Sterilization Infected Root Canal Dentin by Tri-mix

8/4/2019 Sterilization Infected Root Canal Dentin by Tri-mix

1/8

International Endodontic Journal {1996) 2 9 , 1 1 8 - 1 2 4

Sterilization of infected root-canal den tine by topical applicatioof a m ixture of ciprofloxacin, metronidazole and minocyclinein situI . SATO**, N . ANDO-KURIHARA**, K. KOTA**, M. IWAKU** &E. H O SH I N O * t ,*Cariology Research Unit, fDepartment of Oral Microbiology and tDepartment of Operative Dentistry and EndodontiNiigata University School of Den tistry, Niiga ta, Japan

Summ ary ; , .The aim of this study was to observe the potential of amixture of ciprofloxacin, metronidazole and minocy-cline to kill bacteria in the deep layers of root canaldentine in situ. After the crowns of extracted teeth hadbeen removed, the drug combination (0.5 mg of eachdrug), or sterile saline, as the control, was placed in theroot canals which had been previously irrigated ultra-sonically with 0. 4 M EDTA. The penetration a nd bacteri-cidal efficacy were estimated by various procedures asfollows. (1) A cell suspension of E. coli w as placed intosmall cavities prepared parallel to the root canals on thecut planes of nine single-rooted teeth. The teeth werethen entirely covered with blue inlay wax. At time 0,and at 5h, 2 4 h and 48 h after the drug combination hadbeen applied, cells of E. coll were recovered from thecavities by w ash ing the cavities several times with sterilesaline solution, and were cultured on the surfaces ofheart-infiision (HI) agar plates. Total colony-foriningunits were tuen counted. Bacterial recoveries decreasedwith time, and no bacteria were recovered 4 8 h afterapplication of the drug combination, while bacteriasurvived in all cases with th e controls. (2) After the drugcombination or sterile saline had been placed into andsealed in the root canal with blue inlay wax, the teethwere placed into HI agar plates where cells of E. coli ha dbeen inoculated. After culturing, a clear zone caused bythe inhibition of bacterial growth was observed aroundthe teeth, but not in the control experiment. (3) Aftersampling infected root dentine of 12 freshly extractedteeth as positive controls, the drug combination (0.5 mgeach) was placed in the root canals. No bacteria wererecovered from the infected dentine of the root canalCorre sponde nce: Professor Hoshino Etsu ro, Deptirtm ent of OralMicrohiology, Niigata Un iversity School of Dentistry, G akkocho-dori 2,

wall 24 h after application of the drug combinaexcept in one case in which a few bacteria were rered. On the basis of these results, penetration thrdentine and antibacterial efficacy of the drug comtion can be expected against bacteria infectingdentine of the root canal wall in situ when the dwere placed in root canals which had been irriultrasonically.Keywords: ciprofloxacin, disinfection, infected dentine, metronidazole, minocycline

Introduction ~-Bacteria present in root canals may be removed byor by chemical irrigation during conv entional root trea tment. However, bacteria in the deeper layeinfected root dentine may sometimes remain evenconventional root ca nal treatm ent (Ando & Ho1990), and may occasionally cause periapicai comtions (Yamata 1973, BystromeUi. 1987). Such bashould be eliminated to ensure a successful outcVarious medicam ents, including non-specific antisand antibiotics, have been used in root canal trea tmand each of them has both advantages and disatages (Grossman 19 65 , 1 972 , Haapasalo & Or1987, Morse 1987, Seltzer 1988, Abbott et al 1Safavi et al 199 0, Spangberg 199 4). Besides the unon-specific antiseptics (Haapasalo & Orstavik 1Safavi et al 199 0), the application of antibacterial may represent a way to eradicate bacteria duringcanal treatment. The selection of these antibacdrugs should be based on th e m ost up-to-date micrlogical know ledge.

With the technology for culturing obligate anaerincluding the use of an anaerobic glove box, ba


2/8

Sterilization of root-canal dentine 11

& Hoshino 199 0). Metronidazole has aet al 1975), even against

et al, 1988) and infected rootal, 199.') were not recovered in the

xg ml~ i m etronidazole in in-vitro experi-mi"^, could not kifl afl the bacteria

al 199.?), indicating th at oth er drug s may be

The aim of the present study was to observe thede ntin e of infected n ecro tic pulp s in situ,

als and metho dsorganisms

eoli strain NIHJ-JC-2 was obtained from the

al conce ntra tion of 10^ cells m l-^ ). The cells of E, coli

xg ml -i each).

