An SEM study on the effect of 9.3-µm CO2 laser on dentinal tubules for hypersensitivity treatment.

OBJECTIVES: In-vitro studies were performed on dentin of extracted human molars to investigate the effectiveness of 9.3 µm CO2 laser irradiation to occlude dentinal tubules. The observed occlusion of dentinal tubules with the irradiation was compared with application of three reagents: 2% Sodium Fluoride gel, an aqueous solution of hydroxyapatite nanoparticles and an equal mix of the two. We show that 9.3 µm CO2 laser irradiation occludes dentinal tubules, and the use of laser irradiation produces better occlusion of the opened tubules compared to the use of topical reagents. METHODS: Nine extracted and cleaned human molars were cut to obtain dentin disks of thickness of 3-5 mm. Each disc was divided into four quarters, and each quarter served as two samples corresponding to irradiated and non-irradiated group counterparts. Five disks were used to study the effect of various laser irradiation energies on the dentinal tubules to find a good pulse fluence for occlusion of the dentinal tubules, and four disks were used for studying the effects of reagents and irradiation at the pulse fluences found in the first part of the study. The samples were irradiated with a beam diameter of 1 mm (1/e2) at 15 Hz pulse repetition rate, scanned automatically using a set of scanning mirrors. Samples were imaged using Scanning Electron Microscope (SEM) which were processed to determine tubule diameter. Safety of the irradiation treatment was investigated on 6 samples by measuring pulpal temperature rise. The effect of three topical reagents corresponding to 2% Sodium Fluoride gel (F), Hydroxyapatite nanoparticles (HA) and an equal mix of F and HA (HAF) on dentinal tubule occlusion was evaluated and compared with the laser irradiation. RESULTS: In all examined cases, laser irradiation at a fluence of 0.81 J/cm2 resulted in a temperature increase less than 3 °C which is safe, and no surface cracking was observed. There is a threshold pulse fluence of 0.27 J/cm2 above which, laser produced surface melting. At a pulse fluence of 0.81 J/cm2 a layer of recast of melted dentin was formed. Under this layer, peritubular dentin melting and occluding of the dentinal tubules was observed. Application of either F or HA or HAF did not produce visible occlusion effect on open tubules after washing and microbrushing with excess distilled water. CONCLUSIONS: 9.3 µm CO2 laser irradiation on extracted human molar dentin at pulse fluence of 0.81 J./cm2 resulted in tubule area reduction by 97% without rising pulpal temperatures to unsafe levels.

Combined Effects of Glutaraldehyde-based Desensitizer and Nd: YAG Laser on Dentinal Tubules Occlusion.

AIM: The aim of this study was to assess the impact of Nd:YAG laser, glutaraldehyde-based desensitizer (GD), or their combination on occluding dentinal tubules. MATERIALS AND METHODS: Fifty dentin samples were obtained from non-carious human third molars and randomly divided into five groups (n = 10): (1) Control group treated with 37% phosphoric acid, (2) GD group, (3) Nd:YAG laser group (1064 nm, 100 µs, 10 Hz, 300 µm fiber, 1 W power, 100 mJ energy, and 85 J/cm2 energy density), (4) GD followed by Nd:YAG laser group, and (5) Nd:YAG laser followed by GD group. Scanning electron microscopy (SEM) was used to capture five images from each sample for analysis of dentinal tubules using Image J software. SEM/EDX elemental analysis was performed to determine the main mineral contents. Data analyzed using one-way ANOVA and Tukey's post hoc test for statistical comparisons. RESULTS: Laser and combination groups showed a significant decrease in dentinal tubule counts compared with the control and GD groups (p < 0.0001). There were no significant differences in open dentinal tubule counts between the control and GD groups, as well as between the laser and combination groups. However, significant differences were observed in the total area, average size of the tubules, and percentage area between the control group and the treatment groups (GD, laser, GD + laser, laser + GD). No significant difference was found in the Ca/P ratio between the tested groups. CONCLUSION: The use of Nd:YAG laser alone or in combination with GD was more effective in occluding dentinal tubules compared to GD alone. CLINICAL SIGNIFICANCE: This study has shown that Nd:YAG laser alone and in combination with GD has superior dentinal tubule occlusion in vitro. Its clinical use in the treatment of dentinal hypersensitivity may overcome the drawback of conventional treatment approaches for dentin hypersensitivity needing repeated applications to achieve continuous relief from pain since acidic diet and toothbrushing result in the continuing elimination of precipitates and surface coatings. How to cite this article: Alzarooni AH, El-Damanhoury HM, Aravind SS, et al. Combined Effects of Glutaraldehyde-based Desensitizer and Nd: YAG Laser on Dentinal Tubules Occlusion. J Contemp Dent Pract 2024;25(1):52-57.

