Effects of calcium silicate-based materials on collagen matrix integrity of mineralized dentin.

INTRODUCTION: Calcium silicate-based materials (CSMs) are used in various endodontic procedures. The present study examined whether prolonged contact of mineralized dentin with recently commercialized versions of these materials adversely affects dentin collagen matrix integrity. METHODS: Dentin slabs prepared from extracted human third molars (7 × 3 × 0.3 mm) were divided into 3 groups on the basis of the material to which dentin was exposed (MTA Plus, Biodentine, untreated control dentin slabs) and the time period of exposure (24 hours, 1, 2, and 3 months; n = 6). Hydroxyproline assay was performed on each group's supernatant to quantify the collagen extraction amounts of each group per time period. Data were analyzed with two-factor repeated-measures analysis of variance and Holm-Sidak pair-wise comparisons (α = 0.05) to determine the effects of material and aging time on collagen extraction. Dentin slabs from the 3 months of aging group were demineralized for transmission electron microscopy examination of collagen matrix ultrastructural changes. RESULTS: Material (P = .002), aging time (P < .001), and their interactions (P = .007) significantly affected the amount of hydroxyproline (pg/mg of mineralized dentin) extracted from mineralized dentin and were significantly correlated by power regression models. Collagen degradation was identified from the surface of dentin slabs that were in direct contact with CSMs. CONCLUSIONS: Prolonged contact of mineralized dentin with CSMs has an adverse effect on the integrity of the dentin collagen matrix. However, the amount of collagen extracted was limited to the contact surface. Clinicians can continue to apply CSMs in endodontic procedures; however, caution is advised when these materials are applied to thin dentinal walls.

Effects of calcium silicate-based materials on the flexural properties of dentin.

INTRODUCTION: Prolonged exposure of root dentin to calcium hydroxide alters the fracture resistance of dentin. Calcium silicate-based materials (CSMs) used in endodontics release calcium hydroxide on setting. This study examined whether prolonged contact of dentin with CSMs adversely affects its mechanical properties. METHODS: Dentin beams prepared from extracted human molars (7 × 3 × 0.3 mm) were divided into 3 groups on the basis of the material to which dentin was exposed (Biodentine, MTA Plus, and untreated control beams). Three-point flexure to failure was performed for each beam at designated exposure times (24 hours, 1, 2, and 3 months; n = 10). Data were analyzed with 2-factor repeated-measures analyses of variance to determine the effects of material and aging time on flexural modulus, flexural strength, and modulus of toughness (α = 0.05). RESULTS: For flexural modulus, there was no significant difference for material (P = .947) or aging time (P = .064) when compared with baseline control. For flexural strength, significant differences were associated with aging time (P < .001) but not with material (P = .349). Flexural strength of dentin exposed to Biodentine decreased significantly after 2 and 3 months, whereas that exposed to MTA Plus decreased significantly after 3 months of aging (P < .05). For modulus of toughness, significant declines were observed for both material (P < .004) and aging time (P < .001). CONCLUSIONS: Both CSMs alter material toughness more than the strength and stiffness of dentin after aging in 100% relative humidity. Because dentin toughness is attributed to its collagen matrix, the amount of collagen extracted from mineralized dentin and changes in collagen ultrastructure should be further examined after exposure of dentin to CSMs.

Impact of the quality of coronal restoration versus the quality of root canal fillings on success of root canal treatment: a systematic review and meta-analysis.

INTRODUCTION: Thorough cleaning and shaping of root canals are essential for periapical healing. Restoration of endodontically treated teeth is also required for them to function and prevent coronal leakage. This study compared the impact of the quality of root canal treatment versus the quality of coronal restoration in treatment outcomes. METHODS: Literature search was conducted using the search terms "coronal restoration," "root canal," "periapical status," and "quality." Articles that evaluated the effect of the quality of root filling and coronal restoration or both on the success of root canal treatment were selected. Nine articles were identified and were reviewed by 3 investigators. Data were collected based on predetermined criteria. Percentages of teeth without apical periodontitis were recorded for each category: adequate root canal treatment (AE), inadequate root canal treatment (IE), adequate restoration (AR), and inadequate restoration (IR). Data were analyzed using meta-analysis for odds ratios (ORs). RESULTS: After adjusting for significant covariates to reduce heterogeneity, the results were combined to obtain pooled estimates of the common OR for the comparison of AR/AE versus AR/IE (OR = 2.734; 95% confidence interval [CI], 2.61-2.88; P < .001) and AR/AE versus IR/AE (OR = 2.808; 95% CI, 2.64-2.97; P < .001). CONCLUSIONS: On the basis of the current best available evidence, the odds for healing of apical periodontitis increase with both adequate root canal treatment and adequate restorative treatment. Although poorer clinical outcomes may be expected with adequate root filling-inadequate coronal restoration and inadequate root filling-adequate coronal restoration, there is no significant difference in the odds of healing between these 2 combinations.

