Evaluation of Antimicrobial Efficacy of Antibiotics and Calcium Hydroxide against Enterococcus faecalis Biofilm in Dentine

8 pages
17 views

Please download to get full document.

View again

of 8
All materials on our website are shared by users. If you have any questions about copyright issues, please report us to resolve them. We are always happy to assist you.
Share
Description
The objective of this study was to investigate the antimicrobial efficacy of erythromycin, oxytetracycline and calcium hydroxide [Ca(OH)(2)] against Enterococcus faecalis biofilm in dentine. E. faecalis ATCC 29212 (American type culture collection) was inoculated into standard tooth sections and incubated in aerobic atmosphere at 37 degrees C for 21 days. The infected tooth sections were then exposed to the test agents for 5 and 10 min. The colony forming units (CFU) after the exposure periods at three different depths
Tags
Transcript
  Sains Malaysiana 42(1)(2013): 73–80 Evaaton of Antcroba Efcacy of Antbotcs and Cac Hydroxde against  Enterococcus faecalis Bo n Dentne (Kajan Keberkesanan Antbotk Antkrob dan Kas Hdroksda ke AtasBoe  Enterococcus faecalis d Daa Dentn) W . L . C HAi*, H . H AmimAH & M . A BDullAHABSTRACT The objective of this study was to investigate the antimicrobial efcacy of erythromycin, oxytetracycline and calciumhydroxide [Ca(OH) 2 ] against  Enterococcs faecas biolm in dentine. E. faecas ATCC  29212 (American type culture collection) was inoculated into standard tooth sections and incubated in aerobic atmosphere at 37°C for 21 days. Theinfected tooth sections were then exposed to the test agents for 5 and 10 min. The colony forming units ( CFU  ) after the exposure periods at three different depths <100 µm, 100-350 µm and 350-500 µm were enumerated. After 5 min of exposure, both antibiotics had signicantly lower CFU  count than Ca(OH) 2 solution at three dentinal depths. Comparingwith the oxytetracycline, the CFU  count of the erythromycin was signicantly ( p <0.05) lower at the depth of 100-500 μm. Similarly, after 10 min of exposure, erythromycin had signicantly lower CFU  count ( p <0.05) at three dentinaldepths. Oxytetracycline showed signicantly lower CFU  count than Ca(OH) 2 at 100 μm depth. Comparing with the twoexposure times, the erythromycin and Ca(OH) 2 groups showed signicant lower CFU  counts after 10 min of exposure in the antimicrobial agents to 5 min. In conclusion, both antibiotics show better antimicrobial activity than Ca(OH) 2 inremoving the E. faecas biolm in dentine.Keywords: Antimicrobials; biolm; calcium hydroxide; dentinal tubules; Enterococcs faecas ABSTRAK Tujuan kajian ini adalah untuk mengkaji keberkesanan antimikrob eritromisin, okstetrasilisin dan kalsium hidroksida [Ca(OH) 2 ] ke atas biolem Enterococcs faecas di dalam dentin. E. faecas    ATCC  29212 (American Type CultureCollection) diguna untuk menjangkiti keratan piawai akar gigi manusia dalam keadaan atmosfera aerobik pada 37°C  selama 21 hari. Keratan akar gigi yang telah dijangkiti didedahkan kepada ejen ujian selama 5 dan 10 min. Selepas tempoh pendedahan, kiraan unit pembentukan koloni ( CFU  ) pada tiga kedalaman dentin, iaitu <100 µm, 100-350 µm dan 350-500 µm yang berbeza dibandingkan. Selepas pendedahan selama 5 min, kedua-dua jenis antibiotik didapati mempunyai kiraan CFU  yang kurang namun signikan berbanding Ca(OH) 2 pada ketiga-tiga kedalaman dentin. Berbanding denganokstetrasilisin, eritromisin menunjukkan kiraan CFU  yang jauh lebih kurang berbanding okstetrasilisin dan signikan ( p <0.05) pada kedalaman dentin 100-500 μm. Selepas pendedahan selama 10 min, eritromisin mempunyai kiraan CFU     yang paling rendah dan signikan ( p <0.05) pada ketiga-tiga kedalaman dentin.   