Variousindustrialandtoxicologylaboratoriesareactivelyseekingalternativestoexpensiveclinicalorwholeanimaltesting.Oralcare,personalcare,andpharmaceuticalcompanieshaveinitiatedinvitrotoxicologytestingtoevaluatetheirrawmaterialsandfinalproductformulations.Technology:MatTek’sEpiOralandEpiGingivaltissuesconsistofnormal,human-derivedoralepithelialcells.Thecellshavebeenculturedtoformmultilayered,highlydifferentiatedmodelsofthehumanbuccal(EpiOral)andgingival(EpiGingival)phenotypes.Thetissues areculturedonspeciallypreparedcellcultureinsertsusingserumfreemediumandattainlevelsofdifferentiationonthecuttingedgeofinvitrocellculturetechnology.TheEpiOralandEpiGingivaltissuemodelsexhibitinvivo-likemorphologicalandgrowthcharacteristicswhichareuniformandhighlyreproducIBLe.Figure1:Immuno-stainingof:A)EpiOralandB)EpiGingivaltissueforcytokeratinK13andK14.Figure2:Immuno-stainingoftheORL-200tissueforhumanbetadefensins(HBDs).Figure3:Immuno-stainingoftheGIN-100tissueforhumanbetadefensins(HBDs).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.Figure4:EffectofSDSsolutions(40L)onEpiOral(ORL-200)tissueviABIlityfollowingexposurefor1hour.SDSconcentrationswerechosentobeinrangenormallypresentintoothpaste(0.0–3.0%). Table1:Lipidanalysisofinvivoandinvitrotissues.Theinvivobuccalandgingivalvaluesareaveragesfor14tissuesamples.Theinvitrotissuevaluesarefromasinglelot.Applications:MucosalandGingivalIrritationMeasureoralorgingivalepithelialtissueviabilityandcytokinereleaseusingastraightforwardMTTassayorcommerciallyavailableELISAkit,andobtaindefinitiveresultsinafewdays.EpiOral&EpiGingivalIrritationProtocolOralIrritationReferenceDrugDeliveryEpiOralandEpiGingivalareusefulindeterminingtheabsorptionpotentialoffinalformulationsinvitro.EpiOralAbsorptionProtocolEpiGingivalDrugAbsorptionProtocolOralDrugDeliveryReferenceModelforOralPathologiesEpiOralhasbeenusedtostudyoralcandidiasis,oralmucositis,andtheanti-microbialpropertiesoftheoralmusosa.OralCandidiasisPublicationOralMucositisPublicationOralAnti-MicrobialPeptidesPublicationSearchourreferencelibraryformoreinformation.a[rel~="mtli_filesize29269kB"]:after {content:" (292.69 kB)"}a[rel~="mtli_filesize19392kB"]:after {content:" (193.92 kB)"}a[rel~="mtli_filesize15019kB"]:after {content:" (150.19 kB)"}TechSpecs: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:EpiOralType:Normalhumanoralkeratinocytes(NHOK)aredifferentiatedintotissueswithanon-cornified,buccalphenotype,partnumberORL-200.Geneticmake-up:Singledonor.Derivedfrom:Non-diseased,adulthumanoraltissueobtainedfrompatientsundergoingtoothextractions.Screenedfor:HIV,Hepatitis-B,Hepatitis-C,mycoplasma,bacteria,yeast,fungi.EpiGingivalAlternatedonorsavailable.EpiGingivalType: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&EpiGingivalBasemedium:Dulbecco’sModifiedEagle’sMedium(DMEM)Growthfactors/hormones:EpidermalgrowthfactorandotherproprietaryfactorsSerum:NoneAntibiotics:Gentamicin5µg/mlAnti-fungalagent:AmphotericinB0.25µg/mlpHIndicator:PhenolredOtheradditives:ProprietaryAlternatives:Phenolred-free,antibiotic-free,anti-fungal-freemediumandtissueareavailable.Agentsareremoved3dayspriortoshipmentMaintenancemedium:ORL-200-MMorGIN-100-MMQualityControlandSterility: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.
MatTek公司是麻省理工学院两个化学工程教授于1985成立的科技公司。1991年,该公司利用其核心的聚合物表面改性技术进入新兴的组织工程领域,目前已成长为的组织工程技术公司,致力于生产创新性的3D重构人体组织模型,该模型相较于传统2D细胞模型具有更可靠的实验结果。其生产的重构皮肤,眼部和呼吸道组织模型已广泛应用于化妆品,化工,医药和家用产品等行业的毒理学监控(OECD,欧盟的指导方针)及毒理学研究。
MatTek公司的基于人体细胞的3D皮肤模型EpiDerm于1993年研发上市,立即获得巨大的成功。从此MatTek通过以下方式彻底改变了基于人体细胞的体外模型市场:(1)低成本(降低50-80%的成本);(2)通过发表质量控制标准(同时具备阳性和阴性对照)以确保可为行业及监管机构接受的高可重复性水平;(3)优化研究人员的实验结果。通过创造性使用非动物的体外测试,融合降低(直接/间接)成本的强大协同效应及显著提高的产品质量,MatTek极大的降低产品成本、提高实验室效率及高质量的高可重复性产品,减少实验动物的使用,具有无可超越的长期组织可重复性;3-D仿真结构;高度分化的组织模型;代谢和分裂活性;源于正常的人体细胞;培养、操作简便;无血清培养;可定量的客观检测;物美价廉的优点。因而使得MatTek人细胞来源的体外组织模型被包括美国、加拿大、欧洲、日本和东亚的众多跨国公司与研究机构采用。
目前MatTek公司的产品主要包括EpiDerm(皮肤模型)、MelanoDerm(黑色素模型)、EpiOcular(眼周表皮模型)、EpiAirway呼吸道上皮模型)、EpiDermFT (全厚的皮肤模型) 、EpiVaginal(阴道上皮模型)、EpiOral(口腔上皮模型)、 EpiGingivalTM(牙龈上皮模型)、Psoriasis tissue(牛皮癣组织模型)及树突细胞。