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陨灶贼允韵责澡贼澡葬造皂燥造熏灾燥造援9熏晕燥援5熏May18,圆园16www.ijo.cn栽藻造押8629原愿圆圆源缘员苑圆8629-82210956耘皂葬蚤造押ijopress岳员远猿援糟燥皂
窑BasicResearch窑
1DepartmentofOphthalmology,theFourthAffiliatedHospitalofChinaMedicalUniversity,EyeHospitalofChinaMedicalUniversity,KeyLensResearchLaboratoryofLiaoningProvince,Shenyang110005,LiaoningProvince,China2DepartmentofPharmaceutics,SchoolofPharmacy,ShenyangPharmaceuticalUniversity,Shenyang110016,LiaoningProvince,ChinaCorrespondenceto:Jin-SongZhang;JunKong.
DepartmentofOphthalmology,theFourthAffiliatedHospitalofChinaMedicalUniversity,EyeHospitalofChinaMedicalUniversity,KeyLensResearchLaboratoryofLiaoningProvince,No.11XinhuaRoad,Shenyang110005,LiaoningProvince,China.76450751@qq.com;kongjun@sina.comReceived:2015-04-09Accepted:2015-11-18
Abstract·AIM:Todesignandinvestigatetheefficacyofamodifiednanostructuredlipidcarrierloadedwithgenistein(Gen-NLC)toinhibithumanlensepithelialcells(HLECs)proliferation.
·METHODS:Gen-NLCwasmadebymeltemulsificationmethod.Themorphology,particlesize(PS),zetapotentials(ZP),encapsulationefficiency(EE)andreleasewerecharacterized.Theinhibitioneffectofnanostructuredlipidcarrier(NLC),genistein(Gen)andGen-NLConHLECsproliferationwasevaluatedbycellcountingkit-8(CCK-8)assay,geneandproteinexpressionoftheproliferationmarkerKi67wereevaluatedwithreal-timequantitativepolymerasechainreaction(RT-qPCR)andimmunofluorescenceanalyses.
·RESULTS:ThemeanPSofGen-NLCwas80.12依1.55nmwithameanpolydispersityindexof0.11依0.02.ThemeanZPwas-7.14依0.38mVandtheEEofGeninthenanoparticleswas92.3%依0.73%.TransmissionelectronmicroscopyshowedthatGen-NLCdisplayedspherical-shapedparticlescoveredbyanouter-layerstructure.releaseexperimentsdemonstratedaprolongeddrugreleasefor72h.TheCCK-8assayresultsshowedtheNLChadnoinhibitoryeffectonHLECsandGen-NLCdisplayedamuchmoreprominentinhibitoryeffectoncellulargrowthcomparedtoGenofthesameconcentration.ThemRNAandproteinexpressionofKi67inLECsdecreasedsignificantlyinGen-NLCgroup.
·CONCLUSION:SustaineddrugreleasebyGen-NLCsmayimpedeHLECgrowth.·KEYWORDS:posteriorcapsularopacification;genistein;
nanostructuredlipidcarrier;humanlensepithelialcellsDOI:10.18240/ijo.2016.05.01
LiuJL,ZhangWJ,LiXD,YangN,PanWS,KongJ,ZhangJS.Sustained-releasegenisteinfromnanostructuredlipidcarriersuppresseshumanlensepithelialcellgrowth.2016;9(5):643-649
INTRODUCTIONCataractisoneofthemostcommondiseasesthataffect
theelderlypeopleandcataractsurgeryisthemostfrequentlyperformedocularprocedureintheworld.Despiteahighsuccessrate,displacementofintraocularlens(IOL),posteriorcapsuleopacification(PCO)andanteriorcapsulecontractionarehighlylikelytoreducethevisualqualityandevenresultinsecondvisionloss[1].Duringthefirstfewmonthsaftercataractsurgery,theresiduallensepithelialcells(LECs)begintomigrate,proliferate,andundergoepithelial-to-mesenchymaltransition(EMT)bywhichresultsincollagendepositionandfibrosisofthelenscapsule.Thisprocesscannotonlycausecontractionandnarrowingoftheanteriorcapsuleopening,butalsoresultsinPCOwhichleadstopoorvisualacuity[1].AlthoughNd:YAGlasercapsulotomyisthemostcommonmethodtotreatPCO,itsrelativecomplicationsandincreasedfinancialburdentothehealthcaresystemarethemainconcerns[2].OthermethodshavebeenemployedtopreventPCOsuchasnewIOLmaterials,square-edgeIOLdesign,andimprovedsurgicalprocedures[3-5],.However,therateofPCOwasnotreducedaspresumption.Theoretically,thedrugswhichcouldpreventLECsfromproliferationandmigrationcanbeusedtopreventPCO[6-8].Pharmacologicaltherapiesareanotherwaybroughtaboutrecentlyandsomeanti-inflammationandanti-metabolicagentshavebeenprovedtobesafeandeffectiveprophylacticstrategies.Genistein(Gen),(4',5,7-trihydroxyisoflavone),apotenttyrosinekinaseinhibitor,isaphytoestrogenwithawidevarietybiologicalfunctionssuchasanti-oxidant[9],phyto-oestrogenicandtyrosinekinaseinhibitoractivities[10]
andhasbeenshowntobeusefulagainstbreastandprostatecancers[11],cardiovasculardiseasesandpost-menopausal
Sustained-releasegenisteinfromnanostructuredlipidcarriersuppresseshumanlensepithelialcellgrowth
643ailments.Genhasbeenreportedtoprotectagainstlensopacityinhumanlensepithelialcells(HLECs)andinrateyes[12]anditssafetyalsohasbeenprovedasanintravitrealdrugintherabbitmodel[13].AlthoughGenhasbeenprovedtobeeffective,theshortresidencetimeinocular,especiallyinanteriorchamber,hindersitsperformanceinclinics.Duetothemultipleconstraintsimposedbytheeyeagainstthepenetrationofdrugs,theoculardeliveryandtargetingareparticularlyproblematic.Themajorchallengeinoculardrugtherapeutictreatmentwaspoorintraocularpenetrationandrapidocularelimination[14].Oneofthepromisingapproachestoimproveoculardrugeffectivenessisnanostructuredlipidcarrier(NLC).NLC,asthenewgenerationoflipidnanoparticledrugcarriersystem,havemanyadvantagesforenhancementofdrugpermeability,controlledrelease,targetingandsoon[15-16].Inthisstudy,wedesignedandmodifiedaninnovativeNLCfordrugdeliveryofGenbasedonourpreviousstudytoprovidehigherdrugloading,sustaineddrugreleaseandbetterbiocompatibility.Theeffectivenessofinhibitoryeffectofnanostructuredlipidcarrierloadedwithgenistein(Gen-NLC)onHLECsgrowthwasalsoevaluated.MATERIALSANDMETHODS