EDTA-treated Dentin Research Articles

Effects of sodium hypochlorite and ethylenediaminetetraacetic acid on proliferation, osteogenic/odontogenic differentiation, and mechanosensitive gene expression of human dental pulp stem cells

Sodium Hypochlorite (NaOCl) and Ethylene Diamine Tetraacetic Acid (EDTA) can change the biochemical and biophysical properties of dentin. However, the response of human dental pulp stem cells (hDPSCs) to NaOCl and EDTA-treated dentin remains unknown. This study was conducted to investigate the effect of NaOCl and EDTA on cell proliferation, osteogenic/odontogenic differentiation, and the response to mechanosensitive gene expression in hDPSCs. Dentin slices were treated with 5.25% NaOCl, 17% EDTA, and saline (0.9% NaCl) separately. The cell viability and osteogenic/odontogenic differentiation of hDPSCs were analyzed using scanning electron microscopy, cell counting assay, alkaline phosphatase (ALP) staining, and quantitative polymerase chain reaction (qPCR). Besides, the hardness was measured by a Vickers microhardness tester. The expression of mechanosensitive genes was detected by the qPCR assay. All the irrigant-treated dentin allowed cell attachment. The EDTA-treated dentin significantly boosted the ALP and osteogenic/odontogenic differentiation, followed by NaCl and NaOCl groups. Remarkably, these trends were similar to the expression of mechanosensitive genes but were different from the trends of hardness values. The effect of irrigant-treated dentin on regulating hDPSCs differentiation might correlate with mechanosensitive signals. Whereas, the hardness changes between groups might not produce significant roles in regulating osteogenic/odontogenic differentiation of stem cells on dentin surfaces.

Morphological and elemental analysis of silver penetration into sound/demineralized dentin after SDF application

ObjectiveThe aim of this study was to evaluate the penetration depth of silver into sound and demineralized dentin after application of silver diamine fluoride (SDF). MethodsTwo hundred and eighty-eight dentin specimens were used. The specimens were divided into 3 groups: (1) sound dentin (control), (2) 30min EDTA-treated dentin; and (3) 13h EDTA-treated dentin. SDF was applied to all specimens. Each group was divided into 3 subgroups according to storage time into: 24h, 2 weeks and 1-year storage time. Each subgroup was further divided into four subgroups (n=8) according to different examinations as optical microscope (OM) observation, scanning electron microscopic (SEM) observation, elemental analysis with energy dispersive spectroscopy (EDS) and Micro-PIXE test. ResultsThe OM showed discoloration in the superficial layer after 24h and keep extending deeper after 2 weeks and 1-year. SEM showed silver crystals within dentinal tubules after 2 weeks and 1-year. EDS analysis can detect silver penetration only in the 1-year group reaching around 1200μm inside dentin. Micro-PIXE test detected silver at all time intervals, confirming the EDS depth results. SignificanceIt can be concluded that silver ions can completely infiltrate the demineralized dentin lesion with further penetration into the underlying mineralized dentin.

Effect of EDTA-treated dentin on the differentiation of mouse iPS cells into osteogenic/odontogenic lineages in vitro and in vivo.

To investigate the effect of EDTA-treated dentin on the differentiation of mouse induced pluripotent stem (iPS) cells. Dentin discs were prepared from bovine incisors and treated with 17% EDTA. Embryoid bodies (EBs) formed from mouse iPS cells were seeded on the dentin discs for the experiment. The roughness of the EDTA-treated dentin surface, Sa and Sdr, was higher and collagen fibrillike structures were observed by the scanning electron microscopy (SEM) in vitro. In RT-PCR, the mRNA levels of the osteoblast markers Bsp and Ocn were significantly higher in the experimental group. Expression of the DMP1, DSP, and BSP proteins were more notable in the experimental group by immunofluorescence (ICF) study. In vivo study, cartilage and bone-like tissue were observed adjacent to the EDTA-treated dentin. The study demonstrates that the dentin treated with 17% EDTA induces mouse iPS cells to differentiate into the osteo/odontogenesis.