Setting properties and cytotoxicity evaluation of a premixed bioceramic root canal sealer.

INTRODUCTION: This study investigated the setting time and micohardness of a premixed calcium phosphate silicate-based sealer (EndoSequence BC Sealer; Brasseler USA, Savannah, GA) in the presence of different moisture contents (0-9 wt%). The moisture content that produced the most optimal setting properties was used to prepare set EndoSequence BC Sealer for cytotoxicity comparison with an epoxy resin-based sealer (AH Plus; Dentsply Caulk, Milford, DE). METHODS: Standardized disks were created with BC Sealer, AH Plus, Pulp Canal Sealer EWT (positive control) (SybronEndo, Orange CA), and Teflon (Small Parts Inc., Miami Lakes, FL; negative control). Disks were placed in Transwell Inserts, providing indirect contact with MC3T3-E1 cells. Succinate dehydrogenase activity of the cells was evaluated over a 6-week period using MTT ((3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. Cytotoxicity profiles of BC Sealer and AH Plus were fitted with polynomial regression models. The time for 50% of the cells to survive (T(0.5)) was analyzed using the Wald statistic with a two-tailed significance level of 0.05. RESULTS: BC Sealer required at least 168 hours to reach the final setting using the Gilmore needle method, and its microhardeness significantly declined when water was included in the sealer (P = .004). All set sealers exhibited severe cytotoxicity at 24 hours. The cytotoxicity of AH Plus gradually decreased and became noncytotoxic, whereas BC Sealer remained moderately cytotoxic over the 6-week period. A significant difference (P < .001) was detected between T(0.5) of BC Sealer (5.10 weeks; 95% confidence interval [CI], 4.69-5.42, standard error [SE] = 0.09) and T(0.5) of AH Plus (0.86 weeks; 95% CI, 0.68-1.05; SE = 0.18). CONCLUSIONS: Further studies are required to evaluate the correlation between the length of setting time of BC Sealer and its degree of cytotoxicity.

Effects of different exposure times and concentrations of sodium hypochlorite/ethylenediaminetetraacetic acid on the structural integrity of mineralized dentin.

INTRODUCTION: This study tested the null hypothesis that there is no difference between the use of 1.3% NaOCl/17% ethylenediaminetetraacetic acid (EDTA) and 5.25% NaOCl/17% EDTA irrigation regimens on the collagen degradation and flexural strength reduction in mineralized dentin. METHODS: Dentin powder and mineralized dentin sections were immersed in 1.3% or 5.25% NaOCl for 10-240 minutes and then rinsed with 17% EDTA as the final irrigant for 2 minutes. Untreated mineralized dentin powder/sections served as controls in the respective experiments. Dentin powders were examined by using Fourier transform infrared (FT-IR) spectroscopy to analyze their relative subsurface intact collagen content with the apatite/collagen ratio. Hydrated dentin sections were subjected to 3-point flexure under water for determining their flexural strengths. RESULTS: Collagen degradation was significantly increased and the flexural strength of mineralized dentin was significantly reduced after the use of 5.25% NaOCl as the initial irrigant for more than 1 hour (P < .05). Conversely, changes were insignificant when 1.3% NaOCl was used as the initial irrigant for up to 4 hours (Kruskal-Wallis analysis of variance, n = 10, P < .05). CONCLUSIONS: The null hypothesis was rejected. The deleterious effects attributed to the use of NaOCl on dentin are concentration-dependent and time-dependent and are not associated with the demineralization caused by the use of EDTA as the final active irrigant.

The self-etching potential of RealSeal versus RealSeal SE.

