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Sau npe rau koj cov ntsiab lus tshwj xeeb thiab cov tshuaj tshwj xeeb uas txaus siab, thiab peb yuav phim cov ntaub ntawv koj muab nrog cov ntawv hauv peb cov ntaub ntawv dav dav thiab xa koj daim ntawv PDF ntawm email raws sijhawm.
Puas yog me me nanoparticles ib txwm zoo dua?Nkag siab txog cov teebmeem lom ntawm qhov loj me-nyob ntawm kev sib sau ntawm cov nyiaj nanoparticles nyob rau hauv cov xwm txheej lom lom.
Sau: Bélteky P, Rónavári A, Zakupszky D, Boka E, Igaz N, Szerencsés B, Pfeiffer I, Vágvölgyi C, Kiricsi M, Kónya Z
Péter Bélteky, 1, * Andrea Rónavári, 1,* Dalma Zakupszky, 1 Eszter Boka, 1 Nóra Igaz, 2 Bettina Szerencsés, 3 Ilona Pfeiffer, 3 Csaba Vágvölgyi, 3 Mónika Kiricsi ntawm Environmental Chemistry ntawm Kev Tshawb Fawb, Hung , University of Szeged;2 Department of Biochemistry thiab Molecular Biology, Kws Qhia Ntawv ntawm Kev Tshawb Fawb thiab Cov Ntaub Ntawv, University of Szeged, Hungary;3 Department of Microbiology, Kws Qhia Ntawv ntawm Kev Tshawb Fawb thiab Cov Ntaub Ntawv, University of Szeged, Hungary;4MTA-SZTE Reaction Kinetics thiab Surface Chemistry Research Group, Szeged, Hungary* Cov kws sau ntawv no tau pab sib npaug rau txoj haujlwm no.Kev sib txuas lus: Zoltán Kónya Department of Applied and Environment Chemistry, Kws qhia ntawv ntawm Science thiab Informatics, University of Szeged, Rerrich Square 1, Szeged, H-6720, Hungary Xov tooj +36 62 544620 Email [Email tiv thaiv] Lub hom phiaj: Silver nanoparticles (AgNPs) yog ib qho ntawm feem ntau kawm nanomaterials, tshwj xeeb tshaj yog vim lawv cov kev siv biomedical.Txawm li cas los xij, vim yog kev sib sau ua ke ntawm nanoparticles, lawv cov cytotoxicity zoo heev thiab cov tshuaj tua kab mob feem ntau cuam tshuam rau hauv cov xov xwm lom neeg.Hauv kev ua haujlwm no, kev coj tus cwj pwm sib sau ua ke thiab kev ua haujlwm lom neeg muaj feem cuam tshuam ntawm peb qhov sib txawv citrate-txiav nyiaj nanoparticle kuaj nrog qhov nruab nrab txoj kab uas hla ntawm 10, 20, thiab 50 nm tau kawm.Txoj kev: Siv cov tshuab hluav taws xob xa hluav taws xob los ua ke thiab ua tus yam ntxwv ntawm nanoparticles, ntsuas lawv tus cwj pwm sib sau ua ke ntawm ntau yam pH qhov tseem ceeb, NaCl, qabzib thiab glutamine concentrations los ntawm dynamic light scattering thiab ultraviolet-pom spectroscopy.Tsis tas li ntawd, nyob rau hauv lub cell kab lis kev cai nruab nrab Cheebtsam xws li Dulbecco txhim kho tus cwj pwm aggregation nyob rau hauv Eagle Medium thiab Fetal Calf Serum.Cov txiaj ntsig: Cov txiaj ntsig tau pom tias acidic pH thiab cov ntsiab lus physiological electrolyte feem ntau ua rau cov micron-scale aggregation, uas tuaj yeem kho los ntawm kev tsim ntawm biomolecular corona.Nws yog tsim nyog sau cia tias cov khoom loj dua tuaj yeem ua rau muaj kev cuam tshuam sab nraud ntau dua li lawv cov khoom me me.Hauv vitro cytotoxicity thiab tshuaj tua kab mob tau ua los ntawm kev kho cov hlwb nrog nanoparticle aggregates nyob rau theem sib txawv.Xaus: Peb cov txiaj ntsig tau nthuav tawm qhov kev sib raug zoo ntawm colloidal stability thiab toxicity ntawm AgNPs, vim tias kev sib sau hnyav ua rau ua tiav kev ua haujlwm lom lom.Qhov siab dua ntawm kev tiv thaiv kev sib sau ua ke pom rau cov khoom loj dua muaj qhov cuam tshuam loj rau hauv vitro toxicity, vim tias cov qauv no khaws ntau cov tshuaj tiv thaiv kab mob thiab cov tsiaj ntawm tes.Cov kev tshawb pom no ua rau kev txiav txim siab tias, txawm hais tias muaj kev xav dav dav hauv cov ntaub ntawv muaj feem cuam tshuam, tsom rau qhov tsawg tshaj plaws nanoparticles yuav tsis yog qhov zoo tshaj plaws ntawm kev ua.Keywords: noob-mediated kev loj hlob, colloidal stability, loj-dependent aggregation cwj pwm, aggregation puas toxicity
Raws li qhov kev thov thiab cov khoom tsim tawm ntawm nanomaterials txuas ntxiv mus, ntau thiab ntau cov kev saib xyuas tau them rau lawv cov biosafety lossis lom lom.Silver nanoparticles (AgNPs) yog ib qho ntawm feem ntau synthesized, tshawb fawb thiab siv cov neeg sawv cev ntawm cov ntaub ntawv no vim lawv zoo heev catalytic, kho qhov muag thiab lom zog.1 Nws feem ntau ntseeg tau tias cov yam ntxwv tshwj xeeb ntawm nanomaterials (xws li AgNPs) feem ntau yog los ntawm lawv qhov chaw loj tshwj xeeb.Yog li ntawd, qhov teeb meem inevitably yog cov txheej txheem uas cuam tshuam rau qhov tseem ceeb no, xws li particle loj, txheej txheej los yog aggregation, seb nws puas yuav ua rau cov khoom ntawm nanoparticles tseem ceeb heev rau cov ntawv thov tshwj xeeb.
