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Kostićisar./Promenenahelaćelijskojkulturiuprisustvuakrilatazabazuproteze
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PROMENE NA HELA ĆELIJSKOJ KULTURI U PRISUSTVU AKRILATA ZA BAZU PROTEZE
CHANGES ON HELA CELL CULTURE IN PRESENCE ON ACRILIC RESINS
Milena Kostić1, Nebojša Krunić1,2, Stevo Najman2,3, Jelena Kocić2,3, Milena Veselinović2,3
KliniKazastomatologiju,odeljenjezastomatološKuProtetiKu,niš,srbija; 2medicinsKifaKultet,niš,srbija;3institutzabiologijuihumanugenetiKu,niš,srbija
1clinicofstomatology,dePartmentofProsthodontics,nis,serbia;2medicalfaculty,nis,serbia; 3institutofbiomedicalresearch,nis,serbia
actastomatologicanaissi decembar/december2008,vol.24,br./num.58str./p793-800
Kratak sadržaj
Akrilati su često upotrebljavani materijali u stomatoprotetskoj praksi, posebno u izradi baza zubnih proteza. U pojedinim slučajevima uočava se reakcija oralnih tkiva na mestima kontakta sa bazom akrilatnih pro-teza. Materijal za ispitivanje je obuhvatao četiri različita akrilata za bazu proteze. Ispitivan je uticaj ekstrakata akrilata različitih koncentracija na vijabilnost HeLa ćelija, kao i reverzibilnost nastalih promena na ćelijskoj kulturi. Kao kontrola je služila kultura koja je rasla u medi-jumu bez ekstrakata. Procena vijabilnosti HeLa ćelija vršena je MTT testom.Sa porastom koncentracije ekstrakata ispitivanih akrilata vijabilnost HeLa ćelija značajno opada, a i usporeniji je njihov oporavak. Potpuni oporavak HeLa ćelija nije primećen ni u jednoj od ispitivanih koncen-tracija.
Ključne reči: akrilati, baza proteze, HeLa ćelijska kultura, MTT test
Uvod Poli(metil metakrilat) (Pmma) predstav-
ljadanasnajčešćekorišćenimaterijalzaizradubazeproteze,opturatorimaksilofacijalnihpro-teza, ortodontskih aparata, kao i za potrebenjihovih podlaganja i reparatura1,2,3. razloziza njihovu značajnu primenu jesu korektnefizičke i mehaničke osobine, transparentnostkao i relativno laka manipulacija 4. u sva-kodnevnoj praksi, najčešće se upotrebljavajutoplo ihladnopolimerizujućiakrilatikoji se, sobziromnaulogumorfološkogifunkcionalnogsupstituentauusnojduplji,svrstavajuugrupubiomaterijala5.
NAUČNI RADSCIENTIFIC ARTICLE
Abstract
Acrylic resins are materials often used in dental practice, especially in denture base manufacturing. In some cases, a reaction of the oral tis-sues appears on the contact spots with the base of acrylic dentures.Testing materials considered of four different acrylic resins. The in-fluence of differently concentrated acrylic extracts on the viability of HeLa cells was examined, together with the reversibility of the changes which appeared on cells’ culture. A culture that grew in an extract free medium was used as control. The estimation of HeLa cells’ viability was done by the MTT test.As the concentration of examined acrylic extracts grows, the viability of HeLa cells considerably declines, and their recovery is slower. Com-plete recovery of HeLa cells has not occurred in any concentration of all examined.
Key words: acrylic resins, denture base, HeLa cell culture, MTT test
IntroductionPoly (methylmethacrilate) (Pmma) is, at
present, the most frequently used material inmanufacturing denture bases, obturator andmaxillofacial dentures, orthodontic devices,and also, for their relining and reparation1,2,3.therearenumerousreasonsforitsapplicabili-ty,suchasitsphysicalandmechanicalfeatures,transparency and the possibility of relativelyeasymanipulation4.heat-curedandcold-curedacrylicresinsaremostcommonlyusedineverydaypractice.astheyplaytheroleofamorpho-logicalandfunctionalsubstituteinoralcavity,theyareplacedintothegroupofbiomaterials5.
