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Int Poster J Dent Oral Med 13 (2011), No. 2     15. June 2011

Int Poster J Dent Oral Med 2011, Vol 13 No 2, Poster 528

A comparative Evaluation of PH, surface tension and antimicrobial efficacy of Oxum, Vancomycin, Doxorubicin drugs vs. Sodium Hypochlorite – an in vitro study

Language: English
 

Authors:
Assoc. Prof. Dr. K. Madhu Varma, Dr. M. Sita Ram Kumar, Dr. Suresh P, Prof. Dr. Girija S Sajjan,
Department of Conservative Dentistry and Endodontics, Vishnu Dental College and Hospital, Vishnupur, Bhimavaram, West Godavari District, Andhra Pradesh, India

Date/Event/Venue:
15/01/2011
ICDRO World Congress 2011
Pune, India

Poster Award
"BEST POSTER" presentation in endodontic pavillion

Introduction

The presence of microorganisms and derivative toxins in the infected root canals cause the development of apical periodontitis. a number of studies (1,2) show that the endodontic infection is poly-microbial, and that in teeth with necrotic pulp it is represented by obligatory and facultative anaerobes, microaerophilic bacteria and fungi. An important stage in the successful endodontic therapy is the treatment of the microflora with endodontic irrigants, both during the biomechanical preparation of the root canal and between the visits. Sodium hypochlorite (naocl) in various concentrations is a frequently recommended endodontic irrigant in contemporary endodontics (3). Super oxidized water (sow) is an alternative root canal irrigant. during the last decade it has become widely used in endodonic practice as an effective antimicrobial means for irrigating the root canal(4,5,6). studies have shown that the local application of antibiotics (root canal irrigants)(7) within the root canal system may be a more effective mode for delivering such drugs than systemic routes of administration. the surface tension is defined as "the force between molecules that produces a tendency for the surface area of a liquid to decrease"(8). this force tends to limit the ability of the liquid to penetrate a capillary tube.the irrigants for endodontic use should have very low surface tension.the aim of the present study has been formulated on the basis of the above sources and previous results of our research.
 

Objectives

The aim in this study has been to evaluate PH, surface tension and antimicrobial efficacy of Oxum(SOW), Vancomycin, Doxorubicin drugs as potential root canal irrigants in comparison with Sodium Hypochlorite.
 

Material and Methods

Enterococcus faecalis, Streptococcus pyogenes , Candida abicans were obtained from the department of microbiology , Vishnu Dental College , Bhimavaram . samples were obtained and maintained by using nutrient agar medium for e.faecalis , c.albicans and serum agar medium for Streptococcus pyogenes. The test solutions vancomycin, doxorubicin, sodium hypochlorite and superoxidised water were prepared in distilled water with a 3% concentration (Fig 1). A single pure colony of the test organism from the nutrient agar is inoculated in 4-5ml of nutrient both and incubated at 37°c for 6-8hrs. An agar plate is cultured with the required number of organism by keeping 0.5 ml of broth on the surface of the agar medium by swabbing with sterile swab. The cultured plate were allowed to dry for 15 minutes at room temperature and four wells are made with a 6mm diameter in the agar with proper distance between the wells. The wells are numbered and required antibiotics were filled with the standard amount of test solution i.e (0.5 ml of test solution) and incubated at 37°c for 24 hrs. Around each well the inhibitory zones were measured (in mm) and results were recorded (Fig 5). The largest diameter was recorded in mm (Fig 4). The data was recorded and statistically analysed with f test .Colony counting procedure: In this study viable count procedure (or) pour plate method was used. The plates were incubated at 37°c for 24 hours (Fig 6). The colonies were calculated with colony counting meter ( Shakthi Scientific, India) (Fig 7) and the result was recorded by group and was statistically analysed. The ph of test solutions were measured using ph meter (L1 120, Elico,India) (Fig 2). The surface tension of the test solutions were analysed with stalagmometer (JSGW, India) (Fig 3).

