Impact of collagen cross-linking agents on release of hydroxyproline from radicular dentin in endodontically treated mandibular premolars with bio-ceramic sealers: An in vitro analysis

Abstract Context: Bio ceramic sealers are effective in sealing root canals but their high alkalinity may degrade radicular dentin collagen, compromising tooth strength. Collagen cross-linking agents have the potential to protect and stabilize collagen. Cross-linking agents are used to create chemical bonds between collagen molecules. This process stabilizes the collagen structure, making it more resistant to degradation by enzymes like collagenase. A reduction in the amount of hydroxyproline (HYP) released indicates that the cross-linking agent is effectively protecting the collagen from breakdown. Aims: To evaluate the effect of collagen cross-linking agents on collagen degradation by measuring HYP release. Methodology: Forty mandibular premolars were decoronated, instrumented to ProTaper F2, and irrigated with 5.25% NaOCl and 17% ethylenediaminetetraacetic acid. Samples were divided into four groups: Group 1 (6.5% proanthocyanidin [PA]), Group 2 (cashew nut shell liquid [CNSL]), Group 3 (chlorhexidine), and Group 4 (saline). Canals were obturated with gutta-percha and bio-ceramic sealer, then stored in artificial saliva. HYP release was measured on days 7, 14, and 21 using spectrophotometry. Statistical Analysis Used: The amount of HYP released was analyzed using one-way analysis of variance, Kruskal–Wallis test, and Tukey honestly significant difference test. Results: Groups 1 and 2 exhibited significantly lower HYP release than Groups 3 and 4, indicating reduced collagen degradation. Conclusions: Collagen cross-linking agents, especially PA and CNSL, reduce collagen breakdown and may improve dentin durability following bio-ceramic sealer application.