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Potencijalno toksične supstance iz akrilatavremenom se oslobađaju iz površnih slojevaprotezne baze i difunduju u pljuvačku i, neretko,izazivajuinflamatorneaređeialergijskereakcijemekihtkivasakojimadolazeukontakt6,7.
Patološkepromene sekliničkimanifestujukao kontaktni stomatit (stomatitis protetica),stomatodinije i sor (candidiasis)8,9. smatra sedačak17%korisnikapločastihzubnihprotezaispoljava preosetljivost na akrilate1. Veomaretkomogunastatiisistemskaoštećenja10.
Procena tolerancije nekog materijala odstraneživogtkivamožeseizvršiti,izmeđuos-talog i na osnovu testiranja njihove biokom-patibilnostiin vitrometodama.In vitro testovicitotoksičnosti predstavljaju preliminarne tes-tove za procenubiokompatibilnostimaterijala2,3,11.najzastupljenijibiološkisistemizatesti-ranjetoksičnihefekatastomatološkihmaterijalajesućelijskekulture.Prednostispitivanjain vi-tro,upoređenjusastudijamanaeksperimental-nimživotinjama, jeumogućnostiponavljanjapod identičnim uslovima, strogoj kontroli posvakom parametru i ekonomskoj isplatljivosti12.
cilj radabiojedaseispitaefekatrazličitihkoncentracijaekstrakataakrilatana vijabilnostHeLaćelijskekulture,kaoimogućnostinjenogoporavka.
Materijal i metode
materijal za ispitivanje su činila četirirazličitaakrilatnamaterijala(tabela1).
napravljenojepo5uzorakaodsvakegrupe(n=20)istihdimenzija(15x10x2mm).
u istraživanju je korišćenaHeLa s3 ćelij-ska linija (americantypeculturecollection,rockville,md,sad)kojasesmatraanalogomepitelnihćelijaoralnemukoze.
ekstrakti akrilata dobijeni su inkubacijomudmem-u(dulbecco’smodifiedeagle’smi-nimal essential medium, Paa laboratoriesgmbh)uodnosu0.2grakrilatau5mlmediju-ma (iso 10993: 1998)13. ekstrakcija uzorakavršena jeuzatvorenimplastičnimepruvetamanatemperaturiod37±10cuvodenomkupatilu,u trajanju od 3 dana.napravljene su koncen-tracijeekstrakataod10%,25%,50%i100%.efektivnekoncentracijeekstrakatasubiledvo-strukomanje,jersuekstraktidodavaninaistu
Potentionally toxic substances from theacrylic resins are being leached from the sur-facelayersofthedenturebaseanddiffundedinthe saliva, which often causes inflammatoryreactions,and rarely,allergic reactionsof softtissues towhich the toxic substances come incontact6,7.
Pathological changes are clinically mani-festedasstomatitisprotetica,stomatodyniaandcandidiasis8,9.itisconsideredthateven17%ofdentures’usersaresensitivetoacrylicresins1.systematicdamagescanappear, too,butveryrarely10.
tolerance that a live tissue may createagainstsomematerialcanbeestimatedaccord-ingtothetestingoftheirbiocompatibilityusingin vitromethods.In vitrocytotoxicitytestsrep-resentpreliminarytestsforestimationofmate-rials’ biocompatibility2,3,11.most applied bio-logicalsystemsforexaminingthetoxiceffectsofdentalmaterialsarecellcultures.Whencom-paredtostudiesonexperimentalanimals,thein vitromethodhasamajoradvantagewhichliesin the possibility of repetition under identicalconditions,precisecontrolofeveryparameterandfinancialbenefit11.
the purpose of the studywas to examinetheeffectofdifferentconcentratedacrylicex-tractsontheviabilityofHeLacellculture,andthepossibilityofitsrecovery.
Material and methods
thetestingmaterialconsistedoffourdiffer-entacrylicresins(table1).
five samples with same dimensions(15x10x2mm) were made from each group(n=20).
