Original Article The effect of agitation techniques with increasing natrium hypochlorite temperature on bovine pulp tissue dissolution Abstract Michell Wijaya Koesuma1.
Bernard Ongki Iskandar2 Student of Faculty of Dentistry.
Universitas Trisakti.
Jakarta.
Indonesia.
Department of Conservative Dentistry.
Faculty of Dentistry.
Universitas Trisakti Jakarta.
Indonesia Received: 23-Feb-2025 Revised: 27-Sep-2025 Accepted: 24-Nov-2025 Published: 27-Feb-2025 Background: Root canal irrigation is one of the most determining factors that affect the success of a root canal treatment.
Natrium hypochlorite (NaOC.
is the most commonly used root canal irrigants, which, in high concentrations, increases the risk of irritation to the periradicular tissues and oral mucosa if extrusion occurs.
Low concentration NaOCl with increased temperature and agitation techniques could reduce the risk of Objective: To evaluate the effect of agitation techniques with the increased temperature of natrium hypochlorite on bovine pulp tissue dissolution capacity.
Method: Twenty-five pulp tissues of bovine mandibular incisors .
= 5 per grou.
were weighed and then placed individually in Eppendorf test tubes containing the following irrigants:
NaCl 0.
NaOCl 5% at 25AC.
NaOCl 5% at 60AC.
Group 1 NaCl 0.
received no further treatment.
Group 2 NaOCl 5% at 25AC agitated with Group 3 NaOCl 5% at 60AC agitated with sonic.
Group 4 NaOCl 5% at 25AC agitated with ultrasonic.
Lastly.
Group 5 NaOCl 5% at 60AC agitated with ultrasonic.
Dissolution speed was calculated by dividing the difference between initial pulp weight and after-treatment pulp weight .
by the period of time .
Result: NaOCl 5% at 60AC agitated with the ultrasonic group and showed the best result at dissolving pulp tissue.
Conclusion: Increased temperature of natrium hypochlorite showed no significant difference toward bovine pulp tissue dissolution, while sonic and ultrasonic agitation techniques showed a significant difference toward bovine pulp tissue dissolution.
Keywords: NaOCl, agitation techniques, temperature, tissue dissolution Background oot canal treatment is a treatment aimed at teeth with a microorganism infected pulp, either on a vital tooth or non-vital tooth.
1 The success of the root canal treatment is directly tied to the root preparation, which involves cleaning and shaping the root canal, irrigation, which involves the cleaning of necrotic tissue and dentin debris that mechanical instruments cannot reach, and also obturation, which involves the filling of the prepared root canal.
2Ae4 Address for correspondence: Michell Wijaya Koesuma.
Faculty of Dentistry.
Universitas Trisakti Ae Indonesia.
Email: michelwijaya@gmail.
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php/dental DOI :
How to cite this article: Koesuma MW.
Iskandar BO.
The effect of agitation techniques with increasing natrium hypochlorite temperature on bovine pulp tissue dissolution.
Sci Dent J 2025.
8:5-14.
Scientific Dental Journal | Volume 8 | Nomor 2 | 2025.
Februari - April Koesuma MW, et.
The effect of agitation techniques with increasing natrium hypochlorite temperature on bovine pulp tissue dissolution Irrigation is the key to the success of a root canal treatment, because it is the only way to reach the wall area of the root canal that cannot be reached during the mechanical preparation phase.
Natrium Hypochlorite solution (NaOC.
is an irrigation solution that is most commonly used and the most recommended in root canal treatment because of its high antimicrobial activity and capacity to dissolve organic tissues.
5 The efficacy of NaOCl in dissolving tissue is affected by several factors, which is concentration, temperature, volume, contact time with the tissue, agitation, and effective refreshment.
The usage of NaOCl at a high concentration as an irrigant in root canal treatment will increase its antimicrobial properties.
however, it will also increase its toxicity.
5 The capability of NaOCl to dissolve tissue can be maximized with refreshing and thorough agitation3.
Among the tools of agitation, sonic and ultrasonic equipment has been Sonic activation results in mechanical agitation, especially on the tip of the file, and the activation ultrasonic equipment results in an acoustic streaming and cavitation7.
Sonic agitation works on frequencies between 190 HzAe6kHz, and ultrasonic agitation works on frequencies between 25Ae30 kHz.
According to previous research, the NaOCl group that works at 25AC can dissolve 96.
94% of tissue with an average time of 985 seconds.
The reaction that happens is slower in comparison with NaOCl at 60AC, where it can dissolve 98.
31% of tissue at the rate of 389 seconds or 7 minutes.
9 This proves that the temperature increase in NaOCl influences the ability to dissolve tissue.
Research performed by Stoijicic et al.
significantly higher tissue dissolution time after a simultaneous action of temperature and agitation in comparison with one simultaneous action6.
therefore, if NaOCl is used in maximum concentration and maximum temperature and continously moved by agitation, it will increase its ability to dissolve pulp tissue and reduce the risk of complication due to root canal treatment.
