蠶絲負(fù)載軟骨細(xì)胞體外構(gòu)建組織工程軟骨.pdf

1、西北農(nóng)林科技大學(xué)碩士學(xué)位論文蠶絲負(fù)載軟骨細(xì)胞體外構(gòu)建組織工程軟骨姓名：尹寶英申請學(xué)位級別：碩士專業(yè)：臨床獸醫(yī)學(xué)指導(dǎo)教師：張涌20050601FORMATIONOFTISSUEENGTNEEREDCARTILAGEWITHSILKANDARTICULARCHONDROCYTES，Ⅳ明豫0ABSTRACTLimitedregenerativecapacityofcartilagetissuemakestreatmentofchondrall

2、esionsdifficultThecurrentmethodstOtreatcatilagelesionmayrelievesymptoms，butcannotregeneratetheinjuredtissueTissueengineeringtechniquesisnewresolventofsuffererformcartilagelesionItincludeseedcell，thecocultureofscaffoldand

3、cells，implantationofcellandscaffoldcompoundItistheprerequisitetOharvestseedcellsmaintainingstablephenotype，highproliferousactivityandwithbroadextensivesourcetoconstructengineeringcartilageinvitro。andamajorchallengetofind

4、aappropriatescaffoldconfromingtissueengineeringresearchersInthisworkwestudiedthemethodsofchondrocytesisolationandtheprotentialofsilkasscaffoldofengineeredcartilage，1Inthisstudyrabbitarticularchondrocyteswereharvestedbyme

5、thodsoftissueblockandenzymedigestionbystagesChondrocytesbytissueblockshewshortshuttle—like，madmaintainedsuchshapeafter7passages，whilechondrocytesofenzymedigestionpresenteddifferentshapes，andbecamefibre—likelittlebylittle

6、2Chondrocyteswereisolatedwith02％collengasentypeIIconfectedbyserumfreeHamF12mediumandPBSfleeCa2、MgpBythemethodofdigestionbystages，thelostofcellsdecreased，andcellsmmntmnedhigherviabilitythechondrocytesisolatedwithcollengas

7、etypeconfectedbyselMmfreemediumweremoreactativethanthatofphosphatebufferfree—Ca2、M92TherewassignificantdifferencebetweentWOgroups3Furthermore，weusedsilkscaffoldtoconstructengineeredcartilageThegrowthandproliferationofcho

8、ndrocytesinsilkscaffoldwerecomparedwiththatofchondrpcytesincollagenThesilksweredigestedbytrypsineandcoatedbycollagentypeIIChondrocytesaccachedsilkscaffoldduringtheprocessofsurgeThegrowthofcellswereobservedundermicroscope

9、，AndthecompoundofchondrocytesandcollagengelwereobservedinhistologysliceTheresu!tsshewsilkspresentedfavourableattachmenttOchondrocytesandkeptcellsgrowandproliferate。Thereweremuchextracellularsurroundingchondrocytes。Ininit

10、ialculture，chondrocytesincollagengelgrewbetterthanthatinsilkscaffoldHowevercollagenscattoldsdegradedandlosttheirstructure，silkconstructsretainedtheirstructurethroughoutthecultureaftertwoweeksofincubationThereforeaboveoft