EDTA Enhances Stromal Cell–derived Factor 1α–induced Migration of Dental Pulp Cells by Up-regulating Chemokine Receptor 4 Expression

IntroductionIn regenerative endodontics, irrigation is an important step to ensure the success of treatment. EDTA as a common irrigant has been recommended in the American Associations of Endodontists guidelines. It has been suggested that EDTA-treated dentin slices could increase the attachment, differentiation, and migration of dental pulp stem cells. However, no information is available about the effect of EDTA on the migration of dental pulp cells (DPCs). The aim of this study was to explore how EDTA affects the migration of DPCs. MethodsCells were obtained from human premolars or third molars, and cell counting kit-8 was used to evaluate the influence of EDTA on cell proliferation at various concentrations and time points. Real-time polymerase chain reaction was used to detect the messenger RNA expression levels of transforming growth factor beta (TGF-β) and chemokine receptor 4 (CXCR4). Protein expression was tested by the enzyme-linked immunosorbent assay and Western blot, respectively. In addition, the transwell migration assay was performed to investigate the role of EDTA pretreatment in stromal cell–derived factor 1α (SDF-1α)-induced DPC migration. ResultsStimulation with 12% EDTA enhanced SDF-1α–induced migration of DPCs. Both expressions of TGF-β1 and CXCR4 were increased by 12% EDTA in a time-dependent manner. After silencing CXCR4, EDTA-enhanced migration was decreased. Furthermore, the transcriptional regulation of CXCR4 by EDTA was found to be mediated by TGF-β1/ERK1/2 and TGF-β1/Smad2/3 signal pathways. ConclusionsOur results showed that 12% EDTA could promote SDF-1α–induced migration of DPCs by up-regulating CXCR4 expression in which TGF-β1 signal pathways were involved.

EDTA Treatment for Sodium Hypochlorite–treated Dentin Recovers Disturbed Attachment and Induces Differentiation of Mouse Dental Papilla Cells

IntroductionThe disturbance of cellular attachment to dentin by sodium hypochlorite (NaOCl) may hamper pulp tissue regeneration. The aims of this study were to examine the recovering effect of EDTA on the attachment/differentiation of stemlike cells and to address the mechanisms of EDTA-induced recovery under the hypothesis that attachment to the exposed dentin matrix and the subsequent activation of integrin/phosphatidylinositol 3-kinase (PI3K) signaling play a crucial role. MethodsMouse dental papilla (MDP) cells were cultured on bovine dentin disks treated with NaOCl (0%, 1.5%, or 6%) followed by EDTA (0%, 3%, or 17%). Cell attachment was evaluated by cell density, viability, and scanning and transmission electron microscopy. Odonto-/osteoblastic gene expression in attached MDP cells was analyzed with or without a pan-PI3K inhibitor (LY294002) using real-time polymerase chain reaction. ResultsNaOCl treatment (1.5%, 10 minutes) significantly diminished attached MDP cells (P < .00001), but EDTA treatment (3% and 17%, ≥10 minutes) of NaOCl-pretreated dentin induced a significant increase in attached cells (P < .05). Ultrastructurally, MDP cells on EDTA-treated dentin showed attachment to exposed collagen fibers. MDP cells cultured on EDTA-treated disks (with or without 1.5% NaOCl pretreatment) showed significant up-regulation of alkaline phosphatase, dentin matrix protein 1, and dentin sialophosphoprotein messenger RNAs (P < .05). Alkaline phosphatase expression was down-regulated by LY294002 (P < .05). ConclusionsUnder the present experimental conditions, 10 minutes of EDTA treatment was sufficient to recover attachment/differentiation of MDP cells on 1.5% NaOCl-pretreated dentin. EDTA-induced exposure of collagen fibers and subsequent activation of integrin/PI3K signaling may contribute, at least partly, to the recovery.

