Abstract
The temporomandibular joint disc plays a fundamental role in daily activities, and when it is compromised, severely disturbs oral health and quality of life. Decellularization is gaining interest in tissue engineering (TE) applications, but requires maintaining the native structure and composition to mimic human disc properties. This study characterizes the native ovine disc and compares conservation protocols to preserve its morphology, biochemical content (sulfated glycosaminoglycans, total and soluble collagen), and mechanical and thermal behavior. Three storage protocols were tested: (i) freezing at -20°C in phosphate-buffered saline (PBS) and thawing at 4°C (PBS + 4°C); (ii) freezing at -20°C in PBS and thawing at room temperature (RT) (PBS + RT); and (iii) wrapping the discs in PBS-embedded gauze, freezing at -20°C, and thawing at RT (Gauze + RT). Protocols were evaluated at 1, 7, and 14 days, and compared with a native disc, and a collagenase-treated discs. All conservation protocols caused changes, though less pronounced than degradation. The PBS + 4°C and PBS + RT protocols maintained original morphology, yet highlighted, a contrasting biochemical and mechanical outcome based on the thawing method. Thermal analysis revealed collagen structure changes within the first 7 days of freezing. The Gauze + RT protocol showed no significant biochemical changes over time, but the disc became dehydrated and with a higher compression modulus. For TE approaches involving decellularization, it is crucial to consider these alterations. For powdered tissue applications, the Gauze + RT method for 14 days is recommended due to minimal structural impact.