Though the mineral distribution of the dentine carious lesion varies largely from tooth to tooth and from patient to patient, there are two main distribution profiles that characterize natural carious lesions in dentine. These profiles include softened and subsurface lesion types. The mineral distribution relationship between the starting profile and the profile after remineralization is not known. In order to study the relational aspects, we have produced demineralized dentine samples in vitro with mineral profiles similar to those of typical natural carious lesions, and subsequently remineralized the samples in a remineralizing solution with various fluoride concentrations (0, 2 and 10 ppm F). The mineral distributions were obtained by using an improved microradiographic technique. In addition, the nature of deposited mineral was analyzed by diamond–coupled total internal reflectance spectroscopy. Definite relationship was observed between the original lesion mineral distribution and the mineral distributions following remineralization. The amount of mineral present in approximately the first 50 μm of the lesion influenced the overall mineral profile after remineralization, possibly through influencing ion transport. If the amount was high (> approximately 10 vol%), the deposited mineral was confined to the surface (0–50 μm). The original mineral at those depths acted like a nucleus of mineral regrowth when the amount of residual mineral was intermediate, and like a transport barrier when the surface layer was well mineralized. If a surface barrier was not present, mineral was deposited at deeper depths in the lesion. Fluoride effect on dentine remineralization was dependent on the original mineral content and its distribution in the lesion. Although a high concentration of fluoride was very effective in low–mineral lesions, it produced hyperremineralization on well–mineralized subsurface lesions so that it prevented effective remineralization especially in deeper lesions.

The remineralization of whole human dentine treated with a neutral EDTA solution was investigated: the treatment periods were 15, 30 and 120 min. From the literature it is known that EDTA removes noncollagenous proteins (NCPs) from dentine powders. In order to extract more phosphoproteins from dentine lesions, in part of this work samples were also treated with 1 M NaCl or 4 M guanidine chloride solutions. All the dentine samples after the treatments mentioned were immersed subsequently in a remineralizing solution without fluoride for 2 weeks and microradiographed. To investigate the effects of fluoride, samples treated with EDTA for 120 min were also remineralized with 2ppm fluoride in solution. The results presented show that: (1) Measurable remineralization did not occur when fluoride was free in the remineralizing solution. Because remineralization occurred neither at the lesion front nor in the nonmineral part of the surface–softened tissue, presumably the NCPs (inhibitors) of underlying dentine diffused into the tissue during the remineralization period. (2) 2ppm fluoride caused remineralization at the lesion front. In this case we assume fluoride either acted as nucleating agent or overcame the NCP effects at the lesion front.

Thin sections of natural white spot enamel lesions (WS) and of artificial in vitro lesions (VL) were remineralized simultaneously in vitro. The sections, clamped in a PMMA holder, were microradiographed at baseline and after remineralization in a calcium– and phosphate–containing solution (pH = 7.0; 1 ppm F) after 2 and 4 weeks. All data were analyzed with respect to baseline. The results show that the lesion depth values did not change significantly during 2 and 4 weeks of remineralization. The mineral accumulation (change in ΔZ), however, was substantial and significant in WS and VL. In WS the change in mineral accumulation was roughly proportional to the amount of mineral at baseline. The WS accumulated more than two times the amount of mineral than VL in the same periods of remineralization. After 4 weeks of remineralization the maximum mineral value V_max in the surface layer of the WS was nearly up to the sound enamel level ≈87 vol%. This study shows that the technique and calculation procedure described make this single section method attractive for longitudinal demineralization–remineralization studies in vitro or in situ. Both WS and VL samples obviously remineralized in vitro similarly with respect to the baseline. Furthermore, this in vitro work indicates that remineralization inhibitors present in saliva, and previously penetrated into the enamel tissue, do not influence remineralization later on.

Dentine consists simplified of mineral and of several organic components. Sodium hypochlorite (NaOCl) is a well-known nonspecific proteolytic agent capable to remove organic material. The aim of this study was to investigate the influence of organic material removal from artificial dentine lesions by means of NaOCl pretreatment on subsequent remineralization with and without fluoride. Human root dentine samples were demineralized in an acidic gel (pH = 5) at 37°C for 2 weeks. After 2 min of pretreatment with a 0.4, 2 or 10% NaOCl solution, the samples were remineralized in a 20 mM HEPES buffer (pH = 7) containing 1.5 mMCa²⁺ and 0.9 mM phosphate with or without addition of 10 ppm F^– as NaF at 37°C for 8 days. Mineral profiles were assessed by means of transversal micro-radiography after diol treatment to avoid shrinkage caused by drying. In a separate experiments the dentine contraction caused by 10% NaOCl was assessed. The contraction (negligible for sound dentine) was found to be about 12% for the lesions. The remineralization results showed that pretreatment with a 10% NaOCl solution for 2 min, increased lesion remineralization. After NaOCl treatment, the amount of accumulated mineral increased by about 27% without F in the remineralization solution, and by about 4% with 10 ppm in solution. The in vitro results suggest that removal of organic materials from dentine lesions is an interesting approach to enhance remineralization.

The effects of a 10% NaOCl treatment for 2 min on demineralized human root dentin were investigated by means of: microradiography (MR), scanning electron microscopy (SEM), confocal laser scanning microscopy (CLSM) and secondary ion mass spectroscopy (SIMS). MR measurements revealed that NaOCl caused a tissue contraction not related to water loss but to removal of organic substance(s), resulting in reductions of the lesion depth and mineral loss values by 15% and 42%, respectively. CLSM observations on wet dentin showed that the dentinal tubules underneath the surface are clearly observable and not deformed substantially by the NaOCl, except near the outermost surface. This indicates the importance of wet as well as of dried (high vacuum) observations. SEM micrographs (high vacuum) showed definite changes in the outer dentin surface structure; 85% of the originally open dentinal tubules were closed after NaOCl treatment. No marked changes were observed in the dentin ultrastructure inside lesions, as shown by SEM on fractured surfaces. SIMS data, pertaining to samples in high vacuum, showed a remarkable increase of chlorine (Cl) content in the entire lesion due to the NaOCl, indicating deep penetration of the original OC1 ions. The results suggest that the 2-min treatment of demineralized dentin by NaOCl solutions removes and/or changes part of the dentin matrix in nearly the whole lesion. As a consequence the mineral is somewhat redistributed, the outermost surface of a few mu m is changed, but the main dentin structure and element composition are still intact. These findings indicate that NaOCl treatments are of interest in remineralization and hyper-remineralization studies of dentin.