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Na li-nanoparticles tse nyane li lula li le betere?Utloisisa litlamorao tsa bioloji tsa ho kopanya ho itšetlehileng ka boholo ba li-nanoparticles tsa silevera tlas'a maemo a amanang le biologically
Bangoli: 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 oa Environmental Chemistry, Hungary, Hungary, Hungary, Hungary , Univesithi ea Szeged;2 Lefapha la Biochemistry le Biology ea Limolek'hule, Lefapha la Saense le Boitsebiso, Univesithi ea Szeged, Hungary;3 Lefapha la Microbiology, Lefapha la Saense le Boitsebiso, Univesithi ea Szeged, Hungary;4MTA-SZTE Reaction Kinetics and Surface Chemistry Research Group, Szeged, Hungary* Bangoli bana ba kentse letsoho ka ho lekana mosebetsing ona.Puisano: Zoltán Kónya Lefapha la Applied and Environment Chemistry, Faculty of Science and Informatics, University of Szeged, Rerrich Square 1, Szeged, H-6720, Hungary Phone +36 62 544620 Email [Tsireletso ea lengolo-tsoibila] Sepheo: Li-nanoparticles tsa silevera (AgNPs) li teng e 'ngoe ea li-nanomaterials tse ithutoang hangata, haholo ka lebaka la ts'ebeliso ea tsona ea biomedical.Leha ho le joalo, ka lebaka la ho bokellana ha li-nanoparticles, ts'ebetso ea tsona e ntle ea cytotoxicity le antibacterial hangata li senyeha mecheng ea litaba tsa baeloji.Mosebetsing ona, ho ile ha ithutoa boitšoaro ba ho bokellana le mesebetsi e amanang le likokoana-hloko ea lisampole tse tharo tse fapaneng tse felisitsoeng tsa silevera tsa nanoparticle tse nang le bophara ba 10, 20 le 50 nm.Mokhoa: Sebelisa microscope ea elektronike ea phetisetso ho kopanya le ho tšoaea li-nanoparticles, ho lekola boits'oaro ba tsona ba ho bokellana maemong a fapaneng a pH, NaCl, glucose le glutamine concentrations ka ho hasanya ha leseli le ultraviolet-spectroscopy.Ntle le moo, ka har'a likarolo tse mahareng tsa setso sa sele joalo ka Dulbecco e ntlafatsa boitšoaro ba ho bokellana ho Eagle Medium le Fetal Calf Serum.Liphetho: Liphetho li bonts'a hore acidic pH le physiological electrolyte content ka kakaretso li susumetsa micron-scale aggregation, e ka tsamaellanang le ho thehoa ha biomolecular corona.Ke habohlokoa ho hlokomela hore likaroloana tse khōloanyane li bontša khanyetso e phahameng ho litšusumetso tsa ka ntle ho feta tse nyenyane tsa tsona.Liteko tsa in vitro cytotoxicity le antibacterial li entsoe ka ho phekola lisele tse nang le nanoparticle aggregates ka mekhahlelo e fapaneng ea ho kopanya.Qetello: Liphetho tsa rona li senola kamano e tebileng pakeng tsa botsitso ba colloidal le chefo ea li-AgNP, kaha ho kopanya ho feteletseng ho lebisa tahlehelong e feletseng ea mesebetsi ea likokoana-hloko.Tekanyo e phahameng ea anti-aggregation e bonoang bakeng sa likaroloana tse kholo e na le tšusumetso e kholo ho chefo ea in vitro, hobane lisampole tse joalo li boloka tšebetso e ngata ea lisele tsa antimicrobial le mammalian.Liphuputso tsena li lebisa qetong ea hore, ho sa tsotellehe maikutlo a akaretsang libukeng tse amehang, ho lebisa tlhokomelo ea li-nanoparticles tse nyenyane ka ho fetisisa e ka 'na ea se ke ea e-ba mokhoa o molemo ka ho fetisisa oa ho nka khato.Mantsoe a bohlokoa: kholo e lipakeng tsa peo, botsitso ba colloidal, boits'oaro bo kopaneng bo itšetlehileng ka boholo, chefo ea tšenyo e kopaneng.
Ha tlhoko le tlhahiso ea li-nanomaterials li ntse li eketseha, tlhokomelo e eketsehileng e lefshoa ho ts'ireletso ea bona ea tlhaho kapa ts'ebetso ea likokoana-hloko.Li-nanoparticles tsa silevera (AgNPs) ke e 'ngoe ea baemeli ba atisang ho etsoa, ​​​​ho batlisisoa le ho sebelisoa ba sehlopha sena sa thepa ka lebaka la thepa ea bona e babatsehang ea ho thusa, ea mahlo le ea likokoana-hloko.1 Ka kakaretso ho lumeloa hore litšobotsi tse ikhethang tsa li-nanomaterials (ho kenyeletsoa le li-AgNP) li hlahisoa haholo ke sebaka sa tsona se seholo se ikhethileng.Ka hona, bothata bo ke keng ba qojoa ke ts'ebetso leha e le efe e amang tšobotsi ena ea bohlokoa, e kang boholo ba likaroloana, ho roala holim'a metsi Kapa ho kopanya, ho sa tsotellehe hore na ho tla senya thepa ea nanoparticles e bohlokoa haholo ho likopo tse itseng.
