Basic Research—Technology
The Effects of Sodium Hypochlorite (5.25%), Chlorhexidine
(2%), and Glyde File Prep on the Bond Strength of
MTA-Dentin
Ping Yan, DDS, PhD,* Bin Peng, DDS, PhD,* Bing Fan, DDS, PhD,*
Mingwen Fan, DDS, PhD,* and Zhuan Bian, DDS, PhD*†
Abstract
The purpose of this study was to evaluate the effects of
sodium hypochlorite (5.25%), chlorhexidine (2%), and
Glyde File Prep on the bond strengths of MTA-dentin in
vitro. Standardized dentin disks were prepared and the
central hole in each disk was filled with mineral trioxide
aggregate (MTA). The specimens were randomly di-
vided into four groups to be immersed in saline, 5.25%
NaOCl, 2% chlorhexidine, and Glyde File Prep for 2
hours. The bond strengths of MTA-dentin were mea-
sured with a material testing system (MTS) and the
fractured surfaces on the root walls were observed by
scanning electron microscopy. Compared with the con-
trol group, the bond strengths were significantly lower
in Glyde File Prep group (p ⬍ 0.05) and there was no
significant difference in the chlorhexidine group or in
the NaOCl group (p ⬎ 0.05). This study suggested that
Glyde File Prep could negatively affect the bond
strengths of MTA-dentin. (J Endod 2006;32:58 – 60)
Key Words
Bond strengths, mineral trioxide aggregate, scanning
electron microscopy
From the *Department of Endodontics, School of Stoma-
tology; †Key Lab for Oral Biomedical Engineering of Ministry of
Education, Wuhan University, Wuhan, China.
Address requests for reprint to Professor Zhuan Bian, Key
Lab for Oral Biomedical Engineering of Ministry of Education,
Wuhan University, Luoyu Road 237, Wuhan City, China
430079. E-mail address: kqyywjtx@public.wh.hb.cn.
0099-2399/$0 - see front matter
Copyright © 2006 by the American Association of
Endodontists.
doi:10.1016/j.joen.2005.10.016
58 Yan et al.
M ineral trioxide aggregate (MTA; ProRoot, Dentsply/Tulsa Dental, Tulsa, OK) has
been widely used as a promising biomaterial to repair root perforations because
of its excellent biocompatibility, superior sealing abilities, and ability to set in the
presence of blood (1– 4). In previous studies, different treatment strategies were ap-
plied for sealing perforations with MTA (5, 6). An alternative treatment sequence was to
place MTA into the perforation defects after complete instrumentation and obturation of
the canals with gutta-percha apical to the perforation sites (7). However, there were
some disadvantages in selecting such a treatment sequence. Firstly, some irrigants may
cause severe irritation of the periodontal tissue through the perforated channel during
the cleaning and shaping of root canals (8). Secondly, the sealer and gutta-percha can
be pushed into the periodontal defect during compaction. Thirdly, the root canal space
can be contaminated by the ingress of tissue fluids containing bacteria from the perfo-
ration sites (9). Therefore, most clinicians suggested that the perforation defects should
be repaired before proceeding with definitive endodontic treatment (10).
After the repair of perforations with MTA, nonsurgical endodontic therapy must be
performed with various medications such as 5.25% sodium hypochlorite, 2% chlo-
rhexidine, or Glyde File Prep to clean the root canals (11–13). However, the question
as to whether these medications would potentially initiate chemical reactions to degrade
MTA or interfere its bond to dentin has not been addressed. The purpose of this study
was to evaluate the effects of sodium hypochlorite (5.25%), chlorhexidine (2%), and
Glyde File Prep on the bond strengths of MTA-dentin in vitro.
Materials and Methods
Sound human premolars extracted for orthodontic purposes were used in the
present study. The coronal parts were removed and mid-root dentin was horizontally
sectioned into 1.0 mm thick slices with a diamond saw microtome (Leica saw SP1600
microtome). The canals were instrumented with Gates Glidden burs #3 through #5
(Mani, Tochigi, Japan) to a standardized diameter of 1.3 mm. Dentin disks were thor-
oughly irrigated with 2% sodium hypochlorite for 5 min and then washed with distilled
water.