INTRODUCTION: This study evaluated the true self-etching potential of RealSeal and RealSeal SE to ensure that they are aggressive enough to etch through endodontic smear layers and demineralize the intact radicular dentin without adjunctive use of ethylenediaminetetraacetic acid (EDTA). METHODS: Mixed RealSeal and RealSeal SE were applied to (1) instrumented canal wall dentin that was irrigated with water as the final rinse to preserve smear layers, (2) fractured radicular dentin that was devoid of smear layers, and (3) instrumented canal wall dentin that was irrigated with EDTA as the final rinse to remove smear layers. Sealer-bonded specimens were processed for transmission electron microscopy. RESULTS: Unlike RealSeal, RealSeal SE could hardly demineralize the fractured dentin and was unable to etch beyond smear layers into the underlying intact dentin. EDTA dissolved smear layers completely and partially demineralized the intact dentin.. CONCLUSIONS: RealSeal SE might not be aggressive enough to achieve optimal dentin bonding to root canal walls in locations that are not reachable by calcium chelating irrigants. Conversely, RealSeal possesses mild etching ability on the fractured and smear layer-covered radicular dentin.

Bonding of self-adhesive (self-etching) root canal sealers to radicular dentin.

The latest generation of methacrylate resin-based sealers has eliminated the use of separate self-etching primers by incorporating acidic resin monomers in the sealers to render them self-adhesive to dentin. This study examined the adhesive strengths, interfacial ultrastructure, and tracer penetration of a nonetching (EndoREZ; Ultradent, South Jordan, UT) and two self-adhesive methacrylate resin-based sealers (MetaSEAL; Parkell, Farmington, NY, and RealSeal SE; SybronEndo, Orange, CA) when they were applied to radicular dentin following the manufacturers' recommended use of EDTA as the active final rinse. A modified push-out testing design was used to evaluate the dislodgement of core-free sealers. The mixed sealers were placed in dimensionally identical, artificially created canal spaces prepared in the coronal, middle, and apical thirds of radicular dentin. After setting, each sealer-filled cavity was subjected to compressive loading until failure. Additional specimens were prepared for transmission electron microscopy to examine the ultrastructure and nanoleakage within the sealer-radicular dentin interface. The two self-adhesive sealers MetaSEAL and RealSeal SE exhibited higher push-out strengths than the nonetching sealer EndoREZ when EDTA was used as the active final rinse. All three sealers showed a 1- to 1.5-microm thick zone of partially demineralized dentin, with the EDTA dentin demineralization effect masking the true self-etching potential of MetaSEAL and RealSeal SE. The true self-etching potential of self-adhesive sealers is a clinically important attribute that should be further investigated. Incomplete smear layer removal from the apical third of instrumented canal walls may jeopardize the performance of self-adhesive sealers should they fail to self-etch without the adjunctive use of calcium chelating irrigants.

The impact of dental devices on neurostimulators.

Spinal cord stimulation has been a therapeutic option for chronic pain for over 40 years. The neurostimulator (NS) is a device consisting of three primary components: an electrode array configured either as a paddle or wire; an implantable pulse generator (IPG) consisting of a minicomputer, a transceiver/antenna, an electrical generator, and a battery; and insulated wiring connecting the electrode to the IPG. The electrode array can be implanted into the epidural space overlying the dorsal spinal cord or along a peripheral nerve. The device generates pulsed electrical signals that stimulate the underlying dorsal columns of the spinal cord resulting in the perception of paresthesia by the patient. When overlapped with painful areas, the paresthesia can help decrease the patient's level of pain. The increased applications and indications for this technology enhance the likelihood that the NS patient will be seen in the dental practice. Therefore, the purpose of this study was to investigate whether electromagnetic interference of the NS occurred during the operation of the apex locator, the electric pulp tester (EPT), or the electrocautery unit. An NS was implanted into the epidural space of a human cadaver. The dental devices were used intraorally, and the implant's circuitry was tested after each trial. Two apex locators, two EPTs, and one electrocautery unit were tested. Seventy trials were used by each dental device on each tissue. Using the exact binomial method of determining confidence intervals, the probability of damage to the NS by any of the devices was negligible.