Liphello tsa boholo ba likaroloana le li-stabilizers ke lihlooho tse 'nileng tsa ngoloa hantle ka har'a lingoliloeng.Ka mohlala, pono e amoheloang ka kakaretso ke hore li-nanoparticles tse nyenyane li chefo ho feta li-nanoparticles tse kholoanyane.2 Tumellanong le lingoliloeng tse akaretsang, lithuto tsa rona tse fetileng li bonts'itse ts'ebetso e itšetlehileng ka boholo ba nanosilver ka lisele tsa mammalian le likokoana-hloko.3– 5 Ho roala holim'a sefahleho ke tšobotsi e 'ngoe e nang le tšusumetso e pharaletseng ho thepa ea nanomatadium.Ka ho eketsa kapa ho fetola li-stabilizer holim'a eona, li-nanomaterial tse tšoanang li ka ba le litšobotsi tse fapaneng tsa 'mele, tsa lik'hemik'hale le tsa tlhaho.Ts'ebeliso ea li-capping agents hangata e etsoa e le karolo ea nanoparticle synthesis.Mohlala, li-nanoparticles tsa silevera tse felisitsoeng ka citrate ke e 'ngoe ea li-AgNP tse bohlokoa ka ho fetisisa lipatlisisong, tse entsoeng ka ho fokotsa letsoai la silevera ka tharollo e khethiloeng ea stabilizer e le mokhoa oa ho arabela.6 Citrate e ka sebelisa monyetla oa litšenyehelo tsa eona tse tlase, ho fumaneha, ho lumellana ha lintho, le kamano e matla ea silevera, e ka bontšoang litšebelisanong tse fapa-fapaneng tse reriloeng, ho tloha ho adsorption ea holimo e ka fetoloang ho ea ho litšebelisano tsa ionic.Limolek'hule tse nyane le li-ion tsa polyatomic tse haufi le 7,8, joalo ka li-citrate, li-polymers, polyelectrolytes, le li-biological agents le tsona li sebelisoa ka tloaelo ho tsitsisa nano-silevera le ho etsa mesebetsi e ikhethang ho eona.9-12
Le hoja monyetla oa ho fetola ts'ebetso ea li-nanoparticles ka ho khabisa ka holimo ka boomo ke sebaka se thahasellisang haholo, karolo e ka sehloohong ea ho roala holim'a metsi ha e na thuso, ho fana ka botsitso ba colloidal bakeng sa tsamaiso ea nanoparticle.Sebaka se seholo se ikhethileng sa li-nanomaterials se tla hlahisa matla a maholo a holim'a metsi, a sitisang matla a thermodynamic a sistimi ho fihlela matla a eona a tlase.13 Ntle le botsitso bo nepahetseng, sena se ka lebisa ho agglomeration ea nanomatadium.Aggregation ke sebopeho sa likaroloana tsa likaroloana tsa libopeho le boholo bo fapaneng tse etsahalang ha likaroloana tse hasantsoeng li kopana 'me litšebelisano tsa morao-rao tsa thermodynamic li lumella likaroloana ho khomarelana.Ka hona, li-stabilizers li sebelisetsoa ho thibela ho kopana ka ho hlahisa matla a lekaneng a lekaneng a lelekang pakeng tsa likaroloana ho loantša ho khahloa ha tsona ke thermodynamic.14
Le hoja taba ea boholo ba likaroloana le sekoahelo sa holim'a metsi e 'nile ea hlahlojoa ka ho feletseng molemong oa taolo ea eona ea mesebetsi ea likokoana-hloko e bakoang ke nanoparticles, ho kopanya likaroloana ke sebaka se hlokomolohuoang haholo.Hoo e ka bang ha ho na thuto e feletseng ea ho rarolla botsitso ba colloidal ba nanoparticles tlas'a maemo a amanang le baeloji.10,15-17 Ho phaella moo, monehelo ona ha o fumanehe ka ho khetheha, moo chefo e amanang le ho bokellana le eona e 'nileng ea ithutoa, esita le haeba e ka baka liphello tse mpe, tse kang vascular thrombosis, kapa tahlehelo ea litšobotsi tse lakatsehang, tse kang chefo ea eona, e bontšitsoeng ho Setšoantšo sa 1.18, 19 se bontšitsoeng.Ha e le hantle, e 'ngoe ea mekhoa e seng mekae e tsejoang ea ho hanyetsa nanoparticle ea silevera e amana le ho kopanya, hobane mefuta e itseng ea E. coli le Pseudomonas aeruginosa ho tlalehoa hore e fokotsa kutlo ea bona ea nano-silver ka ho hlahisa protheine flagellin, flagellin.E na le kamano e phahameng bakeng sa silevera, kahoo e susumetsa ho bokellana.20
Ho na le mekhoa e mengata e fapaneng e amanang le chefo ea li-nanoparticles tsa silevera, 'me ho bokellana ho ama mekhoa ena kaofela.Mokhoa oo ho buisanoang ka oona ka ho fetisisa oa tšebetso ea likokoana-hloko ea AgNP, eo ka linako tse ling e bitsoang "Trojan Horse" mochine, e nka AgNPs e le bajari ba Ag+.1,21 The Trojan horse mechanism e ka etsa bonnete ba hore ho na le keketseho e kholo ea sebaka sa Ag + sa libaka, se lebisang ho moloko oa ROS le membrane depolarization.22-24 Aggregation e ka ama ho lokolloa ha Ag +, kahoo e ama chefo, hobane e fokotsa sebaka se sebetsang se sebetsang moo li-ion tsa silevera li ka bang oxidized le ho qhibiliha.Leha ho le joalo, li-AgNP li ke ke tsa bonts'a chefo feela ka ho lokolloa ha ion.Litšebelisano tse ngata tse amanang le boholo le mofoloji li tlameha ho nahanoa.Har'a bona, boholo le sebōpeho sa nanoparticle ke litšobotsi tse hlalosang.4,25 Pokello ea mekhoa ena e ka aroloa e le "mekhoa ea chefo e bakoang."Ho na le likarolo tse ngata tsa mitochondrial le membrane ea holim'a metsi tse ka senyang li-organelles le ho baka lefu la lisele.25-27 Kaha ho thehoa ha lihlopha ka tlhaho ho ama boholo le sebōpeho sa lintho tse nang le silevera tse amoheloang ke litsamaiso tse phelang, litšebelisano tsena le tsona li ka ameha.
Pampiring ea rona e fetileng mabapi le ho bokellana ha li-nanoparticles tsa silevera, re bontšitse mokhoa o sebetsang oa ho hlahloba o nang le liteko tsa lik'hemik'hale le tsa in vitro tsa bioloji ho ithuta bothata bona.19 Dynamic Light Scattering (DLS) ke mokhoa o ratoang bakeng sa mefuta ena ea tlhahlobo hobane thepa e ka hasanya li-photon ka bolelele bo lekanang le boholo ba likaroloana tsa eona.Kaha lebelo la ho sisinyeha ha Brownian la likaroloana sebakeng sa mokelikeli le amana le boholo, phetoho ea matla a khanya e hasaneng e ka sebelisoa ho fumana hore na bophara ba hydrodynamic bo boholo (Z-bolela) ba sampole ea mokelikeli.28 Ho phaella moo, ka ho sebelisa motlakase ho sampuli, bokhoni ba zeta (ζ bokhoni) ba nanoparticle bo ka lekanyetsoa ka mokhoa o tšoanang le oa karolelano ea Z.13,28 Haeba boleng bo feletseng ba bokhoni ba zeta bo phahame ka ho lekaneng (ho ea ka litataiso tse akaretsang> ± 30 mV), e tla hlahisa matla a matla a electrostatic pakeng tsa likaroloana ho loantša ho bokellana.Characteristic surface plasmon resonance (SPR) ke ts'ebetso e ikhethang ea optical, haholo-holo e hlahisoang ke nanoparticles ea bohlokoa ea tšepe (haholo-holo Au le Ag).29​ Ho ipapisitsoe le li-oscillation tsa elektroniki (li-plasmons tse kaholimo) tsa lisebelisoa tsena ho nanoscale, hoa tsebahala hore li-AgNP tse chitja li na le tlhoro ea ho monya ha UV-Vis haufi le 400 nm.30 Ho fetoha ha matla le maqhubu a maqhubu a likaroloana ho sebelisoa ho tlatsetsa liphetho tsa DLS, kaha mokhoa ona o ka sebelisoa ho lemoha nanoparticle aggregation le adsorption ea holim'a li-biomolecules.
