Cell adhesion and mechanical properties of a flexible scaffold for cardiac tissue engineering(5)

3.1.HL-1adhesiontoPOC lms

HL-1celladhesionwasassessedonthePOC lms,bothuncoatedandcoatedwith bronectin,collagenorlaminin,followingovernightincubation.MonolayercultureofHL-1cellsonPOCshowedapoorattachmenttothesurfaceofuncoated lms,coveringnomorethan10%ofthearea.CellsseededonPOC lmscoatedwith bronectinattachedtothesurfaceandproliferatedmorerapidlythanthoseseededoncollagen-andlaminin-coated lms.Fibronectinusedataconcentrationof400lgcmÀ2showeda90±5%HL-1cellareacoverage,incontrastto32±4%withthesameproteincoatedat40lgcmÀ2andto11±1%whentheconcentrationofthe bronectinwasthelowest,at4lgcmÀ2.Ofparticularnote,laminindidnotsupportcompletedi erentiationintothecardiomyo-cytemorphologyasafterthistimesomeofthecellspossessedaroundedmorphologyratherthanthecharac-teristicellipticalshape.Weobservedthat,becausethecellsinlaminin-coated lmsdidnotspreadtoanormalphysiol-ogyasthecellsintheothertwosurfaces,theirtotalareacoveragewasa ected.Inourexperiments,cellcoveragealsodecreasedastheproteinconcentrationreduced,reach-ingthesamelevelsofattachmentasuncoated lmswiththelowestconcentrations(Fig.1a).

3.2.CharacteristicsofPOCprocessedsca olds

POCprocessedbysalt-leachingresultedinporousstruc-tureswithinter-connectedpores(Fig.1bandc)intherangeof340–380lm,withameanporesizeof360±20lm(asdeterminedbySEMandmicrocomputedtomography)forallthreeporosities.Porosity,evaluatedwithmicrocomputedtomographyandheliumpycnometry,wascon rmedtobe60±3%,70±4%or80±3%(±SD).POCsca oldsdegraded50±6%in3monthsandreportedalossofdry-weightofatleast90%after6months.

Mechanicaltestswereperformedatroomtemperatureonsca oldspreviouslystoredatÀ20°Candatroomtem-perature;eachsetincludedsamplesofthreedi erentporos-ities:60%,70%and80%.WefoundthattheYoung’smodulusofthesamplesstoredatroomtemperaturerangedfrom0.2to3.3kPa,comparedwiththeYoung’smodulusofPOCsca oldsstoredatÀ20°C,whichrangedfrom0.1to1.7kPa(Fig.2aandb),dependingontheporosity

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