Contemporary methacrylate resin-based root canal sealers exhibit different degrees of ex vivo cytotoxicity when cured in their self-cured mode.

The cytotoxicity of four methacrylate resin-based sealers was investigated by the 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-tetrazolium bromide assay, which measures cell viability by assessing its succinate dehydrogenase activity. The sealers were polymerized in the self-cured mode to simulate the setting condition upon their extrusion into periradicular tissues. Disks were prepared from EndoREZ (Ultradent, South Jordan, UT), RealSeal (SybronEndo, Orange, CA), MetaSEAL (Parkell, Farmington, NY), and RealSeal SE (SybronEndo) together with positive and negative controls. After setting, they were placed in direct contact with rat osteosarcoma (ROS 17/2.8) cells and for 5 succeeding weeks after immersing in simulated body fluid (SBF). All sealers exhibited severe toxicity initially (week 0). EndoREZ and RealSeal remained severely toxic after five cycles of SBF immersion. Toxicity of the two self-etching resin-based sealers MetaSEAL and RealSeal SE decreased gradually over time. Transmission electron microscopy of cells exposed to RealSeal SE showed variable degrees of cell injury that reflect its toxicity status. Cells with intact mitochondria were identifiable after the sealer became noncytotoxic at week 5.

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.

Resistance of a 4-META-containing, methacrylate-based sealer to dislocation in root canals.

The dislocation resistance of root fillings created with MetaSEAL, a self-adhesive 4-META-containing methacrylate resin-based sealer, was evaluated. Forty-six incisors were cleaned and shaped using NaOCl and EDTA as irrigants. They were filled with gutta-percha/MetaSEAL or gutta-percha/AH Plus sealer using either a single-cone technique or warm vertical compaction (n = 10). The roots were sectioned at the coronal and middle thirds to obtain thin slices, which were subjected to compressive loading to displace the set sealer/filling toward the coronal side of the slice. The remaining six teeth were filled with gutta-percha/MetaSEAL and cryofractured for scanning electron microscopic examination. The push-out strength of AH Plus was significantly higher than MetaSEAL irrespective of filling techniques (p < 0.05). A minimal hybrid layer was seen in radicular dentin, and resin tags were inconsistently identified from canal walls in the MetaSEAL-filled canals. The lower dislocation resistance in MetaSEAL-filled canals challenges the use of a self-adhesive bonding mechanism to create continuous bonds inside root canals.

In vitro evaluation of a Ceramicrete-based root-end filling material.

Ceramicrete is an impervious inorganic binder widely used for encapsulating radioactive and hazardous wastes. This study evaluated the feasibility of using a radiopaque Ceramicrete-based material for root-end fillings. Apical seals of root-end preparations filled with Super EBA (Harry J. Bosworth Co, Skokie, IL), White ProRoot MTA (Dentsply Tulsa Dental Specialties, Tulsa, OK), or Ceramicrete-D were evaluated using a computerized fluid filtration approach after the fillings were immersed in phosphate-containing fluid (PCF). The Ceramicrete-D fillings exhibited significantly (P < 0.05) better seals than the other two commercially available, frequently advocated root-end-filling materials. Scanning electron microscopy and x-ray diffraction of Ceramicrete-D after setting revealed a relatively nonporous KMgPO4.6H2O matrix that binds other incompletely reacted and new reaction phases such as CaHPO4.2H2O. Polished dentin slabs filled with Ceramicrete-D and immersed in PCF for 72 hours revealed depositions of acicular-shaped, apatite-like crystallite clusters on the material surface as the pH of the PCF increased with immersion time. The experimental Ceramicrete-based material is potentially bioactive in the presence of PCF.

Calcium phosphate phase transformation produced by the interaction of the portland cement component of white mineral trioxide aggregate with a phosphate-containing fluid.