Ho ipapisitsoe le tlhaiso-leseling e fumanoeng, ho sebetsa ha sele (MTT) le litlhahlobo tsa antibacteria li etsoa ka mokhoa oo chefo ea AgNP e hlalosoang e le ts'ebetso ea boemo ba ho bokellana, ho fapana le (ntho e sebelisoang haholo) nanoparticle concentration.Mokhoa ona o ikhethang o re lumella ho bonts'a bohlokoa bo tebileng ba boemo ba ho bokellana mosebetsing oa likokoana-hloko, hobane, ka mohlala, li-AgNP tse felisitsoeng ke citrate li lahleheloa ke mosebetsi oa tsona oa likokoana-hloko ka mor'a lihora tse 'maloa ka lebaka la ho bokellana.19
Mosebetsing oa hona joale, re ikemiselitse ho atolosa haholo menehelo ea rona ea pele ho botsitso ba li-colloid tse amanang le li-bio le phello ea tsona mosebetsing oa likokoana-hloko ka ho ithuta phello ea boholo ba nanoparticle ho nanoparticle aggregation.Ha ho pelaelo hore ena ke e 'ngoe ea lithuto tsa nanoparticles.Pono ea boemo bo phahameng le 31 Ho batlisisa taba ena, mokhoa oa ho hōla oa peo o ile oa sebelisoa ho hlahisa li-AgNP tse felisitsoeng ka citrate ka mefuta e meraro e fapaneng ea boholo (10, 20, le 50 nm).6,32 e le o mong oa mekhoa e tloaelehileng haholo.Bakeng sa li-nanomaterials tse sebelisoang ka bongata le ka tloaelo lits'ebetsong tsa bongaka, li-AgNP tse felisitsoeng ka citrate tsa boholo bo fapaneng li khethoa ho ithuta ho itšetleha ka boholo bo ka bang teng ba thepa ea tlhaho e amanang le ho bokellana ea nanosilver.Kamora ho kopanya li-AgNP tsa boholo bo fapaneng, re ile ra tšoaea lisampole tse hlahisitsoeng ke transmission electron microscopy (TEM), ebe re hlahloba likaroloana tseo re sebelisa mokhoa o boletsoeng ka holimo.Ho feta moo, boteng ba in vitro cell cultures Dulbecco's Modified Eagle's Medium (DMEM) le Fetal Bovine Serum (FBS), boitšoaro ba ho bokellana bo itšetlehileng ka boholo le boitšoaro ba eona bo ile ba hlahlojoa ka litekanyetso tse fapaneng tsa pH, NaCl, glucose, le glutamine.Litšobotsi tsa cytotoxicity li khethoa tlasa maemo a felletseng.Tumellano ea saense e bontša hore ka kakaretso, likaroloana tse nyenyane li molemo;lipatlisiso tsa rona li fana ka sethala sa lik'hemik'hale le tsa baeloji ho fumana hore na ho joalo.
Li-nanoparticles tse tharo tsa silevera tse nang le mefuta e fapaneng ea boholo li ile tsa lokisoa ke mokhoa oa ho hōla o entsoeng ke peo o hlahisitsoeng ke Wan et al., Ka liphetoho tse fokolang.6 Mokhoa ona o thehiloe ho fokotsa lik'hemik'hale, ho sebelisa nitrate ea silevera (AgNO3) e le mohloli oa silevera, sodium borohydride (NaBH4) e le moemeli oa ho fokotsa, le sodium citrate e le stabilizer.Ntlha ea pele, lokisetsa 75 mL ea 9 mM citrate aqueous solution ho tloha sodium citrate dihydrate (Na3C6H5O7 x 2H2O) le mocheso ho 70 ° C.Ka mor'a moo, 2 mL ea 1% w/v AgNO3 tharollo e ile ea eketsoa sebakeng sa karabelo, ebe tharollo e sa tsoa lokisoa ea sodium borohydride (2 mL 0.1% w/v) e tšeloa ka har'a motsoako oa dropwise.Ho emisoa ho hlahisitsoeng ke mosehla-sootho ho ne ho bolokoa ho 70 ° C ka ho tsosoa ka matla bakeng sa hora ea 1, ebe e pholile mocheso oa kamore.Sampole e hlahisoang (e bitsoang AgNP-I ho tloha joale ho ea pele) e sebelisoa e le motheo oa kholo ea lipeo mohatong o latelang oa ho kopanya.
Ho kopanya ho emisoa ha likaroloana tse mahareng (tse hlalosoang e le AgNP-II), mocheso 90 mL 7.6 mM citrate tharollo ho 80 ° C, kopanya le 10 mL AgNP-I, ebe o kopanya 2 mL 1% w / v The AgNO3 Tharollo. e ne e bolokoa tlas'a ts'ebetso e matla ea mochine bakeng sa hora ea 1, ebe sampuli e pholile mocheso oa kamore.
Bakeng sa karolo e kholo ka ho fetisisa (AgNP-III), pheta mokhoa o tšoanang oa ho hōla, empa tabeng ena, sebelisa 10 mL ea AgNP-II e le ho emisoa ha peo.Ka mor'a hore lisampole li fihle mocheso oa kamore, li beha mahloriso a tsona a Ag ho latela kakaretso ea AgNO3 ho 150 ppm ka ho eketsa kapa ho ntša mouoane o eketsehileng ho 40 ° C, 'me qetellong ba li boloke ho 4 ° C ho fihlela li sebelisoa hape.
Sebelisa FEI Tecnai G2 20 X-Twin Transmission Electron Microscope (TEM) (FEI Corporate Headquarters, Hillsboro, Oregon, USA) ka 200 kV acceleration voltage ho hlahloba litšobotsi tsa morphological tsa nanoparticles le ho hapa paterone ea bona ea electron diffraction (ED).Bonyane litšoantšo tse 15 tse emelang (~ 750 particles) li ile tsa hlahlojoa ho sebelisoa software ea ImageJ, 'me li-histograms tse hlahisitsoeng (le kerafo eohle thutong eohle) li entsoe ho OriginPro 2018 (OriginLab, Northampton, MA, USA) 33, 34.
Karolelano ea bophara ba hydrodynamic (Z-average), bokhoni ba zeta (ζ-potential) le tšobotsi ea sefahleho sa plasmon resonance (SPR) ea lisampole e ne e lekantsoe ho bontša thepa ea bona ea pele ea colloidal.Karolelano ea bophara ba hydrodynamic le bokhoni ba zeta ba sampole bo ile ba lekanyetsoa ke sesebelisoa sa Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK) se sebelisa lisele tsa capillary tse lahliloeng ho 37±0.1 ° C.Ocean Optics 355 DH-2000-BAL UV-Vis spectrophotometer (Halma PLC, Largo, FL, USA) e ile ea sebelisoa ho fumana litšobotsi tsa SPR ho tsoa ho UV-Vis e monyang lisampole tsa 250-800 nm.
Nakong ea teko eohle, mefuta e meraro e fapaneng ea tekanyo e amanang le botsitso ba colloidal e ile ea etsoa ka nako e le 'ngoe.Sebelisa DLS ho lekanya bophara ba karolelano ea hydrodynamic (Z karolelano) le bokhoni ba zeta (ζ bokhoni) ba likaroloana, hobane karolelano ea Z e amana le boholo bo tloaelehileng ba likaroloana tsa nanoparticle, 'me bokhoni ba zeta bo bontša hore na ho reteleha ha electrostatic tsamaisong. e matla hoo e ka felisang khoheli ea Van der Waals lipakeng tsa nanoparticles.Litekanyo li etsoa ka makhetlo a mararo, 'me ho kheloha ho tloaelehileng ha Z mean le zeta ho baloa ke software ea Zetasizer.Sebopeho sa SPR sa likaroloana se hlahlojoa ke UV-Vis spectroscopy, hobane liphetoho tsa tlhōrō ea matla le bolelele ba maqhubu li ka bontša ho kopanya le ho sebelisana holimo.29,35 Ha e le hantle, resonance ea holim'a plasmon ka tšepe ea bohlokoa e na le tšusumetso e kholo hoo e lebisitseng mekhoa e mecha ea ho hlahloba li-biomolecules.29,36,37 Mahloriso a AgNP ka motsoako oa liteko ke hoo e ka bang 10 ppm, 'me morero ke ho beha matla a ho kenngoa ha SPR ho ea pele ho 1. Teko e ile ea etsoa ka mokhoa o itšetlehileng ka nako ho 0;1.5;3;6;Lihora tse 12 le 24 tlasa maemo a fapaneng a amanang le baeloji.Lintlha tse ling tse hlalosang teko li ka bonoa mosebetsing oa rona o fetileng.19 Ka bokhutšoanyane, litekanyetso tse fapaneng tsa pH (3; 5; 7.2 le 9), sodium chloride e fapaneng (10 mM; 50 mm; 150 mM), tsoekere (3.9 mM; 6.7 mM) le glutamine (4 mm) mahloriso, le e boetse e lokiselitse Dulbecco's Modified Eagle Medium (DMEM) le Fetal Bovine Serum (FBS) (ka metsing le DMEM) e le litsamaiso tsa mohlala, mme a ithuta litlamorao tsa tsona boitšoarong ba ho bokellana ha li-nanoparticles tsa silevera tse kopaneng.pH Litekanyetso tsa, NaCl, glucose, le glutamine li hlahlojoa ho ipapisitsoe le maemo a 'mele, ha lipalo tsa DEM le FBS li lekana le maemo a sebelisitsoeng tekong eohle ea in vitro.38-42 Litekanyo tsohle li entsoe ka pH 7.2 le 37 ° C ka letsoai le sa khaotseng la letsoai la 10 mM NaCl ho felisa litšebelisano leha e le life tsa sebaka se selelele (ntle le liteko tse itseng tse amanang le pH le NaCl, moo litšobotsi tsena e leng mefuta e sa tšoaneng. boithuto).28 Lethathamo la maemo a fapaneng le akaretsoa ho Lethathamo la 1. Teko e tšoailoeng ka † e sebelisoa e le sesupo 'me e lumellana le sampuli e nang le 10 mM NaCl le pH 7.2.