HeLa s3cell line(americantypeculturecollection,rockville,md,usa)wasusedinresearch,acelllineconsideredtobeanalogoustoepithelialcellsoforalmucosa.
acrylic extractswere obtained by incuba-tion indmem (dulbecco’smodified eagle’sminimalessentialmedium,Paalaboratoriesgmbh) in proportion of 0.2g of acrylic resinin 5ml of medium (iso 10993:1998)13. theextractionofsampleswasperformedinclosedplasticvials,at37±10c,inwaterbathfor3days.10%,25%,50%and100%extracts’concentra-tions was made. the effective concentrationsof the extractswere being added to the same
Kostićetal./changesonhelacellcultureinpresenceonacrilicresins
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zapreminumedijumasaćelijama.sviekstraktisusterilisanifiltracijomkroz0,2µmfilter.
Ćelije su zasađene u tri sterilne ploče zakultivacijusa96mesta.usvakopojedinačnomestosađenojepo2x104ćelijau50µldmem-a sa dodatkom2mm l-glutamina, 100 iu/mlpenicilina, 100 gu/ml streptomicina (Paalaboratories gmbh) i 10% fetalnog goveđegseruma(gibco,uK),nakojejedodatojoš50µlekstrakatačetiriakrilatazabazuzubneprotezerazličitih koncentracija. Kontrola je sadržala2x104 ćelija u 100 µl dmem-a. ispitivanjeza svaki od materijala vršeno je u kvadrip-likatu.Ćelijesuinkubirane3dana,uatmosferizasićenojvodenomparom,sa5%co2na37ºc.Potomjeutrađenmtttest.
da bi se ispitala reverzibilnost nastalihpromena nakon navedenog vremenskog peri-oda, iz jedne od ploča za kultivaciju ćelijaizvučen jemedijum sa ekstraktimamaterijalaizsvihpojedinačnihmestaizamenjensvežimdmem-om.udrugojpločizamenajeizvršenasapo100µlekstrakatačetiriakrilatazabazuproteze odgovarajućih koncentracija. sledilajetrodnevnainkubacija.nakonukupno6danainkubacijeurađenjemtttest.
mtt test zasniva se na aktivnosti enzimasukcinat-dehidrogenaze koji je sastavni deomitohondrijalnog respiratornog lanca vijabil-nihćelija.navedenienzimredukuježututetra-
volumeofmediumwithcells.allextractsweresterilizedbyfiltrationthrougha0.2µmfilter.
thecellswereplacedinthreesteriletissuecultureplateswith96wells.ineachindividualwell,2x104cellsin50µlofdmemwereplaced,withadditionof2mml-glutamine,100ui/mlpenicillin,100gu/mlstreptomycin(Paalab-oratoriesgmbh)and10%of fetalbovine se-rum(gibco,uK),towhichanother50µloffouracrylic resins’ extractswere added, all of dif-ferentconcentrations.thecontrolconsistedof2x104cellsin100µldmem.theexperimentsforeachmaterialweredoneinaquadruplicate.thecellswereincubatedfor3daysinafullyhumidifiedatmospherewith5%co2at37
0c.mtttestfollowed.
to examine the reversibility of the newlyformed changes after the appointed time, themediumwithmaterials’extractswasremovedfrom all thewells in one tissue culture plate,and replaced by a freshdmem. in a secondplate,thereplacementwasdonewith100µlofthe fouracrylic resins’extractseach,andcor-responding concentration. 3 days’ incubationperiodfollowed.aftertotalincubationtimeof6days,anmtttestwasperformed.
mtttestisbasedontheactivityofsucci-nate-dehyrogenasesenzyme,whichisconstitu-entpartofthemitochondrialrespiratorycycleofviablecells.theenzymementionedreducestheyellowtetrazoliumsalt((3-(4,5-dimethyltia-zolil-2)-2, 5-diphenyltetrazolijum bromide-mtt)
Tabela 1. Ispitivani akrilati za bazu proteze Table 1. Tested acrylic resins
Naziv akrilata i proizvođač Resins’ name and manu-facture
Način polimerizacije Polymerization type
Sastav Components
Odnos praha i tečnosti (g/ml) Powder/liquid
ratio (g/ml)polimer polymer
monomer monomer
Simgal-R galenika,srbija
15-18min.na220C 15-18min.at220C
Pmma MMAdodavatiprahdo
zasićenja Powdertosaturation
Triplex Cold ivoclarVivadent,lihtenštajn
15min.na220C 15min.at220C
Pmma MMA, EGDMA 13:10
Biocryl - RN galenika,srbija
30minna70°c,30minna100°c 30minat70°c,30minat100°c
Pmma MMA 20:10
Triplex Hot ivoclarVivadent,lihtenštajn
45minna100°c 45mina100°c
Pmma MMA, EGDMA 23,4:10
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zolijumovuso((3-(4,5-dimetiltiazolil-2)-2,5-di-feniltetrazolijum bromid-mtt) do formazana,jedinjenja plave boje koje se u vidu kristalataložiućelijama.medijumukomesuinkubi-ranećelijeizvučenjepozavršetkušestodnevneinkubacije, ćelije su isprane sa 100 µl Pbs-a(Phosphatebufferedsaline) idodatojepo20µlmtt-a.nakon4hinkubacijena37ºc,nastalikristaliformazanarastvorenisudodatkom100µlizopropanola.spektrofotometrijskomerenjeredukcijemtt-avršenojenaoptičkojgustiniod540nm,navišekanalnomfotometru(multi-skanascentno354,thermolabsystems,fins-ka).spektrofotometrijskiočitanintenzitetplaveboje,nakonekstrakcijeformazana,direktnojeproporcionalanbrojuvijabilnihćelija.
