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Table of Contents   
ORIGINAL RESEARCH  
Year : 2011  |  Volume : 22  |  Issue : 1  |  Page : 22-27
Effect of endodontic irrigating solutions on the microhardness and roughness of root canal dentin: An in vitro study


Department of Conservative & Endodontics, K.L.E's. V. K. Institute of Dental Sciences, Belgaum, Karnataka, India

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Date of Submission01-Nov-2009
Date of Decision28-Apr-2010
Date of Acceptance10-Nov-2010
Date of Web Publication25-Apr-2011
 

   Abstract 

Context: To evaluate the effect of widely used endodontic irrigating solutions on root dentin microhardness and surface roughness.
Materials and Methods: One hundred twenty, non-carious extracted human permanent incisor teeth were selected. The crowns of the teeth were sectioned and the roots were separated longitudinally to get 240 specimens. These specimens were then divided into six groups according to the irrigating solutions used. The solutions used were 5% and 2.5% NaOCl solutions, 3% H2 O2 , 17% EDTA solution, 0.2% chlorhexidine gluconate, and distilled water. Then, the specimens were subjected to microhardness and roughness testing. The data were analyzed using ANOVA and Tukey's multiple comparison tests.
Results: The results of this study indicated that all irrigation solutions, except 0.2% chlorhexidine gluconate, decreased the microhardness of root dentin, and 3% H2 O2 and 0.2% chlorhexidine gluconate had no effect on surface roughness.
Conclusions: Within the limitation of this study, it is concluded that 0.2% chlorhexidine gluconate seems to be an appropriate irrigation solution, because of its harmless effect on the microhardness and surface roughness of root canal dentin.

Keywords: Irrigating solutions, microhardness, root dentin, surface roughness

How to cite this article:
Patil CR, Uppin V. Effect of endodontic irrigating solutions on the microhardness and roughness of root canal dentin: An in vitro study. Indian J Dent Res 2011;22:22-7

How to cite this URL:
Patil CR, Uppin V. Effect of endodontic irrigating solutions on the microhardness and roughness of root canal dentin: An in vitro study. Indian J Dent Res [serial online] 2011 [cited 2020 Jan 25];22:22-7. Available from: http://www.ijdr.in/text.asp?2011/22/1/22/79969
Chemomechanical preparation is one of the important factors for successful endodontic treatment. Chemomechanical debridement of root canal system is achieved by the use of instruments and effective irrigating solutions. The aim of instrumentation and irrigating is to prepare a clean, debris-free canal for subsequent obturation. [1]

An instrument thrust into the canal is likely to force noxious materials like necrotic pulp or shreds of mummified tissue with bacteria into it, through the apical foramen, with resulting periradicular pathology. During each instrumentation procedure, the canals are simultaneously washed out or irrigated with a solution capable of disinfecting them and dissolving organic matter. [2]

In addition to the debriding action, irrigation serves the purpose of facilitating instrumentation by lubricating canals and by floating out dentinal filings. [2]

Thus, the goals of irrigation are [2] :

  1. lavage of debris;
  2. tissue dissolution;
  3. antibacterial action; and
  4. lubrication.
Various irrigating solutions have been used, such as: 5% and 2.5% sodium hypochlorite, 3% hydrogen peroxide, 17% EDTA, and 0.2% chlorhexidine.

Among these, one of the most popular irrigating solutions is sodium hypochlorite. [2] An in vitro study has demonstrated that 2.6% NaOCl has excellent predentin and tissue solvent action. [3]

The 3% hydrogen peroxide alone also effectively "bubbles out" and mildly disinfects the canal by release of nascent oxygen. [2]

Investigators at Lomba Linda University found that 5.25% NaOCl was not effective against anaerobic bacteria as 0.2% chlorhexidine gluconate or 3% hydrogen peroxide. [2]

Mechanically prepared dentin surfaces are always covered with a so-called smear layer, a loosely bonded amorphous layer of organic and inorganic debris. In this respect, several endodontic irrigating solutions have been used to remove such smear layer with varying degrees of success. They have ranged from acids and chelates, to those intended to dissolve organic debris. [4]