Kankere ea tšoelesa ea senya ea motho (DU145) le keratinocyte ea motho e sa shoeng (HaCaT) e fumanoe ho tloha ATCC (Manassas, VA, USA).Lisele li hlahisoa ka tloaelo ho Ntsu ea bohlokoa ea Dulbecco (DMEM) e nang le tsoekere ea 4.5 g/L (Sigma-Aldrich, Saint Louis, MO, USA), e tlatsitsoeng ka 10% FBS, 2 mM L-glutamine, 0.01 % Streptomycin le 0.005% Penicillin (Sigma-Aldrich, St. Louis, Missouri, USA).Lisele li entsoe ka 37°C ka tlas'a 5% CO2 le 95% ea mongobo.
E le ho hlahloba liphetoho ho AgNP cytotoxicity e bakoang ke ho kopanya likaroloana ka mokhoa o itšetlehileng ka nako, ho ile ha etsoa tlhahlobo ea MTT ea mehato e 'meli.Ntlha ea pele, ho sebetsa ha mefuta e 'meli ea lisele ho ile ha lekanyetsoa ka mor'a phekolo ea AgNP-I, AgNP-II le AgNP-III.Ho fihlela sena, mefuta e 'meli ea lisele e ile ea kenngoa ka har'a lipoleiti tsa seliba tse 96 ka bongata ba lisele tsa 10,000 / hantle' me li tšoaroa ka mefuta e meraro e fapaneng ea li-nanoparticles tsa silevera ka ho eketseha ha likhahla ka letsatsi la bobeli.Ka mor'a lihora tse 24 tsa phekolo, lisele li ile tsa hlatsuoa ka PBS 'me tsa kenngoa ka 0.5 mg / mL MTT reagent (SERVA, Heidelberg, Germany) e hlapolotsoeng ka mokhoa oa setso bakeng sa hora ea 1 ho 37 ° C.Likristale tsa Formazan li ile tsa qhibiliha ho DMSO (Sigma-Aldrich, Saint Louis, MO, USA), 'me ho kenngoa ho ile ha lekanyetsoa ho 570 nm ho sebelisa sebali sa poleiti ea Synergy HTX (BioTek-Hungary, Budapest, Hungary).Boleng ba ho monya sa sampole ea taolo e sa phekoloang bo nkuoa e le sekhahla sa ho pholoha sa 100%.Etsa bonyane liteko tse 3 u sebelisa likopi tse 'ne tse ikemetseng tsa baeloji.IC50 e baloa ho tloha ho tekanyo ea karabo ea litekanyetso ho latela liphetho tsa matla.
Ka mor'a moo, mohato oa bobeli, ka ho kenya likaroloana ka 150 mM NaCl ka nako e fapaneng (0, 1.5, 3, 6, 12, le 24 lihora) pele ho phekolo ea lisele, linaha tse fapaneng tsa ho bokellana ha li-nanoparticles tsa silevera li ile tsa hlahisoa.Ka mor'a moo, tlhahlobo e tšoanang ea MTT e ile ea etsoa joalokaha e hlalositsoe pele ho hlahloba liphetoho tsa ho sebetsa ha lisele tse anngoeng ke ho kopanya likaroloana.Sebelisa GraphPad Prism 7 ho lekola sephetho sa ho qetela, bala lipalo-palo tsa bohlokoa ba teko ka tlhahlobo ea t e sa sebetseng, 'me u tšoaee boemo ba eona e le * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001) ) Le **** (p ≤ 0.0001).
Mefuta e meraro e fapaneng ea li-nanoparticles tsa silevera (AgNP-I, AgNP-II le AgNP-III) li ne li sebelisetsoa ho hlaseloa ke likokoana-hloko ho Cryptococcus neoformans IFM 5844 (IFM; Setsi sa Lipatlisiso sa Pathogenic Fungi le Microbial Toxicology, Univesithi ea Chiba) le Bacillus Test megaterium SZMC 60 (SZMC: Szeged Microbiology Collection) le E. coli SZMC 0582 ho RPMI 1640 medium (Sigma-Aldrich Co.).Bakeng sa ho lekola liphetoho tsa ts'ebetso ea antibacteria e bakoang ke ho bokellana ha likaroloana, pele, khatello ea bona e fokolang ea inhibitory (MIC) e ile ea khethoa ke microdilution ka poleiti ea 96-well microtiter.Ho 50 μL ea ho emisoa ha lisele tse emeng (5 × 104 lisele / mL ka har'a RPMI 1640 medium), eketsa 50 μL ea ho emisoa ha nanoparticle ea silevera 'me u hlapolle ka makhetlo a mabeli mahloriso (ka mokhoa o boletsoeng ka holimo, mefuta e fapaneng ke 0 le 75 ppm, ke hore, sampole ea taolo e na le 50 μL ea ho emisoa ha sele le 50 μL ea mahareng ntle le nanoparticles).Ka mor'a moo, poleiti e ne e kenngoa ka 30 ° C bakeng sa lihora tse 48, 'me boima ba optical ba setso bo ne bo lekanngoa ho 620 nm ho sebelisoa SPECTROstar Nano plate reader (BMG LabTech, Offenburg, Germany).Teko e entsoe ka makhetlo a mararo ka makhetlo a mararo.
Ntle le hore 50 μL ea lisampole tse kopaneng tsa nanoparticle li ne li sebelisoa ka nako ena, mokhoa o tšoanang le o hlalositsoeng pejana o ile oa sebelisoa ho lekola phello ea ho kopanya tšebetsong ea antibacteria ho mefuta e boletsoeng ka holimo.Likarolo tse fapaneng tsa li-nanoparticles tsa silevera li hlahisoa ka ho kenya likaroloana ka 150 mM NaCl ka linako tse fapaneng (0, 1.5, 3, 6, 12, le 24 lihora) pele ho ts'ebetso ea lisele.Ho emisoa ho tlatsitsoeng ka 50 μL ea RPMI 1640 medium e ne e sebelisoa e le taolo ea kgolo, ha e le ho laola chefo, ho emisoa ka li-nanoparticles tse sa kopanngoeng li ne li sebelisoa.Teko e entsoe ka makhetlo a mararo ka makhetlo a mararo.Sebelisa GraphPad Prism 7 ho lekola sephetho sa ho qetela hape, u sebelisa tlhahlobo ea lipalo-palo e tšoanang le tlhahlobo ea MTT.
