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Madiki nanoparticles anogara ari nani here?Nzwisisa mhedzisiro yebhayoloji yehukuru-inotsamira kuunganidzwa kwesirivheri nanoparticles pasi pemamiriro ehupenyu akakodzera
Vanyori: 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 of Environmental Chemistry, Hungary, Hungary, Hungary , Yunivhesiti yeSzeged;2 Dhipatimendi reBiochemistry uye Molecular Biology, Faculty of Science uye Information, University of Szeged, Hungary;3 Dhipatimendi reMicrobiology, Faculty of Science uye Information, University of Szeged, Hungary;4MTA-SZTE Reaction Kinetics uye Surface Chemistry Research Group, Szeged, Hungary* Vanyori ava vakapa zvakaenzana mubasa iri.Kukurukurirana: Zoltán Kónya Department of Applied and Environment Chemistry, Faculty of Science and Informatics, University of Szeged, Rerrich Square 1, Szeged, H-6720, Hungary Phone +36 62 544620 Email [Email protection] Chinangwa: Silver nanoparticles (AgNPs) zviri imwe yeanonyanya kudzidzwa nanomatadium, kunyanya nekuda kwekushandisa kwavo biomedical.Nekudaro, nekuda kwekuunganidzwa kwenanoparticles, yavo yakanakisa cytotoxicity uye antibacterial basa rinowanzo kukanganiswa mubiological media.Mubasa iri, maitiro ekuunganidza uye ane chekuita nebiological zviitiko zveatatu akasiyana citrate-akagumiswa sirivha nanoparticle samples ane avhareji dhayamita ye10, 20, uye 50 nm zvakadzidzwa.Method: Shandisa kutapurirana maikorosikopu erekitironi kugadzira uye kuratidza nanoparticles, ongorora maitiro adzo ekuunganidza pamhando dzakasiyana dzepH, NaCl, glucose uye glutamine concentrations nekupararira kwechiedza uye ultraviolet-inooneka spectroscopy.Uye zvakare, musero tsika yepakati zvikamu zvakaita seDulbecco inovandudza maitiro ekuunganidza muEagle Medium uye Fetal Mhuru Serum.Mhedzisiro: Mhedzisiro yacho inoratidza kuti acidic pH uye physiological electrolyte zvemukati zvinowanzokonzera micron-scale aggregation, iyo inogona kuyananisirwa nekuumbwa kweiyo biomolecular corona.Zvakakosha kuziva kuti zvimedu zvakakura zvinoratidzira kupikisa kwakanyanya kune zvekunze pesvedzero pane zvidiki zvidiki.In vitro cytotoxicity uye antibacterial bvunzo yakaitwa nekurapa maseru ane nanoparticle aggregates pamatanho akasiyana ekubatanidza.Mhedziso: Mhedziso yedu inoratidza kuwirirana kwakadzama pakati pekugadzikana kwecolloidal uye chepfu yeAgNPs, sezvo kuunganidzwa kwakanyanya kunotungamira mukurasikirwa kwakakwana kwezviitiko zvebhayoloji.Iyo yepamusoro dhigirii yeanti-aggregation inocherechedzwa yezvimedu zvihombe ine chekuita kwakakura pane in vitro toxicity, nekuti masampuli akadai anochengeta mamwe antimicrobial uye mammalian cell chiitiko.Izvi zvakawanikwa zvinotungamirira kumhedziso yokuti, pasinei nemafungiro evanhu vakawanda mumabhuku akakodzera, kutarisa zviduku zvingaita nanoparticles zvingasava nzira yakanakisisa yekuita.Keywords: mbeu-yakapindirana kukura, colloidal kugadzikana, saizi-inotsamira aggregation maitiro, aggregation kukuvadza chepfu
Sezvo kudiwa uye kubuda kwemananomaterials kuri kuramba kuchiwedzera, kutarisisa kwakawanda kunobhadharwa kune yavo biosafety kana biological chiitiko.Silver nanoparticles (AgNPs) ndeimwe yeanonyanya kugadzirwa, kutsvagirwa uye kushandiswa vamiriri veiyi kirasi yezvinhu nekuda kweakanakisa catalytic, optical uye biological zvimiro.1 Zvinowanzo davirwa kuti akasiyana hunhu emananomaterials (kusanganisira maAgNPs) anonyanya kuverengerwa kune yakakura chaiyo nzvimbo.Nokudaro, dambudziko risingadzivisiki nderimwe nzira inobata ichi chinhu chinokosha, chakadai sekukura kwechidimbu, kuvhara kwepamusoro Kana kuunganidza, kana ichakuvadza zvakanyanya zvinhu zve nanoparticles zvakakosha kune mamwe maitiro.