Irrigation of root canal with 10 ml of 17% EDTA followed by 10 ml of 5% NaOCl has been recommended as an effective method to remove smear layer. [5]

The most common chelating agent used for irrigating includes 17% EDTA solution (pH of 5-9). The authors have shown that optimal time of EDTA is 15 min, after which time no chelating action can expected. [2]

It is important to test the effect of the irrigating solutions on all dentin tissues, as they may come in contact during irrigating procedures. These irrigating solutions cause alterations on dentin and enamel surfaces and affect their interactions with materials used for obturation and coronal restorations. [4]

Studies on modes of action and efficiency of various chemical irrigating solutions have shown their direct effect on both organic and inorganic components of root canal dentin. In turn, the mechanical, chemical, and physical properties of dentin structure changes.

It has been noted that microhardness and roughness are sensitive to composition and surface changes of tooth structures. [6]

A similar correlation can be made between microhardness and roughness of root dentin and irrigating solutions. [4] Thus, it is of interest to investigate to what extent the dentin of the root canal is affected by the use of various irrigating solutions.

Therefore, this study was designed to evaluate the effect of widely used irrigating solutions on the microhardness and roughness of root canal dentin.


   Materials and Methods Top


Sample selection

One hundred and twenty non-carious, non-hypoplastic, extracted human intact permanent maxillary and mandibular incisor teeth from patient with age groups of 35-45 years were selected. The teeth were stored at 37°C in buffered saline.

Specimen preparation

The crowns of the teeth were sectioned at cemento enamel junction (CEJ)using a high-speed diamond point under water-cooling. Then, the roots were separated longitudinally using a diamond disc under water-cooling. Thus, 240 specimens were obtained. These specimens were then examined under stereomicroscope to eliminate the teeth with cracks and other specimens were added to compensate for them.

The specimens were then ground-polished with water-cooled carborandum disc. Final polishing was carried out in felt cloth and buff by using 0.05 μm size aluminium oxide powder mixed with distilled water.

A plastic ring was then taken, and it was poured with a mixture of cold cure resin. And the specimens were embedded on the resin with polished surface facing outside. After curing of the resin, the ring was removed and repolishing of specimens was done to remove the excess material present on the tooth surface [Figure 1]a.
Figure 1: (a) Specimens (b) Specimens immersed in solutions (c) Microhardness tester (d) Microhardness testing of specimens (e) Surface roughness-testing machine (f) Surface roughness testing of specimens

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Grouping

A total of 240 specimens were then divided into six groups, with 40 specimens in each group according to the irrigating solutions used.

Exposed dentin surfaces were immersed in plastic jar containing irrigating solutions [Figure 1]b as follows:

Group 1: 5 ml, 5.0% NaOCl for 15 min

Group 2: 5 ml, 2.5% NaOCl for 15 min

Group 3: 5 ml, 3% Hydrogen peroxide for 15 min

Group 4: 5 ml, 17% EDTA solution for 15 min

Group 5: 5 ml, 0.2% Chlorhexidine gluconate for 15 min

Group 6: 5 ml, Distilled water for 15 min (control).

At the end of active treatment period (15 min), the samples were rinsed with distilled water and dried. Every group was then divided into two subgroups of 20 each.

Group 1a, 2a, 3a, 4a, 5a, 6a were used to determine the microhardness of root dentin. And Group 1b, 2b, 3b, 4b, 5b, and 6b were used to determine the surface roughness of root dentin.

Microhardness testing

The specimens were mounted on stage of Vickers microhardness tester [Figure 1]c. The midroot portion is halfway from the outer surfaces was focused for testing. Indentations were made with Vickers diamond indenter using 300 gm load with a dwell time of 20 s [Figure 1]d. These indentations were measured and converted into Vickers hardness number (VHN) values by the monitor.