The aim of this study was to evaluate the effect of agitation techniques with the increased temperature of natrium hypochlorite on bovine pulp tissue dissolution capacity.
MATERIALS AND METHODS
The sample in this research study uses pulp tissue from the mandibular incisor of the bovine.
Sample retrieval is done immediately after the bovine is slaughtered at the Cakung Slaughterhouse.
North Jakarta.
The lower bovine incisor is prepared as a negative control (NaCl 0.
9%) and NaOCl solution with 5% concentration as the active group, and it is continued with a 25AC and 60AC water bath.
The pulp tissue is inserted in an Eppendorf tube that is filled with NaCl 0.
9%, with a volume of 1 ml using Scientific Dental Journal | Volume 8 | Nomor 2 | 2025.
Februari - April tweezers, and the initial mass is weighed.
Observation is performed to observe the changes that happen to the sample for 7 minutes.
9 Then, they were extracted, and an outline was created using a mini grinder with a cutting disk with 2.
2 cm An outline is created without contact with the pulp tissue.
After the outline was created using a cutting disk (Fig .
, the division of the tooth was performed using chisel and mallet.
The pulp was extracted as a whole, and then, it was inserted on the sample bottle that was filled with fixation fluid Phosphate Buffered Saline (PBS) and frozen at a temperature of -22AC to prevent tissue damage.
Sample mass equalization was performed by drying the pulp tissues and whatman filtration paper no.
1 for 5 minutes, and the pulp tissue was weighed and The sample was cut using scalpel and blade no 11, until a mass of A150Ae170 mg is achieved, and then, the pulp tissue is placed in the fixation fluid PBS and frozen with a temperature of -22AC to prevent tissue damage.
Figure 1.
Bovine Pulp Tissue Extraction Research was performed in a laboratorium by increasing the sample temperature with room temperature .
AC).
Several experimental methods used for measuring the dissolution capacity, such as measuring the time of tissue dissolution and visual Afterwards, the remaining tissue that remained undissolved was weighed.
As reported in other studies, the initial and final weights of the sample were compared.
Koesuma MW, et.
The effect of agitation techniques with increasing natrium hypochlorite temperature on bovine pulp tissue dissolution Eppendorf tube that was filled with 1 ml 5% NaOCl solution was inserted in a water bath by dividing into 2 treatment groups with temperatures of 25AC and 60AC, and the temperature of the NaOCl solution within the Eppendorf tube was conditioned with temperatures of 25AC and 60AC, and the solution was measured with a thermometer.
After, the 5% NaOCl solution with temperatures of 25AC and 60AC pulp tissue was inserted within the Eppendorf tube using tweezers, and the initial mass was weighed.
Samples of each temperatures were divided into 2 groups, which uses sonic agitator using EndoActivatorA .
-3kH.
and IrrisafeA for ultrasonic .
Observational changes were performed on the sample for 7 minutes, according to previous study used stopwatch.
9 The quantification of pulp tissue dissolution has been carried out in several ways, for example by the evaluation of weight loss.
12,13 The remainder of the tissue within the Eppendorf tube that was unable to be dissolved dried using whatman filtration paper no.
1 and Research documentation was performed using Nikon DSLR D7000 camera using Nikkor 105 mm f/2,8 micro lens.
Lighting was performed using softbox and light emitting diode (LED) lamps, using 5,500K temperature in order to get constant lighting.
Table 1.
Pulp Mass Loss in NaCl 0.
9% and NaOCl 5% Solution at Different Temperature with Different Agitation Techniques NaCl 0.
Initial Final Pulp Mass .
NaOCl 5% Sonic Agitation 25AC Initial Final Initial Final NaOCl 5% Ultrasonic Agitation 25AC Initial Final Initial Final Table 2.
Mean dissolution speed .
within the groups (Tissue Dissolution A Standard Deviatio.
Sample Group Treatment NaCl 0,9% NaOCl 5% 25AC Sonic Agitation NaOCl 5% 60AC Sonic Agitation NaOCl 5% 25AC Ultrasonic Agitation NaOCl 5% 60AC Ultrasonic Agitation Mass Difference .
Time .
Dissolution speed .
cUIA SD) 09 A 0.
26 A 0.
28 A 0.
34 A 0.
39 A 0.
RESULT
A comparison of mass after and before treatment with each group can be seen on table 1.
The dissolving rate of the pulp tissue can be viewed on Table 2.
The normality test of this research is performed using Shapiro-Wilk tests, in which the p value is p > 0.
05, and the data distribution is considered to be normal.
The homogenity test of this research study was performed using LeveneAos Test of Equality of Error Variances, in which the value is p > 0,05 and data distribution was considered to be Two-Way ANOVA tests showed no significant differences between each group of increasing temperatures in dissolving rate.
it showed significant differences in groups using sonic and ultrasonic agitation methods.