Bond strength and bioactivity of Zn-doped dental adhesives promoted by load cycling.

The purpose of this study was to evaluate if mechanical loading influences bioactivity and bond strength at the resin-dentin interface after bonding with Zn-doped etch-and-rinse adhesives. Dentin surfaces were subjected to demineralization by 37% phosphoric acid (PA) or 0.5 M ethylenediaminetetraacetic acid (EDTA). Single bond (SB) adhesive—3M ESPE—SB+ZnO particles 20 wt% and SB+ZnCl2 2 wt% were applied on treated dentin to create the groups PA+SB, SB+ZnO, SB+ZnCl2, EDTA+SB, EDTA+ZnO, and EDTA+ZnCl2. Bonded interfaces were stored in simulated body fluid for 24 h and tested or submitted to mechanical loading. Microtensile bond strength (MTBS) was assessed. Debonded dentin surfaces were studied by high-resolution scanning electron microscopy. Remineralization of the bonded interfaces was assessed by atomic force microscope imaging/nanoindentation, Raman spectroscopy/cluster analysis, and Masson's trichrome staining. Load cycling (LC) produced reduction in MTBS in all PA+SB, and no change was encountered in EDTA+SB specimens, regardless of zinc doping. LC increased the mineralization and crystallographic maturity at the interface; a higher effect was noticed when using ZnO. Trichrome staining reflected a narrow demineralized dentin matrix after loading of dentin surfaces that were treated with SB-doped adhesives. This correlates with an increase in mineral platforms or plate-like multilayered crystals in PA or EDTA-treated dentin surfaces, respectively.

The use of traditional and novel techniques to determine the hardness and indentation properties of immature radicular dentin treated with antibiotic medicaments followed by ethylenediaminetetraacetic acid.

Objective:The objective was to investigate the effect of intracanal antibiotic medicaments followed by ethylenediaminetetraacetic acid (EDTA) on the indentation properties and hardness of radicular dentin using a BioDent reference point indenter and a traditional microhardness technique, respectively.Materials and Methods:Specimens with intact root canal dentin surfaces and polished radicular dentin specimens were obtained from immature human premolars. Each type of specimen was randomly assigned (n = 10 per group) and treated with either double antibiotic paste (DAP) for 4-week followed by EDTA for 5 min, triple antibiotic paste (TAP) for 4-week followed by EDTA for 5 min, EDTA for 5 min or Hank's balanced salt solution (control). The BioDent reference point indentor and Vickers microhardness tester were used to measure the indentation properties of root canal surfaces and the hardness of polished dentin specimens, respectively. One-way ANOVA followed by Fisher's protected least significant differences were used for statistical analyses.Results:Both types of radicular dentin treated with antibiotic pastes and/or EDTA had a significant increase in the majority of indentation properties and a significant reduction in hardness compared to the untreated dentin. Furthermore, treatment of dentin with antibiotic pastes and EDTA caused significant increases in indentation properties and a significant reduction in hardness compared to EDTA-treated dentin. However, the RPI technique was not able to significantly differentiate between DAP + EDTA and TAP + EDTA-treated dentin.Conclusion:Dentin treated with antibiotic medicaments followed by EDTA had a significant increase the indentation properties and significantly reduction in hardness of radicular dentin.