MTA powder was mixed with sterile water on a glass slab with a cement spatula
according to the manufacturer’s directions at a powder to liquid ratio of 3:1. When the
mixture exhibited a putty consistency, it was immediately placed into root canals with a
Dovgan carrier (G. Hartzell & Son, Concord, CA) and compacted with pluggers
(Dentsply, Maillefer, Switzerland). Saline-moistened gelfoam was used as a matrix
material to simulate clinical conditions. Excess material was trimmed from the surface
of the specimens with a scalpel. All specimens were examined using a microscope at 16
times magnification. Any specimens found to have cracks, defects, or gaps between the
material and dentin walls were excluded from this study. These specimens were
wrapped in wet gauze and sealed in a plastic bag, which was placed in an incubator and
allowed to set for 7 days at 37°C. Each dentin disk was tested with the head of gutta-
percha point to ensure the set of the MTA material before the displacement test. After the
7-day period for initial set, 32 specimens were randomly divided into four groups to be
immersed in different medications in glass plates for 2 hours as follows: 5.25% NaOCl
(Xilong Chemical Factory, Shantou, China), 2% chlorhexidine (Medicine Company,
Wuhan, China), Glyde File Prep (Dentsply, Maillefer, Ballaigues, Switzerland), and
JOE — Volume 32, Number 1, January 2006

saline (control group, Wuhan, China). After being immersed for 2
hours, all specimens were removed from the solutions and rinsed with
distilled water before testing.
A plexiglass block was placed underneath each specimen, and a
hole was drilled in the block to accommodate the displacement of MTA.
The bond strengths were tested using a MTS testing machine. The cyl-
inder-shaped end of a 2/4 hand plugger with 1 mm diameter (Dentsply,
Maillefer, Switzerland) was used as a force probe, mounted on the
moving head of the MTS. The force probe, traveling at a speed of 2
mm/min, applied pressure to the surface of MTA in each specimen until
the material was dislodged. The maximum force applied to MTA before
dislodgment occurred was recorded in N of force.
All data were entered into the computer and analyzed by means of
the SPSS 11.0 system for Windows. A one-way ANOVA was performed to
compare the differences in dislodgment forces among the four groups.
Scheffer’s multiple-comparison was used to compare the significant
differences found between the groups. The level of statistical significant
was set at 0.05.
One specimen in each group was randomly chosen for scanning
electronic microscopy (SEM) examination. After the dislodgment of
MTA, the specimens were longitudinally split from the center of the
canals. The dentin walls were sputter-coated with gold and observed
using a SEM (JSM-5610LV, Japan) for fractographic analysis.
Results
Push-Out Test
Table 1 shows the mean and standard deviations for dislodgement
force (N) in the different groups. A statistically significant difference in
the mean dislodgement force was found between the Glyde File Prep
group and the control group (p ⬍ 0.05). Although the mean dislodge-
ment force of MTA-dentin decreased in both the 2% chlorhexidine
group and the 5.25% NaOCl group, there was no significant difference
when compared to the control group (p ⬎ 0.05). No difference was
observed among the 5.25% NaOCl group, 2% chlorhexidine group, and
Glyde File Prep group (p ⬎ 0.05).
SEM Examination
Under the SEM evaluation at a magnification of 5000⫻, de-bond-
ing was mainly observed at the MTA-dentin interface, and the dentin
walls of root canals were almost entirely covered with a consistent
interfacial layer that was firmly attached to the dentin walls in the spec-
imens from each group.
Gelatinous hydrated products of MTA with needle-shaped residue
and floc covered the interfacial layer of dentin walls in the control group
(Fig. 1A). The SEM photographs of 5.25% NaOCl group were similar to
those of control group (Fig. 1B). In 2% chlorhexidine group, the nee-
dle-shaped residue of MTA on the dentin walls increased significantly
and the amount of floc was reduced (Fig. 1C). The regular flake-shaped
material on the dentin surface was observed in Glyde File Prep group
(Fig. 1D).
TABLE 1. Mean and standard deviations of dislodgement force in different
groups (ANOVA)
Groups Mean ⴞ SD (N)
Saline 48.0 ⫾ 21.0
5.25% NaOCl 36.6 ⫾ 12.7
2% Chlorhexidine 37.9 ⫾ 13.4
Glyde File Prep 21.6 ⫾ 11.2*
*p ⬍ 0.05.
