varied topographically with undulating mineral thickness. (4) Conclusions: This imaging approach could be used to monitor tidemark remodeling in live explant cultures, toward identifying new treatments that inhibit tidemark advancement and slow osteoarthritis progression.