Cov teebmeem ntawm particle loj thiab stabilizers yog cov ntsiab lus uas tau raug txheeb xyuas zoo hauv cov ntaub ntawv.Piv txwv li, qhov pom pom dav dav yog tias cov nanoparticles me me yog cov tshuaj lom ntau dua li cov nanoparticles loj.2 Raws li cov ntaub ntawv dav dav, peb cov kev tshawb fawb yav dhau los tau qhia txog qhov loj me-nyob ntawm kev ua haujlwm ntawm nanosilver ntawm mammalian hlwb thiab kab mob.3- 5 Cov txheej txheej saum npoo yog lwm tus cwj pwm uas muaj kev cuam tshuam dav rau cov khoom ntawm nanomaterials.Tsuas yog los ntawm kev ntxiv lossis hloov cov stabilizers ntawm nws qhov chaw, tib lub nanomaterial tej zaum yuav muaj qhov sib txawv ntawm lub cev, tshuaj, thiab cov khoom lom.Daim ntawv thov ntawm capping agents feem ntau yog ua ib feem ntawm nanoparticle synthesis.Piv txwv li, citrate-terminated silver nanoparticles yog ib qho tseem ceeb tshaj plaws AgNPs hauv kev tshawb fawb, uas yog synthesized los ntawm kev txo cov nyiaj ntsev nyob rau hauv ib tug xaiv stabilizer tov raws li cov tshuaj tiv thaiv nruab nrab.6 Citrate tuaj yeem yooj yim ua kom zoo dua ntawm nws tus nqi qis, muaj, biocompatibility, thiab muaj zog affinity rau nyiaj, uas tuaj yeem cuam tshuam los ntawm ntau qhov kev sib tham, los ntawm kev thim rov qab ntawm qhov adsorption mus rau ionic kev sib cuam tshuam.Cov molecules me me thiab polyatomic ions nyob ze 7,8, xws li citrates, polymers, polyelectrolytes, thiab cov tshuaj lom neeg kuj tau siv los ua kom ruaj khov nano-silver thiab ua haujlwm tshwj xeeb ntawm nws.9-12
Txawm hais tias muaj peev xwm hloov pauv qhov kev ua ntawm nanoparticles los ntawm kev txhob txwm nto capping yog ib qho chaw nthuav dav, lub luag haujlwm tseem ceeb ntawm cov txheej txheej no yog qhov tsis txaus ntseeg, muab colloidal stability rau nanoparticle system.Qhov loj tshwj xeeb ntawm thaj chaw ntawm nanomaterials yuav tsim lub zog loj, uas cuam tshuam lub peev xwm ntawm lub cev kom ncav cuag nws lub zog yam tsawg kawg nkaus.13 Yog tias tsis muaj kev ruaj ntseg zoo, qhov no tuaj yeem ua rau agglomeration ntawm nanomaterials.Aggregation yog tsim los ntawm aggregates ntawm ntau cov duab thiab ntau thiab tsawg uas tshwm sim thaum dispersed hais sib ntsib thiab tam sim no thermodynamic kev sib cuam tshuam cia cov hais mus rau ib leeg.Yog li ntawd, stabilizers yog siv los tiv thaiv kev sib sau los ntawm kev qhia lub zog txaus txaus ntshai ntawm cov khoom los tiv thaiv lawv qhov kev nyiam thermodynamic.14
Txawm hais tias cov ntsiab lus ntawm qhov loj me thiab qhov chaw npog tau raug tshawb xyuas meej hauv cov ntsiab lus ntawm nws cov kev cai ntawm cov dej num lom neeg uas tshwm sim los ntawm nanoparticles, particle aggregation yog ib qho chaw tsis saib xyuas ntau.Yuav luag tsis muaj kev kawm zoo los daws cov colloidal stability ntawm nanoparticles nyob rau hauv cov xwm txheej lom.10,15-17 Tsis tas li ntawd, qhov kev pab cuam no yog qhov tshwj xeeb tshaj yog tsis tshua muaj, qhov twg cov tshuaj lom neeg cuam tshuam nrog kev sib sau ua ke kuj tau kawm, txawm tias nws yuav ua rau muaj kev cuam tshuam tsis zoo, xws li vascular thrombosis, lossis poob ntawm cov yam ntxwv xav tau, xws li nws cov toxicity, xws li qhia hauv daim duab 1.18, 19 qhia.Qhov tseeb, ib qho ntawm ob peb lub tswv yim ntawm cov nyiaj nanoparticle tsis kam yog cuam tshuam nrog kev sib sau ua ke, vim qee yam E. coli thiab Pseudomonas aeruginosa strains tau tshaj tawm kom txo lawv cov nano-silver rhiab heev los ntawm kev qhia cov protein flagellin, flagellin.Nws muaj affinity siab rau nyiaj, yog li inducing aggregation.20
Muaj ntau ntau yam sib txawv uas cuam tshuam txog kev lom ntawm cov nyiaj nanoparticles, thiab kev sib sau ua ke cuam tshuam rau tag nrho cov txheej txheem no.Txoj kev sib tham tshaj plaws ntawm AgNP lom neeg ua haujlwm, qee zaum hu ua "Trojan Horse", suav txog AgNPs li Ag + cov neeg nqa khoom.1,21 Lub Trojan nees mechanism tuaj yeem ua kom muaj kev nce ntxiv hauv lub zos Ag + concentration, uas ua rau lub cim ntawm ROS thiab membrane depolarization.22-24 Kev sib sau ua ke tuaj yeem cuam tshuam rau kev tso tawm Ag +, yog li cuam tshuam rau toxicity, vim tias nws txo qis qhov ua haujlwm zoo ntawm cov nyiaj ions yuav oxidized thiab yaj.Txawm li cas los xij, AgNPs yuav tsis tsuas pom muaj tshuaj lom los ntawm kev tso tawm ion.Ntau qhov loj thiab morphology-txog kev sib cuam tshuam yuav tsum tau xav txog.Ntawm lawv, qhov loj thiab cov duab ntawm nanoparticle nto yog cov yam ntxwv uas txhais tau.4,25 Cov kev sau los ntawm cov txheej txheem no tuaj yeem raug cais raws li "kev ua kom muaj kuab lom lom zem."Muaj peev xwm ua tau ntau yam mitochondrial thiab nto membrane cov tshuaj tiv thaiv uas tuaj yeem ua rau cov organelles puas tsuaj thiab ua rau cov cell tuag.25-27 Txij li thaum tsim cov aggregates ib txwm muaj feem xyuam rau qhov loj thiab cov duab ntawm cov khoom uas muaj nyiaj uas pom tau los ntawm cov kab ke nyob, cov kev sib cuam tshuam no kuj tseem cuam tshuam.
Hauv peb daim ntawv dhau los ntawm kev sib sau ua ke ntawm cov nyiaj nanoparticles, peb tau pom cov txheej txheem tshuaj ntsuam xyuas zoo uas muaj cov tshuaj lom neeg thiab hauv vitro kev sim tshuaj lom neeg los kawm txog qhov teeb meem no.19 Dynamic Light Scattering (DLS) yog cov txheej txheem nyiam rau cov hom kev tshuaj xyuas no vim tias cov khoom tuaj yeem tawg photons ntawm lub wavelength piv rau qhov loj ntawm nws cov khoom.Txij li thaum Brownian motion ceev ntawm hais nyob rau hauv cov kua nruab nrab muaj feem xyuam rau qhov luaj li cas, qhov kev hloov nyob rau hauv qhov kev siv ntawm tawg tawg lub teeb yuav siv tau los txiav txim qhov nruab nrab hydrodynamic txoj kab uas hla (Z-txhais) ntawm cov kua qauv.28 Tsis tas li ntawd, los ntawm kev siv ib qho hluav taws xob rau cov qauv, qhov zeta muaj peev xwm (ζ muaj peev xwm) ntawm nanoparticle tuaj yeem ntsuas zoo ib yam li Z tus nqi nruab nrab.13,28 Yog tias tus nqi tiag tiag ntawm lub peev xwm zeta yog siab txaus (raws li cov lus qhia dav dav> ± 30 mV), nws yuav ua kom muaj zog electrostatic repulsion ntawm cov khoom los tiv thaiv cov aggregation.Cov yam ntxwv saum npoo plasmon resonance (SPR) yog qhov tshwm sim tshwj xeeb kho qhov muag, feem ntau yog los ntawm cov hlau muaj nuj nqis nanoparticles (tsuas yog Au thiab Ag).Raws li cov hluav taws xob oscillations (surface plasmons) ntawm cov ntaub ntawv no ntawm nanoscale, nws paub tias spherical AgNPs muaj tus yam ntxwv UV-Vis absorption ncov ze 400 nm.30 Kev siv zog thiab kev hloov pauv ntawm cov khoom siv los ntxiv cov txiaj ntsig DLS, vim tias txoj kev no tuaj yeem siv los txheeb xyuas nanoparticle aggregation thiab nto adsorption ntawm biomolecules.