Mhedzisiro yehukuru hwezvikamu uye stabilizers inyaya dzakanyatsonyorwa mumabhuku.Semuenzaniso, maonero anogamuchirwa ndeokuti madiki nanoparticles ane chepfu kupfuura hombe nanoparticles.2 Zvichienderana nemabhuku ezvinyorwa, zvidzidzo zvedu zvekare zvakaratidza basa rinoenderana nehukuru hwe nanosilver pamasero emammalian uye microorganisms.3– 5 Surface coating ndohumwe hunhu hune simba rakawanda pazvinhu zvenanomaterials.Nekungowedzera kana kugadzirisa zvinodzikamisa pamusoro payo, iwo manomaterial akafanana anogona kunge akasiyana zvachose emuviri, makemikari, uye biological zvimiro.Iko kushandiswa kwecapping agents kunowanzoitwa sechikamu che nanoparticle synthesis.Semuenzaniso, citrate-yakapera sirivheri nanoparticles ndeimwe yeanonyanya kukosha maAgNP mukutsvagisa, ayo anogadzirwa nekudzikisa sirivheri munyu mune yakasarudzwa yakagadzikana mhinduro seyekuita svikiro.6 Citrate inogona kutora mukana wemutengo wayo wakaderera, kuwanikwa, biocompatibility, uye hukama hwakasimba hwesirivheri, iyo inogona kuratidzwa mukudyidzana kwakasiyana kwakarongwa, kubva pakudzoreredzwa kwepasi adsorption kusvika kune ionic interactions.Mamorekuru madiki uye polyatomic ions padyo ne7,8, senge citrate, mapolima, mapolyelectrolytes, uye biological agents anoshandiswawo kudzikamisa nano-silver uye kuita mashandiro akasiyana pairi.9-12
Kunyange zvazvo mukana wekushandura basa re nanoparticles nekuzvipira kwepamusoro capping inzvimbo inonakidza zvikuru, basa guru rekuputira pamusoro apa harina basa, rinopa colloidal kugadzikana kwe nanoparticle system.Iyo hombe yakananga nzvimbo yenanomaterials inoburitsa yakakura pamusoro pesimba, iyo inotadzisa iyo thermodynamic kugona kweiyo system kusvika yakaderera simba.13 Pasina kudzikamiswa kwakakodzera, izvi zvinogona kutungamira mukuunganidzwa kwemananomaterials.Aggregation ndiko kuumbwa kwezvimedu zvezvimiro zvakasiyana uye hukuru hunoitika kana zvidimbu zvakaparadzirwa zvasangana uye kupindirana kwazvino kwethermodynamic kunobvumira zvimedu kuti zvibatane.Naizvozvo, stabilizers anoshandiswa kudzivirira kuunganidzwa nekuunza yakakura zvakakwana inosemesa simba pakati pezvimedu kurwisa kukwezva kwavo thermodynamic.14
Kunyangwe iyo nyaya yehukuru hwechikamu uye kufukidzwa kwepasi kwakanyatso kutariswa mumamiriro ekugadzirisa kwayo kwezviitiko zvipenyu zvakakonzereswa nanoparticles, particle aggregation inzvimbo isingatariswe zvakanyanya.Ikoko kunenge kusina kunyatsodzidza kugadzirisa colloidal kugadzikana kwe nanoparticles pasi pemamiriro ezvinhu ehupenyu akakodzera.10,15-17 Mukuwedzera, mupiro uyu unonyanya kushoma, apo huturu hunobatanidza nekuunganidza hwakadzidzwawo, kunyange kana hunogona kukonzera maitiro akashata, akadai sevascular thrombosis, kana kurasikirwa kwemaitiro anodiwa, akadai sehuputi hwayo, se inoratidzwa Mufananidzo 1.18, 19 inoratidzwa.Kutaura zvazviri, imwe yenzira shomanana dzinozivikanwa dzesirivha nanoparticle resistance ine chokuita nekuunganidza, nokuti mamwe maE. coli nePseudomonas aeruginosa strains anonzi anoderedza nano-silver senitivity kuburikidza nekutaura puroteni flagellin, flagellin.Iyo ine hukama hwepamusoro kune sirivheri, nekudaro ichikurudzira kuunganidzwa.20
