Differences in the microstructure and fatigue properties of dentine between residents of North and South America.

UNLABELLED: Spatial variations in the microstructure of dentine contribute to its mechanical behaviour. OBJECTIVE: The objective of this investigation was to compare the microstructure and fatigue behaviour of dentine from donors of two different countries. METHODS: Caries-free third molars were obtained from dental practices in Colombia, South America and the US to assemble two age-matched samples. The microstructure of the coronal dentine was evaluated at three characteristic depths (i.e. deep, middle and superficial dentine) using scanning electron microscopy and image processing techniques. The mechanical behaviour of dentine in these three regions was evaluated by the fatigue crack growth resistance. Cyclic crack growth was achieved in-plane with the dentine tubules and the fatigue crack growth behaviour was characterized in terms of the stress intensity threshold and the Paris Law parameters. RESULTS: There was no difference in the tubule density between the dentine of patients from the two countries. However, there were significant differences (p≤0.05) in the tubule lumen diameters between the two groups in the deep and peripheral regions. In regards to the fatigue resistance, there was a significant increase (p≤0.05) in threshold stress intensity range, and a significant decrease in fatigue crack growth coefficient with increasing distance from the pulp in teeth from the US donors. In contrast, these properties were independent of location for the dentine of teeth from the Colombian donors. CONCLUSIONS: The microstructure of dentine and its mechanical behaviour appear to be a function of patient background, which may include environmental factors and/or ethnicity.

Cysteine cathepsins in human carious dentin.

Matrix metalloproteinases (MMPs) are important in dentinal caries, and analysis of recent data demonstrates the presence of other collagen-degrading enzymes, cysteine cathepsins, in human dentin. This study aimed to examine the presence, source, and activity of cysteine cathepsins in human caries. Cathepsin B was detected with immunostaining. Saliva and dentin cysteine cathepsin and MMP activities on caries lesions were analyzed spectrofluorometrically. Immunostaining demonstrated stronger cathepsins B in carious than in healthy dentin. In carious dentin, cysteine cathepsin activity increased with increasing depth and age in chronic lesions, but decreased with age in active lesions. MMP activity decreased with age in both active and chronic lesions. Salivary MMP activities were higher in patients with active than chronic lesions and with increasing lesion depth, while cysteine cathepsin activities showed no differences. The results indicate that, along with MMPs, cysteine cathepsins are important, especially in active and deep caries.

Ethanol wet-bonding challenges current anti-degradation strategy.

The long-term effectiveness of chlorhexidine as a matrix metalloproteinase (MMP) inhibitor may be compromised when water is incompletely removed during dentin bonding. This study challenged this anti-bond degradation strategy by testing the null hypothesis that wet-bonding with water or ethanol has no effect on the effectiveness of chlorhexidine in preventing hybrid layer degradation over an 18-month period. Acid-etched dentin was bonded under pulpal pressure simulation with Scotchbond MP and Single Bond 2, with water wet-bonding or with a hydrophobic adhesive with ethanol wet-bonding, with or without pre-treatment with chlorhexidine diacetate (CHD). Resin-dentin beams were prepared for bond strength and TEM evaluation after 24 hrs and after aging in artificial saliva for 9 and 18 mos. Bonds made to ethanol-saturated dentin did not change over time with preservation of hybrid layer integrity. Bonds made to CHD pre-treated acid-etched dentin with commercial adhesives with water wet-bonding were preserved after 9 mos but not after 18 mos, with severe hybrid layer degradation. The results led to rejection of the null hypothesis and highlight the concept of biomimetic water replacement from the collagen intrafibrillar compartments as the ultimate goal in extending the longevity of resin-dentin bonds.

Tubular occlusion optimizes bonding of hydrophobic resins to dentin.

Although hydrophobic resins may be bonded to acid-etched dentin with an ethanol wet-bonding technique, the protocol is sensitive to moisture contamination when bonding is performed in deep dentin. This study tested the hypothesis that the use of oxalate or poly(glutamic) acid-modified, diluted ceramicrete (PADC) for dentinal tubule occlusion prevents fluid contamination and improves the bonding of an experimental hydrophobic adhesive to acid-etched, ethanol-dehydrated dentin. Mid-coronal and deep acid-etched moist dentin pre-treated with oxalate or PADC was dehydrated by ethanol wet-bonding and infiltrated with the experimental three-step etch-and-rinse hydrophobic adhesive under simulated pulpal pressure. Tensile bond strengths to deep dentin without pre-treatment were severely compromised. Conversely, oxalate and PADC pre-treatments reduced dentin permeability, prevented water contamination, and improved bond strengths. Minimal nanoleakage was identified within hybrid layers created in the oxalate- and PADC-pre-treated deep dentin. The use of tubular occluding agents optimized bonding of hydrophobic resins to dentin.

Chlorhexidine preserves dentin bond in vitro.