eksperimentjeponovljendvaputa.za statističku obradu podataka korišćena
je analiza varijanse (anoVa). statističkiznačajnimsmatranisunivoiodp<0,05.
rezultati suprezentiranikaoprocenatvi-jabilnosti ćelija. Vijabilnost ćelija kontrolnegrupepredstavljenjekao100%.
Kvantitativnepromeneuvijabilnostićelijanakon inkubacije u ekstraktima akrilata pred-stavljenesuiopisno:14
1.netoksičniakrilati:vijabilnostćelija>90%kontrole,
2. blago citotoksični akrilati: vijabilnostćelija60-90%kontrole,
3.umerenocitotoksičniakrilati:vijabilnostćelija30-59%kontrole,i
4.ozbiljnocitotoksičniakrilati:vijabilnostćelija<30%kontrole.
Rezultati nakon kultivacije HeLa ćelija u uzor-
cima akrilata dobijenih trodnevnom ekstrak-cijomudmem-u uocena je negativnakore-lacijaizmeđukoncentracijeekstrakataakrilataivijabilnostićelija(grafikon1).srazmernosaporastomkoncentracije ekstrakatadošlo jedopadaćelijskeproliferacije, štogovoriuprilognjihovoj toksičnosti. statistički značajan padvijabilnosti ćelija prisutan je kod svih tipovaakrilata(p<0,01),osimkodbiocryl-arn.
Procenti vijabilnostiHeLa ćelija određenimtt testom, prilikom ispitivanja reverzibil-nostinastalihpromena,prikazanisunagrafiko-nima2-5.
statistički značajna razlika u vijabilno-sti kultura kojima su nakon tri dana ekstrakti
toformasan,abluechemicalcompoundwitchmakesacrystal-likelayerinsidethecells.themediumwiththeincubatedcellswastakenoutafter 6 days’ long incubation. the cells werewashed out with 100 µl of Pbs (Phosphatebuffered saline) and with 20 µlmtt addedto each.after another 4 hours of incubationat 370c, the newly created formasan crystalsweresolutedwith100µlofizopropanol.spec-trophotometricalmeasuringofmttreductionwas performed at an optical density of 540nm,onamultichannelphotometer (multiskanascentno354,thermolabsystems,finland).spectrophotometricallyviewed,theintensityofblue,afterformasanextraction,isindirectpro-portiontothenumberofviablecells.
theexperimentwasperformedtwice.the analysis of variance (anoVa) was
used for statistical data examining. levels ofp<0.05weretreatedasstatisticallysignificant.
the resultswerepresentedas thepercent-ageofcells’viability.thecellviabilityofthecontrolgroupwaspresentedas100%.
Quantitativechangesincells’viabilityafterincubationintheacrylicextractsarepresenteddescriptively14:
1.non-cytotoxicacrylicresins:cellviability>90%inrelationtocontrol;
2. slightly acrylic resins: cell viability 60-90%inrelationtocontrol;
3. moderately acrylic resins: cell viability30-59%inrelationtocontrol,and
4. severely acrylic resins: cell viability<30%inrelationtocontrol.
