• Restorative Dentistry and Endodontics
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• v.39 no.4
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• pp.258-264
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• 2014

Objectives: This in vitro study aimed to investigate the ability of Candida albicans (C. albicans) and Enterococcus faecalis (E. faecalis) to penetrate dentinal tubules of instrumented and retreated root canal surface of split human teeth. Materials and Methods: Sixty intact extracted human single-rooted teeth were divided into 4 groups, negative control, positive control without canal instrumentation, instrumented, and retreated. Root canals in the instrumented group were enlarged with endodontic instruments, while root canals in the retreated group were enlarged, filled, and then removed the canal filling materials. The teeth were split longitudinally after canal preparation in 3 groups except the negative control group. The teeth were inoculated with both microorganisms separately and in combination. Teeth specimens were examined by scanning electron microscopy (SEM), and the depth of penetration into the dentinal tubules was assessed using the SMILE view software (JEOL Ltd). Results: Penetration of C. albicans and E. faecalis into the dentinal tubules was observed in all 3 groups, although penetration was partially restricted by dentin debris of tubules in the instrumented group and remnants of canal filling materials in the retreated group. In all 3 groups, E. faecalis penetrated deeper into the dentinal tubules by way of cell division than C. albicans which built colonies and penetrated by means of hyphae. Conclusions: Microorganisms can easily penetrate dentinal tubules of root canals with different appearance based on the microorganism size and status of dentinal tubules.

• Restorative Dentistry and Endodontics
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• v.21 no.1
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• pp.267-279
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• 1996

Microorganisms are implicated the endodontic treatment failures. Persistent endodontic infection may be the result of retention of microorganisms in the dentin of the root canal walls. Dentinal tubules of the root canal walls have been shown to harbor microorganisms. The purpose of this study was to investigate the invasion of microorganism into the root dentin and dentinal tubules. The effects of irrigation solutions and smear layer on bacterial colonization of root canal were evaluated using a scanning electron microscopy. Canals of extracted human teeth with single and straight canals were stepback prepared using normal saline. Tooth samples were divided into four groups according to the irrigation solutions -5 % sodium hypochlorite and normal saline-and smear layer treatment. The smear layer was removed by 5% NaOCl and 20% EDTA for 10 min respectively. After sterilization, they were incubated with each strains of Streptococcus sanguis, Enterococcus faecalis, Staphylococcus aureus and Escherichia coli. Sodium hypochlorite solution reduced the adhesion of microorganisms effectively compared to normal saline. The smear layer inhibited colonization of E. faecalis, S. aureus and E. coli in the root canals due to their blocking of dentianl tubules. But S. sanguis invaded dentinal tubules in the root canals without smear layer. It was suggested that bacterial attachment might be different according to the strains. Sodium hypochlorite inhibited bacterial attachment in the dentinal tubules dramatically. The absence or presence of smear layer affected bacterial invasion of the dentinal tubules.

• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.508-526
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• 1995

This experiment was performed to study mechanisms of desensitization by chemical desensitizing agents in hypersensitive dentin and compare effects of these agents by measuring the activity of intradental nerves and observing their occluding aspects on dentinal tubules with SEM over time after application of chemical desensitizing agents to the exposed dentinal surfaces. Canines of adult cats weighing 2-3 kg were cross-sectioned at 1.5 mm from incisal apex, and the smear layer of the exposed dentinal susface was removed by 32 % $H_3PO_4$ for 15 sec. Chemical desensitizing agents such as 10% $SrCl_2$, 5% $KNO_3$ and 30% $K_2C_2O_4$, were applied to the exposed dentin surfaces for 2 minutes. Intradental nerve activity was measured immediately after application of the agents, at 15 minutes and at 30 minutes by stimulating with 4M NaCl. To compare occluding ability of desensitizing agents on dentinal tubules in vivo and in vitro, the structures of the exposed dentinal surfaces of nonvital and vital teeth were morphologically observed by SEM. The results obtained were as follows : 1. Intradental nerve activity was decreased immediately after the application of 10 % $SrCl_2$, 5% $KNO_3$ and 30% $K_2C_2O_4$. (p<0.01), among which 30% $K_2C_2O_4$. showed the highest desensitizing effect(p<0.01). 2. The immediately decreased intradental nerve activity after application of 10 % $SrCl_2$ and 5% $KNO_3$ was increased over time. 10% $SrCl_2$ and 5% $KNO_3$ showed no desensitizing effect respectively at 30 minutes and at 15 minutes after application. 3. The immediately decreased intradental nerve activity after application of 30 % $K_2C_2O_4$ was persistently continued during the period of observation (p<0.01). 4. Precipitates of $SrCl_2$ and $KNO_3$ were not noted on the exposed dentinal surfaces and within dentinal tubules by SEM examination. On the other hand, 30 % $K_2C_2O_4$ produced precipitates on the exposed dentinal surfaces and openings of dentinal tubules without any formed preciptates within dentinal tubules. 5. Ten percent $SrCl_2$, 5 % $KNO_3$ and 30 % $K_2C_2O_4$ showed no differences in their occluding aspects on dentinal tubules either in vivo or in vitro studies and either immediately following application or at 30 minutes. These results suggest that the desensitizing effect of $SrCl_2$ and $KNO_3$ is resulted from their reducing effect on the intradental nerve activity rather than from their precipitates' occluding the dentinal tubules. However, desensitizing effect of 30 % $K_2C_2O_4$, is probably resulted from its precipitates' occluding the openings of the dentinal tubules as well as from it's reducing effect on the intradental nerve actibity.