; C, v i,The crowns of 14 single-rooted

depth and 1 m m in diameter were prepared on the cu t

4M EDTA solution (pH 7.6). ^ ^ ^ '- '-

1 mm3 mm _ ^_ 1 mm

Fig. 1 Preparation of artificial cavities. The crown of the tooth waremoved. Four cavities of depth 3 mm and diameter 1 mm werprepared a t sites 1 mm apart from and parallel to the root canal on thcut surface.

Sttrvival of test organisms. The suspensions of E. coli (1 Jwere placed into the cavities, and a mixture of ciprofloxacin, metronidazole and minocycline (0,5 mg eachor sterile saline as the control, was placed into the roocanals . The teeth were then completely covered m^tblue inlay wax. At time 0, and at 5h, 24h and 48h aftethe drug combination had been applied, cells oE. coli were recovered from the cavit ies by washing thcavities several tim es wi th sterile saline. Organism s w ercultured on the surfaces of HI ag ar p lates for 1 week. Thtotal numb er of colony-forming un its was determineand moni tored after incu batio n for 3 weeks.Experimental procedure 2Penet ra t ion of the drug mixture from the root canal tthe dent ine-cementum junct ion through the dent inatubules was also determined. For this experiment, throot surfaces were carefully checked for cracks ancollateral bran ches of root canals . Following removal othe crown and cemen tum, the root canals were carefullprepared as described previously. A mixture of ciprofloxacin, metronidazole and minocycline (0.5 mg eachwas placed in the root canals after the ultrasonic treament and the openings of the root canals were closewith blue inlay wax. To avoid leakage of the drugth rough apical ramifications, the apical one-fifth of throot surface was also covered with blue inlay wax. Tallow the drug combination to penetrate into thdentine, the teeth were allowed to stand for 6h. Thewere then placed on HI aga r plates on which E. coli w ainoculated. The plates were incubated for 24h at 37CPreliminary experiments had shown that E. coli grewwell on HI agar plates. As a control , the drug combination was replaced by ster ile sa lin e. Jf=f -it .+ ; !' ;: .


3/8

procedure 3extracted teeth with infected root dentine, which

afl single-rooted teeth and extracted because of14 patients. The

and notches were made on thea sterile diamond disc in order to maketo split the root. The root canals were enlargedultrasonically as described previously.into an anaerobic glove boxAZ-Hard, Hirasawa, Tokyo, Japan) containing% Nj, 10% H2 and 10% CO2, While in the box, thewas split in half with forceps. Infected root dentine

to represent time 0 from the split surface of(Fig. 2a), using a sterile excavator or a dentalr at low speed (


4/8

sterilization of root-canal dentitie 1 2Bacterial recovery by experimen tal procedure 1*

^ -

Time-0

5.435.205.935.625,465.495.115.865.795.595.935.005.115.72

After5 h 24 hlog (colony-forming units m g'M

5.005.304.864.934.085.114.584.304.665.625.895.265.685.36

2.302.002.002.00002.30004.784 . 935.664 . 344 . 70

4 8 h

0**000000002 . 302.455.644 . 755.18

Antibacterial efficacy of the drug combination in situagainst bacteria in infected root dentine

E. coli from cavities prepared artificially 1 mm from the

-13 ) w as placed. The root canals were treated (samples 1-9) or notNo bacter ia were recove red. ' ^

erent from th ose of time-0 sam ples (paired t-test; P< 0.0 00 5. while

Substantial numhers of bacteria (lOi-lOS) occurred inthe time-0 samples taken from the infected root canadentine (Table 2; samples 1 to 12). However, none warecovered after a 24-h application of a mixtu re (0.5 mgeach) of ciprofioxacin, metronidazole and minocyclin(Table 2; samples 1 to 3 and 5 to 12), except for one cas(Table 2; sample 4), from which 21 hacteria were recovered even after a 24-h app lication of the d rug comhination. However, all the isolates from the sample wersensitive to the drug com bination, because they did nogrow on BHI-hlood agar plates in the presence of thdrug mixture (25 |xg ml~i each). This may indicate th athe bactericidal efficacy of the topically applied drumixture is potent enough to sterilize root dentine withi48h.Discussion , , . > > ,Bacteria in the main root canals and superficial layers oinfected root canal walls may be easily removed bconventional root can al trea tment. Bacteria, whicrema in in the deep layers of root canal den tine, may leaout to periapical regions and cause complication(Yawata 1973, Bystrom et al 1987). It has bee

!], :;;.%/-H

Fig 3 The inhibitory zone of bacterial growth, (a) surface view of the inhibition, (b) The transversesection. . , .