ResultsafterthecultivationofHeLacellsinacryl-
icsamplesobtainedafter3days’extractionindmem,ithasbeenobservedthatthereexistedanegativecorrelationbetween theacrylicex-tracts’concentrationandcells’viability(graph1). Proportionally, as the concentration of theextracts increased, the cell proliferation de-creased,witchindicatestheir toxicity.statisti-callysignificantdeclineofcellviabilityispres-entwithallacrylictypes(p<0.01),expectwithbiocrylrn.
the percentage ofHeLa cell viability, de-termined bymtt testwhile testing the new-ly formed changes’ reversibility, is shown ingraphs2-5.
statistically significant difference betweenthe viability of cultures to which acrylic ex-tractswere replaced, after 3 days, bydmemandthose,towhichtheextractswerereplaced
Kostićisar./Promenenahelaćelijskojkulturiuprisustvuakrilatazabazuproteze
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akrilata zamenjeni dmem-om i onih kojimasu ekstrakti zamenjeni istim, 5% ekstraktimaakrilatanijeuočenanikodjednogodispitiva-nihuzoraka.
sviispitivaniakrilatikoncentracije5%po-kazalisublagicitotoksičniefekat.
Vijabilnost ćelija veća je u slučaju kad su12,5%ekstraktiakrilatazamenjeničistimme-dijumom, osim u slučajutriplexhot-a. ni ujednomodispitivanihslučajevanemastatističkiznačajnerazlike.
sviispitivani12,5%ekstraktiakrilatapoka-zalisublagicitotoksičniefekat.
Pri koncentraciji akrilata od 25%, vijabil-nost ćelija takođe je veća u slučaju zamenemedijuma dmem-om sa izuzetkom triplexhot-a.statističkaznačajnostpostojikoduzora-kabiocrylrn-a.
svi akrilati, osimtriplexcold-a, pokazalisublagicitotoksičniefekatprikoncentracijiod25%.ekstraktitriplexcold-aispoljilisuume-
bysame5%acrylicextracts,wasnotdetectedwithanyofexaminedsamples.
alltestedacrylicresinswithconcentrationof5%showedoslightcytotoxiceffect.
cellviabilityisathigherrateincaseswhere12.5%acrylicextractsarereplacedbypureme-dium,exceptinthecaseoftriplexhot.thereisnotstatisticallysignificantdifferenceinanyofthesituationstested.
alltestedacrylicextractsof12.5%showedaslightcytotoxiceffect.
atacrylicconcentrationof25%,cellviabil-ityisatahigherdegreeincaseswhentheme-diumisreplacedbydmem,withtheexception
p<0,01
Grafikon 1. Vijabilnost ćelija nakon trodnevne inkubacije u ekstraktima akrilata
Graph. 1. Cell viability after 3 days’ incubation in acrylic extracts
Grafikon 2. Vijabilnost HeLa ćelija nakon inkubacije u 5% ekstraktima akrilata
Graph. 2. HeLa cell viability after incubation in 5% acrylic extracts
Grafikon 3. Vijabilnost HeLa ćelija nakon inkubacije u 12, 5% ekstraktima akrilata
Graph. 3. HeLa cell viability after incubation in 12. 5% acrylic extracts
Grafikon 4. Vijabilnost HeLa ćelija nakon inkubacije u 25% ekstraktima akrilata
Graph. 4. HeLa cell viability after incubation in 25% acrylic extracts
Grafikon 5. Vijabilnost HeLa ćelija nakon inkubacije u 50% ekstraktima akrilata
Graph. 5. HeLa cell viability after incubation in 50% acrylic extracts
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reni toksični efekat, što govori u prilog većetoksičnostihladnopolimerizujućihakrilata.
Pri najvišim ispitivanim koncentracijamaakrilata(50%)utvrđenjevećiprocenatvijabilnihćelijauslučajuzameneekstrakatadmem-om,kodsvihtipovaakrilata.statističkaznačajnostpostoji kod svih vrsta akrilata osimsimgal-a.uslučajutriplexcold-aonajenajvišegnivoa(p<0,001).
ekstrakti svih ispitivanih akrilata koncen-tracije 50% pokazali su umereni citotoksičniefekatnavijabilnostHeLaćelija.