Raws li cov ntaub ntawv tau txais, cell viability (MTT) thiab tshuaj tua kab mob tau ua raws li AgNP toxicity tau piav qhia raws li kev ua haujlwm ntawm qib sib sau ua ke, tsis yog (qhov feem ntau siv ntau tshaj) nanoparticle concentration.Txoj kev tshwj xeeb no tso cai rau peb los ua kom pom qhov tseem ceeb ntawm qib sib sau ua ke hauv kev ua haujlwm lom neeg, vim hais tias, piv txwv li, citrate-terminated AgNPs kiag li poob lawv cov kev ua ub no hauv ob peb teev vim kev sib sau ua ke.19
Hauv kev ua haujlwm tam sim no, peb tsom kom nthuav dav peb cov kev koom tes yav dhau los hauv kev ruaj ntseg ntawm bio-txog colloids thiab lawv cov kev cuam tshuam rau kev ua haujlwm lom neeg los ntawm kev kawm txog cov txiaj ntsig ntawm nanoparticle loj ntawm nanoparticle aggregation.Qhov no yog undoubtedly ib qho ntawm cov kev tshawb fawb ntawm nanoparticles.Txoj kev xav ntau dua thiab 31 Los tshawb xyuas qhov teeb meem no, ib txoj kev loj hlob ntawm cov noob tau siv los tsim cov citrate-terminated AgNPs hauv peb qhov sib txawv (10, 20, thiab 50 nm).6,32 raws li ib qho ntawm feem ntau txoj kev.Rau cov khoom siv nano uas tau dav thiab niaj hnub siv hauv kev siv kho mob, citrate-terminated AgNPs ntawm ntau qhov sib txawv raug xaiv los kawm txog qhov muaj peev xwm loj ntawm kev sib sau ua ke txog kev lom neeg ntawm nanosilver.Tom qab kev sib txuas AgNPs ntawm ntau qhov sib txawv, peb pom cov qauv tsim los ntawm kev xa hluav taws xob microscopy (TEM), thiab tom qab ntawd tshuaj xyuas cov khoom siv cov txheej txheem tshuaj ntsuam tau hais tseg.Tsis tas li ntawd, nyob rau hauv lub xub ntiag ntawm in vitro cell kab lis kev cai Dulbecco's Modified Eagle's Medium (DMEM) thiab Fetal Bovine Serum (FBS), qhov loj-dependent aggregation cwj pwm thiab nws tus cwj pwm raug soj ntsuam ntawm ntau yam pH qhov tseem ceeb, NaCl, qabzib, thiab glutamine concentrations.Cov yam ntxwv ntawm cytotoxicity yog txiav txim siab raws li kev ua tiav.Qhov kev pom zoo ntawm kev tshawb fawb qhia tias feem ntau, cov khoom me me yog qhov zoo dua;peb qhov kev tshawb nrhiav muab cov tshuaj lom neeg thiab tshuaj lom neeg los txiav txim seb qhov no puas yog.
Peb cov nyiaj nanoparticles nrog ntau qhov sib txawv tau npaj los ntawm txoj kev loj hlob ntawm cov noob los ntawm Wan li al., nrog kev hloov me ntsis.6 Txoj kev no yog los ntawm kev txo cov tshuaj lom neeg, siv nyiaj nitrate (AgNO3) ua cov nyiaj, sodium borohydride (NaBH4) ua tus txo qis, thiab sodium citrate ua tus stabilizer.Ua ntej, npaj 75 mL ntawm 9 mM citrate aqueous tov los ntawm sodium citrate dihydrate (Na3C6H5O7 x 2H2O) thiab kub rau 70 ° C.Tom qab ntawd, 2 mL ntawm 1% w / v AgNO3 tov tau ntxiv rau qhov nruab nrab cov tshuaj tiv thaiv, thiab tom qab ntawd cov tshuaj tshiab tau npaj sodium borohydride (2 mL 0.1% w / v) tau nchuav rau hauv qhov sib tov dropwise.Qhov tshwm sim daj-xim av raug tshem tawm tau khaws cia ntawm 70 ° C nrog lub zog nplawm rau 1 teev, thiab tom qab ntawd txias rau chav tsev kub.Cov qauv uas tau tshwm sim (hu ua AgNP-I txij tam sim no mus) yog siv los ua lub hauv paus rau kev loj hlob ntawm cov noob hauv cov txheej txheem synthesis tom ntej.
Txhawm rau ua kom muaj qhov nruab nrab qhov nruab nrab qhov nruab nrab (lub npe hu ua AgNP-II), cua sov 90 mL 7.6 mM citrate tov rau 80 ° C, sib tov nrog 10 mL AgNP-I, thiab ces sib tov 2 mL 1% w / v Cov tshuaj AgNO3 tau khaws cia nyob rau hauv lub zog txhua yam nplawm rau 1 teev, thiab tom qab ntawd cov qauv tau txias rau chav sov.
Rau qhov loj tshaj particle (AgNP-III), rov ua tib txoj kev loj hlob, tab sis nyob rau hauv cov ntaub ntawv no, siv 10 mL ntawm AgNP-II raws li cov noob raug tshem tawm.Tom qab cov qauv mus txog qhov kub thiab txias, lawv tau teeb tsa lawv cov nominal Ag concentration raws li tag nrho AgNO3 cov ntsiab lus rau 150 ppm los ntawm kev ntxiv lossis evaporating ntxiv hnyav ntawm 40 ° C, thiab thaum kawg khaws cia ntawm 4 ° C kom txog thaum siv ntxiv.
Siv FEI Tecnai G2 20 X-Twin Transmission Electron Microscope (TEM) (FEI Corporate Headquarters, Hillsboro, Oregon, USA) nrog 200 kV acceleration voltage los tshuaj xyuas cov yam ntxwv morphological ntawm nanoparticles thiab ntes lawv cov electron diffraction (ED) qauv.Yam tsawg kawg 15 tus neeg sawv cev duab (~750 hais) tau soj ntsuam siv ImageJ software pob, thiab cov txiaj ntsig histograms (thiab tag nrho cov duab hauv tag nrho txoj kev tshawb fawb) tau tsim hauv OriginPro 2018 (OriginLab, Northampton, MA, USA) 33, 34.
Qhov nruab nrab hydrodynamic txoj kab uas hla (Z-nruab nrab), zeta muaj peev xwm (ζ-muaj peev xwm) thiab tus yam ntxwv nto plasmon resonance (SPR) ntawm cov qauv raug ntsuas los qhia lawv cov thawj colloidal zog.Qhov nruab nrab hydrodynamic txoj kab uas hla thiab zeta muaj peev xwm ntawm cov qauv raug ntsuas los ntawm Malvern Zetasizer Nano ZS ntsuas (Malvern Instruments, Malvern, UK) siv cov folded capillary hlwb ntawm 37 ± 0.1 ° C.Dej hiav txwv Optics 355 DH-2000-BAL UV-Vis spectrophotometer (Halma PLC, Largo, FL, USA) tau siv los muab cov yam ntxwv SPR los ntawm UV-Vis absorption spectra ntawm cov qauv hauv thaj tsam ntawm 250-800 nm.