Kune akati wandei akasiyana masisitimu ane hukama nehupfu hwesirivheri nanoparticles, uye kuunganidzwa kunokanganisa ese aya maitiro.Nzira inonyanya kukurukurwa yeAgNP biological chiitiko, dzimwe nguva inonzi "Trojan Horse", inotora maAgNP seAg+ vatakuri.1,21 Iyo Trojan bhiza meshini inogona kuve nechokwadi chekuwedzera kukuru kwenzvimbo yeAg + concentration, inotungamira kuchizvarwa cheROS uye membrane depolarization.22-24 Aggregation inogona kukanganisa kuburitswa kweAg +, nekudaro ichikanganisa chepfu, nekuti inoderedza inoshanda inoshanda nzvimbo uko sirivheri ions inogona kuve yakasvibiswa uye yakanyungudika.Nekudaro, maAgNP haangoratidzi chepfu kuburikidza nekuburitswa kweion.Saizi yakawanda uye morphology-inoenderana nekudyidzana kunofanirwa kutariswa.Pakati pavo, saizi uye chimiro che nanoparticle pamusoro ndiwo anotsanangura maitiro.4,25 Kuunganidzwa kwemaitiro aya anogona kuiswa muchikamu se "induced toxicity nzira."Iko kune akawanda mitochondrial uye pamusoro membrane maitiro anogona kukuvadza organelles uye kukonzera sero kufa.25-27 Sezvo kuumbwa kweaggregates kunokanganisa hukuru uye chimiro chezvinhu zvine sirivheri zvinozivikanwa nemararamiro ehupenyu, kusangana uku kunogonawo kukanganisa.
Mupepa redu rapfuura pamusoro pekuunganidzwa kwesirivheri nanoparticles, takaratidza maitiro anoshanda ekuongorora anosanganisira kemikari uye in vitro biological kuyedza kudzidza dambudziko iri.19 Dynamic Light Scattering (DLS) ndiyo nzira inosarudzirwa yemhando idzi dzekuongorora nekuti zvinhu zvacho zvinogona kuparadzira mafotoni paurefu hunofananidzwa nehukuru hwezvikamu zvawo.Sezvo iyo Brownian inofamba kumhanya kwezvikamu zviri mukati memvura ine hukama nehukuru, shanduko mukusimba kwechiedza chakapararira inogona kushandiswa kuona avhareji hydrodynamic dhayamita (Z-kureva) yemuenzaniso wemvura.28 Mukuwedzera, nekushandisa voltage kune sampuli, iyo zeta inogona (ζ inogona) ye nanoparticle inogona kuyerwa zvakafanana neZ yevhareji kukosha.13,28 Kana iyo yakakwana kukosha kweiyo zeta yakakwira zvakakwana (maererano neyakajairwa mirayiridzo> ± 30 mV), inoburitsa simba rakasimba rekuti electrostatic repulsion pakati pezvimedu kurwisa kuunganidzwa.Hunhu hwepamusoro plasmon resonance (SPR) chinhu chakasarudzika chemaziso, chinonyanya kuverengerwa kune yakakosha simbi nanoparticles (kunyanya Au uye Ag).29 Zvichienderana nemagetsi oscillations (maplasmons epamusoro) ezvishandiso izvi pane nanoscale, zvinozivikanwa kuti spherical AgNPs ine hunhu UV-Vis absorption peak pedyo ne400 nm.30 Kusimba uye wavelength kuchinja kwezvimedu zvinoshandiswa kuwedzera mhedzisiro yeDLS, sezvo nzira iyi inogona kushandiswa kuona nanoparticle aggregation uye pamusoro adsorption yebiomolecules.