Surface roughness testing

The specimens were placed on the flat table surface and the needle of roughness tester was on the mid root region of the tooth surface. The machine was then made to record the surface roughness values of root dentin [Figure 1]d-f. The values were displayed digitally on the screen of the roughness tester. These values were expressed as Ra (μm). The Ra parameter describes the overall roughness of the surface and can be defined as the arithmetical average value of all absolute distances of the roughness profile from the centre line within the measuring length.

Statistical analysis used

The teeth were analyzed statistically using One-way analysis of variance (ANOVA) and the comparison of means was conducted using Tukey's multiple comparison test. The testing was performed at the 95% level of confidence (P < 0.05).


   Results Top


Results of study were evaluated and tabulated as follows.

[Table 1] showed the mean of microhardness values in VHN of all groups. ANOVA test showed that there was a significant difference between each group. When Tukey's multiple comparison test was performed, Group Va did not show any significant difference, compared to Group VIa (control). Between Group Ia and Group IIa, there was no significant difference observed. The Group IIa showed a maximum decrease in microhardness compared with all other groups. The Group IIIa did not show a significant difference with IVa, but showed a significant difference with Group Va and Group VIa, respectively.
Table 1: Mean and standard deviation of VHN values

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[Table 2] showed the mean roughness values in Ra (μm) of all the groups. ANOVA test showed that there was a significant difference in the values among all groups. When Tukey's multiple comparison test was performed, except Group Vb and Group IIIb all other groups showed a significant difference with Group VIb (control). Group IVb showed maximum increase in roughness value. Group Ib and Group IIb showed a significant difference compared with Group VIb. Between Group Vb and Group IIIb, there was not any significant difference present.
Table 2: Mean and standard deviation of roughness values

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   Discussion Top


The success of root canal therapy depends on the method and quality of instrumentation, irrigation, disinfection, and three-dimensional obturation of root canal. Irrigation is presently the best method for removal of tissue remnants and dentin debris during instrumentation. During irrigation, radicular and coronal dentins were exposed to solution deposited in the pulp chamber. This caused alterations on dentin surface. NaOCl, H2 O2, EDTA, and chlorhexidine are the widely used irrigating solutions.

In the present study, anterior teeth were selected to help in ease of separating these single-rooted teeth longitudinally. The roots were separated longitudinally to expose the root dentin surfaces for testing. These sections were embedded in acrylic resin to have a proper base and support for further testing. The sections were cut under water-cooling to prevent desiccation of teeth while sectioning. The sections were ground polished to have even and polished surfaces for microhardness and roughness testing.

These specimens were then viewed under stereomicroscope to check for cracks and those having cracks were eliminated to prevent false results.

The authors reported that the microhardness of dentin declined when tested from superficial to deep regions. The increased number of widely opened dentin tubules free of peritubular dentin near the pulp offered little resistance to the microhardness testing indenter. Thus, Pashley proposed an inverse correlation between dentine microhardness and tubular density. [7]

Thus, in this study midroot region was used for testing, approximately halfway between the central lumen, and root cementum where the dentin surface of root was uniform. This was also done to minimize the effect of the structural variations of different teeth and to establish a reasonable baseline for evaluation. [7]

In this study, the endodontic irrigating solutions were used on root canal dentin surface for 15 min, to obtain optimum results. [2],[8]

Surface changes evaluation of dental hard tissues for alteration in Ca/P ratio of dental tissue has been done by many methods. Such as microhardness measurement, micro-radiographic assessments, scanning election microscopic methods, energy dispersive spectrometric analysis. [9] Also micro-multiple internal reflectance Fourier transform infrared spectroscopy (micro-MIR FTIR) and surface-roughness testing. [10]

Dentin microhardness depends on the amount of calcified matrix per mm 2 (Pashley et al.[7] ) and its determination provides indirect evidence of mineral loss or gain in the dental hard tissues. [9]

The microhardness measurement was one of the simplest non-destructive mechanical characterization methods. Previous investigations have shown the suitability and practicality of VHN microhardness test for evaluating surface changes of dental hard tissues treated with chemical agents. Thus, this method was adopted in this study. [4],[7]

