Although it is generally proposed that dentin bonding results from adhesive infiltration of superficially demineralized dentin, it is not clear how well the resin monomers seal the dentin collagen fibrils under wet bonding conditions. The aim of this study was to determine the quality and molecular structure of adhesive/dentin (a/d) interfaces formed with wet bonding as compared with adhesive-infiltrated demineralized dentin (AIDD) produced under controlled conditions (optimum hybrid). From each extracted, unerupted human 3rd molar, one fraction was demineralized, dehydrated, and infiltrated with Single Bond (SB) adhesive under optimum conditions; the remaining, adjacent fraction was treated with SB by wet bonding. AIDD and a/d interface sections were stained with Goldner’s trichrome; corresponding sections were analyzed with micro-Raman spectroscopy. The histomorphologic and spectroscopic results suggest that, under wet bonding, the a/d interface is a porous collagen web infiltrated primarily by the hydrolytically unstable HEMA.

Fibroblastic growth factor 23 (FGF23) regulates renal phosphate reabsorption and 1α-hydroxylase activity. Ablation of FGF23 results in elevated serum phosphate, calcium, and 1,25-dihydroxyvitamin D3 [1,25(OH)₂D] levels; vascular calcifications; and early death. For determination of the independent roles of hyperphosphatemia and excess vitamin D activity on the observed phenotypic abnormalities, FGF23 null mice were fed a phosphate- or vitamin D–deficient diet. The phosphate-deficient diet corrected the hyperphosphatemia, prevented vascular calcifications, and rescued the lethal phenotype in FGF23 null mice, despite persistent elevations of serum 1,25(OH)₂D and calcium levels. This suggests that hyperphosphatemia, rather than excessive vitamin D activity, is the major stimulus for vascular calcifications and contributes to the increased mortality in the FGF23-null mouse model. In contrast, the vitamin D–deficient diet failed to correct either the hyperphosphatemia or the vascular calcifications in FGF23 null mice, indicating that FGF23 independently regulates renal phosphate excretion and that elevations in 1,25(OH)₂D and calcium are not sufficient to induce vascular calcifications in the absence of hyperphosphatemia. The vitamin D–deficient diet also improved survival in FGF23 null mice in association with normalization of 1,25(OH)₂D and calcium levels and despite persistent hyperphosphatemia and vascular calcifications, indicating that excessive vitamin D activity can also have adverse effects in the presence of hyperphosphatemia and absence of FGF23. Understanding the independent and context-dependent interactions between hyperphosphatemia and excessive vitamin D activity, as well as vascular calcifications and mortality in FGF23 null mice, may ultimately provide important insights into the management of clinical disorders of hyperphosphatemia and excess vitamin D activity.

To date, the dentin/adhesive (d/a) bond has primarily been studied by morphologic analysis in conjunction with bond strength measurement. Although these analyses have enhanced our understanding, numerous questions about the chemistry have not been answered. The purpose of this study was to determine, at the molecular level, quantitative differences in the composition of the d/a interface formed under "wet" bonding conditions. The occlusal one-third of the crown was removed from 10 extracted, unerupted human third molars. The prepared dentin surfaces were treated, per manufacturers' instructions, with either Single Bond (3M) or One-Step adhesive (Bisco). Three-micron-thick sections of the d/a interface were cut and stained with Goldner trichrome for light microscopy. Companion slabs were analyzed with micro-Raman spectroscopy; the sample was placed at the focus of a 100x microscope objective, and spectra were acquired at 1-μm intervals across the d/a interface. Reference spectra were collected on model compounds of type I collagen and adhesive; the relative ratios of the integrated intensities of spectral features from adhesive and collagen were determined and plotted as a function of wt% adhesive. The same ratios were determined for the interface samples; by comparing these ratios with the calibration curve generated from the model compounds, we determined the percent of adhesive as a function of spatial position across the d/a interface. The relative percent of Single Bond adhesive was < 50% throughout more than half of the hybrid layer; One Step adhesive was ≥ 50% throughout most of the hybrid. The results from this study provide the first direct chemical evidence of phase separation in a dentin adhesive and its detrimental effect on the dentin/adhesive bond.