Antimicrobial effects of agitational irrigation on single- and multispecies biofilms in dentin canals.

This study aimed to compare the antibacterial effects of different agitation devices on single- and multispecies biofilms in dentin canals using confocal laser scanning microscopy (CLSM). Dentin blocks were prepared from human root dentin. Enterococcus faecalis and multiple species were introduced into the dentinal tubules via centrifugation and incubation. Two infected dentin samples were placed at 8 and 16 mm in a customized model. Samples were randomly divided into eight groups according to the agitation device used: syringe needle irrigation, EndoActivator, passive ultrasonic irrigation (PUI), and EDDY, at 2.5% or 6% NaOCl concentrations. The samples were stained and observed using CLSM. Statistical analysis was performed using an independent sample t test and analysis of variance. Linear models were used to assess the joint impact of the experimental groups on the proportion of biofilms killed. No significant differences were observed between the killing rates of the single- and multispecies biofilms. Both concentrations of NaOCl significantly increased the percentage of dead bacteria compared with the control. Biofilms in dentin tubules was more effectively killed when NaOCl was agitated; however, the difference between PUI and EDDY was not significant. Significantly more bacteria were killed in dentin blocks placed at 8 mm than at 16 mm (p < 0.05). In conclusion, EDDY was as effective as PUI when combined with NaOCl. However, the apical portion, which had a low antimicrobial efficiency, remains a concern. Mechanical instrumentation is incapable of completely eradicating bacteria, and additional research is required to improve the efficacy of root canal disinfection.

[Antibacterial effect of low-temperature plasma on Enterococcus faecalis in dentinal tubules in vitro].

OBJECTIVE: To construct a model of Enterococcus faecalis (E. faecalis) infection in dentinal tubules by gradient centrifugation and to evaluate the antibacterial effect of low-temperature plasma on E. faecalis in dentinal tubules. METHODS: Standard dentin blocks of 4 mm×4 mm×2 mm size were prepared from single root canal isolated teeth without caries, placed in the E. faecalis bacterial solution, centrifuged in gradient and incubated for 24 h to establish the model of dentinal tubule infection with E. faecalis. The twenty dentin blocks of were divided into five groups, low-temperature plasma jet treatment for 0, 5 and 10 min, calcium hydroxide paste sealing for 7 d and 2% chlorhexidine gel sealing for 7 d. Scanning electron microscopy and confocal laser scanning microscope were used to assess the infection in the dentinal tubules and the antibacterial effect of low-temperature plasma. RESULTS: The results of scanning electron microscopy and confocal laser scanning microscopy showed that after 24 h of incubation by gradient centrifugation, E. faecalis could fully enter the dentinal tubules to a depth of more than 600µm indicating that this method was time-saving and efficient and could successfully construct a model of E. faecalis infection in dentinal tubules. Low-temperature plasma could enter the dentinal tubules and play a role, the structure of E. faecalis was still intact after 5 min of low-temperature plasma treatment, with no obvious damage, and after 10 min of low-temperature plasma treatment, the surface morphology of E. faecalis was crumpled and deformed, the cell wall was seriously collapsed, and the normal physiological morphology was damaged indicating that the majority of E. faecalis was killed in the dentinal tubules. The antibacterial effect of low-temperature plasma treatment for 10 min exceeded that of the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d. These two chemicals had difficulty entering deep into the dentinal tubules, and therefore only had a few of antibacterial effect on the bacterial biofilm on the root canal wall, and there was also no significant damage to the E. faecalis bacterial structure. CONCLUSION: Gradient centrifugation could establish the model of E. faecalis dentin infection successfully. Low-temperature plasma treatment for 10 min could kill E. faecalis in dentinal tubules effectively, which is superior to the calcium hydroxide paste sealing for 7 d and the 2% chlorhexidine gel sealing for 7 d.

A SEM Evaluation of the Permeability of Different Desensitizing Methods on Occlusion of Dentinal Tubules: An In-vitro Study.

