Collagen intrafibrillar mineralization as a result of the balance between osmotic equilibrium and electroneutrality.

Mineralization of fibrillar collagen with biomimetic process-directing agents has enabled scientists to gain insight into the potential mechanisms involved in intrafibrillar mineralization. Here, by using polycation- and polyanion-directed intrafibrillar mineralization, we challenge the popular paradigm that electrostatic attraction is solely responsible for polyelectrolyte-directed intrafibrillar mineralization. As there is no difference when a polycationic or a polyanionic electrolyte is used to direct collagen mineralization, we argue that additional types of long-range non-electrostatic interaction are responsible for intrafibrillar mineralization. Molecular dynamics simulations of collagen structures in the presence of extrafibrillar polyelectrolytes show that the outward movement of ions and intrafibrillar water through the collagen surface occurs irrespective of the charges of polyelectrolytes, resulting in the experimentally verifiable contraction of the collagen structures. The need to balance electroneutrality and osmotic equilibrium simultaneously to establish Gibbs-Donnan equilibrium in a polyelectrolyte-directed mineralization system establishes a new model for collagen intrafibrillar mineralization that supplements existing collagen mineralization mechanisms.

Effect of Ultrasonic Activation of Irrigants on Smear Layer Removal.

INTRODUCTION: The objective of this study was to evaluate the efficacy of passive ultrasonic irrigation (PUI) with 17% EDTA and 1% NaOCl solutions on smear layer removal. METHODS: Root canal preparations of 32 human teeth were performed with the ProTaper system. Next, they were longitudinally fractured to permit quantitation of smear layer creation from the cervical, middle, and apical thirds of the roots by using scanning electron microscopy. After reassembling the fractured tooth halves, they were divided into 4 groups according to different final irrigation protocols: group1, EDTA + NaOCl; group 2, EDTA with PUI + NaOCl; group 3, EDTA + NaOCl with PUI; and group 4, EDTA + NaOCl, both with PUI. After irrigation, the tooth halves were separated to permit imaging the same areas by scanning electron microscopy, and a percentage of opened dentinal tubules in irrigated areas as a percent of the total area was obtained. The results were submitted to Kruskal-Wallis, analysis of variance, and Bonferroni tests (α = 0.05). RESULTS: The cervical third of the samples from all groups showed higher percentage of smear layer removal and open dentinal tubule areas, followed by the middle and apical thirds. Among the irrigation groups, there were statistically significant differences in cervical third between group 2 and group 4 samples, with the highest and lowest percentage of smear layer removal, respectively. CONCLUSIONS: PUI by using 1% NaOCl and ultrasonic tip placed within 1 mm of the apical foramen did not show higher efficacy in smear layer removal compared with conventional irrigation.

Effects of different sonic activation protocols on debridement efficacy in teeth with single-rooted canals.

OBJECTIVES: The effects of different EndoActivator® (EA) sonic activation protocols on root canal debridement efficacy were examined. METHODS: Root canals in 48 single-rooted teeth were instrumented, irrigated initially with NaOCl and divided into 6 groups (N=8) based on the application time of QMix (antimicrobial calcium-chelating irrigant), and the time and sequence of EA irrigant activation - Positive Control: 90s QMix; Negative Control: 90s saline; Group 1A: 15s QMix+15s QMix with EA-activation; Group 1B: 30s QMix+30s of QMix with EA-activation; Group 2A: 15s QMix with EA-activation+15s QMix; Group 2B: 30s QMix with EA-activation+30s QMix. Split roots were examined with scanning electron microscopy for assignment of smear and debris scores in locations along the coronal, middle and apical thirds of the canals. The overall cleanliness of pooled canal locations in the Positive Control and the 4 experimental groups were compared with chi-square tests. RESULTS: Significant differences were detected among the 5 groups (P<0.001). Post hoc pairwise comparisons indicated that the overall canal cleanliness was in the order (from best to worst): 1B=2B>2A>1A>Positive Control. Completely clean canals could not be achieved due to the absence of continuous irrigant flow for EA to clear intraradicular debris. CONCLUSIONS: Irrespective of the sonic activation sequence, irrigant activation for 30s during a 60-s period of QMix application appears to maximise the smear layer and debris removal potential of the EndoActivator® system. CLINICAL SIGNIFICANCE: Sonic activation of root canal irrigants produces cleaner root canals and reduces the time required for final delivery of a canal wall smear later-removing irrigant when compared to the use of needle irrigation alone.