Boemo ba ho bokellana ba likaroloana tse nyenyane ka ho fetisisa (AgNP-I) bo khetholloa, 'me liphello li ile tsa hatisoa ka mokhoa o itseng mosebetsing oa rona o fetileng, empa bakeng sa papiso e ntle, likaroloana tsohle li ile tsa hlahlojoa ka botlalo.Lintlha tsa liteko li bokelloa 'me li tšohloa likarolong tse latelang.Mefuta e meraro ea AgNP.19
Litekanyo tse entsoeng ke TEM, UV-Vis le DLS li netefalitse katleho ea lisampole tsa AgNP (Setšoantšo sa 2A-D).Ho ea ka mola oa pele oa Setšoantšo sa 2, karoloana e nyenyane ka ho fetisisa (AgNP-I) e bonts'a morphology e tšoanang ea spherical e nang le bophara ba hoo e ka bang 10 nm.Mokhoa oa ho hōla oa peo o boetse o fana ka AgNP-II le AgNP-III ka mefuta e fapaneng ea boholo bo nang le bophara ba likaroloana tse ka bang 20 nm le 50 nm, ka ho latellana.Ho ea ka ho kheloha ho tloaelehileng ha kabo ea likaroloana, boholo ba mehlala e meraro ha e kopane, e leng ea bohlokoa bakeng sa tlhahlobo ea bona ea ho bapisa.Ka ho bapisa karo-karolelano ea likaroloana le bosesaane ba likhakanyo tsa 2D tse thehiloeng ho TEM, ho nahanoa hore sphericity ea likaroloana e hlahlojoa ke plug-in ea sebopeho sa ImageJ (Figure 2E).43 Ho latela tlhahlobo ea sebopeho sa likaroloana, karo-karolelano ea tsona (lehlakore le leholo / le khuts'oane la khutlonnetsepa e nyane) ha e amehe ke kholo ea likaroloana, 'me karo-karolelano ea bona e tšesaane (sebaka se lekantsoeng sa selikalikoe / sebaka sa thuto. ) ea fokotseha butle-butle.Sena se fella ka likaroloana tse ngata tsa polyhedral, tse pota-potiloeng ka mokhoa o phethahetseng, tse tsamaellanang le tekanyo ea bosesane ea 1.
Setšoantšo sa 2 Sets'oants'o sa 2 Transmission electron microscope (TEM) (A), electron diffraction (ED) pattern (B), size distribution histogram (C), characteristic ultraviolet-visible (UV-Vis) light absorption spectrum (D), le mokelikeli o tloaelehileng oa Citrate. -li-nanoparticles tsa silevera tse felisitsoeng tse nang le bophara ba mochine (Z-average), bokhoni ba zeta, karolelano ea likarolo le boholo ba boholo (E) li na le mefuta e meraro ea boholo bo fapaneng: AgNP-I ke 10 nm (moleng o ka holimo), AgNP -II ke 20 nm (mola o bohareng ), AgNP-III (mola o ka tlase) ke 50 nm.
Le hoja mokhoa oa cyclic oa mokhoa oa ho hōla o ile oa ama sebōpeho sa likaroloana ho isa bohōleng bo itseng, e leng se ileng sa fella ka hore ho be le sekhahla se senyenyane sa li-AgNP tse khōloanyane, mehlala eohle e meraro e ile ea lula e le quasi-spherical.Ho phaella moo, joalokaha ho bontšitsoe ka mokhoa oa electron diffraction ho Setšoantšo sa 2B, nano The crystallinity ea likaroloana ha e amehe.Ring e hlahelletseng ea diffraction-e ka amahanngoang le (111), (220), (200), le (311) li-indices tsa Miller tsa silevera-e lumellana haholo le lingoliloeng tsa mahlale le menehelo ea rona e fetileng.9, 19,44 Ho arohana ha selikalikoe sa Debye-Scherrer sa AgNP-II le AgNP-III se bakoa ke taba ea hore setšoantšo sa ED se nkoa ka mokhoa o ts'oanang oa ho hōlisa, e le hore boholo ba likaroloana bo ntse bo eketseha, palo ea likaroloana tse fapaneng ka nako e le 'ngoe. yuniti e eketseha le ho fokotseha.
Boholo le sebōpeho sa nanoparticles li tsejoa ho ama ts'ebetso ea likokoana-hloko.3,45 Ts'ebetso e itšetlehileng ka sebopeho le ts'ebetso ea likokoana-hloko e ka hlalosoa ke taba ea hore libopeho tse fapaneng li atisa ho atisa lifahleho tse itseng tsa kristale (ho ba le li-indices tse fapaneng tsa Miller), 'me lifahleho tsena tsa kristale li na le mesebetsi e fapaneng.45,46 Kaha likaroloana tse lokiselitsoeng li fana ka liphello tse tšoanang tsa ED tse tsamaellanang le litšoaneleho tsa kristale tse tšoanang haholo, ho ka nkoa hore litekong tsa rona tse latelang tsa botsitso ba colloidal le tsa likokoana-hloko, phapang leha e le efe e hlokometsoeng e lokela ho hlahisoa ke boholo ba Nanoparticle, eseng thepa e amanang le sebōpeho.
Liphetho tsa UV-Vis tse akarelitsoeng ho Setšoantšo sa 2D li boetse li totobatsa sebopeho se makatsang sa AgNP e kopaneng, hobane litlhōrō tsa SPR tsa lisampole tsena tse tharo li haufi le 400 nm, e leng boleng ba boleng ba li-nanoparticles tsa silevera tse chitja.29,30 Pono e hapuoeng e boetse e netefalitse kholo e atlehileng ea peo ea nanosilver.Ha boholo ba likaroloana bo ntse bo eketseha, bolelele ba maqhubu bo lumellana le khanya e phahameng ea khanya ea AgNP-II-ho feta ka mokhoa o hlahelletseng-Ho ea ka lingoliloeng, AgNP-III E na le phihlelo ea redshift.6,29
Mabapi le botsitso ba pele ba colloidal ea tsamaiso ea AgNP, DLS e ne e sebelisetsoa ho lekanya bophara ba hydrodynamic bophara le bokhoni ba zeta ba likaroloana ho pH 7.2.Liphetho tse bontšitsoeng ho Setšoantšo sa 2E li bontša hore AgNP-III e na le botsitso bo phahameng ba colloidal ho feta AgNP-I kapa AgNP-II, hobane tataiso e tloaelehileng e bontša hore matla a zeta a 30 mV absolute a hlokahala bakeng sa botsitso ba nako e telele ea colloidal. boleng bo tloaelehileng ba Z (bo fumanoeng e le bophara ba hydrodynamic ba likaroloana tse sa lefelloeng le tse kopantsoeng) bo bapisoa le boholo ba karoloana e fumanoeng ke TEM, hobane ha litekanyetso tsena tse peli li le haufi haholo, tekanyo ea Bokella sampoleng e bobebe haholo.Ha e le hantle, karolelano ea Z ea AgNP-I le AgNP-II e phahame ka mokhoa o utloahalang ho feta boholo ba likaroloana tse lekantsoeng tsa TEM, kahoo ha li bapisoa le AgNP-III, lisampole tsena ho boleloa esale pele hore li ka 'na tsa kopana, moo ho nang le monyetla o moholo oa zeta. e tsamaisana le boholo bo haufi The Z karolelano ea boleng.