Prilikomzameneekstrakataakrilatakoncen-tracije5%medijumom,nijedošlodooporavkaHeLa ćelija. dobijeni rezultati za veće kon-centracijeekstrakatapokazalisureverzibilnostnastalihpromenakod biocryl-arn,simgal-a itriplexcold-a. efekatreverzibilnostinijeuočenkodtriplexhot-a,osimzakoncentrac-iju50%.
Diskusija
Kakojeproteznabazaukontaktusavelikompovršinom oralne sluzokože i u toku dužegvremenskog perioda, postoji rizik za njenooštećenje,bezobziranauputstvaproizvođačaonjihovoj indiferentnosti. zbog toga je ispiti-vanje bioloških osobina akrilata od izuzetnogznačajasobziromnanjihovumasovnuupotre-bu u svakodnevnoj stomatoprotetskoj praksi.Pri tome, ispitivanje citotoksičnosti akrilatnihmaterijalauuslovimain vitropredstavljaveo-mainteresantnuoblast.utomsmislu,kontinu-irane ćelijske kulture, kao biološki sistemi zatestiranje,imajuznačajnuuloguizvišerazloga.Ćelije je lako gajiti u kulturi, nema varijabil-nostikojabisevezivalazarazličitedonoretki-vaapostojiimogućnostponavljanjatestapodidentičnimuslovima1,15.
uovomraduispitivanjeefekatekstrakataakrilatazabazuprotezenapermanentnućelijskuliniju, HeLa.ovećelijesuodabranezbognji-hove sličnosti sa epitelnim ćelijama oralnemukoze.Kako jeproteznabazaukontaktusasluzokožom usne duplje efekti na ispitivanućelijskukulturumogusesmatratiklinički rel-evantnim. zahvaljujući njihovoj izuzetnoj os-etljivosti,eventualnihemijskiuticajakrilatanavijabilnostigustinućelijamogaosepreciznoregistrovati16.
oftriplexhot.statisticsignificanceexistsonlywithbiocrylrnsamples.
all acrylic resins, except triplex cold,showed a slight cytotoxic effect at concentra-tion of 25%.triplexcold extracts revealed amoderate cytotoxic effect, which proves thefact that cold/polymerized acrylic resins aremoretoxic.
atthehighestacrylicconcentrationstested(50%), more viable cells were noticed an allacrylic resins’ types, when the extracts werereplacedbydmem.statistic significance ex-istswithallacrylicresins,withtheexceptionofsimgal.thehighestlevelisincaseoftriplexcold(p<0.001).
theextractsofalltestedacrylicresinswithconcentration of 50% showedmoderate cyto-toxiceffectonHeLacellviability.
While replacing 5% concentrated extractsofacrylicresinsbyamedium,HeLa cells’re-coverydidnotoccur.theobtainedresults forhigher concentrations of extracts showed re-versibility of the changes that occurred withbiocrylrn,simgalandtriplexcold.theef-fectofreversibilitywasnotdetectedwithtri-plexhot,exceptfor50%concentration.
Discussionasthedenturebaseisinlonglastingcontact
withalargesurfaceoforalmucousmembrane,thereisariskofdamaging,despitethemanu-facturers’ instructions about the base’ s indif-ference.duetothat,testingthebiologicalfea-turesofacrylicresinsisexterminalyimportant,considering theiroftenuse ineverydaydentalpractice.inaddition,cytotoxicitytestingoftheacrylicresinsunderinvitroconditionsbelongsto a very interesting researchingfield. in thatcontext,continuingcellcultures,viewedasbio-logical testing systems,playa significant roleforvariousreasons.cellsareeasytobebredinaculture;thereisnovariabilitytobeconnectedtodifferent tissuesdonors;andthere isapos-sibilityofre-performingthetestunderidenticalconditions1,15.
the study has examined the effect whichtheacrylicresins’extractshaveontheperma-nentHeLa cell line. these cells were chosenfortheirsimilaritytotheepithelialcellsoftheoralmucosa.asdenturesareincontactwiththemucousmembraneoftheoralcavity,theeffectsonthetestedcellculturecanbetreatedasclini-
Kostićetal./changesonhelacellcultureinpresenceonacrilicresins
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ispitivani materijali ispoljili su blagu doumerenucitotoksičnost,štojeusaglasnostisanalazimahuangisar.iliuisar15,17.takođesuinalazitsuchiyaisar.,cimpanisar.,campan-chaisar.ukazalinacitotoksičniefekatakrila-ta korišćenjem drugih in vitro testova 18,19,20.nasuprot ovim rezultatima, jorge i sar. nisuutvrdilicitotoksičnostakrilata2.razlikeudobi-jenimrezultatimamogusepripisati razlikamau eksperimentalnom dizajnu koje obuhvatajuizabranitipakrilatazatestiranje,izborćelijskelinije i primenu različitih testova za analizucitotoksičnosti.
