Annalisa Mazzoni

MMP Activity in the Hybrid Layer Detected with in situ Zymography

Dentinal proteases are believed to play an important role in the degradation of hybrid layers (HL). This study investigated the HL gelatinolytic activity by in situ zymography and functional enzyme activity assay. The hypotheses were that HLs created by an etch-and-rinse adhesive exhibit active gelatinolytic activity, and MMP-2 and -9 activities in dentin increase during adhesive procedures. Etched-dentin specimens were bonded with Adper Scotchbond 1XT and restored with composite. Adhesive/dentin interface slices were placed on microscope slides, covered with fluorescein-conjugated gelatin, and observed with a multi-photon confocal microscope after 24 hrs. Human dentin powder aliquots were prepared and assigned to the following treatments: A, untreated; B, etched with 10% phosphoric acid; or C, etched with 10% phosphoric acid and mixed with Scotchbond 1XT. The MMP-2 and -9 activities of extracts of dentin powder were measured with functional enzyme assays. Intense and continuous enzyme activity was detected at the bottom of the HL, while that activity was more irregular in the upper HL. Both acid-etching and subsequent adhesive application significantly increased MMP-2 and -9 activities (p < 0.05). The results demonstrate, for the first time, intrinsic MMP activity in the HL, and intense activation of matrix-bound MMP activity with both etching and adhesive application.

Effect of UVA-activated Riboflavin on Dentin Bonding

Recent studies have reported collagen cross-linking after exposure to riboflavin followed by ultraviolet-A (UVA) exposure. This study is the first to investigate the effect of a riboflavin-containing primer on adhesive interface stability and dentinal matrix metalloproteinase activity. Human dentin was etched with 35% phosphoric acid, treated with 0.1% riboflavin, exposed to UVA for 2 min, and bonded with a two-step etch-and-rinse adhesive. Adhesive was applied to control specimens without riboflavin/UVA. Specimens were subjected to microtensile bond strength tests and pulled to failure after storage for 24 hrs, 6 mos, or 1 yr. Interfacial nanoleakage was evaluated by light and transmission electron microscopy. To investigate dentinal matrix metalloproteinase activity, we performed correlative zymographic assays on protein extracts obtained from phosphoric-acid-etched dentin powder with or without riboflavin/UVA treatment and XP Bond. Ultraviolet-activated riboflavin treatment increased the immediate bond strength to dentin at all aging intervals (p < 0.05 vs. control) and decreased interfacial nanoleakage in aged specimens (1 yr; p < 0.05). Zymograms revealed that riboflavin/UVA pre-treatment inhibited dentinal matrix metalloproteinase activity (especially MMP-9). In conclusion, dentinal collagen cross-linking induced by riboflavin/UVA increased immediate bond strength, stabilized the adhesive interface, and inhibited dentin matrix metalloproteinases, thereby increasing the durability of resin-dentin bonds.

Influence of preliminary etching on the stability of bonds created by one‐step self‐etch bonding systems

We evaluated the effects of preliminary etching of dentine on the stability of the bond created by one-step self-etch adhesives under different storage conditions. Adper Easy Bond (3M ESPE) and iBond Self-Etch (iBond SE; Heraeus Kulzer) were applied with an etch-and-rinse (i.e. after preliminary phosphoric acid etching for 15 s) or a self-etch approach. Resin-dentine bonded specimens were sectioned perpendicularly to the adhesive interface according to the non-trimming technique. Beams were stored in artificial saliva for 24 h, 6 months, or 1 yr at 37°C, or in 10% NaOCl for 5 h at room temperature, and then stressed until failure; the microtensile bond strengths were calculated. Interfacial nanoleakage of additional teeth was evaluated using light microscopy or transmission electron microscopy. Adper Easy Bond showed higher bond strength than iBond SE, regardless of the dentine treatment. Similar microtensile bond strength results were obtained for teeth subjected to artificial ageing in 10% NaOCl for 5 h at room temperature and for teeth stored in artificial saliva for 6 months at 37°C. The additional etching step increased the microtensile bond strength for Adper Easy Bond and iBond SE. This study supports the use of one-step adhesives on etched dentine because of the increased bond strength compared with their application onto smear-layer-covered dentine, regardless of storage conditions.

Interfacial nanoleakage and internal cement thickness of three esthetic crown systems

OBJECTIVESnTo evaluate interfacial nanoleakage expression of the combination of different cementation procedures and different crown systems.nnnMETHODSnForty-five human premolars prepared to receive single crowns were randomly divided into three groups (n=15) based on the materials to be used for crown fabrication and cementation: group 1: Zirc (Ivoclar-Vivadent) cemented with Multilink Automix; group 2: Ivoclar disilicate IPS Empress 2 luted with Excite DSC in combination with Variolink II; group 3: AAdva Zirconia (GC) cemented with G-Cem Automix. The specimens were then assessed for interfacial nanoleakage expression and the amount of silver deposits along the interface was quantified. The thickness of the cement was assessed at 5 different levels: cervical margins, midway between the cervical margin and the occlusal wall along the axial walls and at the occlusal wall. The nanoleakage scores and the cement thickness were analyzed with Kruskall-Wallis non-parametric Analysis of Variance and Dunns Multiple-range post hoc test.nnnRESULTSnGroup 2 showed significantly less nanoleakage expression than group 1 (p<0.05). The groups can be ranked in the following order 3<1<2 with regard to the cement thickness.nnnCONCLUSIONSnThere was no correlation among combination of different cementation procedures and different crown systems and interfacial nanoleakage. Also the cement thickness and the degree of nanoleakage cannot be related. The amount of cement found at the cervical margins of all groups showed discrepancies within a clinical acceptable range.

Electrical properties of resin monomers used in restorative dentistry

OBJECTIVESnThe application of an electric field has been shown to positively influence the impregnation of the resin monomers currently used in dentin bonding systems during hybrid layer formation. This study presents an experimental characterization of the electrical properties of these monomers with the aim of both correlating them to their chemical structures and seeking an insight into the mechanisms of the monomer migration under an applied electric field.nnnMETHODSnSome common monomers examined were TEGDMA (triethyleneglycoldimethacrylate), HEMA (2-hydroxyethyl methacrylate), UDMA (urethane dimethacrylate), 2-MP (bis[2-(methacryloyloxy)ethyl] phosphate, TCDM di(hydroxyethyl methacrylate) ester of 5-(2,5-dioxotetrahydrofurfuryl)-3-methyl-3-cyclohexenyl-1,2-dicarboxylic anhydride) and Bis-GMA [2,2-bis(4-2-hydroxy-3-methacryloyloxypropoxyphenyl)propane]. A customized cell produced for the measurement of the electrical properties of monomers was manufactured and electrical conductivity and permittivity of resin monomers were measured.nnnRESULTSnThe permittivity of the tested monomers is largely affected by electrical frequency. The large values of permittivity and dielectric losses observed as frequency decreased, indicate a dominant effect of ionic polarization, particularly evident in materials showing the highest conductivity. Permittivity and conductivity of the tested monomers showed a similar behavior, i.e. materials with the lowest permittivity also show small values of conductivity and vice versa.nnnSIGNIFICANCEnThe results of the present study revealed a good correlation between electrical properties and Hoy solubility parameters and, in particular, the higher the polar contribution (polar forces plus hydrogen bonding) the higher the permittivity and conductivity. The most relevant outcome of this study is that the electrophoretic mechanism prevails on the electroendoosmotic effect in determining the monomer migration under the application of electric fields.