The bioactivity of mineral trioxide aggregate (MTA) has been attributed to its ability to produce hydroxyapatite in the presence of phosphate-containing fluids. It is known that stoichiometric hydroxyapatites do not exist in biological systems and do not contribute to the osteogenic potential of calcium phosphate-based biomaterials. Because Portland cement is the active ingredient in white MTA, we have characterized the calcium phosphate phases produced when set white Portland cement was immersed in phosphate-buffered saline using pH and turbidity measurements, scanning electron microscopy, energy dispersive X-ray analysis, transmission electron microscopy, electron diffraction, x-ray diffraction, and Fourier transform-infrared spectroscopy. An amorphous calcium phosphate phase was initially formed that transformed to an apatite phase, with the latter consisting of calcium-deficient, poorly crystalline, B-type carbonated apatite crystallites. Amorphous calcium phosphate is a key intermediate that precedes biological apatite formation in skeletal calcification. Thus, the clinical manifestations of bioactivity with the use of MTA may at least be partially attributed to the mineralization induction capacity of its Portland cement component.

Evaluation of the quality of the apical seal in Resilon/Epiphany and Gutta-Percha/AH Plus-filled root canals by using a fluid filtration approach.

This in vitro fluid filtration study compared the apical leakage of root canals that were obturated with Resilon/Epiphany (Resilon Research LLC, Madison, CT) or gutta-percha/AH Plus (GP/AH; DENTSPLY Tulsa, Tulsa, OK) sealer using warm vertical condensation. Fluid flow rate through the filled roots was measured 7 days after the obturations by using a fluid filtration device. Measurements were made before root resection and after 3 to 11 mm of resections. Before any removal of root length, there were no significant differences between the roots filled with Resilon/Epiphany or GP/AH. Analysis of individual root resection results revealed that differences in the fluid flow rate of the two systems occurred only when 9 or 10 mm of the roots were resected with more than half the root length resected when GP/AH Plus leaked more than Resilon/Epiphany. There were no significant differences in the fluid flow rate up to and including 8 mm of root resection. It is concluded Resilon/Epiphany sealed 17-mm root canals as well as gutta-percha and AH Plus sealer and that it does not create a monoblock root filling that does not leak.

Susceptibility of a polycaprolactone-based root canal-filling material to degradation. III. Turbidimetric evaluation of enzymatic hydrolysis.

The susceptibility of Resilon (Pentron Clinical Technologies, Wallingford, CT) to biotic biodegradation by two hydrolases, lipase PS and cholesterol esterase, was investigated with a turbidimetric approach by measuring the optical density reductions in aqueous emulsions containing dissolved, filtered, surfactant-stabilized polymeric components of Resilon. Polycaprolactone, the major polymeric component of Resilon, was also examined in a similar manner using equivalent or a four-fold increase in enzyme concentration. Optical density time plots were characterized by an initially linear steep reduction in optical density, from which the reaction rates were derived. For both enzymes, the rates of hydrolysis for Resilon were much faster than those of polycaprolactone at 1x or even 4x enzyme concentration. Field-emission scanning electron microscopy of air-dried Resilon and polycaprolactone emulsions revealed the presence of spherical polymer droplets that appeared deformed, pitted, or much reduced in dimensions after enzymatic hydrolysis.

Metrologic evaluation of the surface of white MTA after the use of two endodontic irrigants.

This study examined the effects of calcium-depleting endodontic irrigants, ethylenediamine tetra-acetic acid (EDTA) and BioPure MTAD, on the dissolution, surface characteristics, and ultrastructural characteristics of white mineral trioxide aggregate (MTA). The latter was mixed in a 0.35 water-cement ratio, condensed into cylindrical wells created in Plexiglas platforms, and allowed to harden completely before initial treatment with 1.3% NaOCl and final treatments with either 17% EDTA for 5 minutes, or BioPure MTAD for 1, 3, or 5 minutes. Analysis of the mean depths of material removed using three-dimensional profilometry revealed significantly more material removed by BioPure MTAD. Although these minor depth changes (<10 microm) are unlikely to cause clinical concern, the BioPure MTAD-treated MTA surfaces exhibited higher surface roughness and with more calcium extracted when compared with EDTA treatment. Decomposition of particle-binding hydration phases by acid corrosion raises potential concern on the strength and sealing properties of MTA-repaired perforations following final irrigation by BioPure MTAD.

Sealing properties of mineral trioxide aggregate orthograde apical plugs and root fillings in an in vitro apexification model.