AIM: The purpose of the current research was to assess the permeability of three various desensitizing techniques on dentinal tubule occlusion using a scanning electron microscope (SEM). MATERIALS AND METHODS: For this research, 100 human sound premolar teeth that were extracted for orthodontic purposes were gathered. With the aid of a water-cooled diamond saw, the teeth were divided in a mesiodistal (vertically) orientation. A sectioned sample (5 mm long by 5 mm wide by 3.5 mm deep), including the cervical region, was taken from each buccal side. To fully open the dentinal tubules, these samples were then kept in 17% ethylenediaminetetraacetic acid (EDTA) for 40 minutes. The samples were divided into four groups (n = 25), each receiving the following dentin surface treatments: Group I: Control, Group II: Samples received NaF 5% varnish treatment, Group III: Samples received diode laser treatment, and Group IV: Samples received CPP-ACP treatment. An SEM was used to inspect each specimen at a magnification of about × 3000 and photomicrographs was assessed. RESULTS: The maximum occlusion of dentinal tubules was found in samples were treated with Diode laser (2.96 ± 0.14) followed by samples treated with NaF 5% varnish (3.38 ± 0.10), samples were treated with CPP-ACP (3.42 ± 0.06) and control group (4.26 ± 0.19). There was a statistically significant difference found between the groups. CONCLUSION: In conclusion, all three desensitizing methods used in the present study were successful in the occlusion of dentin tubules. But the application of the Diode laser was effective in the reduction of dentin permeability compared to NaF 5% varnish and CPP-ACP. CLINICAL SIGNIFICANCE: Dentin hypersensitivity (DH) is characterized by a brief period of intense discomfort. One approach to managing DH is to obstruct dentin tubules in order to decrease dentin permeability. There are many substances that can reduce hypersensitivity, but the finest commercially available substance for treating the condition by occluding the tubes should be acknowledged.

Effect of scaling on the invasion of oral microorganisms into dentinal tubules including the response of pulpal cells-an in vitro study.

OBJECTIVES: To investigate how scaling affects the penetration of microorganisms into dentinal tubules, how pulpal cells seeded into the pulp cavity respond to bacterial challenge, and how penetration and inflammatory response may depend on the bacterial composition. MATERIALS AND METHODS: Root canals of 102 extracted human teeth underwent shaping and cleaning. Half of the teeth were subjected to scaling and root planing, the other half remained untreated. Teeth were exposed to either Streptococcus gordonii and Actinomyces oris or S. gordonii and Porphyromonas gingivalis for 10 weeks. Bacterial invasion was assessed in a depth of 1 mm to the root surface. Human pulpal cells were seeded into the cavities to assess the expression of interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1), and matrix metalloproteinase-3 (MMP-3) by real-time polymerase chain reaction and immunoassay. RESULTS: The percentage of teeth with bacteria detected in dentine was higher when teeth received scaling than when they were untreated: 66.6% versus 44.4% when exposed to A. oris/S. gordonii, and 50% versus 25% when exposed to P. gingivalis/S. gordonii (p = 0.043). Scaling had no impact on IL-8 and MMP-3 expression in pulpal cells. P. gingivalis/S. gordonii caused higher levels of IL-8, MCP-1, and MMP-3 than A. oris/S. gordonii (p = 0.003, p = 0.011, p = 0.037). CONCLUSION: Scaling supports the penetration of bacteria into the dentine of extracted human teeth. P. gingivalis may affect the immune response in pulpal cells. CLINICAL RELEVANCE: Root surface debridement with hand instruments may facilitate bacterial penetration. Other kinds of mechanical instrumentation in this experimental setting should be investigated.

Novel Management of Hypersensitive Dentin Using Propolis-based Herbal Desensitizing Agents: An In Vitro Scanning Electron Microscopic Study.

