Are Collagen Protons Visible with the Zero Echo Time (ZTE) Magnetic Resonance Imaging Sequence: A D2O Exchange and Freeze-Drying Study

It is known that ultrashort echo time (UTE) magnetic resonance imaging (MRI) sequences can detect signals from water protons but not collagen protons in short T2 species such as cortical bone and tendons. However, whether collagen protons are visible with the zero echo time (ZTE) MRI sequence is still unclear. In this study, we investigated the potential of the ZTE MRI sequence on a clinical 3T scanner to directly image collagen protons via D₂O exchange and freeze-drying experiments. ZTE and UTE MRI sequences were employed to image fully hydrated bovine cortical bone (<i>n</i> = 10) and human patellar tendon (<i>n</i> = 1) specimens. Then, each specimen was kept in a 30 mL syringe filled with D₂O solution for two days. Fresh D₂O was flushed every 2 h to reach a more complete D₂O-H₂O exchange. Later, the samples were lyophilized for over 40 h and then sealed in tubes. Finally, the samples were brought to room temperature and visualized using the identical 3D ZTE and UTE sequences. All hydrated bone and tendon specimens showed high signals with ZTE and UTE sequences. However, all specimens showed zero signal after the D₂O exchange and freeze-drying procedures. Therefore, similar to UTE imaging, the signal source in ZTE imaging is water. The ZTE sequence cannot directly detect signals from collagen protons in bone and tendons.