• Restorative Dentistry and Endodontics
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• v.18 no.2
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• pp.483-496
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• 1993

Dentin hypersensitivity medicaments such as Gluma, Scotchbond 2, All-Bond 2, which are resin adhesives, were used to compare the sealing effects of dentinal tubule under mechanical stress. Topical application of above medicaments on the dentin surfaces of extracted teeth followed by artificial tooth brushing for 6 weeks was performed for the comparison. The following conclusions on the degree of dentinal tubule exposure versus time by were reached by using polyvinyl siloxane impression material for taking the impression, epoxy resin for the duplication and SEM for observing the surface. 1. SEM was used to compare the accuracy of the duplicated surface, but no differences were found when teeth samples and the duplicated surfaces were observed. 2. After comparing the degree of dentinal tubules exposure with varnish applied contrast group, resin adhesive materials showed much less exposure as time went by. 3. The results indicated that AU-Bond 2 adhesive, under mechanical stress, showed lesser exposure of dentinal tubules comparing with Gluma and Scotchbond 2 adhesives After the results were put together, it was demonstrated that resin replica method is an useful way to evaluate the treatment effects of the dentinal tubule hypersensitivity medicaments. Also, it was noticed that under mechanical stress, All-Bond 2, classified as fourth generation, illustrated the best dentinal tubules sealing effects.

• Journal of Periodontal and Implant Science
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• v.23 no.3
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• pp.564-576
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• 1993

The purpose of this study was to evaluate the effects of Nd:YAG laser treatment on removal of smear layer and exposure of dentinal tubules. The experimental specimens were obtained from root planed surface of 30 human teeth which were extracted due to severe periodontal disease. The specimens were assigned randomly of three groups: root planed group, Tetracycline HCl(100mg/ml, 5min) group, and Nd:YAG laser ($Laser-35^{TM}$, U.S.A. 5 seconds )group. Nd:YAG laser group was divided into 4 subgroups according to 3.0W, 3.5W, 4.0W, 4.5W of power. The surface change of specimens were evaluated by scanning electron microscopy. The number of exposed dentinal tubules and percentage of area occupied by dentinal tubule orifices per unit area between each group was statistically analyzed by paired Student t-test. The results were as follows: 1. By root planing only, dentinal tubule was not exposed, but scale-like smear layer and parallel instrument tracks were resulted. 2. Tetracycline HCl treated surfaces exhibited the small number of partially exposed dentinal tubules with long orifices. The number($3.80{\pm}0.79$) of exposed dentinal tubules on Tetracycline HCl group was significantly less than that in laser groups above 3.5W of power(P<0.001), and the percentage ($0.68{\pm}0.19$) of area occufied by dentinal tubule orifices per unit area($192\;{\mu}m^2$) was significantly lower than that of any laser group(P<0.001). 3. The laser group irradiated with 4.5W of power showed both the most number($10.60{\pm}0.97$) of exposed dentinal tubules among the experimental groups, and the highest percentage($3.75{\pm}0.55\;%$) of area occupied by dentinal tubule orifices. 4. Energy Surge during laser pulsing and overlapping passes of the fiber resulted in melted and resolidified surface textures with lava-like appearance.