Okstetrasilisin menunjukkan kiraan CFU    yang ketara kurang daripada Ca(OH) 2 pada kedalaman 100 μm. Dalam perbandingan masa pendedahan, eritromisin dan Ca(OH) 2 menunjukkan CFU  yang ketara kurang selepas pendedahan selama 10 min berbanding dengan 5 min. Secarakesimpulan, kedua-dua antibiotik menunjukkan aktiviti antimikrob yang lebih berkesan daripada Ca(OH) 2 terhadap biolem E. faecas pada dentin.Kata kunci: Antimikrob; biolem; kalsium hidroksida; Enterococcs faecas ; tubul dentin i NTRODuCTiON  Enterococcus faecalis (  E. faecalis ), a factatve anaerobe,has been coony soated fro persstent root cananfectons and post-treatent nfecton (moander et a.1998; Sndqvst et a. 1998). One of the vrence factorsassocated wth ths persstent presence of   E. faecalis in root canas s ts abty to nvade dentna tbes, adhereto coagen n the dentne (love 2001). An in-vitro stdyhas shown that abet wth the presence of ntracanaedcatons, the  E. faecalis was abe to for bo n theroot canas (Dste et a. 2002). Bo s a hghy organzedstrctre wth cps of bactera bond together by acarbohydrate atrx (Costerton et a. 1994). in addton,the  E. faecalis has the abty to srvve wthot dvdngn the bo (Dste et a. 2002) and ndced the apattereprecptaton, especay n a atre bo (Kshen et a.2006). Hence, these featres expan the ow ssceptbty of   E. faecalis bo towards severa antcroba agentswhen copared to ts other orphotypes sch as thepanktonc and peet fors (Abdah et a. 2005). The  74 poysaccharde coatng of the bo cod be the barrerto ost antcroba agents (Dste et a. 2002).Severa stdes have shown that the  E. faecalis is hghy resstant to the antbactera effect of cachydroxde [Ca(OH) 2 ] (Haapasao & Orstavk 1987;Orstavk & Haapasao 1990; Sqera & de uzeda 1996).Evans et a. (2002) reported that  E. faecalis can srvven an akane envronent as hgh as pH11.1 and codony be ked when pH reached hgher than 11.5. However,the pH of the Ca(OH) 2 n the cana cod ony reached pto 10.3 de to the bfferng effect of the radcar dentn(Evans et a. 2002).Knowng the taton of the standard ntracanaedcaton [Ca(OH) 2 ], varos ethods sch as the seof aser (mehrvarzfar et a. 2011; Sahar-Heft et a. 2011),trasonc (Grndng et a. 2011), dfferent nstrentatonsystes (Gordyss et a. 2011; matos Neto et a. 2011)for the eradcaton of   E. faecalis have been reported. Asost root cana nfectons are of bactera orgn, t snot srprsed that there were new antcroba agentscontanng antbotc sch as the mTAD , a xtre of tetracycne soer, an acd and a detergent (Tong et a.2011; Torabnejad et a. 2003) and Tetracean, a xtre of doxycycne, ctrc acd and poypropyengycoe (Gardnoet a. 2006; Nega et a. 2008) have been deveoped.The presence of antbotcs n these new generatons of antcroba agents sggests that the antbotcs ay payan portant roe n the root cana therapy.Severa stdes have nvestgated the effcacy of varos types of antbotcs sch as cndaycn (Gadet a. 1999; la et a. 2001; ln et a. 2003; moanderet a. 1990), erythroycn (moander & Dahen 2003),etrondazoe (la et a. 2001), tetracycne (Gardnoet a. 2006; ln et a. 2003; moander & Dahen 2003;Tong et a. 2011; Torabnejad et a. 2003) and a xtreof cprooxacn, etrondazoe and nocycne (Hoshnoet a. 1996). in addton, the appcaton of antbotcs ndfferent fors, sch as paste (Hoshno et a. 1996; lnet a. 2003; moander & Dahen 2003; moander et a.1990; Sqera & de uzeda 1996), ge (la et a. 2001),soton (Gardno et a. 2006; Nega et