5/8

2 I Sato etal 'Bactericidal efficacy of a mix ture of ciprofioxacin. m etronid a-

minocy cline in si(u against bacteria of infected root dentin e

p le No . : .

"

'J . Tim e.-0After

24 hlog (colony-forming units mg^i)

3.761.513.713.383.563.204 . 7 6 '2.302.235,983.575.762.384.15

0*001 (21 colonies)0 :000 ':00 . . - ;002.771.15

bacteria were recovered.erile saline wa s applied to the root canal canals.

after 2 4 h (samples 1-12) were significantlyof t ime-0 samples (paired t-test: P


6/8

Sterilization of root-canal dentine 12This research demonstrates that a mixture of

metronidazole and minocycline is usefulr sterilization of infected root dentine, and that themixture can be applied to root canals. The topical

needs only a low dose of the mixture, and isfor only a short period, e.g. 2 days (Tables 1 &

Thus any adverse systemic side-effects could bealthough, as a matter of principle, the appli-of antibiotics should be limited if possible.Minocycline sometimes causes pigmentation,incalcifying teeth, so the bactericidal efficacy

the mixture of ciprofloxacin and metronidazole plus(Mixed-drug I; Sato T et al 1993b), cefaclorII), cefroxadine (Mixed-drug III), fos-IV) or rokitamycin (Mixed-drug V)

been compared and it has been found that thesedrug combinations (100 |xg ml'i each) were able tocarious and endodontic lesions (Sato et alHoshino e a/. 199.?),

authors thank Dr Takuichi Sato, Department of OralNiigata University School of Dentistry, forassistance with the preparation of the manuscript.

investigation was supported in part by TheMinistry of Education, Science, Sport and

under Grants-in-aid for Scientific Research03404055 and 05557092),

(1990) Antibiotics and endodon-tics. AustraliatiDetital Journal 35. 50-60.

N , HOSHINO E (1990) Predominant obligate anacrohes invadingthe deep layers of root canal dentine. Itttertiational Etidodontie Journal23,20-27.

G (1974) Micro-organisms from necrotic pulp oftraumatized teeth. Odontologisk Revij 2S, 347-58.

A. HAPPONEN R- P. SIOCREN U. SUNDQVIST G (1987) Healingof periapicai lesions of pulplcss teeth after endodontic treatmentwith controlled asepsis. Endodontics and Detttal Trawnatologij 3,5 8 - 6 3 .

CAMERON JA (1983) The use of ultrasonic in the removal of the smearlayer: a scanning electron microscope study. Jourtial ofEndodotics 9,289-92.

CsuKAs Z, FERENCZI I, NASZ I. BANOCZY J (1987) Diffusion of metron-idazole through the dentinal tuhulcs of extracted teeth. ActaMicrobiological Hutigarica 34,121-4.

EDWARDSSON S (1974) Bacteriological studies on deep areas of cariousdentine. Odotitoloijisk Revij 25 (Suppl. 32), 1-143.

GROSSMAN LI (1965) Etidodotitic Practice. 6th edn. Philadelphia: Lea &Fehiger, pp. 241-302.

GROSSMAN LI (1972) Sterilization of infected root canals. Journal of theAmerican Dental Association 85, 900-905.

HAAPASALO M , ORSTAVIK D (1987) In vitro infection and disinfectionof dentinal tubules. Journal ofDental Research 66. 1375-9.

HOLDEMAN LV, CATO EP. MOORE WEC (1977) Anaerobe LaboratoManttal, 4th edn. Blacksburg: Virginia Polytechnic Institute andState University.

HOSHINO E (1985) Predominant ohligate anaerobes in human carioudentin. Jotirnal of Dental Research 64. 1195-8.

HOSHINO E (1990) Sterilization of carious lesions by drugs. Journalthe Japanese Association for Detital Science 9, 327 (in Japanese).

HOSHINO E, SATO MICHIKO (1988) Predominant microorganisms oplaque on complete dentures. Jotwnal of the Japan ProstlwdonSociety 32, 763-6.

HOSHINO E. KOTA K.SATO MICHIKO. IWAKU M (1988) Bactericidaefficacy of metronidazole against bacteria of human carious dentiin vitro. Caries Research 22, 280-82.

HOSHINO E, IWAKU M. SATO MICHIKO. ANDO N, KOTA K (1989aBactericidal eflicacy of metronidazole against bacteria of humacarious dentin in vivo. Caries Research 23, 78-80.