One-visit mineral trioxide aggregate (MTA) apexification is gaining in popularity over the use of calcium hydroxide barriers. This study examined the sealing properties of two MTA apexification procedures using an in vitro apexification model. White MTA was introduced into instrumented single-rooted canals with standardized artificially created open apices to form 3- to 5-mm-thick orthograde apical plugs or complete orthograde fillings of the root segments. The remaining canal spaces in the former group were backfilled with thermoplasticized gutta-percha without a sealer. A Flodec fluid filtration device was used to record fluid flow across filled root canals at 48 hours and after 4 weeks of immersion in phosphate-buffered saline (PBS). Although MTA root fillings exhibited a better seal than MTA apical plugs at 48 hours, seals of these two groups were not significantly different after 4 weeks. Interaction of MTA with PBS may result in apatite deposition that improves the seal of MTA apical plugs with time.

Susceptibility of a polycaprolactone-based root canal filling material to degradation. Evidence of biodegradation from a simulated field test.

PURPOSE: To examine if Resilon, a polycaprolactone-based root filling material, was susceptible to microbial biodegradation by using a simulated field test that consisted of incubating the material in wet dental sludge under mesophilic and aerobic conditions. METHODS: Pressed disks prepared from Resilon, polycaprolactone (positive control) and gutta-percha (negative control) were incubated in wet dental sludge for up to 4 months and examined for topographical changes using scanning electron microscopy. RESULTS: Gutta-percha exhibited minimal changes in surface integrity, while polycaprolactone and Resilon exhibited severe surface pitting and erosion. In the latter, disappearance of the polymer matrix was accompanied by exposure of mineral and bioactive glass fillers. Bacteria and hyphae-like structures were present on the disk surfaces.

Reduction in antimicrobial substantivity of MTAD after initial sodium hypochlorite irrigation.

Potential intrinsic tetracycline staining of intraradicular dentin has been observed when BioPure MTAD was employed as the final irrigant after initial rinsing with NaOCl. This study examined the effect of NaOCl-MTAD interaction on the antimicrobial substantivity of MTAD in dentin. Dentin cores previously irrigated with either MTAD, or in conjunction with 1.3% NaOCl as an initial irrigant were placed on blood agar plates inoculated with Escherichia faecalis at 10(5) cfu/ml. Dentin cores irrigated with 1.3% NaOCl only, and autoclaved dentin disks were used as the respective positive and negative controls. After anaerobic incubation, the mean diameter of bacterial inhibition zones formed around the MTAD group was significantly larger than the NaOCl/MTAD group, which, in turn, was not significantly different from the NaOCl positive control. Oxidation of MTAD by NaOCl resulted in the partial loss of antimicrobial substantivity in a manner similar to the peroxidation of tetracycline by reactive oxygen species.

Self-etching adhesives increase collagenolytic activity in radicular dentin.

Endogenous matrix metalloproteinases (MMPs) release from crown dentin and their activation results in degradation of hybrid layers created by dentin adhesives. This study tested the hypothesis that instrumented intraradicular dentin possesses latent collagenolytic activity that is activated by mild self-etching adhesives. Root dentin shavings were produced from 50 cleaned and shaped, saline-irrigated root canals using Gates Glidden drills and rinsed with sodium azide to prevent bacterial growth. Dried dentin powder aliquots were treated with two clinically-relevant MMP inhibitors, 2% chlorhexidine for 10 minutes and 17% EDTA for 1 minute. Additional dentin powder was mixed with Clearfil Liner Bond 2V or Clearfil Tri-S Bond for 1 minute followed by extracting the adhesives with acetone. Dentin powder was also treated with 2% chlorhexidine for 10 minutes before or after adhesive application. Collagenolytic activities of the nine groups were assayed with a fluorometer in 96-well plates, by recording the changes in fluorescence before and after addition of fluorescein-labeled type I collagen. Epoxy resin-embedded powders were examined with TEM for the extent of demineralization. Instrumented, mineralized intraradicular dentin possessed low but detectable collagenolytic activity that was inhibited by chlorhexidine (p < 0.001) and EDTA (p < 0.001). Both adhesives partially demineralized the dentin powder and activated latent MMPs, with 14- to 15-fold increases in collagenolytic activities (p < 0.001) that were significantly (p < 0.001) but incompletely inactivated after 10 min application of chlorhexidine. Mild self-etching adhesives activate latent MMPs without denaturing these enzymes, and may adversely affect the longevity of bonded root canal fillings and posts.