MatTek’sEpiOralandEpiGingivaltissuesconsistofnormal,human-derivedoralepithelialcells.Thecellshavebeenculturedtoformmultilayered,highlydifferentiatedmodelsofthehumanbuccal(EpiOral)andgingival(EpiGingival)phenotypes.Thetissues areculturedonspeciallypreparedcellcultureinsertsusingserumfreemediumandattainlevelsofdifferentiationonthecuttingedgeofinvitrocellculturetechnology.TheEpiOralandEpiGingivaltissuemodelsexhibitinvivo-likemorphologicalandgrowthcharacteristicswhichareuniformandhighlyreproducIBLe.

BothEpiOralandEpiGingivalexpresscytokeratinK13inalllayersexceptthemostapicalonesandweaklyexpresscytokeratinK14intheupperlayersofthetissue(Figure1).Thetissuesalsoproducethenaturallyoccurringantimicrobialpeptidescalledhumanbetadefensins(HBDs).EpiOralconstitutivelyexpressesHBD-1andHBD-3butnotHBD-2(Figure2).EpiGingivalweaklyexpressesHBD-3inalllayersexcepttheuppermostonesand HBD-1 intheapicallayersbutdoesnotexpressHBD-2(Figure3).

LipidanalysisoftheexplanttissuesalongwithEpiGingival(GIN-100)andEpiOral(ORL-200)isshownbelowinTable1. EpiOralandbuccalexplantscontainonlyCeramide2(CeramideNS). EpiGingivalandgingivaltissuecontainCeramides1,2,and3(alsoreferredtoasCeramidesEOS,NS,andEOHP/NP,respectively).

AgrowingbodyofdataindicatesthatEpiOralandEpiGingivaleffectivelyprovideaninexpensive,non-animalmeanstoassessoralirritation,toxicology,andpathology-relatedissues. Arepresentativedose-responsecurvefortheeffectofsodiumdodecylsulfate(SDS),acommonsurfactantusedintoothpaste,isshowninFigure4.

MucosalandGingivalIrritation

MeasureoralorgingivalepithelialtissueviabilityandcytokinereleaseusingastraightforwardMTTassayorcommerciallyavailableELISAkit,andobtaindefinitiveresultsinafewdays.

EpiOral&EpiGingivalIrritationProtocol

OralIrritationReference

DrugDelivery

EpiOralandEpiGingivalareusefulindeterminingtheabsorptionpotentialoffinalformulationsinvitro.

EpiOralAbsorptionProtocol

EpiGingivalDrugAbsorptionProtocol

OralDrugDeliveryReference

ModelforOralPathologies

EpiOralhasbeenusedtostudyoralcandidiasis,oralmucositis,andtheanti-microbialpropertiesoftheoralmusosa.

OralCandidiasisPublication

OralMucositisPublication

OralAnti-MicrobialPeptidesPublication

Searchourreferencelibraryformoreinformation.

Tissue:

Kits:EpiOral(ORL-200)andEpiGingival(GIN-100)kitsconsistof24tissues.ORL-606andGIN-606kitsconsistof6tissues.Tissuekitscontaintissues,culturemedium,toequilibratetissuesandculturethemfor24hours,andplasticware. ContactMatTekforspecifickitcontents.

Substrate:ORL-200andGIN-100:Collagencoated,9mmIDsinglewelltissuecultureinsertsareused.Poresize=0.4µm,InnerDiameter=0.875cm.Surfacearea=0.6cm².ORL-606andGIN-606:Singlewelltissuecultureinserts.Poresize=0.4µm,InnerDiameter=2.5cm.Surfacearea=4.2cm².

Culture:Atair-liquidinterface.

Histology:ORL-200:8-11celllayersofnon-cornifiedtissue.GIN-100:8-11layersofcornifiedtissue.

LotNumbers:Tissuelotsproducedweeklyareassignedaspecificlotnumber.Alltissuekitswithinalotareidenticalwithregardtocells,medium,handling,cultureconditions,etc.

Shipment:At4°Conmediumsupplemented,agarosegels.

Shipmentday:Monday.

Delivery:TuesdaymorningviaFedExpriorityservice(US).OutsideUS:Tuesday-Thursdaydependingonlocation.

Shelflife:Includingtimeintransit,tissuesmaybestoredat4°Cforupto2dayspriortouse.However,extendedstorageperiodsarenotrecommendedunlessabsolutelynecessary.Inaddition,thebestreproducibilitywillbeobtainediftissuesareusedconsistentlyonthesameday,e.g.Wednesdaymorningfollowingovernightstorageat4°C.