HOSHINO E, SATO MICHIKO, SASANO T, KOTA K (1989bCharacterization of bacterial deposits formed in vivo on hydrogenion-sensitive field-effect transistor electrodes and enamel surfaceJapanese Jourtial ofOral Biology 31, 102-6.HOSHINO E, KOTA K, IWAKU M (1990) Sterilization of carious lesions hantibacterial drugs. New attempt to conserve pulp. (Part 1). Tbasic approach. Dental Outlook 75, 1379-86 (in Japanese).

HOSHINO E, ANDON, SATO MICHIKO. KOTA K (1992) Bacterial invasioof non-exposed dental pulp. Intemational Endodontie Journal 22 - 5 .

IMAMURA M, MAC, ATA T, SAITO F. KANEKO D, KOTA K, IWAKU M (198The cleaning effects on the root canal by the ultrasonic devicJapatiese Jourtial ofCotiservative Detististry 32, 769-77 (in Japan

INCHAM HR, SELKON JB, HALE JH (1975) The antibacterial activity ometronidazole. Journal ofAtitiniicrolml Chemotherapy 1,355-61.

KiRYU T, HOSHINO E, IWAKU M (1994) Bacteria invading periapiccementum. Journal ofEtidodotitics 20, 169-72.

MORSE DR (1987) Microbiology and pharmacology. In: Cohen SBurns RC. eds. Pathways of the Pulp,4th edn. St Louis: C.V. MosbCompany, pp. 3 8 0 - 8 1 .

SAFAVI KE, SPANGBERG LSW. LNGELAND K (1990) Root canal dentintubule disinfection. Jorutml ofEndodontics 16, 207-10.

SATO MAKIKO, HOSHINO E, NOMURA S, ISHIOKA K (1993) Salivamicroflora of geriatric edentulous persons wearing dentureMicrobial Ecology in Health attd Disease 6,293-9.

SATO T, HOSHINO E. UEMATSU H, KOTA K, IWAKU M, NODA T (199Bactericidal eflicacy of a mixture of ciprofloxacin, metronidazolminocycline and rifampicin against bacteria of carious anendodontic lesions of human deciduous teeth iti vitro. MicrobEcology in Health and Disease 5,171-7.

SATO T, HOSHINO E, UEMATSU H. NODA T (1993a) Predominaohligate anaerobes in necrotic pulps of human deciduous teetMicrobial Ecology in Health and Disease 6, 269-75 .

SATOT, HOSHINOE, UEMATSU H, NoDAT(1993h) In Wtroantimicrohsusceptihility to comhinations of drugs of bacteria from carioendodontic lesions of human deciduous teeth. Oral MicrobiologyInnnunology 8,172-6.

SELTZER SBA (1988) Microbiological aspects of endodontics. In: SeltzSBA, Farher P, eds. Endodontology, 2nd edn. Philadelphia: LeaFcbiger, pp. 326-56.

SFANGBERG L (1994) Intracanal medication. In; Ingle JL Bakland Leds. Etidodotitics, 4th edn. Malvern: Williams & Wikins. pp. 627-

SuGiTA El (1994) Microbiology of endodontics. In: Ingle JL BaklaLK, eds. Endodontics, 4th edn. Malvern: Willimans & Wikipp . 608-26.


7/8

124 I. Sato et al.SuNDQViST G (1976) Bacteriological Studies of Necrotic Dental Pulps.

Umea. Sweden: Umea University Odontologisk Dissertations.UEMATSU H. HOSHINO E (1992) Predominant obligate anaerobes in

human periodontal pockets. Journal of Periodontal Research 27,15-19.

WITTGOW WC, SABISTON CB (1975) Microorganisms from pulpalchambers of intact teeth with necrotic pulps. Journal ofEndodontics1,168-71.

YAWATA S (1973) On the reliability of endodontic culturing.Histobaeterial examination of the root canal and the dentin

surrounding the root canal which obtained endodontic neculture. Japanese Journal ofConservative Dentistry 16. 79Japanese).

YosHiuA M. FuKUSHiMA H. YAMADA K. OGAWA K. TODA T. SAGA(1987) Correlation between clinical symptoms and microorgaisolated from root canals of teeth with periapical pathosis. JouEndodontics 13, 24^-8.

ZAVISTOSKI K, DZINK J, ONDERDONK A, BARTLETT J (1Quantitative bacteriology of endodontic infections. Oral SuOral Medicine and Oral Pathology 49. 171-4.


8/8

Documents

Sterilization Infected Root Canal Dentin by Tri-mix