Zinc Incorporation Improves Biological Activity of Beta-tricalcium Silicate Resin–based Cement

IntroductionMatrix metalloproteinase (MMP) inhibition may improve endodontic treatment prognosis. The purpose of this study was to determine if zinc incorporation into experimental resin cements containing bioactive fillers may modulate MMP-mediated collagen degradation of dentin. MethodsHuman dentin samples untreated and demineralized using 10% phosphoric acid or 0.5 mol/L EDTA were infiltrated with the following experimental resins: (1) unfilled resin, (2) resin with Bioglass 45S5 particles (OSspray, London, UK), (3) resin with beta-tricalcium silicate particles (βTCS), (4) resin with zinc-doped Bioglass 45S5, and (5) resin with zinc-doped βTCS particles. The specimens were stored in artificial saliva (for 24 hours, 1 week, and 4 weeks) and submitted to radioimmunoassay to quantify C-terminal telopeptide. Scanning electron microscopy analysis was also undertaken on dentin samples after 4 weeks of storage. ResultsCollagen degradation was prominent both in phosphoric acid and EDTA-treated dentin. Resin infiltration strongly reduced MMP activity in demineralized dentin. Resin containing Bioglass 45S5 particles exerted higher and stable protection of collagen. The presence of zinc in βTCS particles increases MMP inhibition. Different mineral precipitation was attained in dentin infiltrated with the resin cements containing bioactive fillers. ConclusionsMMP degradation of dentin collagen is strongly reduced after resin infiltration of dentin. Zinc incorporation in βTCS particles exerted an additional protection against MMP-mediated collagen degradation. However, it did not occur in resin containing Bioglass 45S5 particles, probably because of the formation of phosphate-zinc compounds.

Effect of EDTA on Attachment and Differentiation of Dental Pulp Stem Cells

IntroductionIn regenerative endodontics, it is believed that EDTA induces odontoblast differentiation by releasing growth factors from the dentin matrix. The aim of this study was to evaluate the effect of EDTA on the attachment and differentiation of dental pulp stem cells (DPSCs). We also investigated whether the behavioral changes of DPSCs could be caused by biochemical components released from EDTA-treated dentin. MethodsCells were obtained from human third molars, and the stem-like nature of the cells was investigated by flow cytometric analysis. DPSCs were seeded on EDTA-treated and untreated dentin slices. After 3 days of culture, cell attachment was evaluated by cell density, fibronectin 1 gene expression level using quantitative real-time polymerase chain reaction, and scanning electron microscopy. After 21 days of culture, the expression of differentiation genes was investigated by quantitative real-time polymerase chain reaction, and calcification was observed using alizarin red S staining. To investigate the EDTA-induced growth factor release, DPSCs were cultured with or without direct contact with the EDTA-treated dentin surface. ResultsAfter 3 days of culture, both the cell density and fibronectin expression level were significantly higher in the EDTA-treated dentin group. After 3 weeks, the DPSCs on the EDTA-treated dentin surfaces showed higher expression levels of dentin sialophosphoprotein and dentin matrix protein 1, whereas the DPSCs cultured without direct contact with the EDTA-treated dentin surfaces did not exhibit these findings. ConclusionsOur results showed that EDTA induced cell attachment and odontoblastic/osteoblastic differentiation, which was observed only in the group in which the DPSCs were placed in direct contact with the EDTA-treated dentin surfaces. These findings suggest that EDTA is beneficial for achieving successful outcomes in regenerative endodontics.

