Mechanism of Cellular Uptake of Graphene Oxide Studied by Surface-Enhanced Raman Spectroscopy - Huang - 2012 - Small - Wiley Online Library
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The last few years have witnessed rapid development of biological and medical applications of graphene oxide (GO), such as drug/gene delivery, biosensing, and bioimaging. However, little is known about the cellular uptake mechanism and pathway of GO. In this work, surface-enhanced Raman scattering (SERS) spectroscopy is employed to investigate the cellular internalization of GO loaded with Au nanoparticles (NPs) by Ca Ski cells. The presence of Au NPs on the surface of GO enables detection of enhanced intrinsic Raman signals of GO inside the cell. The SERS results reveal that GO is distributed inhomogeneously inside the cell. Furthermore, internalization of Au-GO into Ca Ski cells is mainly via clathrin-mediated endocytosis, and is an energy-dependent process.二氧化硅在细胞中可降解_图文24-第2页
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二氧化硅在细胞中可降解_图文24-2
FIGURE6.TheSicontentinol；.]；isintherangeof300C430nm.；Itisknownthatmanykindsof；acidic；FIGURE7.TheSicontentince；.]；FIGURE8.Illustrationofa
FIGURE6.TheSicontentinoldmediumrefreshedfromthecellculturesatdifferenttimes(a)andtheaccumulativeSicontent(b)duringdiffer-entperiod.*p&0.05whencomparedwiththevalueatday0.[Color?gurecanbeviewedintheonlineissue,whichisavailableat
isintherangeof300C430nm.BasedonTEMobservation,thediameterofHMSNswasreducedto&200nmeitherincytoplasmorinlysosomesduringthecultureperiod.TheICP-OESresultsfurthershowedthattheSiwasdetectedinthesupernatantofcelllysatesandtheaccumulativeSicon-tentcontinuouslyincreasedinculturemedium.ThesedataclearlydemonstratedthatHMSNsweredegradedtosmallpiecesbyreleasingorexcretingSiionsoutofthecells.Recentresearchhasreportedthata?uorescentmesoporousSNpscanalsobeendocytosedincancercells(HeLa),buttheinternalizedparticlesseemtobemostlyexocytosedfromcellswithin96halmostwithoutdegradation.28Thisresultsuggeststhatnanoparticledegradationinsidethecellsmaypartlydependonthecelltypeandprobablypartlydependontheparticlenature.29
ItisknownthatmanykindsofhydrolyticenzymesresidedinsidethelysosomeatlowpHenvironmenttodigesttheendocytosedsubstrates.30ItwasreportedthatthisenvironmentmayresultincleavageofcovalentbondsbetweentheloadeddrugandSNps,leadingtothecompletereleaseofdrugs.26IronoxidenanoparticleshavebeenshowntobesolubilizedintheabsenceofanyenzymesatapHsimilartothatfoundinendosomesandlysosomes.31Butinthepresentstudy,thehydrolyticenzymeandthe
FIGURE7.TheSicontentincelllysates-5600gandcelllysates-11,000gduringdifferentperiods.After1dayofculture,theSicon-tentswerealmostconstantinlysates-11,000gduringdifferentperiods.Lysates-5600gandlysates-11,000g:thesupernatantsofthecelllysateswhichwerecentrifugedat5600gfor1minandat11,000gfor15min,respectively.[Color?gurecanbeviewedintheonlineissue,whichisavailableat
FIGURE8.IllustrationofapossibleprocessaccountingforHMSNsdegradationinHUVECs.[Color?.]
1402ZHAIETAL.DEGRADATIONOFHOLLOWMESOPOROUSSILICANANOPARTICLESINHUMANCELLS
ORIGINALRESEARCHREPORT
environmentseemlyhadlittleeffectontheHMSNsdegrada-tionafterbeingendocytosedinlysosomesasshowedbylaserscanningconfocalmicroscopy,astheSi-contentinlysates-11,000gfromdegradedHMSNsinsidecellswasrela-tivelyconstant(Figure7).However,furtherexperimentsneedtobedesignedtofullyelucidatetheprocessandthemechanismsoftheHMSNsdegradation.
Thepresentresultalsoshowedthatthedegradedprod-uctcanbeexcretedfromthecells.ThefastaccumulationofSicontentoutsidethecellsinthe?rst2daysmaybeowingtothereleaseofSiionsfromthedegradedHMSNsincyto-plasm,whichmaybeeasilyexcretedthroughcytomem-brane.After2daysofculture,theHMSNswereendocytosedinlysosomes(Figures3and4)andthedegradedproducthadtodefusethroughthelysosomemembrane,thecyto-plasm,andthecytomembranetoexcrete.ThisprocesswasmuchmorecomplexandmightresultinadecreaseoftheSiaccumulationafter2daysofculture.
Basedonourexperiments,HMSNisprovedtobedegradableinHUVEC?rstincytoplasmandlysosomesandsecondlyonlyinlysosomes.Thedegradationproductinsidethecellscanbeexcretedintotheculturemedium.Thedeg-radationrateisfastinthe?rst2daysandsloweddownaf-ter2days.DatafromthisinvestigationprovideaclueforfurtherstudyonthemetabolicwayandcytotoxicityofSNpsasdrugcarrierorfordiagnosticapplications.Furthermore,thepresentresultssuggestedthatHMSNmightbeabletoreleaseloadeddrugsinsidecellsthroughdegradation.
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