Thaum lub sij hawm tag nrho kev sim, peb hom kev ntsuas sib txawv uas muaj feem xyuam rau kev ruaj ntseg colloidal tau ua tib lub sijhawm.Siv DLS los ntsuas qhov nruab nrab hydrodynamic txoj kab uas hla (Z nruab nrab) thiab zeta muaj peev xwm (ζ muaj peev xwm) ntawm cov hais, vim hais tias tus Z nruab nrab muaj feem xyuam rau qhov nruab nrab qhov loj ntawm cov nanoparticle aggregates, thiab lub zeta muaj peev xwm qhia tau hais tias lub electrostatic repulsion nyob rau hauv lub system. muaj zog txaus los cuam tshuam Van der Waals attraction ntawm nanoparticles.Kev ntsuas yog ua nyob rau hauv triplicate, thiab tus qauv sib txawv ntawm Z txhais tau tias thiab zeta peev xwm yog xam los ntawm Zetasizer software.Tus yam ntxwv SPR spectra ntawm cov khoom raug soj ntsuam los ntawm UV-Vis spectroscopy, vim tias kev hloov pauv ntawm qhov siab tshaj plaws thiab lub wavelength tuaj yeem qhia txog kev sib sau ua ke thiab kev sib tshuam ntawm qhov chaw.29,35 Qhov tseeb, qhov chaw plasmon resonance nyob rau hauv cov hlau muaj txiaj ntsig yog qhov muaj txiaj ntsig zoo uas nws tau coj mus rau txoj kev tshiab ntawm kev tshuaj xyuas biomolecules.29,36,37 Qhov concentration ntawm AgNPs nyob rau hauv kev sim sib tov yog hais txog 10 ppm, thiab lub hom phiaj yog los teeb tsa qhov kev siv ntawm qhov siab tshaj plaws thawj zaug SPR nqus rau 1. Qhov kev sim tau ua nyob rau hauv ib lub sij hawm-nyob ntawm seb ntawm 0;1.5;3;6;12 thiab 24 teev nyob rau hauv ntau yam kab mob lom.Ntau cov ntsiab lus piav qhia txog qhov kev sim tuaj yeem pom hauv peb txoj haujlwm dhau los.Nyob rau hauv luv luv, ntau yam pH qhov tseem ceeb (3; 5; 7.2 thiab 9), sib txawv sodium chloride (10 mM; 50 mM; 150 mM), qabzib (3.9 mM; 6.7 mM) thiab glutamine (4 mM) concentration, thiab kuj tau npaj Dulbecco's Modified Eagle Medium (DMEM) thiab Fetal Bovine Serum (FBS) (hauv dej thiab DMEM) ua qauv qauv, thiab kawm txog lawv cov teebmeem ntawm kev coj tus cwj pwm sib sau ntawm cov nyiaj nanoparticles synthesized.pH Cov txiaj ntsig ntawm, NaCl, qabzib, thiab glutamine raug soj ntsuam raws li kev xav ntawm lub cev, thaum cov nyiaj ntawm DMEM thiab FBS yog tib yam li cov qib uas siv tag nrho hauv vitro sim.38-42 Tag nrho cov kev ntsuas tau ua nyob rau ntawm pH 7.2 thiab 37 ° C nrog ib tug tas mus li keeb kwm yav dhau ntsev concentration ntawm 10 mM NaCl kom tshem tawm ib tug ntev-ntev particle sib cuam tshuam (tshwj tsis yog rau tej yam pH thiab NaCl-txog thwmsim, qhov twg cov cwj pwm no yog cov variables nyob rau hauv kawm).28 Cov npe ntawm ntau yam xwm txheej tau sau tseg hauv Table 1. Cov kev sim uas cim nrog † yog siv los siv thiab sib raug rau cov qauv uas muaj 10 mM NaCl thiab pH 7.2.
Human prostate cancer cell line (DU145) thiab immortalized human keratinocytes (HaCaT) tau txais los ntawm ATCC (Manassas, VA, USA).Cov kab mob niaj hnub niaj hnub siv hauv Dulbecco qhov tsawg kawg nkaus qhov nruab nrab Eagle (DMEM) uas muaj 4.5 g / L qabzib (Sigma-Aldrich, Saint Louis, MO, USA), ntxiv nrog 10% FBS, 2 mM L-glutamine, 0.01% Streptomycin thiab 0.005% Penicillin (Sigma-Aldrich, St. Louis, Missouri, USA).Cov hlwb yog kab lis kev cai hauv 37 ° C incubator hauv qab 5% CO2 thiab 95% av noo.
Txhawm rau tshawb nrhiav cov kev hloov pauv hauv AgNP cytotoxicity los ntawm particle aggregation nyob rau hauv lub sij hawm, ib tug ob-kauj ruam MTT xeem tau ua.Ua ntej, qhov muaj peev xwm ntawm ob hom cell raug ntsuas tom qab kho nrog AgNP-I, AgNP-II thiab AgNP-III.Txog rau qhov kawg no, ob hom hlwb tau cog rau hauv 96-zoo daim hlau ntawm qhov ntom ntawm 10,000 hlwb / zoo thiab kho nrog peb qhov sib txawv ntawm cov nyiaj nanoparticles hauv kev nce ntxiv rau hnub thib ob.Tom qab 24 teev ntawm kev kho mob, cov hlwb raug ntxuav nrog PBS thiab incubated nrog 0.5 mg / mL MTT reagent (SERVA, Heidelberg, Lub teb chaws Yelemees) diluted nyob rau hauv kab lis kev cai nruab nrab rau 1 teev ntawm 37 ° C.Formazan crystals tau yaj hauv DMSO (Sigma-Aldrich, Saint Louis, MO, USA), thiab qhov nqus tau ntsuas ntawm 570 nm siv Synergy HTX phaj nyeem ntawv (BioTek-Hungary, Budapest, Hungary).Tus nqi nqus ntawm cov qauv tswj tsis tau kho yog suav tias yog 100% ciaj sia taus.Ua yam tsawg kawg 3 qhov kev sim uas siv plaub qhov kev ywj pheej lom zem dua.IC50 yog xam los ntawm koob tshuaj tiv thaiv kab mob raws li cov txiaj ntsig tseem ceeb.
Tom qab ntawd, nyob rau theem thib ob, los ntawm incubating cov hais nrog 150 mM NaCl rau lub sijhawm sib txawv (0, 1.5, 3, 6, 12, thiab 24 teev) ua ntej kev kho ntawm tes, kev sib xyaw sib txawv ntawm cov nyiaj nanoparticles tau tsim.Tom qab ntawd, tib qhov kev ntsuam xyuas MTT tau ua raws li tau piav qhia yav dhau los los ntsuas cov kev hloov pauv hauv cell viability cuam tshuam los ntawm kev sib sau ua ke.Siv GraphPad Prism 7 los ntsuas qhov kawg tshwm sim, xam qhov tseem ceeb ntawm qhov kev sim los ntawm kev tsis sib xws t-test, thiab kos nws qib li * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001). ) Thiab **** (p ≤ 0.0001).
Peb qhov sib txawv ntawm cov nyiaj nanoparticles (AgNP-I, AgNP-II thiab AgNP-III) tau siv rau cov kab mob tiv thaiv kab mob rau Cryptococcus neoformans IFM 5844 (IFM; Chaw Tshawb Fawb rau Pathogenic Fungi thiab Microbial Toxicology, Chiba University) thiab Bacillus Test megaterium SZMC 603 (SZMC: Szeged Microbiology Collection) thiab E. coli SZMC 0582 hauv RPMI 1640 nruab nrab (Sigma-Aldrich Co.).Txhawm rau ntsuas qhov kev hloov pauv ntawm cov kab mob tua kab mob los ntawm kev sib sau ntawm cov khoom, ua ntej, lawv qhov tsawg kawg nkaus inhibitory concentration (MIC) tau txiav txim siab los ntawm microdilution hauv 96-zoo microtiter phaj.Rau 50 μL ntawm cov txheej txheem kev ncua ntawm tes (5 × 104 hlwb / mL hauv RPMI 1640 nruab nrab), ntxiv 50 μL ntawm cov nyiaj nanoparticle ncua kev kawm ntawv thiab serially dilute ob zaug qhov concentration (hauv qhov nruab nrab tau hais tseg, qhov ntau yog 0 thiab 75 ppm, uas yog, cov qauv tswj muaj 50 μL ntawm kev ncua ntawm tes thiab 50 μL ntawm nruab nrab yam tsis muaj nanoparticles).Tom qab ntawd, lub phaj tau incubated ntawm 30 ° C rau 48 teev, thiab qhov optical ceev ntawm kab lis kev cai tau ntsuas ntawm 620 nm siv SPECTROstar Nano phaj nyeem ntawv (BMG LabTech, Offenburg, Lub teb chaws Yelemees).Qhov kev sim tau ua peb zaug hauv triplicate.