Loss of hybrid layer integrity compromises resin-dentin bond stability. Matrix metalloproteinases (MMPs) may be partially responsible for hybrid layer degradation. Since chlorhexidine inhibits MMPs, we hypothesized that chlorhexidine would decelerate the loss of resin-dentin bonds. Class I preparations in extracted third molars were sectioned into two halves. One half was customarily restored (etch-and-rinse adhesive/resin composite), and the other was treated with 2% chlorhexidine after being acid-etched before restoration. Specimens were stored in artificial saliva with/without protease inhibitors. Microtensile bond strengths and failure mode distribution under SEM were analyzed immediately after specimens' preparation and 6 months later. With chlorhexidine, significantly better preservation of bond strength was observed after 6 months; protease inhibitors in the storage medium had no effect. Failure analysis showed significantly less failure in the hybrid layer with chlorhexidine, compared with controls after 6 months. In conclusion, this in vitro study suggests that chlorhexidine might be useful for the preservation of dentin bond strength.

Chlorhexidine arrests subclinical degradation of dentin hybrid layers in vivo.

The recent paradigm that endogenous collagenolytic and gelatinolytic activities derived from acid-etched dentin result in degradation of hybrid layers requires in vivo validation. This study tested the null hypothesis that there is no difference between the degradation of dentin bonded with an etch-and-rinse adhesive and that in conjunction with chlorhexidine, an MMP inhibitor, applied after phosphoric-acid-etching. Contralateral pairs of bonded Class I restorations in primary molars of clinical subjects were retrieved after a six-month period of intra-oral functioning and processed for transmission electron microscopy. Hybrid layers from the chlorhexidine-treated teeth exhibited normal structural integrity of the collagen network. Conversely, abnormal hybrid layers were seen in the control teeth, with progressive disintegration of the fibrillar network, to the extent that it was beyond detection by collagen staining. Self-destruction of collagen matrices occurs rapidly in resin-infiltrated dentin in vivo and may be arrested with the use of chlorhexidine as an MMP inhibitor.

Adhesive permeability affects coupling of resin cements that utilise self-etching primers to dentine.

OBJECTIVES: To examine the effects of an experimental bonding technique that reduces the permeability of the adhesive layer on the coupling of resin cements to dentine. METHODS: Extracted human third molars had their mid to deep dentin surface exposed flat by transversally sectioning the crowns. Resin composite overlays were constructed and cemented to the surfaces using either Panavia F (Kuraray) or Bistite II DC (Tokuyama) resin cements mediated by their respective one-step or two-step self-etch adhesives. Experimental groups were prepared in the same way, except that the additional layer of a low-viscosity bonding resin (LVBR, Scotchbond Multi-Purpose Plus, 3M ESPE) was placed on the bonded dentine surface before luting the overlays with the respective resin cements. The bonded assemblies were stored for 24 h in water at 37 degrees C and subsequently prepared for microtensile bond strength testing. Beams of approximately 0.8 mm(2) were tested in tension at 0.5 mm/min in a universal tester. Fractured surfaces were examined under scanning electron microscopy (SEM). Additional specimens were prepared and examined with TEM using a silver nitrate-staining technique. RESULTS: Two-way ANOVA showed significant interactions between materials and bonding protocols (p<0.05). When bonded according to manufacturer's directions, Panavia F produced bond strengths that were significantly lower than Bistite II DC (p<0.05). The placement of an additional layer of a LVBR improved significantly the bond strengths of Panavia F (p<0.05), but not of Bistite II DC (p>0.05). SEM observation of the fractured surfaces in Panavia F showed rosette-like features that were exclusive for specimens bonded according to manufacturer's directions. Such features corresponded well with the ultrastructure of the interfaces that showed more nanoleakage associated with the more permeable adhesive interface. The application of the additional layer of the LVBR reduced the amount of silver impregnation for both adhesives suggesting that reduced permeability of the adhesives resulted in improved coupling of the resin cements to dentin. CONCLUSIONS: Placement of an intermediate layer of a LVBR between the bonded dentine surface and the resin cements resulted in improved coupling of Panavia F to dentine.

Effects of HEMA/solvent combinations on bond strength to dentin.

Re-expansion of dried demineralized dentin is required to optimize resin adhesion. This study tested the hypothesis that bond strengths to dentin depend upon the ability of experimental HEMA(2-hydroxy-ethyl-methacrylate)/solvent primers to re-expand the matrix. Dentin surfaces were acid-etched with 37% phosphoric acid for 20 sec, air-dried for 30 sec, primed with either 35/65% (v/v) HEMA/water, HEMA/methanol, HEMA/ethanol, or HEMA/propanol for 60 sec, and bonded with 4-META-TBBO(4-methacryloyloxyethyl trimellitate anhydride-tri-n-butyl borane) adhesive. After storage in water for 1 day at 37 degrees C, the samples were prepared for microtensile bond strength testing. We used transmission electron microscopy to measure the width of interfibrillar spaces in the hybrid layers. The HEMA/ethanol primer and the HEMA/propanol primer produced the highest and the lowest bond strengths, respectively (p < 0.05). Bond strengths were directly correlated with the width of the interfibrillar spaces (p < 0.05). Bond strengths are related to the ability of the primer to maintain the re-expansion of collapsed demineralized dentin matrix.