AIM: The objective of this in vitro study was to assess the efficacy of novel propolis-based varnish against the two conventional varnishes on quantitative and qualitative assessments of occlusion of dentin tubules and resistance to erosive and abrasive wears employing scanning electron microscope (SEM). METHODS: Thirty human premolars free from caries extracted due to orthodontic reasons were included in the study. Experimental group was done based on treatment received and divided into three groups. Group A: ClinProXT Varnish (n = 10), Group B: MI Varnish (n = 10), and Group C: Propolis Varnish (n = 10) were applied. Teeth were cleaned and decoronation of crown was done with dentin disks. Dentin specimens of dimension 4 × 4 × 2 mm were prepared and subjected to finishing and polishing. The sample specimens were submersed in EDTA solution for a period of five minutes to open up the dentinal tubules. This was followed by treatment with varnishes and subjection to acidic-abrasive challenge. The specimens were analyzed with an image analyzer connected to SEM for the verification of the number of opened dentin tubules. The parameter assessed in SEM includes size, topography and surface characteristics of dentinal tubule were assessed. The obliteration potential of dentinal tubules was assessed with SEM images. Additionally, the dentin surface loss and resistance to acidic and abrasive wear were also evaluated with SEM. Data were analyzed with two-way analysis of variance (ANOVA) with post hoc Tukey's test. RESULTS: MI Varnish caused higher obliteration of dentin tubules followed by ClinproXT Varnish. Propolis Varnish showed the least obliteration of dentinal tubules among tested experimental groups. After acidic-abrasive challenge, Propolis Varnish was found to be more efficient with less material loss among the experimental groups tested. There was an insignificant difference among the MI Varnish and ClinProXT Varnish groups. CONCLUSION: Simulation of hypersensitive lesions mimicking the clinical scenario was a challenging task in this in vitro study. All varnishes tested in the study had good efficacy in the management of dentin hypersensitivity (DH). Propolis-based varnish had good resistance to material loss after subjection to acidic-abrasive challenge among the tested materials. The casein phosphopeptide (CPP)-amorphous calcium phosphate (ACP)-based MI Varnish had good efficacy to obliterate the dentinal tubules among the tested materials. It was prudent to select the varnishes with good long-term efficacy to survive in the clinical scenario which still remains a challenging task for the clinicians. CLINICAL SIGNIFICANCE: The stability of the varnish plays a vital role in maintenance of its long-term efficacy. The chemical nature along with the ability of the material to interact with the substrate plays a major role in management of DH.

Avian Eggshell Slurry as a Dentin Desensitizing Agent:An In Vitro Assessment Using Two Techniques.

AIM: This study aims to evaluate the efficacy of avian eggshell slurry as a desensitizing agent compared to casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) containing Tooth Mousse. MATERIALS AND METHODS: Eggshell powder was prepared and characterized using a scanning electron microscope (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDX). Forty dentin disks were immersed in 6% citric acid for 2 min to simulate hypersensitive dentin (baseline). Disks were then divided into two groups (n = 20) according to treatment received: eggshell slurry and commercially available Tooth Mousse desensitizing agent. Each group was further divided into two subgroups (n = 10). In subgroup 1, dentin discs were immersed in artificial saliva for four weeks whereas, in subgroup 2, discs underwent acid challenge. Environmental scanning electron microscope (ESEM) analysis and hydraulic conductance test were utilized to calculate dentinal tubules occlusion and reduction of dentin permeability percentages, respectively. Statistical analysis was performed using SPSS 22.0 at a significance level p ≤ 0.05. RESULTS: After application of desensitizing agents, the eggshell slurry group showed a significantly higher percentage of dentinal tubules occlusion compared to the Tooth Mousse group, whereas no significant difference existed in the percentage reduction of dentin permeability between both groups. After both immersion protocols, the eggshell slurry significantly occluded more dentinal tubules compared to Tooth Mousse. On the contrary, results of percentage reduction of dentin permeability revealed no significant difference between both subgroups after acid challenge. After artificial saliva immersion, Tooth Mousse showed a significantly higher percentage reduction of dentin permeability compared to the eggshell slurry. CONCLUSION: Avian eggshell slurry can effectively occlude open dentinal tubules compared to commercially available Tooth Mousse desensitizing agent. CLINICAL SIGNIFICANCE: Avian eggshell slurry can be considered a promising material for the treatment of dentin hypersensitivity. How to cite this article: El Tahlawy AA, Saba DA, Bakir NG. Avian Eggshell Slurry as a Dentin Desensitizing Agent: An In Vitro Assessment Using Two Techniques. J Contemp Dent Pract 2021;22(5):532-537.

Computer simulations of X-ray phase-contrast images and microtomographic observation of tubules in dentin.