In vitro load-induced dentin collagen-stabilization against MMPs degradation

IntroductionTeeth are continuously subjected to stresses during mastication, swallowing and parafunctional habits, producing a significant reduction of the bonding efficacy in adhesive restorations. The purpose of this study was to evaluate the metalloproteinases (MMPs)-mediated dentin collagen degradation of hybrid layers created by using different demineralization processes, previous resin infiltration, and in vitro mechanical loading. MethodsHuman dentin beams (0.75×0.75×5.0mm) were subjected to different treatments: (1) untreated dentin; (2) demineralization by 37% phosphoric acid (PA) or by 0.5% M ethylenediaminetetraacetic acid (EDTA); (3) demineralization by PA, followed by application of Adper™ Single Bond (SB); (4) demineralization by EDTA, followed by application of SB. In half of the specimens, mechanical loadings (100,000 cycles, 2Hz, 49N) were applied to dentin beams. Specimens were stored in artificial saliva. C-terminal telopeptide (ICTP), determinations (which indicates the amount of collagen degradation) (radioimmunoassay) were performed after 24h, 1 week and 4 weeks. ResultsLoad cycling decreased collagen degradation when dentin was untreated or PA-demineralized and EDTA-treated. ICTP values increased when both PA-demineralized and EDTA-treated and infiltrated with SB dentin beams were loaded, except in samples that were subjected to EDTA treatment and SB infiltration after 4w of storage, which showed similar values of collagenolytic activity than the non loaded specimens. Load cycling preserved the initial (24h) ICTP determination at any time point, in all groups of the study, except in PA-demineralized and SB infiltrated dentin which showed an increased of collagen degradation values, over time. This same trend was observed in all groups without loading. InterpretationMechanical loading enhances collagen's resistance to enzymatic degradation in natural and demineralized dentin. Mild acids (EDTA) lead to a lower volume of demineralized/unprotected collagen to be cleaved by MMPs. Load cycling produced an increase of collagen degradation when PA-demineralized dentin and EDTA-treated dentin were infiltrated with resin, but EDTA-treated dentin showed a constant collagenolytic degradation, over time.

Experimental Resin Cements Containing Bioactive Fillers Reduce Matrix Metalloproteinase–mediated Dentin Collagen Degradation

IntroductionCollagen dentin matrix may represent a suitable scaffold to be remineralized in the presence of bioactive materials. The purpose of this study was to determine if experimental resin cements containing bioactive fillers may modulate matrix metalloproteinase–mediated collagen degradation of etched dentin. MethodsHuman dentin beams demineralized using 10% phosphoric acid or 0.5 mol/L EDTA were infiltrated with the following experimental resins: (1) unfilled resin, (2) resin with Bioglass 45S5 particles (Sylc; OSspray Ltd, London, UK), and (3) resin with β–tricalcium phosphate–modified calcium silicate cement (HCAT-β) particles. The filler/resin ratio was 40/60 wt%. The specimens were stored in artificial saliva, and the determination of C-terminal telopeptide (ICTP) was performed by radioimmunoassay after 24 hours, 1 week, and 4 weeks. Scanning electron microscopic analysis of dentin surfaces after 4 weeks of storage was also executed. ResultsCollagen degradation was prominent both in phosphoric acid and EDTA-treated dentin. Resin infiltration strongly reduced the MMP activity in demineralized dentin. Resin-containing Bioglass 45S5 particles exerted higher and more stable protection of collagen at all tested dentin states and time points. HCAT-β induced collagen protection from MMPs only in EDTA-treated specimens. Dentin remineralization was achieved when dentin was infiltrated with the resin cements containing bioactive fillers. ConclusionsMMP degradation of dentin collagen is strongly reduced in resin-infiltrated dentin. The inclusion of Bioglass 45S5 particles exerted an additional protection of collagen during dentin remineralization.

Assessment of the quality of resin–dentin bonded interfaces: An AFM nano-indentation, μTBS and confocal ultramorphology study

ObjectiveThe aim of this study was to assess by using confocal microscopy (CLSM), AFM nano-indentation and microtensile bond strength test (μTBS) the quality of the resin–dentin interfaces created with selected bonding parameters. MethodsDentin conditioned with H3PO4 or EDTA was bonded in ethanol- or water-wet condition using a HEMA-free or HEMA-containing adhesive. The resin-bonded teeth were stored in distilled water (24h) and sectioned as match-sticks (0.9mm2) for μTBS. Further resin-bonded teeth were sectioned and analyzed using CLSM, and AFM nano-indentation. The AFM imaging and nano-indentation processes were undertaken using a Berkovich diamond indenter. The modulus of elasticity (Ei) and hardness (Hi) across the interface were evaluated with the specimens in a fully hydrated status. The AFM imaging was performed both in dry and wet conditions for evaluating the shrinkage of the hybrid layer on dehydration. ResultsThe HEMA-containing adhesive applied onto H3PO4-etched ethanol or water-wet dentin created hybrid layers with the lowest biomechanical nano-properties (p<0.05); no significant differences in μTBS were found between the two wet-bonding techniques (p>0.05). However, the ethanol-wet bonding reduced the dye penetration into the adhesive layer created with the HEMA-containing adhesive. Hybrid layers with high biomechanical properties, low micropermeability and no shrinkage were only possible when using HEMA-free adhesive applied in ethanol wet-dentin. In particular, a significant increase in Ei and Hi was achieved at the hybrid layer and underneath the resin–dentin interface of ethanol-wet EDTA-treated dentin. SignificanceThe use of HEMA-free adhesives applied onto ethanol-wet dentin may be considered as an alternative and suitable bonding strategy to achieve high quality resin–dentin interfaces.

