Examinationof cytotoxicity and mutagenicity of AH26 and AHPlus sealers I. Miletic¤ 1 , S. Jukic¤ 1 , I. Anic¤ 1 , D. Zí eljezic¤ 2 ,V. Garaj-Vrhovac 2 &M.Osmak 3 1 SchoolofDentistry,UniversityofZagreb, 2 InstituteforMedicalResearchandOccupationalHealth,and 3 RuäerBos›kovic¤ Institute, Croatia Abstract Miletic¤ I, Jukic¤ S, Anic¤ I, Zí eljezic¤ D, Garaj-VrhovacV, Osmak M. Examination of cytotoxicity and mutagenicity of AH26 and AH Plus sealers. International Endodontic Journal , 36, 330^335, 2003. Aim To study invitro the cytotoxic and mutagenic e¡ectsofAH26andAHPlus. Methodology Cytotoxic e¡ects on Chinese hamster V79 cells were determined bycounting viable cells fol- lowing incubation with eluations of AH26 and AH Plus. In one set of experiments, the materials were mixed, set for 1h and then eluted with dimethyl sulphoxide(DMSO)for1h,24hand7days.Intheother set,AH26andAHPlusweremixedandsetfor1h,24h and 7 days in physiological saline then crushed and eluted in DMSO for 24 h.The cytotoxic e¡ects of these eluateswereevaluated.Threeconcentrationswerecho- sento examine the mutagenic e¡ects of AH26 and AH Plus:5.57,16.7and55.7 mgmL 1 .Thestructuralchromo- somal aberration analysis and micronucleus test were performed on human lymphocytes according to stan- dardprocedures. Results Dose^response curves of cell survival were obtained.Bothmaterialswereshowntobecytotoxicin doses larger than 55.7 mgmL 1 , except forAH26, after 7dayssettingtime.AHPluswasalsoshowntobetoxic in concentrations of16.7 mgmL 1 , except after 7 days settingtime.NeitherAH26norAHPlusinducedasigni¢- cant increase of chromosomal aberrations or micronu- cleiinductionatanysettingtimeorconcentration. Conclusion There was no mutagenicity found for AH26 and AH Plus on human lymphocytes in highly controlledconditions invitro. Keywords: AH26, AH Plus, chromosomal aberra- tions, human lymphocytes, micronucleus,V79. Received14June2002;accepted22November2002 Introduction Thepurposeofrootcanaltreatmentistoeliminateinfec- tionintherootcanalandto¢lltherootcanalspace.Var- ious commercial sealers have been developed and used for this purpose. One of them, AH26 sealer (Dentsply, DeTrey,Konstanz,Germany),isfrequentlyusedbecause ofitsexcellentsealingability(Wu etal .1995,Miletic¤ etal. 1999).Ithasbeendemonstrated,however,thatthesealer was cytotoxic duringsetting (Gerosa etal .1995) which can be, to some extent, explained by the release of for- maldehyde(Spangberg etal .1993,Koch1999).Amodi¢ed version of the material AH Plus (Dentsply) was subse- quentlydeveloped. According tothe manufacturer, AH Plus has better physical and clinical properties than AH26andtheformulationnolongerreleasesformalde- hyde. Root¢llingmaterialsareusuallyinclosecontactwith living tissues. Thus, the biological properties of these materials are important as cytotoxic materials can damageperiapicaltissues,andmaterialwithmutagenic potential can induce DNA mutations, possiblycausing malignant transformation of the cells (Bertram 2001). Various tests have been developed for determination of mutagenicity.Themostcommonlyusedandsimplestis theAmestest(Lewis&Chestner1981).However,theposi- tive results of the Ames test alone are not su⁄cient to estimate the carcinogenic risk to a human population (Cross etal . 1983); rather, these are used to detect potentially dangerous chemicals (Lewis & Chestner 1981). The genotoxicity of dental material should also be investigated using other tests, such as theV79/hprt Correspondence:IvicaAnic¤,DepartmentofDentalPathology,Schoolof Dentistry, Universityof Zagreb, Gundulic¤eva 5,10 000 Zagreb, Croatia (e-mail:anic@sfzg.hr). 330 International Endodontic Journal, 36, 330^335, 2003 ß 2003 Blackwell Publishing Ltd DropBooks DropBooks