ENDODONTOLOGY. Chair side disinfection of gutta - percha points - An in vitro comparative study between 5 different agents at different concentrations

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1 Original Research Chair side disinfection of gutta - percha points - An in vitro comparative study between 5 different agents at different concentrations Pradeep.K # K.H Kidiyoor Pavithra Jain Nageshwar Rao ABSTRACT Objectives:. The objectives of present study are: 1) To compare sterilizing effect on gutta-percha points of five disinfectant solutions: Povidone iodine (5%), Sodium hypochlorite (5%), Ethyl alcohol (95%), Hydrogen peroxide (3%) and a combination of Chlorhexidine (1.5%) and Cetrimide (15%) in equal proportions. 2) To find out which disinfectant solution requires less time for effective sterilization of gutta-percha points. Study design: Gutta-percha cones exposed to dental chair-side clinical environment were treated with above mentioned disinfectant solutions for 1, 3, 5, 7 and 10 minutes. They were later cultured into thioglycolate media at 37 0 C and observed after 24, 48 and 72 hours for turbidity. Those showing turbidity were sub-cultured for aerobic bacteria in blood agar, MacConkey s agar and chocolate agar at 37 0 C for hours and for anaerobic bacteria in anaerobic blood agar at 37 0 C for 5 days. Microorganisms were identified by Gram stain and standard microbiological techniques. Results: The results showed that Sodium hypochlorite (5%) and a combination of Chlorhexidine (1.5%) and Cetrimide (15%) were found to be the most effective chemical disinfectants in sterilizing gutta-percha. Conclusion: All the disinfectant agents used in the study were found to be effective in sterilizing the gutta-percha points. However, there were differences in the time taken by these solutions in disinfecting the gutta-percha points. Sodium hypochlorite (5%) and combination of Chlorhexidine (1.5%) + Cetrimide (15%) achieved disinfection of the gutta-percha cones with the immersion time of 1 minute. Keywords: disinfectants, Gutta-percha cones, micro-organisms, routine chair-side decontamination. Introduction Obturation is one of the most important procedures which determine the success of endodontic therapy. The main purpose of the obturation is to promote healing and prevent percolation or ingress of microorganisms into the periapical area. 1 This can be achieved by following strict asepsis protocol during endodontic therapy. The practitioner must be concerned not only with endogenous oral microbial flora, but with exogenous bacterial contamination as well. 2 It is an established fact that the success of endodontically treated teeth depends on the aseptic techniques followed during endodontic procedures. 3 Guttapercha points are the most commonly used endodontic filling material. 4 They become accessible to potential contamination by variety organisms such as cocci, rods, and yeasts present in the air or contacting objects once the box of guttapercha points are exposed to dental chair side clinical environment. 5 Gutta-percha points, due to their chemical and physical properties are not # Dept of Conservative Dentistry and Endodontics, Yenepoya Dental College and Hospital, Mangalore 73

2 PRADEEP.K, K.H KIDIYOOR, PAVITHRA JAIN, NAGESHWAR RAO amenable to the usual methods of physical sterilization in an autoclave or hot air oven. Therefore, chemical sterilization is mandatory for effective sterilization of gutta-percha points. 6 The purpose of the present study is to find out a rapid, reliable, convenient and effective method of sterilization of gutta-percha using readily available germicidal agents, such as, 5% Sodium hypochlorite, 3% Hydrogen peroxide, 95% Ethyl alcohol, 5% Povidone iodine and a combination of 1.5% Chlorhexidine and 15% Cetrimide. Materials and Methods : In the present study, the gutta-percha points were allotted to 5 groups depending upon the type of antibacterial agents used for sterilization. Group I Gutta-percha immersed in Povidone iodine (5%).Group II Gutta-percha immersed in Sodium hypochlorite (5%).Group III Gutta-percha immersed in Ethyl alcohol (95%).Group IV Guttapercha immersed in Hydrogen peroxide (3%).Group V Gutta-percha immersed in combination of Chlorhexidine (1.5%) and Cetrimide (15%) in equal proportions. Each main group was further sub divided into 5 subgroups, A, B, C, D and E, according to the immersion time of gutta-percha into solution. Subgroup A Immersion time of Gutta-percha was 1 minute. Subgroup B Immersion time of Guttapercha was 3 minutes. Subgroup C Immersion time of Gutta-percha was 5 minutes. Subgroup D Immersion time of Gutta-percha was 7 minutes. Subgroup E Immersion time of Gutta-percha was 10 minutes. A total of 216 gutta-percha cones (No. 70 Dentsply maillefer ) were selected for this study, from the packs exposed to dental chair-side clinical environment and from freshly opened sealed pack. For each subgroup, 8 cones were used. So the number of cones in each group was 40. A total of 16 cones were used as controls. The following controls were used. Positive control: A total of eight gutta-percha cones were used as positive control from the pack exposed to dental chair-side clinical environment. The cones were directly transferred to the culture media without disinfection. Negative control: A total of eight gutta-percha cones were taken from freshly sealed pack. The cones were then transferred to culture media without disinfection. Test group: Test group was selected from gutta-percha pack exposed to dental chair side clinical environment. A total of eight gutta-percha cones were included in each sub group. These cones were then treated with each solution of disinfectant for an immersion time of 1, 3, 5, 7 and 10 minutes. All the points were subsequently transferred to thioglycolate media.(photograph 7) The inoculated media were incubated at 37 C and observed after 24, 48 and 72 hrs of incubation for presence of growth of microorganisms by comparing the turbidity with that of the uninoculated medium, which was clear (Photograph 8 A and B). PHOTO GRAPH 1 : GUTTA PERCHA POINTS INOCULATED IN THIOGLYCOLATE MEDIA 74