Resin–dentin bonds to EDTA-treated vs. acid-etched dentin using ethanol wet-bonding. Part II: Effects of mechanical cycling load on microtensile bond strengths

Objective To compare microtensile bond strengths (MTBS) subsequent to load cycling of resin bonded acid-etched or EDTA-treated dentin using a modified ethanol wet-bonding technique. Methods Flat dentin surfaces were obtained from extracted human molars and conditioned using 37% H 3PO 4 (PA) (15 s) or 0.1 M EDTA (60 s). Five experimental adhesives and one commercial bonding agent were applied to the dentin and light-cured. Solvated experimental resins (50% ethanol/50% comonomers) were used as primers and their respective neat resins were used as the adhesives. The resin-bonded teeth were stored in distilled water (24 h) or submitted to 5000 loading cycles of 90 N. The bonded teeth were then sectioned in beams for MTBS. Modes of failure were examined by scanning electron microscopy. Results The most hydrophobic resin 1 gave the lowest bond strength values to both acid and EDTA-treated dentin. The hydrophobic resin 2 applied to EDTA-treated dentin showed lower bond strengths after cycling load but this did not occur when it was bonded to PA-etched dentin. Resins 3 and 4, which contained hydrophilic monomers, gave higher bond strengths to both EDTA-treated or acid-etched dentin and showed no significant difference after load cycling. The most hydrophilic resin 5 showed no significant difference in bond strengths after cycling loading when bonded to EDTA or phosphoric acid treated dentin but exhibited low bond strengths. Significance The presence of different functional monomers influences the MTBS of the adhesive systems when submitted to cyclic loads. Adhesives containing hydrophilic comonomers are not affected by cycling load challenge especially when applied on EDTA-treated dentin followed by ethanol wet bonding.

Effect of dentin etching and chlorhexidine application on metalloproteinase‐mediated collagen degradation

Dentin matrix metalloproteinases (MMPs) are involved in the degradation of collagen in resin-dentin interfaces. This study evaluated whether collagen degradation can be prevented by chlorhexidine digluconate (CHX) after different dentin demineralization procedures. The demineralization of human dentin was performed with phosphoric acid (PA), EDTA or acidic monomers (Clearfil SE Bond and Xeno V). Specimens were stored (for 24 h, or for 1 or 3 wk) in the presence or absence of CHX. In half of the groups, active MMP-2 was incorporated into the storage solution. At the end of each storage period, the C-terminal telopeptide (ICTP) concentration (which indicates the amount of collagen degradation) was measured in the storage solution. Collagen degradation was higher in PA- and EDTA-demineralized dentin. Chlorhexidine digluconate reduced collagen degradation in these groups only for 24 h. When dentin was demineralized with Clearfil SE Bond or Xeno V, collagen degradation was reduced by up to 30%, but the addition of exogenous MMP-2 significantly increased collagen degradation. In self-etchant-treated dentin, the inhibitory effect of CHX on MMPs lasted for up to 3 wk. Treating dentin with EDTA, PA or self-etching agents produces enough demineralization to permit cleavage of the exposed collagen. Monomer infiltration may exert protection on demineralized collagen, probably through immobilization of MMPs. The partial inhibitory action of CHX on MMP activity produced by self-etching adhesives was prolonged compared with the short-acting PA- or EDTA-treated dentin.