Tlhaloso ea ketsahalo ena e ka ba habeli.Ka lehlakoreng le leng, mahloriso a citrate a bolokoa ka mokhoa o ts'oanang mehatong eohle ea ho kopanya, e fana ka palo e batlang e le holimo ea lihlopha tse qosoang ho thibela sebaka se ikhethileng sa likaroloana tse ntseng li hola ho fokotseha.Leha ho le joalo, ho ea ka Levak et al., limolek'hule tse nyenyane tse kang citrate li ka fapanyetsanoa habonolo ke li-biomolecules holim'a nanoparticles.Tabeng ena, botsitso ba colloidal bo tla khethoa ke corona ea li-biomolecules tse hlahisoang.31 Hobane boitšoaro bona bo ne bo boetse bo hlokomeloa litekanyong tsa rona tsa ho kopanya (tse tšohloang ka ho qaqileng hamorao), citrate capping feela e ke ke ea hlalosa ketsahalo ena.
Ka lehlakoreng le leng, boholo ba likaroloana bo lekana ka tsela e fapaneng le mokhoa oa ho kopanya boemong ba nanometer.Sena se tšehetsoa haholo-holo ke mokhoa o tloaelehileng oa Derjaguin-Landau-Verwey-Overbeek (DLVO), moo ho hohelang likaroloana ho hlalosoang e le kakaretso ea matla a khahlehang le a nyonyehang pakeng tsa likaroloana.Ho ea ka He et al., boleng bo phahameng ba lebelo la matla la DLVO bo fokotseha ka boholo ba nanoparticles ho nanoparticles ea hematite, e leng ho nolofalletsang ho finyella bonyane ba matla a mantlha, kahoo ho khothalletsa ho bokellana ho ke keng ha fetoloa (condensation).47 Leha ho le joalo, ho nahanoa hore ho na le lintlha tse ling tse ka nqane ho meeli ea teori ea DLVO.Le hoja matla a khoheli a van der Waals le electrostatic double-layer repulsion li tšoana le ho eketseha ha boholo ba karoloana, tlhahlobo ea Hotze et al.e sisinya hore e na le phello e matla ho kopanyelletsang ho feta DLVO e dumellang.14 Ba lumela hore ho kobeha ha li-nanoparticles ho ke ke ha hlola ho hakanngoa e le sebaka se bataletseng, e leng ho etsang hore khakanyo ea lipalo e se ke ea sebetsa.Ho feta moo, ha boholo ba karoloana bo fokotseha, peresente ea liathomo tse teng holim'a metsi e ba holimo, e lebisang ho sebopeho sa elektroniki le boitšoaro ba tefiso ea holim'a metsi.Le liphetoho holim'a reactivity, tse ka lebisang ho fokotseha ha tefiso ka har'a motlakase habeli le ho khothaletsa ho kopanya.
Ha re bapisa liphetho tsa DLS tsa AgNP-I, AgNP-II, le AgNP-III setšoantšong sa 3, re hlokometse hore lisampole tse tharo kaofela li bonts'itse pH e ts'oanang e susumetsang ho kopanya.Tikoloho e nang le acidic e ngata (pH 3) e fetola bokhoni ba zeta ea sampuli ho 0 mV, e leng se etsang hore likaroloana li thehe li-aggregate tsa boholo ba micron, ha pH ea alkaline e fetola bokhoni ba eona ba zeta ho boleng bo boholo bo bobe, moo likaroloana li etsang likaroloana tse nyenyane (pH 5). ).Le 7.2) ), kapa u lule u sa kopane ka ho feletseng (pH 9).Liphapang tse ling tsa bohlokoa lipakeng tsa lisampole tse fapaneng le tsona li ile tsa bonoa.Nakong eohle ea teko, AgNP-I e ipakile e le eona e amehang ka ho fetisisa ho pH-induced zeta liphetoho tse ka 'nang tsa e-ba teng, hobane matla a zeta a likaroloana tsena a fokotsehile ho pH 7.2 ha a bapisoa le pH 9, ha AgNP-II le AgNP-III e bontšitse AgNP-III feela. phetoho e kholo ho ζ e pota-potile pH 3. Ho phaella moo, AgNP-II e bontšitse liphetoho tse fokolang le bokhoni ba zeta bo itekanetseng, ha AgNP-III e bontšitse boitšoaro bo bonolo ka ho fetisisa ho ba bararo, hobane tsamaiso e bontšitse boleng bo phahameng ka ho fetisisa ba zeta le ho tsamaea butle butle. AgNP-III E hanyetsana haholo le pH-induced aggregation.Liphetho tsena li lumellana le liphetho tse tloaelehileng tsa tekanyo ea bophara ba hydrodynamic.Ha ho nahanoa ka boholo ba likaroloana tsa li-primers tsa tsona, AgNP-I e bonts'itse ho bokellana butle butle maemong ohle a pH, mohlomong ka lebaka la 10 mM NaCl background, athe AgNP-II le AgNP-III li bonts'itse bohlokoa feela ho pH 3 ea ho bokella.Phapang e khahlang ka ho fetesisa ke hore leha e na le boholo ba nanoparticle e kholo, AgNP-III e theha likarolo tse nyane ka ho fetisisa ho pH 3 ka lihora tse 24, e totobatsang thepa ea eona e khahlanong le ho bokellana.Ka ho arola karolelano ea Z ea AgNP ho pH 3 kamora lihora tse 24 ka boleng ba sampole e lokisitsoeng, ho ka hlokomeloa hore boholo bo kopaneng ba AgNP-I le AgNP-II bo eketsehile ka makhetlo a 50, makhetlo a 42 le makhetlo a 22. , ka ho latellana.III.
Setšoantšo sa 3 Liphetho tse matla tsa ho hasanya ha leseli le matla la sampole ea nanoparticles ea silevera ea citrate e felisitsoeng ka boholo bo ntseng bo eketseha (10 nm: AgNP-I, 20 nm: AgNP-II le 50 nm: AgNP-III) e hlahisoa e le bophara ba hydrodynamic (Z karolelano ) (ka ho le letona) Tlas'a maemo a fapaneng a pH, bokhoni ba zeta (ka ho le letšehali) bo fetoha nakong ea lihora tsa 24.
