Effect of iodixanol and propylene glycol as clearing agents in extensor digitorum longus and soleus muscles: mechanical and morphological characterization using the optical coherence tomography technique

Soft tissue engineering and regenerative medicine aim to address the intricate relationship between tissue architecture and biomechanical performance. The traditional technique used to analyze muscular architectures is histology. However, optical coherence tomography is a novel non-destructive, non-invasive imaging tool that provides real-time, high-resolution visualization of tissue microstructure, making it applicable to soft tissues. High-quality images, minimized light scattering, and different clearing agents, such as propylene glycol and iodixanol, have been employed. A stress-relaxation test was performed to characterize the effects of clearing agents on rat extensor digitorum longus and soleus muscles. Additionally, muscle fiber structure images obtained using optical correlation tomography were compared with histological images to corroborate the high precision of the optical method. The results showed that iodixanol is a promising clearing agent for characterizing muscles as it provides good quality images and a satisfactory reversibility process with no permanent damage to the extracellular matrix or muscle fiber structure of the tissue.