Effect of desensitising toothpastes on dentinal tubule occlusion: A dentine permeability measurement and SEM in vitro study

Objectives The purpose of this study was to evaluate the in vitro effectiveness of a new bioglass-containing and two commercial desensitising toothpastes on dentinal tubule occlusion after citric acid challenge or artificial saliva (AS) immersion. Methods One hundred dentin discs from human third molars were used. Specimens were randomly divided into five groups ( n = 20), Group 1: EDTA-treated dentin; Group 2: brushing with distilled water; Group 3: brushing with Novamin; Group 4: brushing with Sensodyne Freshmint; Group 5: brushing with Colgate Sensitive. In each group, samples were then equally split into two subgroups ( n = 10) to test two post-treatments: 6% citric acid challenge or 24 h immersion in artificial saliva. Dentine permeability of each specimen was measured before and after each treatment using a hydrostatic device working at 20 cm H 2O pressure. Data were analysed by two-way repeated measures ANOVA to determine if there were any significant differences within or between groups. Dentine morphology and surface deposits were observed by SEM. Results All three desensitising toothpastes significantly reduced dentine permeability and created precipitates on the treated dentine surfaces. Moreover, the reductions in dentine permeability showed partial recovery after a citric acid and artificial saliva immersion. Sensodyne showed significant resistant to acid attack and Novamin exhibited the lowest permeability after artificial saliva immersion for 24 h. Conclusions The application of the three toothpastes resulted in effective dentinal tubule occlusion. However, the new bioglass-containing toothpaste (Novamin) represented excellent occlusion effects after brushing treatment and AS immersion, while Sensodyne demonstrated more reduction in permeability when citric acid challenged.

Resin–dentin bonds to EDTA-treated vs. acid-etched dentin using ethanol wet-bonding

Objective To compare resin–dentin bond strengths and the micropermeability of hydrophobic vs. hydrophilic resins bonded to acid-etched or EDTA-treated dentin, using the ethanol wet-bonding technique. Methods Flat dentin surfaces from extracted human third molars were conditioned before bonding with: 37% H 3PO 4 (15 s) or 0.1 M EDTA (60 s). Five experimental resin blends of different hydrophilicities and one commercial adhesive (SBMP: Scotchbond Multi-Purpose) were applied to ethanol wet-dentin (1 min) and light-cured (20 s). The solvated resins were used as primers (50% ethanol/50% comonomers) and their respective neat resins were used as the adhesive. The resin-bonded teeth were stored in distilled water (24 h) and sectioned in beams for microtensile bond strength testing. Modes of failure were examined by stereoscopic light microscopy and SEM. Confocal tandem scanning microscopy (TSM) interfacial characterization and micropermeability were also performed after filling the pulp chamber with 1 wt% aqueous rhodamine-B. Results The most hydrophobic resin 1 gave the lowest bond strength values to acid-etched dentin and all beams failed prematurely when the resin was applied to EDTA-treated dentin. Resins 2 and 3 gave intermediate bond strengths to both conditioned substrates. Resin 4, an acidic hydrophilic resin, gave the highest bond strengths to both EDTA-treated and acid-etched dentin. Resin 5 was the only hydrophilic resin showing poor resin infiltration when applied on acid-etched dentin. Significance The ethanol wet-bonding technique may improve the infiltration of most of the adhesives used in this study into dentin, especially when applied to EDTA-treated dentin. The chemical composition of the resin blends was a determining factor influencing the ability of adhesives to bond to EDTA-treated or 37% H 3PO 4 acid-etched dentin, when using the ethanol wet-bonding technique in a clinically relevant time period.