Lengthofexperiments:Culturescanbecontinuedforatleast1weekwithgoodretentionofnormalmorphology.Culturesmustbefedeveryotherdaywith5.0mlofmaintenancemedium(ORL-200-MMorGIN-100-MM).Cellcultureinsertsareplacedatopculturestands(MEL-STND)orwashers(EPI-WSHR)in6-wellplatestoallowtheuseof5.0ml.

Alternativetissues:

ORL-300-FT:FullthicknessORL-200.  Oralepithelialtissueculturedontopofacollagenmatrixcontaininggingivalfibroblasts.

ORL-300-FT-LC:ORL-300-FTcontainingLangerhansCells.

GIN-300-FT:FullthicknessGIN-100. Gingivalepithelialtissueculturedontopofacollagenmatrixcontaininggingivalfibroblasts.

Cells:

EpiOral

Type:Normalhumanoralkeratinocytes(NHOK)aredifferentiatedintotissueswithanon-cornified,buccalphenotype,partnumberORL-200.

Geneticmake-up:Singledonor.

Derivedfrom:Non-diseased,adulthumanoraltissueobtainedfrompatientsundergoingtoothextractions.

Screenedfor:HIV,Hepatitis-B,Hepatitis-C,mycoplasma,bacteria,yeast,fungi.EpiGingival

Alternatedonorsavailable.

EpiGingival

Type:Normalhumanoralkeratinocytes(NHOK)aredifferentiatedintotissueswithacornified,gingivalphenotype,partnumberGIN-100.

Geneticmake-up:Singledonor.

Derivedfrom:Non-diseased,adulthumanoraltissueobtainedfrompatientsundergoingtoothextractions.

Screenedfor:HIV,Hepatitis-B,Hepatitis-C,mycoplasma,bacteria,yeast,fungi.

Alternatedonorsavailable. 

Medium:

EpiOral&EpiGingival

Basemedium:Dulbecco’sModifiedEagle’sMedium(DMEM)

Growthfactors/hormones:Epidermalgrowthfactorandotherproprietaryfactors

Serum:None

Antibiotics:Gentamicin5µg/ml

Anti-fungalagent:AmphotericinB0.25µg/ml

pHIndicator:Phenolred

Otheradditives:Proprietary

Alternatives:Phenolred-free,antibiotic-free,anti-fungal-freemediumandtissueareavailable.Agentsareremoved3dayspriortoshipment

Maintenancemedium:ORL-200-MMorGIN-100-MM

QualityControlandSterility:

Visualinspection:Alltissuesarevisuallyinspectedandifphysicalimperfectionsarenoted,tissuesarerejectedforshipment.

End-usetesting:.Tissuesarealsoexposedto1.0%TritonX-100forvarioustimesandtheexposuretimerequiredtoreducethetissueviabilityto50%(ET-50)isdeterminedusingtheMTTassay. QCspecifications:ORL-200: 34.85<ET-50<105.78minGIN-100: 5.47hr<ET-50<10.42hr.

Sterility:Allmediausedthroughouttheproductionprocessischeckedforsterility.Maintenancemediumisincubatedwithandwithoutantibioticsfor1weekandcheckedforsterility.Theagarosegelfromthe24-wellplateusedforshippingisalsoincubatedfor1weekandcheckedforanysignofcontamination.

Screeningforpathogens:AllcellsarescreenedandarenegativeforHIV,hepatitisBandhepatitisCusingPCR.However,noknowntestmethodcanoffercompleteassurancethatthecellsarepathogenfree.Thus,theseproductsandallhumanderivedproductsshouldbehandledatBSL-2levels(biosafetylevel2)orhigherasrecommendedintheCDC-NIHmanual,“BiosafetyinmicroBIOLOGicalandbiomedicallaboratories,”1998.Forfurtherassistance,pleasecontactyoursiteSafetyOfficerorMatTektechnicalservice.

Notificationoflotfailure:IfatissuelotfailsourQCorsterilitytesting,thecustomerwillbenotifiedandthetissueswillbereplacedwithoutchargewhenappropriate.BecausesomeofourQCandsterilitytestingisdonepost-shipment,notificationwillbemadeassoonaspossible.Undernormalcircumstances,ET-50failureswillbenotifiedbyThursday,5p.m.;sterilityfailureswillbenotifiedwithin8daysofshipment.