• Restorative Dentistry and Endodontics
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• v.6 no.1
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• pp.141-147
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• 1980

The sealing effect of a few cavity liners upon the dentinal tubule were studied in vitro. The materials employed in this study were Silcot (SP$\'{E}$CIALIT$\'{E}$S SEPTODONT, M.-T. GENDRAULT, Pharmacien), Hypo-Cal(Ellman Dental Mfg.Co.Inc.), Cavity Lining(De Trey), and Copaute(Harry J. Bosworth Co.). Freshly extracted human teeth were devided into 5 groups by age-under twenty, twenties, thirties, forties, and over fifty. Class V cavities were prepared routinely. The cavity walls of eight teeth of each group were lined by Silcot, Copalite, Cavity Lining, and Hypo-Cal. Remaining eight were not lined as a control. These specimens were immersed in dye solution (2 gm eosin Y to 800 cc distilled water) for 48 hours to allow maximum dye penetration into dentinal tubules. Each specimen was sectioned longitudinally including Class V cavity floor under water spray. Dye penetration into dentinal tubules were examined and following results were obtained. 1. Liners used on this study showed more or less dye penetration into dentinal tubules. But compared with the teeth without lining, the dye penetration of lined specimens were decreased. 2. Of these liners tested, Silcot was the most effective sealer upon the dentinal tubules. Copalite was the moderate sealer and Cavity Lining showed a tendency similar to Copalite. Hypo-Cal revealed the greatest dye penetration. 3. As the age was increased, the more the dye penetration into the dentinal tubules was decreased.

• Restorative Dentistry and Endodontics
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• v.28 no.5
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• pp.392-401
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• 2003

The purpose of this study was to evaluate the penetration pattern of dentin adhesives according to the orientation of dentinal tubules with confocal laser scanning microscopy. Specimens having perpendicular. parallel and oblique surface to dentinal tubules were fabricated. The primer of dentin adhesives (ALL $BOND^{\circledR}{\;}2,{\;}CLEARFIL^{TM}$ SE BOND and PQ1) was mixed with fluorescent material. rhodamine B isothio-cyanate (Aldrich Cherm. CO., Milw., USA), It was applied to the specimens according to the instructions of manufactures. The specimens were covered with composite resin (Estelite, shade A2) and then cut to a thickness of 500$\mu\textrm{m}$ with low speed saw (Isomet^{TM}, Buehler. USA). The adhesive pattern of dentin adhesives were observed by fluorescence image using confocal laser scanning microscopy. The results were as follows. 1. For the groups with tubules perpendicular to bonded surface. funnel shape of resin tag was observed in all specimen. However. resin tags were more prominent in phosphoric acid etching system (ALL $BOND^{\circledR}$ 2 and PQ1) than self etching system ($CLEARFIL^{TM}$ SE BOND). 2. For the groups with tubules parallel to bonded surface. rhodamine-labeled primer penetrated into peritubular dentin parallel to the orientation of dentinal tubules. But rhodamine-labeled primer of PQ1 diffused more radially into surrounding intertubular dentin than other dentin adhesive systems. 3. For the groups with tubules oblique to bonded surface. resin tags appeared irregular and discontinuous. But they penetrated deeper into dentinal tubules than other groups.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.561-578
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• 1999

The goal of periodontal therapy is the periodontal regeneration by the removal of microorganisms and their toxic products from the periodontally diseased root surface. To achieve periodontal regeneration, root conditioning as an adjunct to root planing has been done. There are low pH etchants such as citric acid, tetracycline-HCl, and EDTA solution which is a neutral chelating agent. The purpose of present study was to examine the effect of root conditioning by citric acid, tetracycline HCl, and EDTA. Total 35 root specimens(6${\times}$3${\times}$2mm) were prepared from the periodontally diseased teeth, scaled and root planed. The specimens were treated with normal saline for 1 minute, saturated citric acid(pH 1) for 3 minutes, 50mg/ml tetracycline-HCl(pH 2) for 5 minutes, 15% EDTA(pH 7) for 5 minutes using rubbing technique. The specimens were examined under scanning electron microscopy at 1000, and 3000 magnification. On the microphotographs taken at 1000 magnification, the numbers of opened and patent dentinal tubules per unit area(10,640${\mu}m^2$) were counted. And the diameters of opened dentinal tubules per unit are (10,640${\mu}m^2$) were measured. The differences of number and diameter among all groups were statistically analyzed by Kruskal Wallis Test. The results were as follows; 1. In the specimens applied with normal saline(control group), the root surface was finely cracked, and was covered by irregular smear layer. Neither exposed dentinal tubules nor any patent dentinal tubules could be seen. 2. In the specimens applied with saturated citric acid(experimental 1 group), the globular collagen fibers were exposed around the peritubular space, and many dentinal tubules were revealed. 3. In the specimens applied with tetracycline-HCl(experimental 2 group), the process-like collagen fibers were exposed around the peritubular space, and some dentinal tubules were revealed. 4. In the specimens applied with 15% EDTA(experimental 3 group), the root surface was covered by the collagenous fibrillar network, and many dentinal tubules were revealed. 5. The numbers of opened and patent dentinal tubules were significantly more in exp. 1 group and exp. 3 group than in exp. 2 group(P<0.05). But there was no significant difference between exp. 1 group and exp. 3 group. In control group, the number of opened and patent dentinal tubules could not be counted because any dentinal tubules couldn't be seen. 6 . The diameter of opened dentinal tubules was significantly smaller in exp. 1 group and exp. 3 group than in exp. 2 group(P<0.05). But there was no significant difference between exp. 1 group and exp. 3 group. In control group, the diameter of opened dentinal tubules could not be measured because any dentinal tubules couldn't be seen. The results demonstrate that root conditioning with citric acid, tetracycline- HCl, and EDTA is more effective in periodontal healing than only root planing, and 15% EDTA solution can replace low pH etching agents such as citric acid, tetracycline-HCl for root conditioning.