Pokello e shebiloeng e itšetlehileng ka pH e boetse e amme sebopeho sa plasmon resonance (SPR) ea lisampole tsa AgNP, joalo ka ha ho pakoa ke pono ea bona ea UV-Vis.Ho ea ka Supplementary Figure S1, ho kopanngoa ha li-suspensions tse tharo tsa silevera tsa nanoparticle ho lateloa ke ho fokotseha ho matla a litlhōrō tsa bona tsa SPR le phetoho e khubelu e itekanetseng.Tekanyo ea liphetoho tsena e le ts'ebetso ea pH e lumellana le tekanyo ea ho bokellana e boletsoeng esale pele ke liphetho tsa DLS, leha ho le joalo, ho hlokometsoe mekhoa e meng e khahlisang.Ho fapana le intuition, hoa fumaneha hore AgNP-II ea boholo bo mahareng ke eona e amehang haholo ho liphetoho tsa SPR, athe lisampole tse ling tse peli ha li na kutloelo-bohloko.Phuputsong ea SPR, 50 nm ke moeli oa boholo ba theoretical particle, o sebelisetsoang ho khetholla likaroloana ho latela thepa ea tsona ea dielectric.Likarolo tse nyane ho feta 50 nm (AgNP-I le AgNP-II) li ka hlalosoa e le li-dielectric dipoles tse bonolo, ha likaroloana tse fihlang kapa tse fetang moeling ona (AgNP-III) li na le thepa e rarahaneng ea dielectric, 'me resonance ea tsona Sehlopha se arohana ka liphetoho tse ngata. .Tabeng ea lisampole tse peli tse nyane, li-AgNP li ka nkuoa e le li-dipoles tse bonolo, 'me plasma e ka kopana habonolo.Ha boholo ba karoloana bo ntse bo eketseha, ho kopana hona ho hlahisa plasma e kholoanyane, e ka hlalosang maikutlo a phahameng a bonoang.29 Leha ho le joalo, bakeng sa likaroloana tse kholo ka ho fetisisa, tekanyo e bonolo ea dipole ha e sebetse ha linaha tse ling tsa ho kopanya li ka boela tsa etsahala, tse ka hlalosang mokhoa o fokotsehileng oa AgNP-III ho bontša liphetoho tsa spectral.29
Tlas'a maemo a rona a liteko, ho pakoa hore boleng ba pH bo na le phello e tebileng ho tsitsipano ea colloidal ea li-nanoparticles tsa silevera tse nang le citrate tsa boholo bo sa tšoaneng.Litsamaisong tsena, botsitso bo fanoa ke lihlopha tse qosoang hampe -COO- holim'a AgNPs.Sehlopha se sebetsang sa carboxylate sa citrate ion se protonated ka palo e kholo ea li-ion tsa H +, kahoo sehlopha sa carboxyl se hlahisitsoeng se ke ke sa hlola se fana ka repulsion electrostatic pakeng tsa likaroloana, joalokaha ho bontšitsoe moleng o ka holimo oa Figure 4. Ho ea ka molao-motheo oa Le Chatelier, AgNP. disampole di kopana ka potlako ho pH 3, empa butle-butle di tsitsa le ho feta ha pH e ntse e eketseha.
Setšoantšo sa 4 Mokhoa oa ho sebelisana ka mokhoa o hlophisitsoeng o hlalosoang ka ho kopanya tlas'a pH e fapaneng (mola o ka holimo), mahloriso a NaCl (mola o bohareng), le li-biomolecules (mola o ka tlaase).
Ho ea ka Setšoantšo sa 5, botsitso ba colloidal ho ho emisoa ha AgNP ka boholo bo fapaneng bo ile ba boela ba hlahlojoa tlas'a ho eketseha ha letsoai.Ho ipapisitsoe le matla a zeta, boholo ba nanoparticle bo ntseng bo eketseha mekhoeng ena ea AgNP e felisitsoeng ka citrate e boetse e fana ka khanyetso e matlafalitsoeng ho litšusumetso tsa kantle tse tsoang ho NaCl.Ho AgNP-I, 10 mM NaCl e lekane ho kopanya hanyenyane, 'me letsoai la 50 mm le fana ka liphello tse tšoanang haholo.Ho AgNP-II le AgNP-III, 10 mM NaCl ha e ame haholo bokhoni ba zeta hobane litekanyetso tsa bona li lula ho (AgNP-II) kapa ka tlase (AgNP-III) -30 mV.Ho eketsa mahloriso a NaCl ho 50 mM 'me qetellong ho 150 mM NaCl ho lekane ho fokotsa haholo boleng bo feletseng ba bokhoni ba zeta ho mehlala eohle, le hoja likaroloana tse kholo li boloka tefiso e mpe haholoanyane.Liphetho tsena li lumellana le tekanyo e lebeletsoeng ea bophara ba hydrodynamic ea AgNPs;karolelano ea mekhoa ea Z e lekantsoeng ho 10, 50, le 150 mM NaCl e bonts'a litekanyetso tse fapaneng, tse ntseng li eketseha butle-butle.Qetellong, li-aggregates tsa boholo ba micron li ile tsa fumanoa litekong tsohle tse tharo tsa 150 mM.
Setšoantšo sa 5 Liphetho tse matla tsa ho hasanya ha leseli le matla la sampole ea nanoparticles ea silevera ea citrate e felisitsoeng ka boholo bo ntseng bo eketseha (10 nm: AgNP-I, 20 nm: AgNP-II le 50 nm: AgNP-III) e hlahisoa e le bophara ba hydrodynamic (Z karolelano ) (ka ho le letona) le matla a zeta (ka ho le letšehali) a fetoha ka hare ho lihora tsa 24 tlas'a maemo a fapaneng a NaCl.
Liphetho tsa UV-Vis ho Supplementary Figure S2 li bonts'a hore SPR ea 50 le 150 mM NaCl lisampoleng tsohle tse tharo e na le phokotso ea hang-hang le e kholo.Sena se ka hlalosoa ke DLS, hobane NaCl-based aggregation e etsahala ka potlako ho feta liteko tse itšetlehileng ka pH, e hlalosoang ke phapang e kholo pakeng tsa litekanyo tsa pele (0, 1.5, le lihora tse 3).Ntle le moo, ho eketsa letsoai ho tla eketsa tumello e lekanyelitsoeng ea sebaka sa liteko, se tla ba le phello e matla holim'a plasmon resonance.29
Phello ea NaCl e akaretsoa moleng o bohareng oa Setšoantšo sa 4. Ka kakaretso, ho ka etsoa qeto ea hore ho eketseha ha motsoako oa sodium chloride ho na le phello e tšoanang le ho eketsa acidity, hobane Na + ions e na le tšekamelo ea ho hokahanya ho pota-pota lihlopha tsa carboxylate, ho hatella li-AgNP tse mpe tsa zeta.Ho feta moo, 150 mM NaCl e hlahisitse li-aggregates tsa boholo ba micron ka lisampole tsohle tse tharo, e leng se bontšang hore motsoako oa physiological electrolyte o senya botsitso ba colloidal ea li-AgNP tse felisitsoeng ke citrate.Ka ho nahana ka "critical condensing concentration" (CCC) ea NaCl lits'ebetsong tse ts'oanang tsa AgNP, liphetho tsena li ka beoa ka bohlale libukeng tse amehang.Huynh et al.e lekantsoe hore CCC ea NaCl bakeng sa li-nanoparticles tsa silevera tse felisitsoeng ka citrate ka bophara ba 71 nm e ne e le 47.6 mM, ha El Badawy et al.e hlokometse hore CCC ea 10 nm AgNPs e nang le citrate coating e ne e le 70 mM.10,16 Ho phaella moo, CCC e phahameng haholo ea hoo e ka bang 300 mM e ile ea lekanngoa ke He et al., E leng se ileng sa etsa hore mokhoa oa bona oa ho kopanya o fapane le oa khatiso e boletsoeng pejana.48 Le hoja tlatsetso ea hona joale e sa rereloa tlhahlobo e pharaletseng ea litekanyetso tsena, hobane maemo a rona a liteko a ntse a eketseha ka ho rarahana ha thuto eohle, motsoako oa biologically NaCl oa 50 mM, haholo-holo 150 mM NaCl, o bonahala o phahame haholo.Induced coagulation, e hlalosang liphetoho tse matla tse fumanoeng.