CHAIR SIDE DISINFECTION OF GUTTA - PERCHA POINTS - AN IN VITRO COMPARATIVE STUDY BETWEEN 5 DIFFERENT AGENTS AT DIFFERENT CONCENTRATIONS Gram positive bacteria: They are those bacteria that resist

A : UNINOCULATED MEDIA ( CLEAR) B: WITH GROWTH(TURBID) Gram negative bacteria: They are those bacteria that are decolorized by organic solvents, therefore take the counter stain (carbol fuschin) and

3 CHAIR SIDE DISINFECTION OF GUTTA - PERCHA POINTS - AN IN VITRO COMPARATIVE STUDY BETWEEN 5 DIFFERENT AGENTS AT DIFFERENT CONCENTRATIONS Gram positive bacteria: They are those bacteria that resist discoloration, retain primary stain (crystal violet) and appear violet. PHOTOGRAPH 2: THIOGLYCOLATE MEDIA AFTER INCUBATION. A : UNINOCULATED MEDIA ( CLEAR) B: WITH GROWTH(TURBID) Gram negative bacteria: They are those bacteria that are decolorized by organic solvents, therefore take the counter stain (carbol fuschin) and appear red. Later, gram stained smears were observed under microscope and the microorganisms were further identified by standard microbiological techniques. PHOTO GRAPH 3: SUBCULTRUE OF THIOGLYCOLATE ON SOLID MEDIA AFTER INCUBATION. The tubes showing turbidity were sub-cultured on Blood agar, MacConkey s agar and Chocolate agar to support the growth of aerobic bacteria (Photograph 9). The culture plates were then incubated at 37 C for hrs in ambient air. For anaerobic bacteria, subcultures were made on Anaerobic blood agar. Anaerobiasis was produced by the use of gas pak anaerobic system. Cultures were incubated at 37 C for hours. The isolates grown from both aerobic and anaerobic cultures were further identified by gram stain. The Gram stain differentiates bacteria into two broad groups: Results : Graph 1 The results of the present study shown in Graph 1. In the present study, sodium hypochlorite 5% (Group 2) and a combination of Chlorhexidine 1.5% + 15% Cetrimide (Group 5) showed the best anti-septic effect and was the most effective disinfectant for chairside decontamination of guttapercha cones. They completely decontaminated the gutta-percha cones in 1 minute. The Povidone iodine 5% (Group 1), Ethyl alcohol 95% (Group 3) and Hydrogen peroxide 3% (Group 4) were also found to be effective in sterilizing gutta-percha cones. Povidone iodine 5% took 3 minutes to achieve sterilization and Hydrogen peroxide 3 % and Ethyl alcohol 95% took 5 minutes each to achieve sterilization.all positive controls showed positive results during the first 24hours. Negative controls 75