An investigation of the problems of X-ray imaging of dentinal tubules is presented. Two main points are addressed. In the first part of this paper, the problem of computer simulating tubule images recorded in a coherent synchrotron radiation (SR) beam has been discussed. A phantom material which involved a two-dimensional lattice of the tubules with parameters similar to those of dentin was considered. By a comparative examination of two approximations, it was found that the method of phase-contrast imaging is valid if the number of tubules along the beam is less than 100. Calculated images from a lattice of 50 × 50 tubules are periodic in free space but depend strongly on the distance between the specimen and the detector. In the second part, SR microtomographic experiments with millimetre-sized dentin samples in a partially coherent beam have been described. Tomograms were reconstructed from experimental projections using a technique for incoherent radiation. The main result of this part is the three-dimensional rendering of the directions of the tubules in a volume of the samples. Generation of the directions is possible because a tomogram shows the positions of the tubules. However, a detailed tubule cross-section structure cannot be restored.

Toothpaste factors related to dentine tubule occlusion and dentine protection against erosion and abrasion.

OBJECTIVES: To investigate the effect of toothpastes on dentine surface loss and tubule occlusion, and the association of toothpaste-related factors to each of the outcomes. MATERIALS AND METHODS: One hundred and sixty human dentine specimens were randomly distributed into 10 groups, according to different toothpastes. The specimens were submitted to artificial saliva (60 min), citric acid (3 min), and brushing abrasion (25 s; totalizing 2 min in toothpaste slurries). This was repeated five times and two outcome variables were analyzed: dentine surface loss (dSL; µm) and tubule occlusion by measurement of the total area of open tubules (Area-OT; µm2). Data were analyzed with Kruskal-Wallis and Mann-Whitney tests (α = 0.05); bivariate and multivariate regressions were used to model the association of the chemical (pH, concentration of F-, Ca2+, and PO43- and presence of Sn2+) and physical (% weight of solid particles, particle size, and wettability) factors of the toothpastes to both outcome variables. RESULTS: Toothpastes caused different degrees of dSL and did not differ in Area-OT. All chemical and physical factors, except the presence of Sn2+, were associated with dSL (p < 0.001). Area-OT was associated only with the presence of Sn2+ (p = 0.033). CONCLUSION: Greater dSL was associated with lower pH, lower concentration of F-, higher concentration of Ca2+ and PO43-, greater % weight of solid particles, smaller particle size, and lesser wettability, whereas tubule occlusion was associated with the presence of Sn2+. CLINICAL RELEVANCE: Depending on their chemical and physical composition, toothpastes will cause different degrees of dentine tubule occlusion and dentine surface loss. This could, in turn, modulate dentine hypersensitivity.

Comparative evaluation of two chelating agents in collagen fiber network modification over dentinal tubules: An in vitro analysis.

OBJECTIVE: To compare effectiveness of ethylenediaminetetraacetic acid (EDTA) and citric acid in removing collagen fiber network covering dentinal tubules of human teeth. MATERIALS AND METHODS: Eighteen dentin discs were divided in three groups; Gp 1: discs received no treatment (control), Gp 2: discs etched with 17% EDTA (pH = 7.1), and Gp 3: discs etched with 6 wt% citric acid (pH = 4.0). Scanning electron microscopy (SEM) was performed to assess collagen fiber removal and X-ray diffraction (XRD) was implemented to analyse crystal peaks of discs. RESULTS: The SEM analysis demonstrated more collagen removal with EDTA treatment compared to citric acid treated specimens. Grade 6 (81% to 100% fiber removal) was mostly achieved for Gp 2 samples whereas grade 2 (1% to 20% fiber removal) was mostly achieved for Gp 3 samples and inter-group comparisons between these groups were statistically significant (p < 0.05). X-ray diffractogram of control and experimental samples demonstrated absence of calcite phase in experimental groups. The change in peak shapes and intensities were observed and citric acid treated samples revealed more intense peaks than EDTA group. CONCLUSION: Our study found 17% EDTA to be more effective in removing collagen fibers when matched with 6% citric acid.

[Morphology of root dentine in teeth extracted because of chronic apical periodontitis in various irrigation protocols]. / Morfologicheskie kharakteristiki dentina kornei zubov, udalennykh u patsientov s khronicheskimi formami periodontita, v zavisimosti ot sposoba medikamentoznoi obrabotki kornevogo kanala.

The purpose of the study was to evaluate the degree of purification of the deltoid root canal branching during endodontic treatment of teeth. Morphological assessment of dentine of 14 extracted incisors and premolars in patients aged from 24 up to 56 years with the diagnosis chronic apical periodontitis was conducted in two experimental groups: main (1 group, 7 teeth) and control (2 group, 7 teeth). In the 2 group conventional root canal treatment was carried out with processing of 3% solution of sodium hypochlorite and final sealing with gutta-percha pins by lateral condensation. In the 1 group root canals were additionally irrigated by 3% sodium hypochlorite solution by means of the RinsEndo device. Based on the study results we recommend using the hydrodynamic method of irrigation of root canals with 3% sodium hypochlorite solution in endodontic treatment of patients with chronic apical periodontitis.