Effect of soft chelating irrigation on the sealing ability of GP/AH Plus root fillings

The purpose of this study was to evaluate the effect of soft chelating irrigant on the sealing ability of root fillings by using a glucose leakage test. A total of 45 single-rooted teeth were selected for the study. The teeth were decoronated leaving a total length of 13mm. The root canals prepared using K3 NiTi rotary instruments to an apical dimension of size 45(0.06 taper). The specimens were then randomly divided into 3 experimental groups of 13 roots each and 2 control groups of 3 roots each. Specimen in each group were prepared with different irrigation protocols : group 1, 2.5% NaOCl; group 2, 2.5% NaOCl and 17% EDTA: group 3, 2.5% NaOCl and 15% HEBP. The root canals were filled with gutta-percha and AH Plus sealer using lateral condensation. After 7 days in , 100% humidity, the coronal-to-apical microleakage was evaluated quantitatively using a glucose leakage model. The leaked glucose concentration was measured with spectrophotometry at 1, 4, 7, 14, 21 and 28 days. There was a tendency of increase in leakage in all experimental groups during experimental period. HEBP-treated dentin showed no significant difference with EDTA-treated dentin during experimental period. From the 21th day onward, HEBP-treated dentin showed significantly lower leakage than smear-covered dentin. HEBP-treated dentin displayed a similar sealing pattern to EDTA-treated dentin and a better sealing ability than smear-covered dentin. Consequently, a soft chelator(HEBP) could be considered as the possible alternative to EDTA.

Root surface texture and early cell and tissue colonization after different etching modalities

The purpose of the present study was to compare the texture of dentin surfaces after etching with citric and phosphoric acids as well as EDTA with respect to mode of application and exposure time, and to assess possible effects on early cell and tissue colonization. It was concluded that EDTA operating at neutral pH was able to selectively remove mineral from a dentin surface, exposing a collagenous matrix. This was in contrast to etching with citric and phosphoric acids, both of which operate at a low pH. They both appeared to remove not only the mineral component but also the collagenous matrix. EDTA-treated dentin surfaces, compared to surfaces etched at low pH, appeared to be more inviting for cellular colonization and subsequent connective tissue formation. The reason for this may have been twofold: Dentin surfaces etched with an etching agent operating at neutral pH, as indicated above, retain more exposed collagen fibers than dentin surfaces etched at low pH. Furthermore, it can not be excluded that a low pH etching agent necrotized an area of the periodontal ligament in the immediate vicinity of the denuded dentin surfaces.

Atomic force microscopy studies of the effects of chemicals on the dentin tissue

Atomic force microscopy (AFM) was used to monitor the effect of several chemicals on the dentin tissue. The action of three different etching agents, namely aqueous solutions of phosphoric acid, polyacrylic acid and EDTA (at clinically relevant concentrations) on dentin was followed in real time. The etched dentin was then treated with an aqueous solutions of glutaraldehyde and hydroxyethylmethacrylate (part of a commercial bonding agent), and the effect of the latter on the dentin morphology was studied as a function of the etching agent used. Results show different mode of actions of the chemicals tested, and that hydroxyethylmethacrylate undergoes a polymerization reaction on EDTA-treated dentin. More generally, it is shown that the use of AFM for real-time imaging of wet samples under normal pressure conditions can give important information on the effects of chemicals on the dentin tissue.

Dentin-polymer bond mediated by glutaraldehyde/HEMA.

A restorative resin was bonded to EDTA-treated dentin by means of a mixture of glutaraldehyde and HEMA. The bond exhibited a tensile strength of 17.5 +/- 1.0 MPa (mean +/- SEM) and was unaffected by water storage at 37 degrees C for up to 6 months. The glutaraldehyde/HEMA mixture was found to be most effective when the pH was between 2 and 5; an application time of 10 s was found to be sufficient for the glutaraldehyde/HEMA mixture as well as for the EDTA-solution.