Tsuas yog tias 50 μL ntawm ib qho kev sib sau ua ke nanoparticles tau siv nyob rau lub sijhawm no, tib txoj kev raws li tau piav qhia yav dhau los tau siv los tshuaj xyuas cov txiaj ntsig ntawm kev sib sau ua ke ntawm cov tshuaj tua kab mob ntawm cov kab mob uas tau hais tseg.Cov xeev sib txawv ntawm cov nyiaj nanoparticles yog tsim los ntawm incubating cov khoom nrog 150 mM NaCl rau lub sijhawm sib txawv (0, 1.5, 3, 6, 12, thiab 24 teev) ua ntej ua haujlwm ntawm tes.Ib qho kev ncua ntxiv nrog 50 μL ntawm RPMI 1640 nruab nrab tau siv los ua kev tswj kev loj hlob, thaum txhawm rau tswj cov tshuaj lom, kev ncua nrog cov nanoparticles tsis sib sau ua ke.Qhov kev sim tau ua peb zaug hauv triplicate.Siv GraphPad Prism 7 los ntsuas qhov kawg tshwm sim dua, siv tib qhov kev txheeb cais raws li MTT tsom xam.
Cov qib sib sau ua ke ntawm cov khoom me me (AgNP-I) tau tshwm sim, thiab cov txiaj ntsig tau ib nrab tau tshaj tawm hauv peb cov haujlwm dhau los, tab sis rau kev sib piv zoo dua, tag nrho cov khoom tau raug kuaj xyuas kom zoo.Cov ntaub ntawv sim tau sau thiab sib tham hauv ntu hauv qab no.Peb qhov ntau thiab tsawg ntawm AgNP.19
Kev ntsuas ua los ntawm TEM, UV-Vis thiab DLS tau txheeb xyuas qhov ua tiav ntawm tag nrho cov qauv AgNP (Daim duab 2A-D).Raws li thawj kab ntawm daim duab 2, qhov tsawg tshaj plaws particle (AgNP-I) qhia tau hais tias ib tug zoo spherical morphology nrog ib tug nruab nrab txoj kab uas hla ntawm txog 10 nm.Txoj kev loj hlob ntawm cov noob qoob loo kuj muab AgNP-II thiab AgNP-III nrog ntau qhov sib txawv nrog qhov nruab nrab particle diameters ntawm kwv yees li 20 nm thiab 50 nm, raws li.Raws li tus qauv sib txawv ntawm qhov kev faib tawm, qhov ntau thiab tsawg ntawm peb cov qauv tsis sib tshooj, uas yog qhov tseem ceeb rau lawv qhov kev sib piv.Los ntawm kev sib piv qhov nruab nrab nam piv thiab nyias qhov piv ntawm TEM-raws li particle 2D projections, nws yog assumed tias lub sphericity ntawm cov hais yog soj ntsuam los ntawm ImageJ cov duab lim plug-in (Daim duab 2E).43 Raws li kev soj ntsuam ntawm cov duab ntawm cov khoom, lawv qhov sib piv (sab loj / luv sab ntawm lub me me bounding duab plaub) tsis cuam tshuam los ntawm particle loj hlob, thiab lawv cov thinness ratio ( ntsuas cheeb tsam ntawm lub voj voog zoo meej / cheeb tsam theoretical. ) maj mam txo.Qhov no ua rau muaj ntau thiab ntau dua polyhedral hais, uas zoo kawg nkaus puag ncig nyob rau hauv txoj kev xav, coj mus rau ib tug thinness ratio ntawm 1.
Daim duab 2 Transmission electron microscope (TEM) image (A), electron diffraction (ED) pattern (B), size distribution histogram (C), yam ntxwv ultraviolet-pom (UV-Vis) lub teeb absorption spectrum (D), thiab nruab nrab kua Citrate -terminated silver nanoparticles nrog mechanical txoj kab uas hla (Z-nruab nrab), zeta muaj peev xwm, nam piv thiab thickness ratio (E) muaj peb txawv loj ntau yam: AgNP-I yog 10 nm (sab saum toj kab), AgNP -II yog 20 nm (nruab nrab kab ), AgNP-III (kab hauv qab) yog 50 nm.
Txawm hais tias qhov xwm txheej ntawm txoj kev loj hlob cuam tshuam rau cov duab particle mus rau qee qhov, ua rau qhov me me ntawm cov AgNPs loj dua, tag nrho peb cov qauv tseem yog quasi-spherical.Tsis tas li ntawd, raws li qhia nyob rau hauv cov qauv electron diffraction hauv daim duab 2B, nano Lub crystallinity ntawm cov khoom tsis cuam tshuam.Lub nplhaib diffraction tseem ceeb-uas tuaj yeem cuam tshuam nrog (111), (220), (200), thiab (311) Miller indices ntawm nyiaj-yog qhov zoo sib xws nrog cov ntaub ntawv tshawb fawb thiab peb cov kev koom tes yav dhau los.9, 19,44 Lub fragmentation ntawm lub nplhaib Debye-Scherrer ntawm AgNP-II thiab AgNP-III yog vim lub fact tias cov duab ED yog ntes nyob rau tib lub magnification, yog li raws li particle loj nce, tus naj npawb ntawm diffracted hais ib. chav tsev nce thiab txo qis.
Qhov loj thiab cov duab ntawm nanoparticles paub tias cuam tshuam rau kev ua haujlwm lom neeg.3,45 Shape-dependent catalytic thiab biological activation tuaj yeem piav qhia los ntawm qhov tseeb tias cov duab sib txawv zoo li nthuav tawm qee lub ntsej muag siv lead ua (muaj qhov sib txawv Miller indices), thiab cov ntsej muag siv lead ua no muaj cov haujlwm sib txawv.45,46 Txij li thaum cov khoom npaj tau muab cov txiaj ntsig zoo sib xws ntawm ED sib haum rau cov yam ntxwv zoo sib xws, nws tuaj yeem xav tias hauv peb cov kev ua haujlwm tom qab colloidal thiab kev sim tshuaj lom neeg, txhua qhov sib txawv yuav tsum raug ntaus nqi rau Nanoparticle loj, tsis zoo li cov khoom muaj feem xyuam.
Cov txiaj ntsig UV-Vis tau sau tseg hauv daim duab 2D ntxiv hais txog qhov xwm txheej loj ntawm qhov sib txuas ntawm AgNP, vim tias SPR peaks ntawm tag nrho peb cov qauv yog nyob ib ncig ntawm 400 nm, uas yog tus yam ntxwv tus nqi ntawm spherical silver nanoparticles.29,30 Lub captured spectra kuj tau lees paub qhov kev vam meej noob-mediated kev loj hlob ntawm nanosilver.Raws li qhov loj me me zuj zus, lub wavelength sib haum mus rau qhov siab tshaj plaws lub teeb nqus ntawm AgNP-II-ntau dua-Raws li cov ntaub ntawv, AgNP-III Muaj kev hloov pauv.6,29 ib
Hais txog qhov pib colloidal stability ntawm AgNP system, DLS tau siv los ntsuas qhov nruab nrab hydrodynamic txoj kab uas hla thiab zeta muaj peev xwm ntawm pH 7.2.Cov txiaj ntsig tau piav qhia hauv daim duab 2E qhia tias AgNP-III muaj kev ruaj ntseg colloidal ntau dua li AgNP-I lossis AgNP-II, vim tias cov lus qhia ib txwm qhia tias zeta muaj peev xwm ntawm 30 mV qhov tseeb yog qhov tsim nyog rau lub sijhawm ntev colloidal stability Qhov kev tshawb pom no tau txhawb ntxiv thaum Z qhov nruab nrab tus nqi (tau txais raws li qhov nruab nrab hydrodynamic txoj kab uas hla ntawm cov dawb thiab aggregated hais) yog muab piv nrog cov thawj particle loj tau los ntawm TEM, vim hais tias qhov ze ntawm ob qhov tseem ceeb, qhov me me ntawm cov degree Sib sau ua ke hauv cov qauv.Qhov tseeb, Z qhov nruab nrab ntawm AgNP-I thiab AgNP-II yog qhov tsim nyog siab dua li lawv qhov tseem ceeb TEM-kev ntsuas particle loj, yog li muab piv nrog AgNP-III, cov qauv no tau kwv yees ntau dua qhov sib sau ua ke, qhov twg muaj qhov tsis zoo zeta muaj peev xwm. yog nrog los ntawm qhov loj me Z qhov nruab nrab tus nqi.