Effects of two disinfection/sterilization methods for dentin specimens on dentin permeability.

OBJECTIVES: To investigate the effects of two disinfection/sterilization methods on the permeability of dentin specimens. MATERIALS AND METHODS: Forty intact human third molars were freshly extracted and cut, close to the pulp chamber, into dentin disks with a 500-µm thickness. The disks were randomized (n = 20 each) into a 70% ethanol group (acid-etched dentin disks soaked in 70% ethanol for 15 min) and a steam autoclaving group (acid-etched dentin disks autoclaved for 25 min). The permeability (Lp) of each dentin disk was measured before and after either treatment using a hydraulic device, and intra- and inter-group differences in values before and after treatment were analyzed using t tests. Field emission scanning electron microscopy (FE-SEM) micrographs of the dentin surface were acquired and examined. FE-SEM samples were prepared using the critical point drying (CPD) method. RESULTS: Immersion in 70% ethanol increased the Lp values of dentin specimens by 17%, which was not statistically significant. Steam autoclaving significantly reduced dentin permeability by 66% because the dentin collagen mesh became compact and collapsed, as detected by FE-SEM. CONCLUSIONS: The disinfection of acid-etched dentin disks using 70% ethanol for 15 min does not significantly affect dentin permeability, whereas sterilization of acid-etched dentin disks via autoclaving significantly reduces dentin permeability. CLINICAL RELEVANCE: Considering the influences of dentin permeability by disinfection/sterilization methods, the disinfection of the acid-etched dentin disks using 70% ethanol for 15 min could be used for the study related to dentin permeability, while the sterilization of autoclaving could not.

Is it really penetration? Part 2. Locomotion of Enterococcus faecalis cells within dentinal tubules of bovine teeth.

OBJECTIVE: The aim of the present vitro study was to examine the question whether devitalized Enterococcus faecalis (E. faecalis) cells can migrate into dentinal tubules and if that process takes place in a time-dependent manner. DESIGN: Sixty bovine root canals were incubated with devitalized and vital streptomycin-resistant E. faecalis strains after root canal enlargement (size 80, taper .02) with 3% NaOCl solution. Incubation times 7, 14, 21, 28, 35, and 42 days. Samples were processed for analysis by scanning electron microscopy (SEM) and DAPI (4',6-diamidino-2-phenylindole) staining. The penetration depth was calculated with the measurement tool of the Axio Vision program (Zeiss, Jena, Germany). Statistical analysis was performed by Kruskal-Wallis (α = 0.05) and Mann-Whitney U test (p < 0.05). RESULTS: Devitalized E. faecalis strains were able to migrate into dentinal tubules. The total number and penetration depth of devitalized E. faecalis cells was lower compared to the vital suspension of E. faecalis. It was noted, that bacterial penetration was not common to all of the dentinal tubules in the vital E. faecalis control and especially in the devitalized control. The migration took place in a time-dependent migration characteristic. CONCLUSIONS: Devitalized E. faecalis cells are still able to migrate into the dentinal tubules due to possible electrokinetic and osmotic processes. Thereby, increased exposure times lead to a time-dependent penetration characteristic. CLINICAL RELEVANCE: Since devitalized bacteria can migrate as well into dentinal tubules, the presence of bacteria within dentinal tubules cannot be interpreted as a failure of tested preparation regimens.

The dentin permeability of anti-inflammatory and antibacterial drugs: In vitro study.