a. 2008; Tonget a. 2011; Torabnejad et a. 2003) or pregnated n adevery vehce (Gad et a. 1999)   have been nvestgated.it was sggested that when the antbotcs were appedocay nto the nfected root canas nstead of adnsteredsysteatcay, t wod redce the rsk of adverse systeceffects (mohaad 2009). The a of ths stdy was toevaate the antcroba effectveness of oca appcatonof two antbotcs and Ca(OH) 2 against  E. faecalis bon dentn. M ATERiAlS AND M ETHODS The ethod of ths stdy was oded fro a prevosstdy (Haapasao & Orstavk 1987). Extracted hananteror teeth (excdng ower ncsors) were dsnfectedovernght n 0.5% sod hypochorte (NaOC). Theapca 5  of the roots was reoved sng a rotatngdaond saw at 1000 rp (isoet® 1000 PrecsonSectonng Saw, Beher ltd., uSA ) nder water coong(Cooet TM , Beher ltd., uSA ). Sbseqenty, the rootswere ct transversey nto sces of 4  thck. The nternadaeter of the root cana was shaped to a standardzedsze of 1.4  wth an iSO sze 014 (No. 4) rond stee bur ( ASH , Aagaated Denta Trade Dstrbtors ltd,Engand) rn n a sow-speed contra-ange handpece. Theceent ayer of the tooth sectons reaned ntact.The sear ayer of the root cana was reoved bytreatng the tooth sectons n an trasonc bath of 17% EDTA (Ethyenedanetetraacetc acd dsod sat, R& m marketng, uK ) (pH7.8) foowed by 5.25% NaOCfor 4 n each. The tooth sectons ersed n bran-heart infusion broth ( BHi ) ( BHi , Schara Chee SA , Span) werethen sterzed n an atocave at 121°C for 20 n. Thesterty of the tooth sectons was checked by observngany changes of the codness of the broth soton after a24 h of ncbaton n an aerobc ncbator.The stere tooth sectons were then nocated wth 1 × 10 8   CFu /mL  E. faecalis   ATCC 29212 (Aercan typectre coecton) and ncbated nder aerobc condtonat 37°C for 21 days. At the end of the 21 days of ncbatonperod, the tooth sectons were washed 3 tes wth freshstere PBS to reove excess ed and panktonc forof bactera. The nfected tooth sectons were dvdednto 6 grops wth 12 tooth sectons n each grop. Theywere exposed to 10 l of the foowng test agents andncbated n an aerobc ncbator at 37°C: Grop 1 & 2tooth sectons were exposed to cac hydroxde soton[Ca(OH) 2 ] (Cac hydroxde p.a., merck, Gerany) atpH12.3 for 5 and 10 n, respectvey; grop 3 & 4 toothsectons were exposed to erythroycn (Erythroycnlactobonate iV, Abbott laboratores, uSA ) for 5 and 10n, respectvey. The erythroycn soton was preparedby xng the 500 g erythroycn powder wth 10 lof water for njecton and Grop 5 & 6 tooth sectons wereexposed to oxytetracyne (Oxy, Atantc laboratoresCorp ltd., Thaand) for 5 and 10 n, respectvey. Theoxytetracyne was avaabe as 500 g n 10 l sotonfor. in each grop, two tooth sectons were sed as acontro, n whch they were exposed to stere PBS for 5and 10 n, respectvey.At the end of each exposre te, the antcrobaactvtes of the test agents were ceased by washng thetooth sectons wth 10 l of stere PBS for 5 tes. Thedentne sapes of the nner srface of the tooth sectonswere coected sng three dfferent daeters of sterestee rond brs [ iSO szes 016 (#5), 021 (#7), 023 (#8)]rn n a sow speed handpece. These brs excavated thedentne sapes at <100 μ, 100-350 μ and 350-500μ depth of root dentne, respectvey. After each drng,the dentne powder on the srfaces of the tooth sectonsand respectve brs was coected by rnsng wth 5 l of  BHi broth nto a stere botte. The bactera were seraydted and nocated on Coba horse bood agarpates (Boeda, utas maj Sdn. Bhd., maaysa) and    75 ncbated n aerobc atosphere at 37°C. The coony-forng nt per tooth secton were enerated after the24 h of ncbaton perod.To deterne