Post-hoc Tukey tests showed that 5% NaOCl that was agitated using IrrisafeA ultrasonic tools exhibited the best capability to dissolve pulp tissue compared to 0.
NaCl without agitation and 5% NaOCl that were agitated using EndoActivatorA sonic appliances.
5% NaOCl that were agitated using EndoActivatorA sonic appliances showed better capability to dissolve Scientific Dental Journal | Volume 8 | Nomor 2 | 2025.
Februari - April Koesuma MW, et.
The effect of agitation techniques with increasing natrium hypochlorite temperature on bovine pulp tissue dissolution pulp tissue compared to 0.
9% NaCl without agitation (Table .
Pulp Tissue Dissolution .
Figure 2.
Pulp Tissue Dissolution Diagram Table 3.
Post Hoc Test Mean Std.
Agitation Technique Difference Error
(I-J)
Without Sonic Agitation Ultrasonic -.
Sonic Confidence Sig.
Interval Lower Upper Bound Bound 000 -.
2509 -.
000 -.
3389 -.
Without Agitation .
Ultrasonic -.
001 -.
1418 -.
Without Ultrasonic Agitation Sonic
DISCUSSION
Tissue dissolution outcome during the process of endodontic irrigation have tested.
However, the capacity and efficiency of organic tissue degradation processes are related to the type, concentration, and how the irrigation solution is agitated.
11 Natrium hypochlorite is the main endodontic irrigant used worldwide, the concentrations ranging from 0.
5% to 6% due to its antimicrobial and tissue-dissolving 14 Research by Estrela et.
al declared that the dissolving rate of pulp tissue can be seen by bubbles that were created by the saponification The saponification reaction happens when the natrium hypcholorite comes in contact with the pulp tissues and destroys the fatty acid structures, which results in soap and glicerol.
15 The bubbles that were formed were caused by oxidation reaction and hydrolisis that happens between NaOCl as an oxidator and strong hydrolisis.
In this study, it was shown that temperatures increase of natrium hypochlorite have no significant difference in dissolving pulp tissue.
Pulp tissue dissolve rate using natrium hypochlorite was influenced with the chlorine solution within the solution itself.
17 The increase of temperature of Scientific Dental Journal | Volume 8 | Nomor 2 | 2025.
Februari - April NaOCl above 25AC causes the chlorine content to demonstrate a tendency to evaporate.
18 The decrease of chlorine solution from 5% NaOCl solution happens significantly at a temperature of 24AC.
According to research by Cunningham and Balekjian .
, data show a decrease in chlorine content from the NaOCl solution at a temperature of 37AC with a concentration of 2.
5% as much as 4%.
however, on 5% concentration as much as 9.
These factors can be linked with one of the factors that causes a temperature increase of NaOCl solution, which shows no significant difference to the dissolving rate of the pulp tissue.
According to the result of pulp dissolution by counted pulp mass and dissolution time, ultrasonic agitation shown significant difference compared with sonic agitation.
Ultrasonic is simple and costeffective irrigant activation method, works by delierivering an acoustic stream that creates shear stress sufficient enough to dislodge the debris of instrumented root canals.
20 Ultrasonic activation of NaOCl has been suggested because it is claimed to accelerate chemical reactions and promote superior cleaning action.
Ultrasonic agitation methods results in vibration that causes in a current change in the NaOCl solution that occurs constantly.
The vibration produced by the ultrasound devices provides continous flow of irrigation solution in root canal.
Those vibrations were directly connected with the acoustic streaming and cavitation that facilitates the increase of capability to clean debris and dissolve the pulp tissue within the root canal.
Possible reason of ultrasonic agitation mechanism is the collapse of bubbles during transient cavitation that produces a pressure-vacuum effect, which sucks the canal content to the inside rather than pushing it further in the canal.
This will be followed by diffusion of the irrigant in the main canal to substitute the space Another possibility is that the streaming around the activated file because of the cohesion between fluid particles inside the accessory canal and the irrigant in the main canal sucks the content of the accessory canals into the main canal with fluid flow toward the main canal.
22 The agitation frequency ultrasonic .
Ae30kH.
is higher in comparison with sonic agitation .
HzAe6kH.
Those occurences can be connected with the speed of NaOCl solution and the friction between the irrigation solution and root canal, which increases the capability to clean debris and increases the dissolving rate of pulp tissue in comparison with sonic agitation.
14 Repetition several cycles of chemomechanical preparation can promote the complete dissolution of the pulp tissue.
CONCLUSION
No significant difference was found between the temperature increase of natrium hypochlorite between bovine pulp tissue.
however, significant Koesuma MW, et.
The effect of agitation techniques with increasing natrium hypochlorite temperature on bovine pulp tissue dissolution differences were found between sonic agitation and ultrasonic agitation compared to the dissolving rate of bovine pulp tissue.
REFERENCES