However, the information provided by microhardness testing alone may of then be complementary and thus use of another method was necessary for comprehensive understanding of the surface changes. Thus, surface roughness measurement had also been included in this study to determine surface changes of dental hard tissues. [11],[12]

The results of the present study indicate that all irrigating solutions except chlorhexidine decreased microhardness of root canal dentin significantly, and 3% H 2 O 2 and 0.2% chlorhexidine gluconate had no effect on surface roughness of root canal dentin (P < 0.05). A significant increase on surface roughness was found in 2.5%, 5% NaOCl, and 17% EDTA treated groups (P < 0.05). These results were in resemblance with previous studies. [4,12]

The distilled water was used as control as it is shown that distilled water does not have any chemical changes on dentin. [13]

Studies showed that NaOCl reduced modulus of elasticity and flexural strength of dentin. [14] NaOCl with PH of 7.4-11.5 caused 70% protein description from the hydroxyapetite surfaces. [15] Dentin contained 22% organic material mainly collagen type I which played major mechanical role. Depletion of the organic phase after NaOCl treatment caused mechanical changes. Other studies also reported that NaOCl decreased dentin microhardness. [16]

The authors reported that hydrogen peroxide caused great decrease in dentin microhardness. [17],[18] H 2 O 2 with pH of 1.7 affected the inorganic parts of dentin through acidic demineralization and attacked the organic. [18] And 3% H 2 O 2 had no effect on surface roughness of root dentin. This result was related to the materials low concentration. [12]

An in vitro study showed that the chelating solutions significantly reduced dentin microhardness. [19] Another study showed that EDTA decreases the microhardness and increases the roughness of root dentin significantly. The authors also showed that EDTA decreases root dentin microhardness. [11]

Hulsmann et al. reveiwed the mode of action of EDTA, using gravimetrical analysis.The authors showed that the properties of EDTA were self-limiting. [20] EDTA with neutral pH (7.3) showed chemically, two co-existing reactions (i) complex formation and (ii) protonation. [21] EDTA was normally present as EDTA HNa 3 . The reactions were

  1. EDTA H 3- + Ca 2+ = EDTACa2- + H
  2. EDTA H3-+ H = EDTAH22-.
The exchange of calcium from the dentin by hydrogen resulted in a subsequent decrease in pH. Because of the release of the acid, the efficiency of EDTA decreased with time; on other hand, the reaction of the acid with hydroxyapetite affected the solubility of dentin. [20]

In this study, 0.2% chlorhexidine did not affect microhardness of root canal dentin. These results were in resemblance with previous study. The authors found that 0.2% chlorhexidine gluconate was more effective, had more residual antibacterial effect and lower toxicity than NaOCl solutions. [22] Thus, 0.2% chlorhexidine gluconate seemed to be an appropriate irrigating solution, because of its harmless effect on the microhardness and roughness of root canal dentin.

The present observations suggested that canal irrigation with these chemical solutions leads to structural changes, as evidenced by reduction of dentin microhardness and augmentation in surface roughness. This effect may be related to the solutions demineralizing effect on root canal dentin. Chemical solutions softening effect on the dentinal walls could be beneficial in the clinic, as it permits rapid preparation and negotiation of tight root canals. However, the degree of softening and demineralization action may have an influence of the physical and chemical properties of this heterogenic structure. [6] These chemicals may also affect the adhesion of sealers and cement to the dentin. [10],[23]

These experimental conditions of the bench test differed substantially from the clinical situations. In this study, it was possible to use large amount of the irrigating solutions in close contact with flat dentin surface. This may not be the case in clinical situations as root canal system has complex morphology, and therefore, more questions need to be answered as to the extent to which these chemical alterations may affect the adhesion of sealers to the treated surfaces.

It is noteworthy that the irrigating solutions used in this study were of limited concentrations. Therefore, the effect of various concentrations and application time on dentin microhardness and roughness can be further evaluated in a wider range of similar studies.

