壳聚糖I型胶原明胶复合材料的生物相容性及神经修复作用精.docx

壳聚糖I型胶原明胶复合材料的生物相容性及神经修复作用精.docx

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壳聚糖I型胶原明胶复合材料的生物相容性及神经修复作用精

NEURALREGENERATIONRESEARCHVolume7,Issue15,May2012

Citethisarticleas:

NeuralRegenRes.2012;7（15）:

1179-1184.

www.nrronline.org

Effectofchitosan/typeIcollagen/gelatincompositesinbiocompatibilityandnerverepair*☆

QingWang1,XiaoleiYang2,MingRen2,YulinHu3,QiangChen2,LeiXing2,ChunyangMeng2,TiemeiLiu1

1China-JapanUnionHospital,JilinUniversity,Changchun130033,JilinProvince,China

2DepartmentofToxicology,SchoolofPublicHealth,JilinUniversity,Changchun130021,JilinProvince,China

3DepartmentofLiver,GallandPancreas,FirstHospitalofJilinUniversity,Changchun130021,JilinProvince,China

Abstract

Chitosan,collagenIandgelatinweremixedinappropriatequantitiestodevelopanewnerverepairmaterial,withgoodarrangementandstructure,aswellasevenaperturesize.Thecomposite

materialwassterilizedby60Coirradiationfor24hourspriortoimplantationintherightthighofratsfollowingsciaticnervedamage.Resultsshowedthatthematerialwasnontoxictothekidneysandtheliver,anddidnotinduceaninflammatoryresponseinthemuscles.Thecompositematerialenhancedtherecoveryofsciaticnervedamageinrats.Theseexperimentalfindingsindicatethatthecompositematerialoffersgoodbiocompatibilityandhasapositiveeffectoninjurednerverehabilitation.

QingWang☆,M.D.,Professor,China-JapanUnionHospital,JilinUniversity,Changchun

130033,JilinProvince,China

Correspondingauthor:

TiemeiLiu,M.D.,Professor,China-JapanUnionHospital,JilinUniversity,Changchun

130033,JilinProvince,Chinaliutiemei777@

Received:

2012-02-03Accepted:

2012-04-24（N20111015003/WLM）

WangQ,YangXL,RenM,HuYL,ChenQ,XingL,MengCY,LiuTM.Effectofchitosan/typeIcollagen/gelatincompositesin

biocompatibilityandnerverepair.NeuralRegenRes.2012;7（15）:

1179-1184.

www.nrronline.org

doi:

10.3969/j.issn.1673-5374.2012.15.009

KeyWords

chitosan;collagenI;gelatin;biomaterials;biocompatibility;nerverepair;neuralregeneration

cartilage,boneandnervetissuerepair[13].Gelatinisbiodegradable;however,itisabrittlesubstance,absorbswaterwellandexpands.Thecombinationofgelatinandchitosancancompensateforthedeficienciesineachofthesetwo

compounds.TypeIcollagenisthemostabundantofallthecollagenproteinsthatundergoesnaturaldegradationinthebody.Collagenhasbeenwidelyusedfordrugdeliverysystems,particularlyinburnrepair,asitprovidesasupportingstructureforcells.Collagenmayenhancetheaffinityofchitosantonervecells[14].Publishedpapershaveusedcombinationsofchitosanandgelatin,chitosanand

collagen,orgelatinandcollagenfornerveortissuerepair[15-17].

However,thereiscurrentlynoresearchusingthecombinationofchitosan,gelatinandcollagenIinnerverepair.Thus,theaimofthisstudyistoexplorethe

biocompatibilityandnerverepairpotentialof

INTRODUCTION

Nervedamageiscommonplaceinclinicsduetoaccidentsandinjuries[1].However,maturenervecellshavealimitedcapacityforregeneration.Injurednerveswilltriggervariousfunctionaldisturbancestothebodyiftheyarenotrepairedinatimelymanner.Anidealnerverepairmaterialshouldbe

biocompatible,biodegradableandenhancenervefunctionrehabilitation.

Chitosanisapolysaccharidewithgreatbiocompatibilityandbiodegradabilitythatfacilitatestissueregeneration;thus,ithasbeenwidelyusedfornerverepair.However,chitosanhasa„crispy‟texturethatmakesitdifficulttosuturetothehosttissues,andtheabsorptionofchitosanisslow,whichaffectsitsefficacyasatreatmentstrategy[2-12].Gelatin,ontheotherhand,isaconstituentofbones,tendons,andconnectivetissuesofanimals,andiswidelyusedinclinicsforskin,

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WangQ,etal./NeuralRegenerationResearch.2012;7（15）:

1179-1184.

achitosan/gelatin/collagenIbiomaterial.

RESULTS

Quantitativeanalysisofexperimentalanimals

Atotalof156ratswereincludedinthisstudy:

96ratswereusedtodeterminethebiocompatibilityofthe

composite,and60ratswereexaminedfornerverepair.Foreachexperiment,theratswereequallydividedintothreegroups:

control（uninjured）,injury（injuredonly）andmaterials（injuredandtreatedwithmaterialplacement）.Overall,156ratswereinvolvedintheresultsanalysis.