Kev piav qhia rau qhov tshwm sim no tuaj yeem ua tau ob npaug.Ntawm qhov tod tes, citrate concentration yog khaws cia nyob rau theem zoo sib xws nyob rau hauv tag nrho cov synthesis cov kauj ruam, muab ib tug kuj siab npaum li cas ntawm cov nqi ntawm cov pab pawg neeg deg los tiv thaiv cov cheeb tsam tshwj xeeb ntawm cov khoom loj hlob los ntawm kev txo qis.Txawm li cas los xij, raws li Levak li al., me me molecules zoo li citrate tuaj yeem hloov tau yooj yim los ntawm biomolecules ntawm qhov chaw ntawm nanoparticles.Hauv qhov no, kev ruaj ntseg colloidal yuav raug txiav txim los ntawm corona ntawm biomolecules tsim.31 Vim hais tias tus cwj pwm no kuj tau pom nyob rau hauv peb cov kev ntsuas sib sau ua ke (sib tham ntxiv tom qab), citrate capping ib leeg tsis tuaj yeem piav qhia qhov tshwm sim no.
Ntawm qhov tod tes, qhov loj me me yog inversely proportional rau aggregation nyiam nyob rau theem nanometer.Qhov no feem ntau yog txhawb nqa los ntawm cov tsoos Derjaguin-Landau-Verwey-Overbeek (DLVO) txoj kev, qhov twg particle attraction tau piav raws li cov txiaj ntsig ntawm kev txaus siab thiab kev xav tsis zoo ntawm cov khoom.Raws li He thiab al., qhov siab tshaj plaws tus nqi ntawm DLVO lub zog nkhaus txo qis nrog qhov loj ntawm nanoparticles hauv hematite nanoparticles, ua kom yooj yim mus txog qhov tsawg kawg nkaus lub zog, yog li txhawb kev sib sau tsis tau (condensation).47 Txawm li cas los xij, nws tau kwv yees tias muaj lwm yam tshaj li qhov txwv ntawm DLVO txoj kev xav.Txawm hais tias van der Waals lub ntiajteb txawj nqus thiab electrostatic ob-txheej repulsion zoo ib yam nrog nce particle loj, kev tshuaj xyuas los ntawm Hotze li al.qhia tias nws muaj kev cuam tshuam rau kev sib sau ntau dua li DLVO tso cai.14 Lawv ntseeg tias qhov curvature ntawm nanoparticles tsis tuaj yeem kwv yees li qhov chaw tiaj tus, ua rau kev kwv yees siv tsis tau.Tsis tas li ntawd, raws li qhov loj me me zuj zus, qhov feem pua ​​​​ntawm cov atoms tam sim no nyob rau saum npoo yuav siab dua, ua rau cov qauv hluav taws xob thiab kev coj tus cwj pwm.Thiab qhov chaw reactivity hloov pauv, uas yuav ua rau txo qis hauv cov nqi hluav taws xob ob txheej thiab txhawb kev sib sau ua ke.
Thaum sib piv DLS cov txiaj ntsig ntawm AgNP-I, AgNP-II, thiab AgNP-III hauv daim duab 3, peb pom tias tag nrho peb cov qauv qhia pom zoo li pH ua kom sib sau ua ke.Ib puag ncig acidic hnyav (pH 3) hloov lub zeta peev xwm ntawm cov qauv mus rau 0 mV, ua rau cov khoom ua rau micron-loj aggregates, thaum alkaline pH hloov nws lub peev xwm zeta mus rau qhov loj dua qhov tsis zoo, qhov twg cov khoom tsim me me aggregates (pH 5. ).Thiab 7.2) ), los yog nyob twj ywm unagregated (pH 9).Qee qhov sib txawv tseem ceeb ntawm cov qauv sib txawv kuj tau pom.Thoob plaws hauv qhov kev sim, AgNP-I tau ua pov thawj tias qhov zoo tshaj plaws rau pH-induced zeta muaj peev xwm hloov pauv, vim tias lub peev xwm zeta ntawm cov khoom no tau txo qis ntawm pH 7.2 piv rau pH 9, thaum AgNP-II thiab AgNP-III tsuas pom A. Kev hloov pauv loj hauv ζ yog nyob ib puag ncig pH 3. Tsis tas li ntawd, AgNP-II pom qhov hloov qeeb qeeb thiab nruab nrab zeta muaj peev xwm, thaum AgNP-III tau pom tus cwj pwm tsis zoo ntawm peb, vim hais tias lub kaw lus pom qhov zoo tshaj plaws zeta tus nqi thiab kev txav qeeb, qhia. AgNP-III Feem ntau resistant rau pH-induced aggregation.Cov txiaj ntsig no zoo ib yam nrog qhov ntsuas qhov nruab nrab hydrodynamic txoj kab uas hla.Xav txog qhov loj me me ntawm lawv cov primers, AgNP-I tau pom qhov sib sau ua ke tsis tu ncua ntawm txhua qhov pH qhov tseem ceeb, feem ntau yuav yog vim 10 mM NaCl keeb kwm yav dhau, thaum AgNP-II thiab AgNP-III tsuas yog pom qhov tseem ceeb ntawm pH 3 ntawm kev sib sau.Qhov sib txawv tshaj plaws yog tias txawm tias nws loj nanoparticle loj, AgNP-III tsim cov aggregates tsawg tshaj plaws ntawm pH 3 hauv 24 teev, qhia txog nws cov khoom tiv thaiv kev sib sau.Los ntawm faib qhov nruab nrab Z ntawm AgNPs ntawm pH 3 tom qab 24 teev los ntawm tus nqi ntawm cov qauv npaj, nws tuaj yeem pom tias cov txheeb ze sib npaug ntawm AgNP-I thiab AgNP-II tau nce 50 npaug, 42 npaug, thiab 22 npaug. , raws.III.
Daim duab 3 Cov txiaj ntsig ntawm lub teeb pom kev zoo ntawm citrate-txiav nyiaj nanoparticles qauv nrog qhov loj me (10 nm: AgNP-I, 20 nm: AgNP-II thiab 50 nm: AgNP-III) yog qhia raws li qhov nruab nrab hydrodynamic txoj kab uas hla (Z nruab nrab. ) (txoj cai) Raws li qhov sib txawv pH, lub peev xwm zeta (sab laug) hloov hauv 24 teev.