 
   References Top

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3.Rosenfeld EF, James GA, Burch BS. Vital pulp tissue response to sodium hypochlorite. J Endo 1978;4:140.  Back to cited text no. 3
    
4.Saleh AA, Ettman WM. Effect of endodontic irrigation solutions on microhardness of root canal dentin. J Dent 1999;27:43-6.  Back to cited text no. 4
    
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9.Arends J, ten Bosch JJ. Demineralization and remineralization evaluation techniques. J Dent Res 1992;71:924-8.   Back to cited text no. 9
    
10.Rotstein I, Dankner E, Goldman A, Heling I, Stabholz A, Zalkind M. Histochemical analysis of dental hard tissues following bleaching. J Endo 1996;22:23-6.   Back to cited text no. 10
    
11.Eldeniz AU, Erdemir A, Belli S. Effect of EDTA and citric acid on the microhardness and roughness of human root canal dentin. J Endo 2005;31:107-10.   Back to cited text no. 11
    
12.Ari H, Erdemir A, Belli S. Evaluation of the effect of endodontic irrigation solutions on the microhardness and roughness of root canal dentin. J Endo 2004;30:792-5.   Back to cited text no. 12
    
13.Cruz-Filho AM, Paula EA, Pécora JD, Sousa-Neto MD. Effect of different EGTA concentrations on dentin microhardness. Braz Dent J 2002;23:188-90.   Back to cited text no. 13
    
14.Sim TP, Knowles JC, Ng YL, Shelton J, Gulabivala K. Effect of sodium hypochlorite on mechanical properties of dentin and tooth surface strain. Int Endod J 2001;34:120-32.   Back to cited text no. 14
    
15.Haikel Y, Gorce F, Allemann C, Voegel JC. In-vitro efficiency of endodontic irrigation solutions on protein desorption. Int Endod J 1994;27:16-20.  Back to cited text no. 15
    
16.Slutzky-Goldberg I, Maree M, Liberman R, Heling I. Effect of sodium hypochlorite on dentin microhardness. J Endo 2004;30:880-2.  Back to cited text no. 16
    
17.Chng HK, Palamara JE, Messer HH. Effect of hydrogen peroxide and sodium perborate on biomechanical property of human dentin. J Endo 2002;28:62-7.   Back to cited text no. 17
    
18.Pécora JD, Cruzfilho AM, Sousaneto MD, Silva RG. In-vitro action of various bleaching agents on microhardnesss of dentin. Braz Dent J 1994;5:129-334.  Back to cited text no. 18
    
19.Cruz-Filho AM, Sousa-Neto MD, Saquy PC, Pécora JD. Evaluation of the effect of EDTAC, CDTA and EGTA on radicular dentin microhardness. J Endo 2001;27:183-4.  Back to cited text no. 19
    
20.Hülsmann M, Heckendorff M, Lennon A. Chelating agents in root canal treatment: mode of action and indication for their use. Int Endod J 2003;36:810-30.  Back to cited text no. 20
    
21.Calvo Pérez V, Medina Cárdenas ME, Sánchez Planells U. The possible role of pH changes during EDTA demoralization. Oral Surg Oral Med Oral Pathol 1989;68:220-2.  Back to cited text no. 21
    
22.Onçað O, Hoþgör M, Hilmioðlu S, Zekioðlu O, Eronat C, Burhanoðlu D. Comparison of antibacterial and toxic effect of various root canal irrigant. J Endo 2003;36:423-32.   Back to cited text no. 22
    
23.Saleh IM, Ruyter IE, Haapasalo M, Ørstavik D. The effect of dentin pretreatment on the adhesion of root canal sealers. Int Endod J2002;35:859-66.  Back to cited text no. 23
    

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Correspondence Address:
Chetan R Patil
Department of Conservative & Endodontics, K.L.E's. V. K. Institute of Dental Sciences, Belgaum, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-9290.79969

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    Tables

  [Table 1], [Table 2], [Figure 1]

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