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.829-844
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• 1997

Dentin hypersensitivity must be one of the most frequent postoperative complaints in periodontal patients. Obliterating the open dentinal tubules or decreasing the diameter of their orifices would, therefore, be an objective of treatment for hypersensitive teeth. The purpose of this study was to evaluate the effect of a pulsed Nd:YAG laser irradiation on obliteration of dentinal tubules and to determine any difference according to irradiation methods. The 45 posterior teeth that had been extracted due to periodontal disease were initially treated with tetracycline HCI(100 mg/ml, 4 min.) to remove the smear layer after root planing. The root surfaces were then irradiated by a pulsed Nd:YAG laser(EL.EN.EN060, Italy) by different laser beam spot size and different exposure condition: ${\cdot}$ group 1: irradiated group by small spot(beam diameter=1mm, lW, 2 sec) ${\cdot}$ group 2: irradiated group by large spot(beam diameter=10mm, 1W, 200 sec) ${\cdot}$ group 3: irradiated group by gradual increase of watt (from 0.3W to 1.0W), beam diameter=4mm ${\cdot}$ group 4: irradiated group by fixed watt(1.0 W), beam diameter=4mm ${\cdot}$ control group: no irradiation but root planing and tetracycline HCI conditioning only. Additionally, the specimens were retreated with tetracycline HCI(100mg/ml, 4min.) to evaluate the stability of obliteration effect by Nd:YAG laser. Specimens were examined under the scanning electron microscope(JEOL, JSM-840A, Japan). Photomicrographs were taken at ${\times}4,000$ magnification and were analyzed statistically. The results were as follows: l. Scanning electron micrographs of root surface treated by tetracycline HCI alone(control group) showed widened, funnel-shaped dentinal tubules, while those of the root surface irradiated by various methods showed partially or completely obliterated dentinal tubules and various surface alterations, eg, flat, multiple pitted, melted and resolidified surface at the same energy density. 2. There was no significant difference in the obliteration effect of dentinal tubules between group 1 and group 2, and between group 3 and group 4(p>0.05). 3. The obliteration effect of dentinal tubules by a Nd:YAG laser irradiation was relatively stable to tetracycline HCI. The results demonstrate that a pulsed Nd:YAG laser irradiation within 1.0W, regardless of irradiation methods, can obliterate dentinal tubules effectively.

• Journal of Periodontal and Implant Science
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• v.43 no.6
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• pp.269-275
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• 2013

Purpose: In this study, the effect of calcium sodium phosphosilicate (NovaMin) desensitizing agent, which is a powder-based system, and hydroxyethyl methacrylate and glutaraldehyde (Gluma desensitizer), which is liquid-based system, on dentinal tubule occlusion was analyzed by scanning electron microscope. The effects of the above two along with one control group were compared to determine the more effective method of sealing the dentinal tubules after initial application. Methods: Twenty specimens were allocated to each of 3 groups: Control, Gluma desensitizer, and NovaMin. Two additional samples were also prepared and treated with Gluma and NovaMin; these samples were longitudinally fractured. The specimens were prepared from extracted sound human premolars and were stored in 10% formalin at room temperature. The teeth were cleaned of gross debris and then sectioned to provide one to two dentin specimens. The dentin specimens were etched with 6% citric acid for 2 minutes and rinsed in distilled water. Control discs were dried, and the test discs were treated with the desensitizing agents as per the manufacturer's instructions. The discs as well as longitudinal sections were later analyzed under the scanning electron microscope. The proportions of completely occluded, partially occluded, and open tubules within each group were calculated. The ratios of completely and partially occluded tubules to the total tubules for all the groups was determined, and the data was statistically analyzed using nonparametric tests and statistical significance was calculated. Results: NovaMin showed more completely occluded tubules ($0.545{\pm}0.051$) while Gluma desensitizer showed more partially occluded tubules ($0.532{\pm}0.075$). The differences among all the groups were statistically significant ($P{\leq}0.05$). Conclusion: Both materials were effective in occluding dentinal tubules but NovaMin appeared more promising in occluding tubules completely after initial application.