BACKGROUND/PURPOSE: Stimuli from the oral cavity may penetrate through exposed dentinal tubules and evoke inflammatory pulp response. Anti-bacterial and anti-inflammatory drugs applied to exposed dentin may infiltrate through the dentinal tubules and cause pulp recovery. This study investigated the dentin permeability of anti-bacterial and anti-inflammation drugs via an in-vitro transwell dentin disc tube model. METHODS: Twenty-seven dentin discs prepared from extracted human molars were collected. Nine kinds of drugs were investigated with three dentin discs in each group. These nine drugs included two anti-bacterial drugs (ampicillin sodium and clindamycin phosphate), two corticosteroids (betamethasone sodium phosphate and hydrocortisone sodium succinate), three non-steroidal anti-inflammatory drugs (NSAIDs, piroxicam, lysine acetylsalicylate, and diclofenac sodium), and two natural extracts with anti-inflammatory effect (Ginsenoside Rg1 and Hinokitol). The drugs were introduced to the transwell dentin disc tube model and the 4-hour cumulative release of the drug was detected and recorded by UV-visible spectroscopy. RESULTS: We found that ampicilin sodium had better dentin permeability than clindamycin phosphate. Betamethasone sodium phosphate revealed better dentin permeability than hydrocortisone sodium succinate. Lysine acetylsalicylate showed the best dentin permeability among the three NSAIDs. Ginsenoside Rg1 had the best dentin permeability among the nine drugs tested. However, Hinokitiol could not penetrate the dentin disc after 4 h. CONCLUSION: Regarding the dentin permeability, Ginsenoside Rg1 is the best among the seven anti-inflammatory drugs tested and ampicilin sodium is the better one between the two anti-bacterial drugs tested. Therefore, these two drugs may have high potential for treating exposed dentinal tubule diseases.

Effect of Dentin Bonding Agent on Intrapulpal Temperature during Fabrication of Provisional Restorations by a Direct Method: An In Vitro Study.

AIM: To determine the effect of three different provisional restorative materials (PRMs) and application of dentin bonding agent (DBA) on intrapulpal temperature rise during fabrication of provisional crowns using a direct method. MATERIALS AND METHODS: Three PRMs-polymethyl methacrylate resin (PMMA), bis-acrylic resin, and dimethyl methacrylate resin (DMMA)-were used in the study. Dentin bonding agent (Single Bond 2, 3M ESPE, Germany) was applied in a single layer or double layer as an insulating material. A total of 45 molars were prepared for complete coverage restoration with chamfer finish line and 1.5 mm axial and 2 mm occlusal reduction. After application of a single or double layer of DBA, provisional materials were polymerized according to the manufacturer's instructions. The temperature increase inside the pulp chamber was measured with a T-type thermocouple wire. The readings were averaged for each group to determine the mean value of temperature rise. RESULTS: Statistical analysis was performed with analysis of variance (ANOVA) followed by post hoc Tukey test (a = 0.05). The temperature rise varied according to the provisional restoration material used (PMMA > DMMA > bis-acrylic resin) (p < 0.001) and the single or double layer of DBA (p < 0.001). Polymethyl methacrylate resin produced a higher intrapulpal temperature. CONCLUSION: Application of a double layer of DBA resulted in a significantly lower intrapulpal temperature.

The evaluation of E. faecalis colonies dissolution ability of sodium hypochlorite in microenvironment by a novel device.

Enterococcus faecalis(E. faecalis) is a common microorganism could be isolated from the infected canals, especially in the case of refractory apical periodontitis. Due to its ability to invade the dentinal tubules and highly resistant to antimicrobial strategies, the thorough debridement of E.faecalis is hard to achieve. And that may be one of the reasons to cause reinfection and therapeutic failure. According to the anatomy of dentinal tubules published before and the results of our team previous work, we designed six types of microtubes with different sizes. By using the method of centrifugation and incubation, a standard infected model mimicking dentinal tubules was established. Sodium hypochlorite (NaClO) is the most popular irrigant applied in root canal treatment. We used three different concentrations with four distinct irrigation duration to observe the antibacterial process of E. faecalis colonies within microtubes dynamically. We concluded that the role of NaClO in the microtubes is concentration dependent and duration dependent. And the interpretation of the results has a certain reference value for clinicians.

Dental properties, ultrastructure, and pulp cells associated with a novel DSPP mutation.