4 PRADEEP.K, K.H KIDIYOOR, PAVITHRA JAIN, NAGESHWAR RAO were followed up for 24 and 72 hours, and no microbial growth was observed in any of the groups tested, demonstrating the efficacy of previous sterilization. Discussion : Sterilization of endodontic instruments and materials is an important step during the endodontic treatment. These commercially available guttapercha cones have several advantages: they do not stain the tooth structure; they are biocompatible, radio opaque, dimensionally stable, easily removed from root canal and are also antibacterial. 1 Even though gutta-percha cones are produced under aseptic conditions and possess potential antimicrobial properties, especially due to their zincoxide component, they can still be contaminated by handling, aerosols and physical sources during the storage process. 7,8 Montgomery in 1971 found that 8% of the commercially available cones when tested showed bacterial growth. 3 Because it is difficult to determine the number of accessory cones to be used during lateral condensation before initiating the procedure, an effective, quick-acting chemical agent is needed to prevent any possible surface contamination by microorganisms. 9,10,11,12,13 The selection of chemical agents for decontamination of gutta percha cones in the present study was based on literature data. 4,9,12,14 Sequeira et al studied the effectiveness of 4 chemical solutions in eliminating B. subtilis spores on guttapercha cones. The solutions tested were 5.25% Sodium hypochlorite, 2% Glutaraldehyde, 2% Chlorhexidine gluconate and 70% Ethyl alcohol. The results showed that 5.25% Sodium hypochlorite was effective in destroying the spores after one minute of immersion time. 1 The study done by Cardoso, Brenda and Nurban ozalp also reveals that Sodium hypochlorite was more effective in sterilization of gutta-percha cones when compared to chlorhexidine and glutaraldehyde. 7,8,9 The antibacterial activity of sodium hypochlorite is mainly due to hypochlorous acid (HClO) in solution which has oxidative action on sulphydryl groups of bacterial enzymes. 12 The combination of Chlorhexidine 1.5% and 15% Cetrimide(group 5) also effective in sterilizing the gutta percha points. 1,4 In the present study 5 % sodium hypo chlorite (group2) and combination of Chlorhexidine 1.5% and 15% Cetrimide(group 5) found to be effective in decontaminating gutta-percha cones in less time. Povidone iodine 5% (Group 1), Ethyl alcohol 95% (Group 3) and Hydrogen peroxide 3% (Group 4) were also found to be effective in sterilizing guttapercha cones, but they took more time compare to other groups (Group 2 and group 5) in this study. In a study evaluating the storage conditions of guttapercha cones, demonstrated that the risk of contamination at the time of opening the containers with sterile gutta-percha cones in a dental clinic was not a source of concern. 10 Our results support these findings and emphasize that simple exposure of the cones to the environment is not of critical importance, but that their handling must respect the basic principles of infection control. Conclusions : Even though gutta-percha cones are usually sterile during storage, they can be easily contaminated if incorrectly manipulated. This study found a 5% concentration of NaOCl and combination of Chlorhexidine 1.5% + 15% Cetrimide to be an effective agent in disinfecting contaminated guttapercha cones at no additional costs. 76

5 CHAIR SIDE DISINFECTION OF GUTTA - PERCHA POINTS - AN IN VITRO COMPARATIVE STUDY BETWEEN 5 DIFFERENT AGENTS AT DIFFERENT CONCENTRATIONS References 1. Siqueira JF Jr., da Silva CH, Cerqueira M das D, Lopes HP, de Uzeda M. Effectiveness of four chemical solutions in eliminating Bacillus subtilis spores on gutta-percha cones. Endod Dent Traumatol 1998; 14: Senia ES, Marraro RV, Mitchell JL. Cold sterilization of gutta-percha cones with formocresol vapors. J Am Dent Assoc 1977; 94: Steve Montgomery. Chemical decontamination of guttapercha cones with polyvinylpyrrolidone-iodine. Oral Surgery 1971; 31: Stabholz A, Stabholz A, Friedman S, Helling I, Sela MN. Efficiency of different chemical agents in decontamination of gutta-percha cones. Int Endod J 1987;20: Linke HAB, Chohayeb AA. Effective surface sterilization of gutta-percha points. Oral Surg Oral Med Oral Pathol 1983;55: Senia ES, Marraro RV, Mitchell JL, Lewis AG, Thomas L. Rapid sterilization of guttapercha cones with 5.25% sodium hypochlorite. J Endod 1975; 1: Nurban ozalp, Zeynep okte, Berrin ozcelik. The Rapid Sterilization of Gutta-Percha Cones with Sodium Hypochlorite and Glutaraldehyde.J Endod 2006;32: Brenda Paula Figueiredo de Almeida Gomes, Morgana Eli Vianna, Carolina Ujissato Matsumoto, Vanessa de Paula e Silva Rossi, Alexandre Augusto Zaia, Caio Cezar Randi Ferraz, Francisco Jose de Souza Filho. Disinfection of Gutta-percha cones with Chlorhexidine and Sodium hypochlorite. Oral Surg Oral Med Oral Patho. Oral Radiol Endod 2005; 100: Cardoso CL, Kotaka CR, Redmerski R, Guilhermetti M, Queiroz AF. Rapid decontamination of gutta-percha cones with sodium hypochlorite. J Endod 1999; 25: Da Motta PG, de Figueiredo CB, Maltos SM, et al. Efficacy of chemical sterilization and storage conditions of guttapercha cones. Int Endod J 2001;34: Lui JN, Sae-Lim V, Song KP, Chen NN. In vitro antimicrobial effect of chlorhexidine-impregnated gutta percha points on Enterococcus faecalis. Int Endod J 2004;37: Gomes BP, Vienna ME, Matsumoto CU, et al. Disinfection of gutta-percha cones with chlorhexidine and sodium hypochlorite. Oral Surg Oral Med Pathol Oral Radiol Endod 2005;100: Frank RJ, Pelleu GB Jr. Glutaraldehyde decontamination of gutta-percha cones. J Endod 1983;9: Pennanchin R, Alvares S. Decontamina-tion of gutta percha cones by ethyl alcohol. Rev Poul Endod.1981; 2:

Evaluation and Comparison of Three Chemical Agents with a New Herbal Agent for Disinfection of Gutta-percha Cones: An in vitro Study

CEJ cej Evaluation and Comparison of Three Chemical Agents with a New Herbal Agent for 10.5005/jp-journals-10048-0009 Disinfection of Gutta-percha Cones Research Article Evaluation and Comparison of Three