Site specific properties of carious dentin matrices biomodified with collagen cross-linkers.

PURPOSE: To assess in non-cavitated carious teeth the mechanical properties of dentin matrix by measuring its reduced modulus of elasticity and the effect of dentin biomodification strategies on three dentin matrix zones: caries-affected, apparently normal dentin below caries-affected zone and sound dentin far from carious site. METHODS: Nano-indentations were performed on dentin matrices of carious molars before and after surface modification using known cross-linking agents (glutaraldehyde, proanthocyanidins from grape seed extract and carbodiimide). RESULTS: Statistically significant differences were observed between dentin zones of demineralized dentin prior to surface biomodification (P < 0.05). Following surface modification, there were no statistically significant differences between dentin zones (P < 0.05). An average increase of 30-fold, 2-fold and 2.2-fold of the reduced modulus of elasticity was observed following treatments of the three dentin zones with proanthocyanidin, carbodiimide and glutaraldehyde, respectively.

A chemical phosphorylation-inspired design for Type I collagen biomimetic remineralization.

OBJECTIVES: Type I collagen alone cannot initiate tissue mineralization. Sodium trimetaphosphate (STMP) is frequently employed as a chemical phosphorylating reagent in the food industry. This study examined the feasibility of using STMP as a functional analog of matrix phosphoproteins for biomimetic remineralization of resin-bonded dentin. METHODS: Equilibrium adsorption and desorption studies of STMP were performed using demineralized dentin powder (DDP). Interaction between STMP and DDP was examined using Fourier transform-infrared spectroscopy. Based on those results, a bio-inspired mineralization scheme was developed for chemical phosphorylation of acid-etched dentin with STMP, followed by infiltration of the STMP-treated collagen matrix with two etch-and-rinse adhesives. Resin-dentin interfaces were remineralized in a Portland cement-simulated body fluid system, with or without the use of polyacrylic acid (PAA) as a dual biomimetic analog. Remineralized resin-dentin interfaces were examined unstained using transmission electron microscopy. RESULTS: Analysis of saturation binding curves revealed the presence of irreversible phosphate group binding sites on the surface of the DDP. FT-IR provided additional evidence of chemical interaction between STMP and DDP, with increased in the peak intensities of the PO and P-O-C stretching modes. Those peaks returned to their original intensities after alkaline phosphatase treatment. Evidence of intrafibrillar apatite formation could be seen in incompletely resin-infiltrated, STMP-phosphorylated collagen matrices only when PAA was present in the SBF. SIGNIFICANCE: These results reinforce the importance of PAA for sequestration of amorphous calcium phosphate nanoprecursors in the biomimetic remineralization scheme. They also highlight the role of STMP as a templating analog of dentin matrix phosphoproteins for inducing intrafibrillar remineralization of apatite nanocrystals within the collagen matrix of incompletely resin-infiltrated dentin.

Hybrid Root SEAL (MetaSEAL) creates hybrid layers in radicular dentin only when EDTA is used as the final rinse.