Mohato o latelang oa teko ea polymerization ke ho sebelisa limolek'hule tse bonolo empa tse amanang le baeloji ho etsisa litšebelisano tsa nanoparticle-biomolecule.Ho ipapisitsoe le DLS (Lipalo 6 le 7) le liphetho tsa UV-Vis (Lipalo tsa Tlatsetso S3 le S4), liqeto tse ling tse akaretsang li ka tiisoa.Tlas'a maemo a rona a liteko, limolek'hule tse ithutoang tsa tsoekere le glutamine li ke ke tsa kenyelletsa ho kopanya tsamaisong efe kapa efe ea AgNP, hobane mokhoa oa Z-mean o amana haufi-ufi le boleng bo lumellanang ba tekanyo ea litšupiso.Leha boteng ba bona bo sa ame ho bokellana, liphetho tsa liteko li bonts'a hore limolek'hule tsena li batla li le ka holim'a li-AgNP.Sephetho se hlahelletseng se tšehetsang pono ena ke phetoho e bonoang ho monyeng ha leseli.Le hoja AgNP-I e sa bontše bolelele ba maqhubu kapa liphetoho tse matla, e ka bonoa ka ho hlaka ka ho lekanya likaroloana tse kholoanyane, tse ka 'nang tsa etsahala haholo ka lebaka la kutloisiso e kholo ea optical e boletsoeng pejana.Ho sa tsotelehe hore na tsoekere e kae, tsoekere e ka baka phetoho e khubelu e kholo kamora lihora tse 1.5 ha e bapisoa le tekanyo ea taolo, e ka bang 40 nm ho AgNP-II le 10 nm ho AgNP-III, e pakang ho ba teng ha litšebelisano tsa sefahleho.Glutamine e bontšitse mokhoa o tšoanang, empa phetoho e ne e sa totobala hakaalo.Ho phaella moo, ho boetse hoa bohlokoa ho bolela hore glutamine e ka fokotsa matla a feletseng a zeta a likaroloana tse mahareng le tse kholo.Leha ho le joalo, kaha liphetoho tsena tsa zeta ha li bonahale li ama boemo ba ho bokellana, ho ka nahanoa hore esita le li-biomolecules tse nyenyane tse kang glutamine li ka fana ka tekanyo e itseng ea ho thibela sebaka pakeng tsa likaroloana.
Setšoantšo sa 6 Liphetho tse matla tsa ho hasanya ha leseli la lisampole tsa nanoparticle tsa silevera ea citrate-termination ka boholo bo ntseng bo eketseha (10 nm: AgNP-I, 20 nm: AgNP-II le 50 nm: AgNP-III) li hlahisoa e le bophara ba hydrodynamic (Z karolelano) (ka ho le letona) Tlas'a maemo a ka ntle a likhahla tse fapaneng tsa tsoekere, matla a zeta (ka ho le letšehali) a fetoha nakong ea lihora tse 24.
Setšoantšo sa 7 Liphetho tse matla tsa ho hasanya ha leseli la sampuli ea nanoparticles ea silevera ea citrate-e felisitsoeng ka boholo bo ntseng bo eketseha (10 nm: AgNP-I, 20 nm: AgNP-II le 50 nm: AgNP-III) e hlahisoa e le bophara ba hydrodynamic (Z karolelano ) (ka ho le letona) Ka pel'a glutamine, bokhoni ba zeta (ka ho le letšehali) bo fetoha nakong ea lihora tsa 24.
Ka bokhutšoanyane, li-biomolecules tse nyane joalo ka tsoekere le glutamine ha li ame botsitso ba colloidal maemong a lekantsoeng: leha li ama bokhoni ba zeta le liphetho tsa UV-Vis ho ea ka likhato tse fapaneng, liphetho tsa karolelano ea Z ha li tsitsane.Sena se bontša hore adsorption holim'a limolek'hule inhibits repulsion electrostatic, empa ka nako e tšoanang fana ka botsitso dimensional.
E le ho hokahanya liphetho tse fetileng le liphetho tse fetileng le ho etsisa maemo a baeloji ka boqhetseke haholoanyane, re khethile tse ling tsa likarolo tse sebelisoang haholo tsa cell culture mme ra li sebelisa e le maemo a liteko bakeng sa ho ithuta botsitso ba AgNP colloids.Tekong eohle ea in vitro, e 'ngoe ea mesebetsi ea bohlokoa ka ho fetisisa ea DMEM e le mokhoa oa ho theha maemo a hlokahalang a osmotic, empa ho ea ka pono ea lik'hemik'hale, ke tharollo e rarahaneng ea letsoai e nang le matla a ionic kaofela a tšoanang le 150 mM NaCl. .40 Ha e le FBS, ke motsoako o rarahaneng oa li-biomolecules-haholo-holo liprotheine-ho tloha sebakeng sa pono ea ho ata ha holim'a metsi, e na le lintho tse ling tse tšoanang le liphello tsa liteko tsa tsoekere le glutamine, ho sa tsotellehe sebopeho sa lik'hemik'hale le mefuta e sa tšoaneng ea thobalano e thata haholo.19 DLS le UV-Liphetho tse bonahalang tse bontšitsoeng ho Setšoantšo sa 8 le Supplementary Figure S5, ka ho latellana, li ka hlalosoa ka ho hlahloba lik'hemik'hale tsa lisebelisoa tsena le ho li hokahanya le litekanyo tse karolong e fetileng.
Setšoantšo sa 8 Liphetho tse matla tsa khanya e matla ea sampole ea nanoparticles ea silevera ea citrate e felisitsoeng ka boholo bo ntseng bo eketseha (10 nm: AgNP-I, 20 nm: AgNP-II le 50 nm: AgNP-III) e hlahisoa e le bophara ba hydrodynamic (Z karolelano ) (ka ho le letona) Boteng ba likarolo tsa setso sa lisele tsa DMEM le FBS, bokhoni ba zeta (ka ho le letšehali) bo fetoha nakong ea lihora tse 24.
Ho hlapolloa ha li-AgNP tsa boholo bo fapaneng ho DEMM ho na le phello e ts'oanang ho botsitso ba colloidal ho e bonoang boteng ba likhakanyo tse phahameng tsa NaCl.Ho qhalakanngoa ha AgNP ho 50 v / v% DMEM ho bontšitse hore kakaretso e kholo e ile ea fumanoa ka ho eketseha ha bokhoni ba zeta le boleng ba Z-average le ho fokotseha ho matla ha matla a SPR.Ke habohlokoa ho hlokomela hore boholo ba kakaretso bo hlahisitsoeng ke DMEM ka mor'a lihora tse 24 bo lekana ka tsela e fapaneng le boholo ba li-nanoparticles tsa primer.
Tšebelisano pakeng tsa FBS le AgNP e tšoana le e bonoang ka pel'a limolek'hule tse nyenyane tse kang glucose le glutamine, empa phello e matla haholoanyane.Karolelano ea Z ea likaroloana e lula e sa amehe, ha keketseho ea bokhoni ba zeta e bonoa.Tlhōrō ea SPR e bontšitse phetoho e khubelu e nyane, empa mohlomong ho khahlisang le ho feta, matla a SPR ha a ka a fokotseha haholo joalo ka tekanyo ea taolo.Liphetho tsena li ka hlalosoa ka ho ata ha tlhaho ha li-macromolecules holim'a nanoparticles (mola o ka tlase ho Setšoantšo sa 4), eo hona joale e utloisisoang e le sebopeho sa biomolecular corona 'meleng.49