Fig. 1: Materials used in the study Fig. 2: PH meter
Fig. 3: Stalagmometer Fig. 4: Vernier Callipers to measure inhibition zones
Fig. 5(I): Inhibition zones of Enterococcus faecalis Fig. 5(II): Inhibition zones of Streptococcus pyogenes
Fig. 5(III): Inhibition zones of Candida albicans Fig. 6(I): Colony forming units of Enterococcus faecalis
Fig. 6(II): Colony forming units of Streptococcus pyogenes Fig. 6(III): Colony forming units of Candida albicans
 
Fig. 7: Colony counter
 
 

Results

Table 1 present the means of the diameters of the activity zones /in mm/ of the studied irrigants in the cases of the different microorganisms. Table II shows the results of test for colony forming units. Table III shows the pH & surface tension values of the four test solutions. As table 1 shows, vancomycin 3%had the strongest influence against all microorganisms (Fig 8). This irrigant has its largest activity zone in the case of streptococcus pyogenes (26 mm) candida albicans(26 mm), and its smallest zone in the case of enterococcus faecalis (24mm) . Doxorubicin 3%had an intermediate impact on enterococcus faecalis(12mm) and streptococcus pyogenes(12mm). Sodium hypochlorite 3%has its largest activity zone in the case of enterococcus faecalis (14 mm) and its smallest zone in the case of candida albicans (6mm) . SOW 3%had the weakest impact on enterococcus faecalis(4mm) and candida albicans(2mm). However SOW 3% had an intermediate impact on streptococcus pyogenes. Statistical analysis using f test shows that there was statistically significant difference between 3% vancomycin and remaining three test solutions.

Agar plates
  E.FAECALIS S PYOGENES C ALBICANS
3% NaOCl 14 mm 8 mm 6 mm
3% SOW 4 mm 12 mm 2 mm
3% VANCOMYCIN 24 mm 26 mm 26 mm
3% DOXORUBICIN 12 mm 12 mm 4 mm
Tab. 1: Inhibition zones
Agar plates
  E.FAECALIS S PYOGENES C ALBICANS
Control 2600-2700 2500-2600 2200-2300
3% NaOCl 100-200 400-500 700-800
3% SOW 1700-1800 1200-1300 1900-2000
3% VANCOMYCIN 20-30 5-10 10-20
3% DOXORUBICIN 200-300 300-400 1000-1100
Tab. 2: Colony forming units
  pH surface tension (dynes/cm)
3% NaOCl 12.34 100.86
3% SOW 5.86 62.72
3% VANCOMYCIN 4.02 95.85
3% DOXORUBICIN 5.82 76.02
Tab. 3
Fig. 8(I): Statistical analysis of Enterococcus faecalis Fig. 8(II): Statistical analysis of Streptococcus pyogenes
 
Fig. 8(II): Statistical analysis of Candida albicans
 
 