uovomradu, izmedjuostalog,utvrđivanojedalisunastalepromenereverzibilnogkara-ktera,odnosnodalinakonprestankadelovanjatoksičnih komponenti iz akrilata za bazu pro-teze dolazi do oporavka opservirane ćelijskelinije.usmisluoporavkaćelijaapoptoza(pro-gramirana ćelijska smrt) predstavlja kaskadninizreakcijaukojemupojedinimtačkamapost-ojimogućnostreverzijeprocesaineophodnijesastavnideoćelijskehomeostazezarazlikuodnekroze,kojajeireverzibilanprocesiukazujena znatno manju kompatibilnost primenjenogmaterijala21,22.
buduća istraživanjamogla bi ići u pravcuodređivanjamehanizmaćelijskesmrti,auciljupoboljšanjakvalitetaovihmaterijala.
Zaključak
sa porastomkoncentracije ekstrakata ispi-tivanih materijala vijabilnost HeLa ćelijaznačajnoopadaismanjijesemogućnostnjiho-vog oporavka. Potpuni oporavak ćelijske kul-turenakonzameneekstrakataakrilatadmem-om nije primećen ni u jednoj od ispitivanihkoncentracija.
Zahvalnica
ovaj rad je urađen u okviru projekta on145072, koji finansira ministarstvo nauke izaštiteživotnesredine.
callyrelevant.duetotheirextremesensitivity,thepossiblechemical acrylic influenceon thecellviabilityanddensitycouldhavebeenpre-ciselynotified16.
thetestedmaterialsdemonstratedslighttomoderatecytotoxicity,whichcorrespondswithresearches ofhuang et al. andliu et al.15,17.also,theresearchesoftsuchiyaetal.,cimpanetal.andcampanchaetal.pointedtothecyto-toxiceffectofacrylicresins,bydoingin vitro experiments18,19,20. contrary to these results,jorgeetal.didn’tdetectcytotoxicityofacrylicresins2.thedifferencesintheobtainedresultsmaybecontributedtothedifferencesoftheex-perimentaldesign, and it includes: the acrylicresin type chosen to be tested, choice of cellline andapplicationofdifferent tests for ana-lyzingcytotoxicity.
thisstudyincluded,amongotherthings,aresearchonwhetherthechangesthatappearedare reversible, i.e. whether the observed cellline recovers when the acrylic resins’ toxiccomponents stopworking. considered as cellrecovery, apoptosis (programmed cell death)represents a successive chain of reactions inwhich there is possibility of a reversible pro-cessatsomeparticularpoints.unlikenecrosis,whichisirreversibleandpointstoasignificantshortage of compatibility of appliedmaterial,apoptosisisanessentialpartofcellhomeosta-sis 21,22.
researchesinthefuturecouldhavethedi-rection of determining themechanism of celldeath.intheway,thequalityoftheacrylicma-terialscouldbeimportant.
Conclusionas the concentration of the acrylic resin
extractsgrows,theHeLa cellviabilityconsid-erablydiminishes and thepossibility for theirrecoveringislessening.acompeterecoveryofthecellculture,after replacing theacrylicex-tractsbydmem,hasn’tbeendetectedinanyoftheconcentrationsexamined.
Acknowledgment this work was supported by a research
grant,on145072,financedbytheministryofscienceofrepublicofserbia.
actastomatologicanaissi,decembar/december2008,vol.24,broj/number58
800
Address of correspondence:milenaKostic,dds.msd.52buld.drzoranaĐinđića
18000nišserbia
Phone+381(0)18226216e-mail:[email protected]
Adresa za korespondenciju:mr.sci.drmilenaKostićbul.drzoranaĐinđića52ništelefon:+381(0)18226216e-mail:[email protected]
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