Clearing Technique and its Potential Applications An In Vitro Study

Meenakshi Verma*

Abstract
Objective : Sectioning, SEM studies and various different methods have been tried out for In Vitro visualization of the complex root canal morphology, but each have their own limitations. A different technique namely the transparent tooth system or the clearing technique was modified and tried out. Material & Methods : 27 teeth extracted for various reasons were randomly selected and were subjected to the process of decalcification, dehydration & clearing, which rendered them transparent. These teeth were then studied for their morphology, micro-leakage, broken instrument etc. Results : The tooth can be rendered transparent in Vitro by a relatively easy & inexpensive method. This can be put to many uses like studying the pulpal anatomy, micro leakage studies, analysis of broken instruments and perforations, homogeneity and adaptation of Guttapercha, etc. Conclusion: The clearing technique allows three dimensional study of the root canal system with a plethora of applications and advantages. Key Words : Clearing technique, Transparent tooth, Three dimensional visualization, Canning electron microscope.

INTRODUCTION
he complete root canal morphology has always daunted the genius of the researcher, teacher & clinician. To study, evaluate & research it in Vitro. Methods like Vertical and Longitudinal sectioning,1 and SEM studies have been tried out however, each have their own shortcomings. Hence a technique called The Clearing Technique2,3 was modified and after some changes tried and carried out successfully.

AIM
The objective of this study was three-dimensional continuous view of the root canal system.

MATERIAL AND METHODS

27 extracted teeth were chosen randomly. Some of them were already root canal treated but extracted due to complications or failures or patients reluctance for re root canal treatment. Some had been extracted due to periodontal reasons or the patient was going in for a complete denture, a few were third molar etc. Technique: A simple inexpensive procedure consisting of decalcification, dehydration and clearing was done.
*Reader, Department of Conservative Dentistry and Endodontia, Bharati Vidyapeeth Dental College & Hospital, Navi Mumbai. 1122

Teeth that were already root canal treated were immersed in pelican ink for 48 hours after removing the coronal restoration to evaluate for microleakage. Teeth whose pulpal anatomy was to be studied were injected with the same ink after coronal access preparation. In some of the teeth the coronal part of the tooth that was not to be made transparent was coated with sticky wax. Decalcification: The teeth were placed in 5% Nitric acid at room temperature, which was changed daily & manually agitated 3 times. Depending on the size of the specimen, they were placed for 48 to 72 hours duration. The end point of decalcification was assured by the needle method, which was inserted in an unimportant part of the tooth. This was followed by a 4 hour running water wash to remove all traces of the acid. Dehydration: Was done in a series of Ethyl alcohol rinses 1. 80% Ethyl alcohol overnight followed by 2. 90% Ethyl alcohol for 3 hours and finally 3. 100% Ethyl alcohol four rinses of 1 hour each. Clearing: The dehydrated teeth were placed in methyl salicylate for 24 hours that rendered them transparent. Preservation of the specimen: The cleared specimens may be stored and displayed in their final clearing agents or permanent mounting in clear plastic too can be done.

RESULTS
The tooth can be rendered transparent in Vitro by a
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Fig. 3 : Maxillary canine Fig. 4 : Microleakage seen Fig. 5 : Perforation in the form of dye that had been RC treated in a mandibular by conventional method. penetration around the GP second molar. point in a mandibular Each GP point can be seen anterior due to loss of clearly. relatively easy & inexpensive method. This can be put to coronal seal. Fig. 1 : Unpredictable Radicular Anatomy and purpal vascular system of a lower third molar. Fig. 2 : Fractured file in a lower mandibular molar.

many uses like studying the pulpal anatomy, micro leakage studies, analysis of broken instruments and perforations, homogeneity and adaptation of Guttapercha, etc (Figs. 1-5).

failures such as broken instruments and perforations.

SUMMARY AND CONCLUSIONS

The methodology of rendering teeth transparent in vitro was studied. 27 teeth extracted for various reasons were randomly selected and were subjected to the process of decalcification, dehydration & clearing, which rendered them transparent. These teeth were then studied for their morphology, micro-leakage, broken instrument etc. The results showed that the Clearing technique is a relatively easy technique to obtain a three dimensional continuous view of the root canal system3. From this study it is also evident that the transparent model system has a number of possible applications2 limited only by imagination. We hope this technique will benefit the student who in due course will become a good researcher or a clinician which will ultimately lead to improved Endodontics.

DISCUSSION
The study of the complex root canal morphology is very important and various techniques have been advocated. Vertical sectioning can be done but this does not allow a continuous view of the entire root canal system. Longitudinal sectioning compensates for it but a three dimensional picture is not possible.1 SEM studies too have a major disadvantage of limited area available. Till 1982 the Clearing procedure was used for anatomical studies of the pulp cavity2. In 1983 Tagger M etal put forward an improved method with methyl salicylate as the clearing agent for three dimensional study on apical leakage.4 The clearing technique or transparent root canal a) Makes three dimensional assessments possible5. b) Maintains the original form of the root canal even to its minute detail.6 c) Failure in preparation of the specimen is rare. d) Specimen can be preserved for a long time. Hence this technique has numerous uses and advantages. 1. Can be used as an important teaching and to demonstrate the Pulpal Vascular System.6,7 2. Can be used for better understanding and appreciation of Instrumentation. 3. Can be used as an important research tool for investigation of micro leakage studies.4 4. In addition to micro leakage, this technique can be used to study, distribution of sealer, filling morphology extent, homogeneity and adaptation of Guttapercha.8 5. Can provide an insight for analysis of clinical
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REFERENCES
1. Weine F.S , et al. Canal configuration in mesio-buccal root of maxillary first molar and its endodontic significance. OS OM OP 1969; 28(3) : 419-25. Tagger Micheal. Clearing of teeth for study and demonstration of pulp. J Dent Edn 1976; 40 (3) : 172-4. Robertson D, et al. A clearing technique for the study of root canal system. J Endo 1980; 5 (2) : 421-24. Tagger M, et al. An improved method of three-dimensional study of apical leakage. Quintessence Int 1983; 10 : 981-6. Robertson DC, Leeb IJ. Evaluation of transparent tooth model system T. M. for evaluation of Endodontically filled tooth. J Endo 1982; 8(7) : 317-21. Al Shalabi RM, et al. Root canal anatomy of maxillary first and second permanent molars. Int Endod J 2000; 33 : 405-14. Blaine M Cleghorn, et al. Root and root canal morphology of permanent maxillary first molars A literature review. J Endod 2006; 32 : 813-21. Evans John, Simon J. Apical Seal produced by injectable thermoplasticised gutta-percha in the absence of a smear layer and root canal sealer. J Endo 1986; 12(3) : 101-7. 1123

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