Kubva pane ruzivo rwakawanikwa, cell viability (MTT) uye antibacterial assays inoitwa nenzira iyo AgNP toxicity inotsanangurwa sebasa rekuunganidza nhanho, pane (inonyanya kushandiswa chinhu) nanoparticle concentration.Iyi yakasarudzika nzira inotibvumira kuratidza kukosha kwakadzama kweaggregation level mubiology chiitiko, nekuti, semuenzaniso, citrate-yakagumiswa AgNPs inorasikirwa zvachose nekuita kwavo kwehupenyu mukati memaawa mashoma nekuda kwekuunganidzwa.19
Mubasa razvino, isu tinovavarira kuwedzera zvakanyanya mipiro yedu yapfuura mukugadzikana kwebio-inoenderana colloids uye maitiro avo pane biological chiitiko nekudzidza maitiro e nanoparticle saizi pane nanoparticle aggregation.Ichi pasina mubvunzo chimwe chezvidzidzo zve nanoparticles.Maonero epamusoro-soro uye 31 Kuti aongorore nyaya iyi, nzira yekukura kwembeu yakashandiswa kugadzira citrate-terinated AgNPs mumatatu matatu akasiyana-siyana (10, 20, uye 50 nm).6,32 seimwe yenzira dzakajairika.Kune nemananomaterials ayo anoshandiswa zvakanyanya uye anogara achishandiswa mukurapa maapplication, citrate-yakamiswa AgNPs ehukuru hwakasiyana inosarudzwa kuti idzidze iyo inokwanisika saizi kutsamira kweaggregation-inoenderana nebiological properties ye nanosilver.Mushure mekugadzira maAgNP ehukuru hwakasiyana, takaratidzira masampula akagadzirwa nekutapurirana electron microscopy (TEM), tichibva taongorora zvimedu tichishandisa nzira yambotaurwa yekuongorora.Uye zvakare, pamberi pe in vitro cell tsika Dulbecco's Modified Eagle's Medium (DMEM) uye Fetal Bovine Serum (FBS), saizi-inotsamira kuunganidza maitiro uye maitiro ayo akaongororwa pamhando dzakasiyana dzepH, NaCl, glucose, uye glutamine concentrations.Maitiro e cytotoxicity anotemerwa pasi pemamiriro akazara.Kubvumirana kwesainzi kunoratidza kuti kazhinji, zvidimbu zvidiki zviri nani;kuferefeta kwedu kunopa makemikari uye zvipenyu chikuva kuti uone kana izvi zviri izvo.
Matatu esirivha nanoparticles ane masaizi akasiyana akagadzirwa nembeu-mediated yekukura nzira yakakurudzirwa naWan et al., ine zvishoma zvigadziriso.6 Iyi nzira inobva pakuderedza makemikari, kushandisa silver nitrate (AgNO3) sesilver source, sodium borohydride (NaBH4) sechinhu chinoderedza, uye sodium citrate seyo stabilizer.Kutanga, gadzirira 75 mL ye9 mM citrate aqueous solution kubva sodium citrate dihydrate (Na3C6H5O7 x 2H2O) uye kupisa kusvika ku70 ° C.Zvadaro, 2 mL ye1% w/v AgNO3 mhinduro yakawedzerwa kune reaction medium, uyezve iyo ichangobva kugadzirwa sodium borohydride solution (2 mL 0.1% w/v) yakadirwa mumusanganiswa unodonha.Iyo yakaguma yero-brown kuturika yakachengetwa pa70 ° C nekukurudzira kwakasimba kweawa 1, ndokuzotonhodzwa kusvika kune tembiricha.Sample inobuda (inonzi AgNP-I kubva zvino zvichienda mberi) inoshandiswa sehwaro hwekukura kwembeu mudanho rinotevera rekubatanidza.