OBJECTIVE: To investigate physical characteristics and behaviours of dental pulp cells of teeth isolated from a dentinogenesis imperfecta (DGI) patient with a novel dentin sialophosphoprotein (DSPP) mutation. SUBJECTS AND METHODS: Whole exome and Sanger sequencing were employed to identify mutations. Physical characteristics of the teeth were examined. Pulp cells' behaviours including cell proliferation, colony-forming unit, osteogenic differentiation, pluripotent markers, and mesenchymal stem cell markers were investigated. RESULTS: The proband had opalescent brown primary teeth with extensive loss of enamel. Mutation analysis revealed a novel heterozygous 4-bp deletion, c.1915_1918delAAGT (p.K639QfsX674), in exon 5 of the DSPP associated with DGI. Analysis of the extracted primary incisor demonstrated a decrease in brightness but an increase in yellow and red chroma. The dentin showed reduced mineral density. The dentinal tubules were present in the predentin, but progressively collapsed in the dentin. The pulp cells exhibited markedly reduced CD105 expression, decreased cell proliferation, and smaller colony-forming units. CONCLUSIONS: We identified a novel mutation in the DSPP gene which disturbed dentin characteristics and pulp cells' behaviours. Our study expands the mutation spectrum and understanding of pathologic dentin phenotypes related to the frameshift deletion in the dentin phosphoprotein (DPP) region of the DSPP gene.

Evaluation of the effects of the solution used for electrochemical dissolution of nickel-titanium endodontic files on dentine structure, microhardness and cell viability.

AIM: To evaluate the effects of the [NaF 12 g L-1  + NaCl 1 g L-1 ] solution used in the electrochemical dissolution process of fractured endodontic files, as well as its NiTi-containing product, on dentine hardness, topography and human fibroblast viability. METHODOLOGY: Sixty single-rooted human teeth were evaluated for dentine microhardness using the Vickers hardness test and the area and number of dentinal tubules by scanning electron microscopy. The samples were divided according to the dentine surface treatment: distilled water; 17% EDTA; [NaF 12 g L-1  + NaCl 1 g L-1 ]; and 17% EDTA + [NaF 12 g L-1  + NaCl 1 g L-1 ]. Thirty-six single-rooted human teeth were divided according to the irrigation protocol: Dulbecco's Modified Eagle's Medium + 10% foetal bovine serum; 5.25% NaOCl; [NaF 12 g L-1  + NaCl 1 g L-1 ]; and [NaF 12 g L-1  + NaCl 1 g L-1  + NiTi]. The extracts in contact with the apical foramen were used in the MTT assay to evaluate human fibroblast viability, with dilutions of 100%, 50%, 25% and 12.5%. Statistical tests used were paired t-tests, one-way anova, Tukey's test, Kruskal-Wallis test and Dunn's post-test. RESULTS: The [NaF 12 g L-1  + NaCl 1 g L-1 ] solution did not modify dentine microhardness or the average dentinal tubule area. However, EDTA induced changes in dentine structure and microhardness (P < 0.05). The [NaF 12 g L-1  + NaCl 1 g L-1 ] solution, and its NiTi-containing product had lower cytotoxicity than NaOCl at dilutions of 25% and 50% (P < 0.01). CONCLUSIONS: The [NaF 12 g L-1  + NaCl 1 g L-1 ] solution did not alter dentine microhardness or damage the dentine structure. It also demonstrated lower cytotoxicity than NaOCl.

Sodium hypochlorite penetration into dentinal tubules after manual dynamic agitation and ultrasonic activation: a histochemical evaluation.

The aim of this study was to compare the effects of Manual Dynamic Agitation and Passive Ultrasonic Irrigation on sodium hypochlorite (NaOCl) penetration into dentinal tubules using its bleaching ability. Thirty-four single-rooted teeth with round-shaped root canals were distributed in two homogeneous groups and one control group, characterized by different NaOCl activation systems: Manual Dynamic Agitation and Passive Ultrasonic Irrigation. After instrumentation, all root canals were stained with 10% copper sulphate solution followed by 1% rubeanic acid alcohol solution under vacuum. Final irrigation was performed with 5 mL of 5.25% NaOCl solution for 1 min and activated with Manual Dynamic Agitation or Passive Ultrasonic Irrigation for another 1 min depending on the treatment group. The teeth were transversely sectioned at the middle portion of the apical, middle, and coronal thirds and observed under light microscope. NaOCl solution penetration was evaluated by measuring the percentage of bleached circumference of the root canal relative to the stained circumference, bleached areas, mean, and maximum penetration depth. No differences in the evaluated parameters were observed between groups (p > 0.05). Within groups, an increase of values was recorded from apical to coronal direction as for percentage of staining, percentage of bleaching and bleached area. NaOCl penetration into dentinal tubules did not significantly vary among the three levels. No significant differences in penetration of sodium hypochlorite into dentinal tubules when activated by means of Manual Dynamic Agitation or Passive Ultrasonic Irrigation were observed in the apical, middle, and coronal thirds of teeth with single straight round root canals.