the foraton of bo, two nfectedtooth sectons were prepared and exaned nder scannngeectron croscope ( SEM ). Brey, at the end of the21days ncbaton, the tooth sectons were xed n 4%gtaradehyde 0.1 m cacodyate bffer soton (AgarScentc ltd., unted Kngdo) and sbseqenty post-xed for 2 h n 2% os tetroxde (Agar Scentcltd., unted Kngdo). Dehydraton was perfored na seres of ascendng ethano concentraton (30%, 50%,70%, 95% and 100%) for 15 n each. The tooth sectonswere then frther dred n a crtca pont dryer (Poaron CPD 7501, Qor Technoogy ltd., unted Kngdo).Foowng that, the tooth sectons were spt nto two andsptter coated wth a ayer of 200Å god paad ( BiO-RAD , Bo-rad mcroscence Dvson, unted Kngdo)before exanaton nder SEM (Phps SEM 515, Phpsindstra & Eectro-acostc Systes, Netherands).The data were entered nto a Statstca Package forSoca Scence ( SPSS ) software (Verson 11.0; SPSS inc,Chcago, il , uSA ). One way ANOVA was sed to coparethe CFu conts of the test grops. independent t-test wassed to anayse the dfference of  CFu conts at the twoexposre tes. A  p -vae of <0.05 was regarded asstatstcay sgncant. R ESulTS There was evdence of coonzaton of   E. faecalis boon the tooth sectons nder SEM exanaton (Fgre 1).The bactera coones encopassed n a dense atrx,appeared as a shroo-shaped coones on the root canasrface were dented.An overa coparson of the antcroba efcacyof the three test agents after 5 and 10 n of exposre tewere shown n Fgres 2 and 3, respectvey.After 5 n of exposre te, the Ca(OH) 2 has thesgncant (  p <0.05) hghest reanng vabe bacteracont at the three dentne depths copared wth theerythroycn and oxytetracyne grops (Fgre 2).Ths sggests that the Ca(OH) 2 was the east effectveantcroba actvty aganst the  E. faecalis bo onthe tooth sectons copared wth the antbotc grops. incontrast, the erythroycn showed a better antcrobaactvty. it had the owest bactera conts and wassgncanty ower copared wth the other 2 test agents nthe dentne depth of 100-500 μ (brs #7 and #8) (Fgre 2).   Sary, after 10 n of exposre, the hghestand owest reanng vabe bactera conts werethe Ca(OH) 2 and erythroycn grops, respectvey(Fgre 3). The CFu conts of Ca(OH) 2 was sgncantyhgher than the erythroycn grop at a three dentnedepths (  p <0.05) bt was not statstcay dfferent when FiGuRE 1. A SEM vew of the root cana srface n an nfected tooth secton. Notea shroo-shaped coony of   E. faecalis ebedded n poysaccharde atrx   andpenetraton of bactera coony nto dentna tbe (Scae bar = 0.1 )  76 copared wth the oxytetracycne n deeper ayers (brs#7 & #8).in genera, the CFu cont was ower after a ongerexposre perod to the test agents (Fgre 4). There werestatstcay ower CFu conts after 10 n of exposren Ca(OH) 2 (Fgre 4(a)) and erythroycn (Fgre4(b)) grops, at sapes taken wth brs #7 and #5,respectvey. No statstcay dfferent of  CFu count in FiGuRE   2. mean og 10 ( CFu +1) of   E. faecalis at three dentne depths after 5 n of exposre te to three test agentsBr sze    m  e  a  n   l  o  g   (   C   F   u   +   1   ) Ca(OH)2 Erythromycin Oxytetracycne FiGuRE 3. mean og 10 ( CFu +1) of   E. faecalis at three dentne depths after 10 n of exposre te to three test agentsBr sze    m  e  a  n   l  o  g   (   C   F   u   +   1   ) Ca(OH)2 Erythromycin Oxytetracycne
Related Search
Similar documents
We Need Your Support
Thank you for visiting our website and your interest in our free products and services. We are nonprofit website to share and download documents. To the running of this website, we need your help to support us.

Thanks to everyone for your continued support.

No, Thanks