Kugadzira yepakati-saizi particle suspension (inonzi AgNP-II), kupisa 90 mL 7.6 mM citrate solution kusvika 80°C, sanganisa ne 10 mL AgNP-I, wozosanganisa 2 mL 1% w/v The AgNO3 solution. yakachengetwa pasi pesimba remagetsi rinokurudzira kweawa 1, uyezve sampu yacho yakatonhodzwa kusvika kukamuri tembiricha.
Pachikamu chikuru (AgNP-III), dzokorora maitiro ekukura akafanana, asi panguva iyi, shandisa gumi mL yeAgNP-II sekumiswa kwembeu.Mushure mekunge masampuli asvika tembiricha yekamuri, vanoisa yavo yezita reAg concentration zvichienderana neiyo yakazara AgNO3 yemukati kusvika 150 ppm nekuwedzera kana kubuda evaporate imwe solvent pa40°C, uye pakupedzisira voichengeta pa4°C kusvikira kumwe kushandiswa.
Shandisa FEI Tecnai G2 20 X-Twin Transmission Electron Microscope (TEM) (FEI Corporate Headquarters, Hillsboro, Oregon, USA) ine 200 kV acceleration voltage kuti uongorore morphological maitiro e nanoparticles uye kutora electron diffraction (ED) yavo pateni.Inosvika mifananidzo gumi neshanu inomiririra (~ 750 zvimedu) yakaongororwa pachishandiswa ImageJ software package, uye zvakazobuda histograms (uye magirafu ese muchidzidzo chose) akagadzirwa muOriginPro 2018 (OriginLab, Northampton, MA, USA) 33, 34.
Avhareji hydrodynamic dhayamita (Z-avhareji), zeta inogona (ζ-inogoneka) uye hunhu hwepamusoro plasmon resonance (SPR) yemasampuli akayerwa kuratidza yavo yekutanga colloidal maitiro.Avhareji ye hydrodynamic dhayamita uye zeta zvingangoita zvemuenzaniso zvakayerwa neMalvern Zetasizer Nano ZS chiridzwa (Malvern Instruments, Malvern, UK) vachishandisa masero akapetwa akapetwa capillary pa 37±0.1 ° C.Ocean Optics 355 DH-2000-BAL UV-Vis spectrophotometer (Halma PLC, Largo, FL, USA) yakashandiswa kuwana hunhu hweSPR kubva kuUV-Vis absorption spectra yemasampuli muhuwandu hwe250-800 nm.
Munguva yese yekuyedza, matatu akasiyana ekuyera marudzi ane chekuita nekugadzikana kwecolloidal akaitwa panguva imwe chete.Shandisa DLS kuyera avhareji hydrodynamic dhayamita (Z avhareji) uye zeta kugona (ζ kugona) kwezvimedu, nekuti avhareji yeZ ine hukama neavhareji saizi ye nanoparticle aggregates, uye mukana we zeta unoratidza kana iyo electrostatic repulsion muhurongwa. yakasimba zvakakwana kugadzirisa Van der Waals kukwezva pakati pe nanoparticles.Zviyero zvinogadzirwa zvakapetwa katatu, uye iyo yakajairwa kutsauka kweZ zvinoreva uye zeta zvinogoneka zvinoverengerwa neZetasizer software.Hunhu hweSPR hunoonekwa hwezvimedu hunoongororwa neUV-Vis spectroscopy, nekuti shanduko mukusimba kwepamusoro uye wavelength inogona kuratidza kuunganidzwa uye kusangana kwepasi.29,35 Kutaura zvazviri, pamusoro peplasmon resonance mumatare anokosha ane simba zvokuti zvakatungamirira kune nzira itsva dzekuongorora biomolecules.29,36,37 Mushonga weAgNPs mumusanganiswa wekuedza inenge 10 ppm, uye chinangwa ndechekugadzirisa kusimba kwepamusoro pekutanga SPR kutora ku 1. Kuedza kwakaitwa nenzira inotarisana nenguva pa0;1.5;3;6;12 uye 24 maawa pasi pezvakasiyana biologically zvinoenderana nemamiriro.Mamwe mashoko anotsanangura kuyedza anogona kuonekwa mubasa redu rekare.19 Muchidimbu, akasiyana pH values ​​(3; 