JOE — Volume 32, Number 1, January 2006
Basic Research—Technology
Figure 1. (A) The SEM photograph of the control group shows gelatinous hy-
drated products of MTA with needle-shaped residue and floc on the interfacial
layer of dentin walls (5000⫻). (B) The SEM photograph in NaOCl group was
similar to those of control group (5000⫻). (C) SEM photograph showed the
increased needle-shaped residue of MTA on the interfacial layer of dentin walls
in chlorhexidine group (5000⫻). (D) Regular flake-shaped material on the
interfacial layer of dentin walls was observed in Glyde File Prep group.
(5000⫻).
Discussion
MTA, a powder consisting of hydrophilic particles of dicalcium
silicate, tricalcium silicate, and tricalcium aluminate, is a mechanical
mixture of Portland cement, bismuth oxide, and gypsum (14). Research
on the physicochemical interaction between MTA and root canal walls
indicated that MTA was a bioactive material and appeared to bond
chemically to dentin via a diffusion-controlled reaction between its apa-
titic surface and dentin, forming an adherent interfacial layer that was
firmly attached to the dentin walls (15). Furthermore, the chemical
adhesion of MTA-dentin was supposed to relate the superior sealing
ability of MTA over conventional filling materials (15).
The NaOCl solution has a nonspecific proteolytic effect that can
effectively remove organic components from root canals and dentinal
tubules (16). Ari et al. indicated that 5.25% NaOCl solutions signifi-
cantly decreased the microhardness of root canal dentin (17). Yamauti
et al. found that the bond strengths of dentin to adhesive resin decreased
significantly with increasing storage time in NaOCl (18). However, in the
present study, though the bond strengths of MTA-dentin presented a
decreased tendency in 5.25% NaOCl group, it was not significantly dif-
ferent when compared with the control group (p ⬎ 0.05). The micro-
structure on the interfacial layer of dentin walls in NaOCl group was
similar to those of control group. It was likely related to the high pH
value and lack of the ability to dissolve inorganic materials of the 5.25%
NaOCl. Conversely, the 2% chlorhexidine group displayed a significantly
morphological alteration on the interfacial layer of dentin walls. Re-
cently, Stowe et al. reported that the change of physical properties was
observed during the experiment of substituting 0.12% chlorhexidine
gluconate for sterile water as a mixing agent, in which the MTA/chlo-
rhexidine mixture seemed to set more rapidly (1–2 min) than the
MTA/water mixture (5– 6 min) and take on a more crumbly texture at
placement (19). Though the bond strengths of MTA-dentin in 2% chlo-
rhexidine had no significant decrease in this study, further research is
needed to clarify the alteration on physical properties of MTA in chlo-
rhexidine solution before advocating the clinical application of chlo-
rhexidine in conjunction with MTA.
Evaluating Bond Strengths of MTA-Dentin In Vitro 59
Basic Research—Technology
Glyde File Prep, a water-soluble acidic gel containing 15% EDTA
and urea peroxide, has the capability to improve chemomechanical
debridement by removing the smear layer when used as a lubricant and
chelating agent during the cleaning and shaping of root canals (20). In
the present study, Glyde File Prep significantly decreased the bond
strengths of MTA-dentin (p ⬍ 0.05). Additionally, it was interesting that
the regular flake-shaped material on the interfacial layer of dentin walls
was observed in Glyde File Prep group. One of the probable reasons was
that Glyde File Prep has the capability to remove the smear layer, which
makes it possible for the Glyde File Prep to infiltrate into the interfacial
layer and interfere the chemical adhesion between MTA and dentin.
Another explanation could be the demineralization effect of Glyde File
Prep on Ca-containing materials. Because of the porous nature of MTA
(21), it was conceivable that this precipitation proceeded internally
within MTA to change the microstructure of MTA and thus led to a
significant decrease in bond strengths of MTA-dentin.
In conclusion, the resistance to displacement of MTA was highly
dependent on the agent in which it was immersed. Glyde File Prep had
an adverse effect on MTA-dentin bond strengths.