Kev soj ntsuam pH-raws li kev sib sau ua ke kuj cuam tshuam rau tus yam ntxwv nto plasmon resonance (SPR) ntawm AgNP cov qauv, raws li pom los ntawm lawv cov UV-Vis spectra.Raws li Daim Duab Ntxiv S1, kev sib sau ua ke ntawm tag nrho peb cov nyiaj nanoparticle ncua kev kawm yog ua raws li kev txo qis ntawm lawv qhov SPR peaks thiab kev hloov pauv liab.Qhov twg ntawm cov kev hloov pauv no raws li kev ua haujlwm ntawm pH yog ua raws li qib ntawm kev sib sau ua ke tau kwv yees los ntawm DLS cov txiaj ntsig, txawm li cas los xij, qee qhov kev nthuav dav tau pom.Contrary to intuition, nws hloov tawm hais tias qhov nruab nrab-qhov loj me AgNP-II yog qhov rhiab tshaj plaws rau SPR kev hloov pauv, thaum lwm qhov ob qho piv txwv tsis tshua nkag siab.Hauv kev tshawb fawb SPR, 50 nm yog qhov theoretical particle loj txwv, uas yog siv los qhia qhov txawv raws li lawv cov khoom dielectric.Cov khoom me dua 50 nm (AgNP-I thiab AgNP-II) tuaj yeem piav qhia tau yooj yim dielectric dipoles, thaum cov khoom uas ncav cuag lossis dhau qhov kev txwv no (AgNP-III) muaj cov khoom siv hluav taws xob ntau dua, thiab lawv cov resonance ntawm pawg sib faib ua ntau yam kev hloov pauv. .Nyob rau hauv rooj plaub ntawm ob qhov qauv me me, AgNPs tuaj yeem suav tias yog cov dipoles yooj yim, thiab cov plasma tuaj yeem sib tshooj yooj yim.Raws li qhov loj me me ntxiv, qhov sib txuas no tseem ceeb ua rau cov ntshav loj dua, uas tuaj yeem piav qhia qhov kev xav tau siab dua.29 Txawm li cas los xij, rau qhov loj tshaj plaws, qhov yooj yim dipole kwv yees tsis siv tau thaum lwm lub xeev coupling kuj tshwm sim, uas tuaj yeem piav qhia qhov txo qis ntawm AgNP-III los qhia txog kev hloov pauv spectral.29
Raws li peb cov kev sim, nws tau ua pov thawj tias tus nqi pH muaj qhov cuam tshuam loj ntawm kev ruaj ntseg colloidal ntawm citrate-coated silver nanoparticles ntawm ntau qhov ntau thiab tsawg.Hauv cov kab ke no, kev ruaj ntseg yog muab los ntawm qhov tsis zoo them nqi -COO- pawg nyob rau saum npoo ntawm AgNPs.Carboxylate functional pawg ntawm citrate ion yog protonated nyob rau hauv ib tug loj tus naj npawb ntawm H + ions, yog li cov generated carboxyl pawg yuav tsis muab electrostatic repulsion ntawm cov hais, raws li qhia nyob rau hauv sab saum toj kab ntawm daim duab 4. Raws li Le Chatelier txoj cai, AgNP. cov qauv sib sau ua ke sai sai ntawm pH 3, tab sis maj mam ua ntau thiab ruaj khov thaum pH nce.
Daim duab 4 Schematic mechanism ntawm kev sib cuam tshuam saum npoo txhais los ntawm kev sib sau ua ke hauv pH sib txawv (kab sab saum toj), NaCl concentration ( kab nruab nrab), thiab biomolecules (kab hauv qab).
Raws li daim duab 5, lub colloidal stability nyob rau hauv AgNP ncua kev kawm ntawv ntawm ntau ntau thiab tsawg kuj tau soj ntsuam nyob rau hauv nce ntsev concentration.Raws li zeta muaj peev xwm, qhov nce nanoparticle loj hauv cov citrate-tawm AgNP systems dua muab kev tiv thaiv rau sab nraud cuam tshuam los ntawm NaCl.Hauv AgNP-I, 10 mM NaCl yog txaus los txhawb kev sib sau me me, thiab cov ntsev ntsev ntawm 50 mM muab cov txiaj ntsig zoo sib xws.Hauv AgNP-II thiab AgNP-III, 10 mM NaCl tsis cuam tshuam rau zeta lub peev xwm vim tias lawv cov txiaj ntsig tseem nyob ntawm (AgNP-II) lossis qis dua (AgNP-III) -30 mV.Ua kom NaCl concentration mus rau 50 mM thiab thaum kawg mus rau 150 mM NaCl yog txaus kom txo tau tus nqi kiag li ntawm zeta muaj peev xwm nyob rau hauv tag nrho cov qauv, txawm hais tias loj dua cov nqi tsis zoo.Cov txiaj ntsig no tau ua raws li qhov xav tau qhov nruab nrab hydrodynamic txoj kab uas hla ntawm AgNPs;Z qhov nruab nrab cov kab ntsuas ntsuas ntawm 10, 50, thiab 150 mM NaCl qhia sib txawv, maj mam nce nqi.Thaum kawg, micron-loj aggregates tau kuaj pom hauv peb qhov kev sim 150 mM.
Daim duab 5 Cov txiaj ntsig ntawm lub teeb pom kev zoo ntawm citrate-txiav nyiaj nanoparticles qauv nrog nce loj (10 nm: AgNP-I, 20 nm: AgNP-II thiab 50 nm: AgNP-III) tau qhia raws li qhov nruab nrab hydrodynamic txoj kab uas hla (Z nruab nrab. ) (txoj cai) thiab zeta muaj peev xwm (sab laug) hloov hauv 24 teev nyob rau hauv qhov sib txawv NaCl ntau.
Cov txiaj ntsig UV-Vis hauv Daim Duab Ntxiv S2 qhia tias SPR ntawm 50 thiab 150 mM NaCl hauv tag nrho peb cov qauv muaj qhov txo qis sai thiab tseem ceeb.Qhov no tuaj yeem piav qhia los ntawm DLS, vim tias NaCl-raws li kev sib sau ua ke tshwm sim sai dua li kev sim pH, uas tau piav qhia los ntawm qhov sib txawv loj ntawm qhov ntsuas thaum ntxov (0, 1.5, thiab 3 teev).Tsis tas li ntawd, ua kom cov ntsev concentration kuj tseem yuav ua rau cov txheeb ze permittivity ntawm qhov kev sim nruab nrab, uas yuav ua rau muaj kev cuam tshuam loj rau saum npoo plasmon resonance.29
Cov nyhuv ntawm NaCl tau sau tseg nyob rau hauv nruab nrab kab ntawm daim duab 4. Feem ntau, nws tuaj yeem txiav txim siab tias kev nce cov concentration ntawm sodium chloride muaj cov nyhuv zoo ib yam li nce acidity, vim Na+ ions muaj kev sib koom ua ke nyob ib ncig ntawm pawg carboxylate, suppressing tsis zoo zeta muaj peev xwm AgNPs.Tsis tas li ntawd, 150 mM NaCl tsim micron-loj aggregates nyob rau hauv tag nrho peb cov qauv, qhia tias lub physiological electrolyte concentration yog detrimental rau lub colloidal stability ntawm citrate-terminated AgNPs.Los ntawm kev txiav txim siab qhov tseem ceeb condensing concentration (CCC) ntawm NaCl ntawm cov tshuab AgNP zoo sib xws, cov txiaj ntsig no tuaj yeem muab tso rau hauv cov ntaub ntawv cuam tshuam.Huynh et al.xam tias CCC ntawm NaCl rau citrate-txiav nyiaj nanoparticles nrog qhov nruab nrab txoj kab uas hla ntawm 71 nm yog 47.6 mM, thaum El Badawy li al.pom tias CCC ntawm 10 nm AgNPs nrog citrate txheej yog 70 mM.10,16 Tsis tas li ntawd, qhov CCC siab heev ntawm txog 300 mM tau ntsuas los ntawm He et al., uas ua rau lawv cov txheej txheem synthesis txawv ntawm cov ntawv tshaj tawm yav dhau los.48 Txawm hais tias qhov kev pab cuam tam sim no tsis yog tsom rau kev tsom xam tag nrho ntawm cov txiaj ntsig no, vim tias peb cov kev sim sim tau nce ntxiv hauv qhov nyuaj ntawm tag nrho cov kev tshawb fawb, cov khoom noj muaj txiaj ntsig ntawm NaCl concentration ntawm 50 mM, tshwj xeeb tshaj yog 150 mM NaCl, zoo li yuav siab heev.Induced coagulation, piav qhia txog cov kev hloov muaj zog uas kuaj pom.