5; 7.2 uye 9), akasiyana sodium chloride (10 mM; 50 mM; 150 mM), glucose (3.9 mM; 6.7 mM) uye glutamine (4 mM) concentration, uye akagadzirirawo Dulbecco's Modified Eagle Medium (DMEM) uye Fetal Bovine Serum (FBS) (mumvura neDMEM) semamodhi masisitimu, uye akadzidza mhedzisiro yavo pakuita kwekuunganidza kweiyo synthesized sirivheri nanoparticles.pH Makoshero e, NaCl, glucose, uye glutamine anoongororwa zvichibva pane physiological concentrations, nepo huwandu hweDMEM neFBS hwakafanana nemazinga anoshandiswa mune yese in vitro kuyedza.38-42 Zviyero zvese zvakaitwa papH 7.2 uye 37 ° C ine munyu unogara uripo we10 mM NaCl kubvisa chero kureba kwechikamu chekudyidzana (kunze kweimwe pH uye NaCl-inoenderana nemiedzo, apo hunhu uhu ndihwo hunhu huri pasi pe. kudzidza).28 Rondedzero yemamiriro akasiyana-siyana inopfupikiswa muTebhu 1. Kuedza kwakanyorwa ne † kunoshandiswa sechirevo uye kunoenderana nemuenzaniso une 10 mM NaCl uye pH 7.2.
Human prostate cancer cell line (DU145) uye immortalized human keratinocytes (HaCaT) yakawanikwa kubva kuATCC (Manassas, VA, USA).Masero anogaro kugadzirwa muDulbecco's minimum essential medium Eagle (DMEM) ine 4.5 g/L glucose (Sigma-Aldrich, Saint Louis, MO, USA), yakawedzerwa ne10% FBS, 2 mM L-glutamine, 0.01 % Streptomycin uye 0.005% Penicillin (Sigma-Aldrich, St. Louis, Missouri, USA).Masero akagadzirwa mu37°C incubator pasi pe5% CO2 uye 95% mwando.
Kuti uongorore shanduko muAgNP cytotoxicity inokonzerwa nekuunganidza chidimbu nenzira inotsamira nguva, nhanho mbiri MTT assay yakaitwa.Kutanga, kugona kwemhando mbiri dzemasero kwakayerwa mushure mekurapwa neAgNP-I, AgNP-II uye AgNP-III.Kuti izvi zviitike, marudzi maviri emasero akadyarwa mu96-tsime mahwendefa pahuwandu hwemasero e10,000 / zvakanaka uye akabatwa nehukuru hutatu hwakasiyana hwesirivha nanoparticles mukuwedzera kuwanda pazuva rechipiri.Mushure memaawa makumi maviri nemana ekurapwa, masero akashambidzwa nePBS uye akaiswa ne 0.5 mg / mL MTT reagent (SERVA, Heidelberg, Germany) yakasvibiswa mutsika yepakati kweawa 1 pa 37 ° C.Makristasi eFormazan akanyungudutswa muDMSO (Sigma-Aldrich, Saint Louis, MO, USA), uye kunyura kwakayerwa pa570 nm uchishandisa Synergy HTX plate reader (BioTek-Hungary, Budapest, Hungary).Iko kunyura kukosha kweiyo isina kubatwa control sampu inoonekwa se100% yekupona mwero.Ita zviedzo zvinokwana zvitatu uchishandisa ina yakazvimirira yebiological replicates.IC50 inoverengerwa kubva kudosi mhinduro curve zvichibva pamhedzisiro yehupenyu.
Mushure meizvozvo, mudanho rechipiri, nekuisa zvimedu zvine 150 mM NaCl kwenguva dzakasiyana (0, 1.5, 3, 6, 12, uye maawa makumi maviri nemana) isati yaitwa maseru kurapwa, kwakasiyana kuunganidzwa nyika dzesirivheri nanoparticles dzakagadzirwa.Mushure meizvozvo, iyo imwechete yeMTT assay yakaitwa sezvakambotsanangurwa kuti iongorore shanduko mukuita kwesero kukanganiswa nekubatanidzwa kwechikamu.Shandisa GraphPad Prism 7 kuongorora chigumisiro chekupedzisira, kuverenga kukosha kwehuwandu hwekuedza neunpaired t-test, uye maka nhanho yayo se * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001) ) Uye **** (p ≤ 0.0001).