Ultrastructureofchitosan-collagenI-gelatincomposite

Thestructureofthenerverepairmaterialwascylindricalanduniform,withgoodflexibilityandstrongelasticity.Themicrostructureofthetransverse,longitudinalanddiagonalsectionsofthesterilizedmaterialunderascanningelectronicmicroscopeshowedthattheouterstructureofthematerialwasfullyenclosed.Theinnerstructureshowedauniformaperture,withparallel

microtubulestravellinguniformly.Thestructurewasnotaffectedbyporesize,andthetrabecularwalloftheporeshadgoodcontinuity,smoothsurfaces,andnofolds;theoutersurfacewasimbricated.Thelongitudinalorientationofthemicrotubuleswasindependentofeachotherandinaclosedstate.Therewerenoexchangesforthebridgeconnectingpipesandtheoverallstructurewasirregular,withaninconsistentdiameter.Thetransversesectionsofthematerialwerepredominantlycircular,witharegularshapeanduniformdiameter（Figure1）.Theinternalstructureofthematerialwasalsoregular,andtheidealmicrostructurewasconducivetotheregenerationoffiber-orientedgrowth.Thebasicdirectionofthe

microtubuleswasparallelandindependent,withauniformdiameter.Thesestructureshelpedtheanatomicalandfunctionalreconstructionofthenerveafterinjury.

Thebiocompatibilityofchitosan/collagenI/gelatincomposite

Aftersurgery,theactivitylevelfortheratsintheinjuryandmaterialgroupsreducedsignificantly.Thefoot

attachedtotheoperatedlimbwaslame,yettherewasnonotableinflammatoryreactionorself-mutilation.Onday2aftersurgery,theoperatedmusclecicatrizedwell.Atdays3,7,14,21aftersurgery,wenotednoobviouschangestotheembeddedmaterial.

Grossandhistologicalchangesofliver,kidneyandmuscles

Underalightmicroscope,thestructuresoftheliverand

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thekidneysintheratsfromthethreegroupsweresimilar,andnoabnormalchangeswerefound.Intheleft

quadricepsfemorismusclesofratsinthematerialsgroup,numerousinflammatorycellshadinfiltratedaroundthematerialondays3and7.Furthermore,thematerialhadbeensubjectedtophagocytosis,degradedandaforeigngranulomahadformed.Aninflammatoryresponsewasalsoseenintheratsoftheinjurygroup.Ondays14and21,theinflammatoryresponseinthethighmusclesofratsfromthematerialgrouphaddisappeared,buttheforeigngranulomawasstillvisible.Forratsintheinjurygroup,theinflammationhadalsodisappeared,similartothatseenintheratsinthecontrolgroup（Figure2）.Therewasnosignificantdifferenceintheorgan:

bodyweightratioinratsamongthesethreegroups（P>0.05;Table1）.

Figure1Surfacesectionsoftherepairmaterialsunderelectronmicroscopy.

（A）Longitudinalsectionofthematerialshowedan

irregularshapeandaninconsistentdiameter（×150）.

（B）Transversesectionofthematerialshowedthatthe

internalstructurewasregular（×601）.

（C）Transversesectionofthematerialshowedthe

trabecularwallsoftheporeshadgoodcontinuity（×1201）.（D）Transversesectionofthematerialshowedthesurfacewassmooth,withnofolds（×2403）.

Changesofbloodroutineandbiochemicalindicatorsinrats

Therewerenosignificantdifferencesintheperipheralhemoglobinandthelevelofbloodurinenitrogenandurinecreatinefromratsamongthethreegroups（P>0.05）.Ondays3and7afterinjury,thenumberof

peripheralwhitebloodcellsinratsfromthematerialandinjurygroupswassignificantlyhigherthanthatfromthecontrolgroup（P<0.05）.Byday14,thenumberof

peripheralwhitebloodcellsdecreasedtonormallevels.Thechangesinglutamicoxaloacetictransaminase,glutamicpyruvictransaminase,lactatedehydrogenaseandcreatinekinaselevelsfromday3today21areshowninTables2-5.

WangQ,etal./NeuralRegenerationResearch.2012;7（15）:

1179-1184.

Liver（×100）Kidney（×100）Muscle（×200）

p

uorglorntoCpuo）rgsylaaidre3（atMpu）osrgyaydru3j（nIpuo）

rgsylaaidre7（atMpu）

osrgyaydru7j（nIpuo）srgyaladi

r4e1（at

M

pu）os

rygaydr

u4j1n（I

p

uo）rsgy

laadire12at（M

p

u）osrygaydru1j2n（IFigure2Hematoxylin-eosinstainedsections.Morphologychangesintheliver,kidneysandmusclesweredeterminedusinglightmicroscopy.

Therewerenochangesinliverandkidneysinratsfromthethreegroups.Inthethighmuscles,numerousinflammatorycellshadinfiltratedaftersurgeryinthematerialsandinjurygroups.Bydays14and21,theinflammationhad

disappeared,butforeigngranulomacouldstillbeobserved

inthematerialsgroup.

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WangQ,etal./NeuralRegenerationResearch.2012;7（15）:

1179-1184.

Chitosan/collagenI/gelatincompositepromotesthefunctionalrehabilitationofsciaticnerve

Onday45aftersurgery,amuscularelectromyogramshowedthattheinjurednerveinthematerialgroup

recoveredfromthelesionquickerthanthenervesintheinjurygroup（P<0.01;Table6）.

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DISCUSSION

Nerveinjuryisverycommonclinically.Whennerveinjuryissevereandthenervefunctionislost,thepatientoftenexhibitsaseriesoffunctionaldisturbancesinotherpartsofthebody,whichissometimespuzzlingforclinicians.Assuch,nerverepairhasattractedalotofattention

withinthemedicalprofession.Varioustissueengineeringmethods