PURPOSE: To test if the hybrid layer formation by Hybrid Root SEAL (Sun Medical Co.), a 4-META-containing auto-adhesive self-etching root canal sealer, is affected by the sequence of irrigants employed for removing canal wall smear layers during root canal treatment. METHODS: Single-rooted teeth were shaped and irrigated with EDTA as initial rinse/NaOCl as active final rinse (Group 1), or NaOCl as initial rinse/EDTA as active final rinse (Group 2). All canals were obturated with Hybrid Root SEAL using a single-cone technique. Root slices derived from the coronal, middle and apical thirds of the roots were processed for transmission electron microscopy after removing the gutta-percha, leaving the sealer intact. Additional filled canals from the two groups were evaluated for fluid leakage. RESULTS: Hybrid layer was absent in Group 1 and was present only when a collagen matrix was produced by EDTA demineralization (Group 2). Significantly more leakage (4.03 +/- 1.94 microL min(-1) vs. 1.50 +/- 0.42 microL min(-1); P < 0.05) was observed in the absence of dentin hybridization.

Changes in stiffness of demineralized dentin following application of collagen crosslinkers.

It is thought that increasing the strength of the dentin matrix using crosslinking agents may improve both the strength and the durability of resin-dentin bonds. The purpose of this study was to evaluate the effect of two collagen crosslinking agents (glutaraldehyde, GD and grape seed extract, GSE) on the modulus of elasticity of demineralized dentin. Sound molar fragments were fully demineralized and divided into five groups according to the type and concentration of crosslinking agents: 2.5% GD; 5% GD, 25% GD; 0.65% GSE; 6.5% GSE. Specimens were immersed in their respective solution and tested at baseline, 10 min, 30 min, 1 h, 2 h, 4 h. The elastic modulus of dentin was significantly affected by the treatment (p < 0.01) and exposure time (p < 0.01). There was a statistically significant interaction between the two factors evaluated (treatment vs. time p < 0.01). Mean baselines values varied between 4.8 and 6.2 MPa in water; after 4 h of treatment the values increased between 34.9 and 242.5 MPa, that were treatment time and agent dependent. The use of these collagen crosslinkers to increase the stiffness of demineralized dentin, was both concentration and time dependent.

Microporous, demineralized collagen matrices in intact radicular dentin created by commonly used calcium-depleting endodontic irrigants.

The use of calcium-depleting solutions in restorative dentistry results in demineralization of intact coronal dentin beneath smear layers. Likewise, application of calcium-depleting irrigants as final rinses might create demineralized collagen matrices in intact radicular dentin after removal of root canal wall smear layers created during canal shaping. These unsupported collagen matrices collapse on desiccation and might be difficult to detect in scanning electron microscopy dehydration techniques traditionally used in endodontic research. Demineralized collagen matrices were identified, by using a hexamethyldisilazane dehydration protocol, on the surface of smear layer-depleted radicular dentin when NaOCl was used as initial rinse, followed by the use of 17% ethylenediaminetetraacetic acid or BioPure MTAD as the final rinse. Compaction of conventional root filling materials over a demineralized collagen matrix in instrumented dentin is analogous to performing the same procedure against the predentin collagen network in the uninstrumented parts of root canals. The potential consequences of this process are, however, unknown.

Have dentin adhesives become too hydrophilic?

This review discusses current trends in the development of dentin adhesives and the possibility that some classes of currently available adhesives are too hydrophilic. Manufacturers have reformulated dentin adhesives to make them more compatible for bonding to intrinsically moist, acid-etched dentin by adding 2-hydroxyethyl methacrylate and other hydrophilic resin monomers. These 3-step adhesives work well but are more time consuming to use and more sensitive to technique than the newer, simplified adhesives. When primers are mixed with adhesives in 2-step single-bottle adhesives and self-etching primers, the adhesives are more permeable to water and hence absorb more water over time than previous generations of adhesives. The most recent single-step self-etching adhesives are even more hydrophilic and hence more permeable to water derived from the underlying bonded dentin. This permeability can lead to a wide variety of seemingly unrelated problems, including incompatibility of chemically or dual-cured composites with simplified adhesives and expedited degradation of resin-dentin bonds.