Saizi nhatu dzakasiyana dzesirivheri nanoparticles (AgNP-I, AgNP-II uye AgNP-III) yakashandiswa kuita antibacterial susceptibility kune Cryptococcus neoformans IFM 5844 (IFM; Research Center yePathogenic Fungi uye Microbial Toxicology, Chiba University) uye Bacillus Test megaterium SZMC 60. (SZMC: Szeged Microbiology Collection) uye E. coli SZMC 0582 muRPMI 1640 yepakati (Sigma-Aldrich Co.).Kuti uongorore kuchinja kwekushanda kweantibacterial kunokonzerwa nekubatanidzwa kwezvikamu, kutanga, zvishoma zvishoma zvekudzivirira kwavo (MIC) zvakagadziriswa ne microdilution mu 96-well microtiter plate.Ku 50 μL yekumisikidzwa kwesero (5 × 104 masero/mL muRPMI 1640 yepakati), wedzera 50 μL yesirivheri nanoparticle kumiswa uye serially dilue kaviri iyo yekumisikidza (mune yambotaurwa svikiro, renji iri 0 uye 75 ppm, kureva, sampu yekudzora ine 50 μL yekumiswa kwesero uye 50 μL yepakati isina nanoparticles).Pashure pacho, ndiro yacho yakanyungudutswa pa 30 ° C kwemaawa 48, uye kusvibiswa kwemaziso kwetsika kwakayerwa pa 620 nm uchishandisa SPECTROstar Nano plate reader (BMG LabTech, Offenburg, Germany).Kuedza kwakaitwa katatu katatu.
Kunze kwekuti 50 μL yeimwe yakaunganidzwa nanoparticle samples yakashandiswa panguva ino, maitiro akafanana sezvakatsanangurwa kare akashandiswa kuongorora maitiro ekuunganidza pane antibacterial chiitiko pane ambotaurwa.Nzvimbo dzakasiyana dzekuunganidza dzesirivheri nanoparticles dzinogadzirwa nekupinza zvimedu ne150 mM NaCl kwenguva dzakasiyana dzenguva (0, 1.5, 3, 6, 12, uye maawa makumi maviri nemana) isati yagadzirwa.Kumiswa kwakawedzerwa ne50 μL yeRPMI 1640 yepakati kwakashandiswa sekudzora kukura, nepo kudzora huturu, kumiswa neasina-akaunganidzwa nanoparticles akashandiswa.Kuedza kwakaitwa katatu katatu.Shandisa GraphPad Prism 7 kuti uongorore mhedzisiro zvakare, uchishandisa ongororo yenhamba yakafanana neyekuongorora kweMTT.
Iyo yekuunganidza nhanho yezvidiki zvidiki (AgNP-I) yakaratidzirwa, uye mhedzisiro yakaburitswa muchikamu chebasa redu rekare, asi kuti tienzanise zvirinani, zvese zvimedu zvakanyatsoongororwa.Iyo data yekuedza inounganidzwa uye inokurukurwa muzvikamu zvinotevera.Saizi nhatu dzeAgNP.19
Zviyero zvakaitwa neTEM, UV-Vis uye DLS zvakasimbisa kuumbwa kwakabudirira kweese masampuli eAgNP (Mufananidzo 2A-D).Zvinoenderana nemutsara wekutanga weMufananidzo 2, diki diki (AgNP-I) inoratidza yunifomu spherical morphology ine avhareji yedhayamita ingangoita gumi nm.Iyo nzira yekukura yembeu inopawo AgNP-II neAgNP-III nehukuru hwakasiyana siyana neavhareji madhayamita anosvika makumi maviri nm uye makumi mashanu nm, zvichiteerana.Zvinoenderana neyakajairwa kutsauka kwekugoverwa kwezvikamu, saizi yemasampuri matatu haaenderani, izvo zvakakosha pakuenzanisa kwavo kuongorora.Nekuenzanisa avhareji yechiyero chechiyero uye mutete reshiyo yeTEM-based particle 2D fungidziro, inofungidzirwa kuti sphericity yezvimedu inoongororwa neImageJ's shape filter plug-in (Mufananidzo 2E).43 Zvinoenderana nekuongororwa kwechimiro chezvimedu, chiyero chazvo (chikamu chikuru / chikamu chipfupi cherectangle diki inosungirirwa) haikanganisike nekukura kwechikamu, uye hutete hwayo (yakayerwa nzvimbo yeinoenderana yakakwana denderedzwa / nzvimbo yetioretical. ) zvishoma nezvishoma inoderera.Izvi zvinoguma nekuwedzera uye kuwanda kwe polyhedral particles, ayo akanyatsotenderera mudzidziso, anoenderana nehutete reshiyo 1.
