Long-Term Influence of Endodontic Irrigants on In Vitro Dentin Biomimetic Remineralization

Endodontic irrigant solutions act as crucial pretreatment conditioning agents in dentin biomimetic remineralization, preparing the collagen scaffold for calcium phosphate infiltration and subsequent tooth structure reconstruction. In this study, root dentin discs were exposed for 10 min to five irrigant solutions: sodium hypochlorite (NaClO, 3%), EDTA (17%), citric acid (CA, 10%), chlorhexidine (CHX, 2%), and an innovative experimental formulation containing citric acid (7%) and surfactants. Samples were then aged in Hank's Balanced Salt Solution (HBSS) at 37 °C for three months to simulate long-term clinical conditions. Physicochemical modifications of the collagen and apatite phases were assessed at each experimental stage using ATR-FTIR spectroscopy, with the A_CaP/A_{Amide I} and A₈₇₀/A_CaP absorbance ratios as markers of the degree of mineralization and apatite carbonate content, respectively. Results indicated that CHX- and EDTA-treated dentin exhibited the highest remineralization after ageing, while NaClO impeded remineralization due to collagen degradation. The experimental irrigant produced the most pronounced demineralization, followed by CA; however, it also facilitated significant remineralization, attributed to citrate-collagen binding and surfactant-enhanced apatite nucleation. NaClO selectively degraded collagen and increased apatite crystallinity; CA inhibited apatite nucleation through adsorbed citrate ions, and CHX and EDTA induced minimal alterations. These findings provide molecular-level evidence linking short-term irrigant effects to the long-term potential for dentin biomineralization, with direct implications for irrigant selection in regenerative endodontic protocols. It should be noted that this study was conducted on dentin discs obtained from a single tooth; all findings should therefore be regarded as preliminary and require confirmation in studies with larger, biologically independent sample sizes.