Acknowledgment
This study was supported by national key technologies R&D
Program of the Tenth-five Year Plan, the Ministry of Science and
Technology, China (2004BA720A24) and National Sciences Foun-
dation of China (30271415)
References
1. Schwartz RS, Mauger M, Clement DJ, Walker WA 3rd. Mineral trioxide aggregate: a
new material for endodontics. J Am Dent Assoc 1999;130:967–75.
2. Apaydin ES, Shabahang S, Torabinejad M. Hard-tissue healing after application of
fresh or set MTA as root-end-filling material. J Endod 2004;30:21– 4.
3. Ferris DM, Baumgartner JC. Perforation repair comparing two types of mineral
trioxide aggregate. J Endod 2004;30:422– 4.
60 Yan et al.
4. Hardy I, Liewehr FR, Joyce AP, Agee K, Pashley DH. Sealing ability of One-Up Bond
and MTA with and without a secondary seal as furcation perforation repair materials.
J Endod 2004;30:658 – 61.
5. Main C, Mirzayan N, Shabahang S, Torabinejad M. Repair of root perforations using
mineral trioxide aggregate: a long-term study. J Endod 2004;30:80 –3.
6. Menezes R, da Silva Neto UX, Carneiro E, Letra A, Bramante CM, Bernadinelli N. MTA
repair of a supracrestal perforation: a case report. J Endod 2005;31:212– 4.
7. Torabinejad M, Chivian N. Clinical applications of mineral trioxide aggregate. J
Endod 1999;25:197–205.
8. Gernhardt CR, Eppendorf K, Kozlowski A, Brandt M. Toxicity of concentrated sodium
hypochlorite used as an endodontic irrigant. Int Endod J 2004;37:272– 80.
9. Alhadainy HA. Root perforations. A review of literature. Oral Surg Oral Med Oral
Pathol 1994;78:368 –74.
10. Ruddle CJ. Nonsurgical endodontic retreatment. In Cohen S, Burns RC, eds. Pathways
of the pulp, 8th ed. St. Louis: CV Mosby, 2002:919.
11. Leonardo MR, Tanomaru Filho M, Silva LA, Nelson Filho P, Bonifacio KC, Ito IY. In
vivo antimicrobial activity of 2% chlorhexidine used as a root canal irrigating solu-
tion. J Endod 1999;25:167–71.
12. Grandini S, Balleri P, Ferrari M. Evaluation of Glyde File Prep in combination with
sodium hypochlorite as a root canal irrigant. J Endod 2002;28:300 –3.
13. Ercan E, Ozekinci T, Atakul F, Gul K. Antibacterial activity of 2% chlorhexidine
gluconate and 5.25% sodium hypochlorite in infected root canal: in vivo study. J
Endod 2004;30:84 –7.
14. Asgary S, Parirokh M, Eghbal MJ, Brink F. Chemical differences between white and
gray mineral trioxide aggregate. J Endod 2005;31:101–3.
15. Sarkar NK, Caicedo R, Ritwik P, Moiseyeva R, Kawashima I. Physicochemical basis of
the biologic properties of mineral trioxide aggregate. J Endod 2005;31:97–100.
16. Andersen M, Lund A, Andreasen JO, Andreasen FM. In vitro solubility of human pulp
tissue in calcium hydroxide and sodium hypochlorite. Endod Dent Traumatol
1992;8:104 – 8.
17. Ari H, Erdemir A, Belli S. Evaluation of the effect of endodontic irrigation solutions on
the microhardness and the roughness of root canal dentin. J Endod 2004;30:792–5.
18. Yamauti M, Hashimoto M, Sano H, et al. Degradation of resin-dentin bonds using
NaOCl storage. Dent Mater 2003;19:399 – 405.
19. Stowe TJ, Sedgley CM, Stowe B, Fenno JC. The effects of chlorhexidine gluconate
(0.12%) on the antimicrobial properties of tooth-colored ProRoot mineral trioxide
aggregate. J Endod 2004;30:429 –31.
20. Hulsmann M, Heckendorff M, Lennon A. Chelating agents in root canal treatment:
mode of action and indications for their use. Int Endod J 2003;36:810 –30.
21. Fridland M, Rosado R. Mineral trioxide aggregate (MTA) solubility and porosity with
different water-to-powder ratios. J Endod 2003;29:814 –7.
JOE — Volume 32, Number 1, January 2006