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1、Downloadedfromhttp://rsif.royalsocietypublishing.org/onAugust13,2018J.R.Soc.Interface(2012)9,2871–2882doi:10.1098/rsif.2012.0269Publishedonline6June2012NanoscalecharacterizationofthebiomechanicalhardeningofbovinezonapellucidaAntonioBoccaccio1,MariaCristinaFrassanito1,Lucian

2、oLamberti1,RobertoBrunelli2,GiuseppeMaulucci3,MaurizioMonaci4,MassimilianoPapi3,CarminePappalettere1,*,TizianaParasassi5,LakamySylla4,FulvioUrsini6andMarcoDeSpirito3,71DipartimentodiIngegneriaMeccanicaeGestionale,PolitecnicodiBari,Bari70126,Italy2DipartimentodiScienzeGineco

3、logico-OstetricheeScienzeUrologiche,Universita`diRoma‘LaSapienza’,Rome00161,Italy3IstitutodiFisica,Universita`CattolicadelSacroCuore,Rome00168,Italy4DipartimentodiPatologia,DiagnosticaeClinicaVeterinaria,Universita`diPerugia,Perugia06126,Italy5IstitutodiFarmacologiaTraslazi

4、onale,ConsiglioNazionaledelleRicerche,Rome00133,Italy6DipartimentodiChimicaBiologica,Universita`diPadova,Padova,Italy7FondazionediRicercaeCura‘GiovanniPaoloII’,Campobasso86100,ItalyThezonapellucida(ZP)isanextracellularmembranesurroundingmammalianoocytes.Theso-calledzonahard

5、eningplaysakeyroleinfertilizationprocess,asitblockspolyspermy,whichmayalsobecausedbyanincreaseinthemechanicalstiffnessoftheZPmembrane.However,structuralreorganizationmechanismsleadingtoZP’sbiomechanicalhardeningarenotfullyunderstoodyet.Furthermore,acorrectestimateoftheelast

6、icpropertiesoftheZPisstilllacking.Therefore,theaimofthepresentstudywastoinvestigatethebio-mechanicalbehaviourofZPmembranesextractedfrommatureandfertilizedbovineoocytestobetterunderstandthemechanismsinvolvedinthestructuralreorganizationoftheZPthatmayleadtothebiomechanicalhar

7、deningoftheZP.Forthatpurpose,ahybridprocedureisdevelopedbycombiningatomicforcemicroscopynanoindentationmeasurements,nonlinear?niteelementanalysisandnonlinearoptimization.TheproposedapproachallowsustodeterminethebiomechanicalpropertiesoftheZPmorerealisticallythantheclassical

8、analy-sisbasedonHertz’scontacttheory,asitaccountsforthenonlinearityof?niteindentat