Conclusions

Vancomycin is a glycopeptide antibiotic primarily acting against gram +ve bacteria. Vancomycin inhibit the synthesis of cell wall in sensitive bacteria(9,10).In the present study when applied to streptococcus pyogenes(26mm),candida albicans(26mm), enterococcus faecalis(24mm), 3%vancomycin produce the largest area of activity. Doxorubicin is a anthracycline antibiotic and an important antitumour agent. The compounds can intercalate with DNA, directly affecting transcription and replication(9,11). In the present study 3%doxorubicin has an intermediate impact on enterococcus faecalis(12mm) and streptococcus pyogenes(12mm). Sodium hypochloride has a wide range of antimicrobial activity. By using different concentrations of sodium hypochloride substantial decrease in bacterial count was achieved(3). Despite being highly effective it does not penetrate well into difficult to access segments of microcanal systems of dentin (8). In addition, it remains in the canal for short period of time. However it is assumed that the use of larger quantity of irrigant leads to extinguishing larger quantities of micro organisms difficulty. In the present study when applied to enterococcus faecalis this endodontic irrigant produced a largest area of activity (14mm) compared to streptococci pyogenes(8mm) and candidia albicans (6mm). Super oxidized water is an alternative root canal irrigant. During the last decade it has been widely studied in the endodontic practice as an effective antimicrobial means for irrigating the root canal. 3% Super oxidized water has demostraded the weakest impact against the tested microorganisms Enterococcus faecalis (4mm) candida albicans(2mm). 3% super oxidized water showed an intermediate impact on streptococcus pyogenes(12mm). However it should be noted that undiluted super oxidized water would have given better results.
Regardless of the intracanal irrigants the bacteria can never be completely eliminated from the root canal. In order to eradicate the bacterial cells from the root canal, the irrigant has to penetrate and reach them. The irrigants for endodontic use should have very little surface tension. The wettability of the solution governs the capability of the penetration both into the lateral canals and dentinal tubules. The results in this study showed that 3%sodium hypochlorite (100.86dynes/cm) had the highest surface tension followed by 3%vancomycin (95.85 dynes/cm), 3% doxorubicin(76.02dynes/cm) and 3%super oxidized water (62.72 dynes/cm).
3%Vancomycin showed more anti microbial efficacy on microorganisms (enterococcus faecalis ,streptococcus pyogenes, candida.albicans) when compared with 3%sodium hypochlorite, 3%doxorubicin and 3%super oxidized water.
 

Literature

  1. Vytaute P, Rasmute M, Estera B, Saulius D, Vygandas R: Microorganisms in root canal infections: A Review. Stomatologija, Baltic Dental and Maxillofacial journal, 2008 10:4-9.
  2. Christine S, Don b. C: Bacterial plasmids in the oral and endodontic microflora. Endodontic Topics 2004, 9, 37-51.
  3. Matthias R: root Canal Irrigants. J Endod 2006;32:389-398.
  4. Marais J. T. & Williams W. P: Antimicrobial effectiveness of electro-chemically activated water as an endodontic irrigation solution. International Endodontic Journal. , 2001, 34, 237-243.
  5. Marais J T: Cleaning efficacy of a new root canal irrigation solution: A preliminary evaluation. International Endodontic Journal, 2000, 33, 320-325.
  6. Landa .S, Gonza'lez .E, Guzma'n .S, Torresc .K, Gutierreza .A.A.: Microcyntm : A novel super-oxidized water with neutral ph and disinfectant activity. Journal of Hospital Infection (2005) xx, 1-9.
  7. Zahed M : An update on the antibiotic-based root canal irrigation solutions. Iranian Endodontic Journal 2008; 3:1-7.
  8. Luciano G, Emanuele A, Carlo B, Lia R,And Marco M: Surface tension comparison of four common root canal irrigants and two new irrigants containing antibiotic. J Endod 2006;32:1091-1093.
  9. Laurence.L.B, John.S.L,Keith.L.P 11th Edition The Pharmacological Basis of Therapeutics.
  10. Farhan. E. A, Essa. M : Antibiotic options for enterococcus faecalis infections. Pak J Med Sci July - September 2006 vol. 22 no. 3 286-290.
  11. Deborah R. D, James B. M, Christopher J. P , Kelly T, Elisabeth A. A, Abbie C. W, Brandie M. J, Barbara E. M, George M. W, and Bruce l. R: Enterococcus faecalis multi-drug resistancetransporters: Application for Antibiotic Discovery. J. Mol. Microbiol. Biotechnol. (2001) 3(2): 179-184.
     

Abbreviations

SOW: Super Oxidized Water
NaOCl: Sodium Hypochlorite
 

This Poster was submitted by Dr. K. Madhu varma.
 

Correspondence address:
Assoc. Prof. Dr. K. Madhu Varma
Vishnu Dental College and Hospital
Department of Conservative Dentistry and Endodontics
Vishnupur, Bhimavaram, West Godavari District
Andhra Pradesh, India
534202