Cov kauj ruam tom ntej hauv kev sim polymerization yog siv cov khoom siv yooj yim tab sis cov khoom siv roj ntsha muaj feem cuam tshuam los simulate nanoparticle-biomolecule kev sib cuam tshuam.Raws li DLS (Figures 6 thiab 7) thiab UV-Vis cov txiaj ntsig (Cov duab ntxiv S3 thiab S4), qee qhov kev txiav txim siab dav dav tuaj yeem lees paub.Raws li peb cov kev sim, cov kev tshawb fawb molecules qabzib thiab glutamine yuav tsis ua kom muaj kev sib sau ua ke hauv AgNP system, vim tias Z-txhais tau zoo muaj feem cuam tshuam nrog qhov sib piv ntsuas tus nqi.Txawm hais tias lawv lub xub ntiag tsis cuam tshuam rau kev sib sau ua ke, cov txiaj ntsig kev sim qhia tau tias cov molecules no yog ib feem adsorbed rau ntawm qhov chaw ntawm AgNPs.Cov txiaj ntsig tseem ceeb tshaj plaws txhawb qhov kev pom no yog qhov pom kev hloov pauv hauv lub teeb nqus.Txawm hais tias AgNP-I tsis pom qhov muaj txiaj ntsig wavelength lossis kev siv zog hloov pauv, nws tuaj yeem pom meej dua los ntawm kev ntsuas cov khoom loj dua, uas feem ntau yuav yog vim qhov pom kev zoo dua qub tau hais ua ntej.Txawm hais tias qhov concentration, qabzib tuaj yeem ua rau muaj kev hloov pauv liab ntau dua tom qab 1.5 teev piv nrog kev ntsuas ntsuas, uas yog kwv yees li 40 nm hauv AgNP-II thiab txog 10 nm hauv AgNP-III, uas ua pov thawj qhov tshwm sim ntawm kev sib cuam tshuam saum npoo.Glutamine pom qhov sib xws, tab sis qhov kev hloov pauv tsis pom tseeb.Tsis tas li ntawd, nws tseem tsim nyog hais tias glutamine tuaj yeem txo qhov zeta lub peev xwm ntawm cov khoom nruab nrab thiab loj.Txawm li cas los xij, txij li cov kev hloov ntawm zeta no tsis zoo li cuam tshuam rau qib sib sau ua ke, nws tuaj yeem xav tias txawm tias cov biomolecules me me xws li glutamine tuaj yeem muab qee qhov kev hloov pauv ntawm qhov nruab nrab ntawm cov khoom.
Daim duab 6 Cov txiaj ntsig ntawm lub teeb pom kev zoo ntawm citrate-terminated silver nanoparticle cov qauv nrog nce loj (10 nm: AgNP-I, 20 nm: AgNP-II thiab 50 nm: AgNP-III) tau qhia raws li qhov nruab nrab hydrodynamic txoj kab uas hla (Z nruab nrab) (txoj cai) Nyob rau hauv cov xwm txheej sab nraud ntawm cov piam thaj sib txawv, lub peev xwm zeta (sab laug) hloov hauv 24 teev.
Daim duab 7 Cov txiaj ntsig ntawm lub teeb pom kev zoo ntawm citrate-terminated silver nanoparticles qauv nrog qhov loj me (10 nm: AgNP-I, 20 nm: AgNP-II thiab 50 nm: AgNP-III) yog qhia raws li qhov nruab nrab hydrodynamic txoj kab uas hla (Z nruab nrab. ) (txoj cai) Nyob rau hauv lub xub ntiag ntawm glutamine, lub zeta muaj peev xwm (sab laug) hloov nyob rau hauv 24 teev.
Nyob rau hauv luv luv, me me biomolecules xws li qabzib thiab glutamine tsis cuam tshuam rau colloidal stability ntawm qhov ntsuas concentration: txawm hais tias lawv cuam tshuam rau zeta muaj peev xwm thiab UV-Vis cov txiaj ntsig mus rau qhov sib txawv, Z qhov nruab nrab cov txiaj ntsig tsis zoo ib yam.Qhov no qhia tau hais tias qhov saum npoo adsorption ntawm molecules inhibits electrostatic repulsion, tab sis tib lub sij hawm muab qhov seem stability.
Txhawm rau txuas cov txiaj ntsig yav dhau los nrog cov txiaj ntsig yav dhau los thiab simulate cov kab mob lom neeg ntau dua, peb tau xaiv qee yam ntawm cov kab lis kev cai feem ntau siv ntawm tes thiab siv lawv los ua kev sim rau kev kawm txog kev ruaj ntseg ntawm AgNP colloids.Hauv kev sim tag nrho hauv vitro, ib qho tseem ceeb tshaj plaws ntawm DMEM ua ib qho nruab nrab yog tsim kom muaj qhov tsim nyog osmotic tej yam kev mob, tab sis los ntawm ib tug tshuaj point of view, nws yog ib tug complex ntsev tov nrog tag nrho cov ionic zog zoo ib yam li 150 mM NaCl. .40 Raws li rau FBS, nws yog ib qho kev sib xyaw ntawm biomolecules-feem ntau cov proteins-los ntawm qhov pom ntawm qhov chaw adsorption, nws muaj qee qhov zoo sib xws nrog cov txiaj ntsig kev sim ntawm cov piam thaj thiab glutamine, txawm hais tias cov tshuaj muaj pes tsawg leeg thiab kev sib txawv ntawm kev sib deev yog ntau qhov nyuaj.19 DLS thiab UV-Cov txiaj ntsig pom tau pom hauv daim duab 8 thiab cov duab ntxiv S5, feem, tuaj yeem piav qhia los ntawm kev tshuaj xyuas cov tshuaj muaj pes tsawg leeg ntawm cov ntaub ntawv no thiab sib txuas nrog cov kev ntsuas hauv ntu dhau los.
Daim duab 8 Cov txiaj ntsig ntawm lub teeb pom kev zoo ntawm citrate-terminated silver nanoparticles qauv nrog nce loj (10 nm: AgNP-I, 20 nm: AgNP-II thiab 50 nm: AgNP-III) yog qhia raws li qhov nruab nrab hydrodynamic txoj kab uas hla (Z nruab nrab. ) (txoj cai) Nyob rau hauv lub xub ntiag ntawm cell kab lis kev cai Cheebtsam DMEM thiab FBS, zeta muaj peev xwm (sab laug) hloov nyob rau hauv 24 teev.
Lub dilution ntawm AgNPs ntawm ntau qhov sib txawv hauv DMEM muaj qhov zoo sib xws ntawm colloidal stability rau qhov uas tau pom nyob rau hauv lub xub ntiag ntawm siab NaCl concentrations.Lub dispersion ntawm AgNP nyob rau hauv 50 v / v% DMEM qhia tau hais tias loj-scale aggregation tau kuaj pom nrog qhov nce ntawm zeta muaj peev xwm thiab Z-nruab nrab tus nqi thiab qhov ntse txo ntawm SPR siv.Nws yog ib nqi sau cia hais tias qhov siab tshaj plaws aggregate loj induced los ntawm DMEM tom qab 24 teev yog inversely proportional rau qhov loj ntawm primer nanoparticles.
Kev sib cuam tshuam ntawm FBS thiab AgNP zoo ib yam li qhov pom ntawm cov khoom me me xws li qabzib thiab glutamine, tab sis cov nyhuv muaj zog dua.Z qhov nruab nrab ntawm cov khoom tseem tsis cuam tshuam, thaum kuaj pom qhov nce hauv zeta.SPR ncov pom qhov hloov pauv me ntsis liab, tab sis tej zaum qhov nthuav dav dua, qhov kev siv SPR tsis txo qis dua li hauv kev ntsuas ntsuas.Cov txiaj ntsig no tuaj yeem piav qhia los ntawm innate adsorption ntawm macromolecules nyob rau saum npoo ntawm nanoparticles (hauv qab kab hauv daim duab 4), uas tam sim no nkag siab tias yog kev tsim ntawm biomolecular corona hauv lub cev.49