Mufananidzo 2 Transmission electron maikorosikopu (TEM) mufananidzo (A), electron diffraction (ED) pateni (B), size distribution histogram (C), characteristic ultraviolet-visible (UV-Vis) light absorption spectrum (D), uye avhareji fluid Citrate. -yakapera sirivha nanoparticles ine mechanical diameter (Z-avhareji), zeta inogona, aspect ratio uye ukobvu reshiyo (E) ine matatu akasiyana saizi renji: AgNP-I iri gumi nm (yepamusoro mutsara), AgNP -II imakumi maviri nm (yepakati mutsara ), AgNP-III (mutsara wezasi) ndeye 50 nm.
Kunyangwe iyo cyclic chimiro chenzira yekukura yakakanganisa chimiro chechikamu kusvika pamwero, zvichikonzera diki sphericity yeAgNPs hombe, ese matatu masampuli akaramba ari quasi-spherical.Mukuwedzera, sezvinoratidzwa mu electron diffraction pattern muFigure 2B, nano The crystallinity of the particles haina kukanganiswa.Iyo yakakurumbira diffraction mhete-iyo inogona kubatanidzwa ne (111), (220), (200), uye (311) Miller indices yesirivheri-inopindirana zvakanyanya nezvinyorwa zvesainzi uye nemipiro yedu yapfuura.9, 19,44 Kupatsanurwa kweDebye-Scherrer mhete yeAgNP-II neAgNP-III imhaka yekuti mufananidzo weED unotorwa pakukura kwakafanana, kuitira kuti ukuru hwechikamu huwedzere, huwandu hwezvikamu zvakasiyana unit nzvimbo inowedzera uye inoderera.
Saizi uye chimiro che nanoparticles anozivikanwa kukanganisa biological chiitiko.3,45 Shape-inotsamira catalytic uye biological chiitiko chinogona kutsanangurwa nenyaya yekuti maumbirwo akasiyana anowanzo wedzera mamwe makristasi zviso (kuva neMiller indices akasiyana), uye zviso zvekristaro izvi zvine mabasa akasiyana.45,46 Sezvo zvimedu zvakagadzirirwa zvinopa zvakafanana ED mhedzisiro inoenderana neyakafanana makristasi maitiro, zvinogona kufungidzirwa kuti mune yedu inotevera colloidal kugadzikana uye biological chiitiko kuedza, chero misiyano inocherechedzwa inofanirwa kuverengerwa kuhukuru hweNanoparticle, kwete maitiro ane hukama.
Mhedzisiro yeUV-Vis yakapfupikiswa muMufananidzo 2D inosimbisa iyo yakawandisa spherical hunhu hweAgNP yakasanganiswa, nekuti iyo SPR yepamusoro yemasamples ese matatu akatenderedza 400 nm, inova kukosha kwehunhu hwespherical sirivheri nanoparticles.29,30 Iyo yakatorwa spectra yakasimbisawo kubudirira kwembeu-mediated kukura kwe nanosilver.Sezvo saizi yechikamu ichiwedzera, iyo wavelength inoenderana neyakakura kunyura kwechiedza kweAgNP-II-yakanyanya mukurumbira-Maererano nezvinyorwa, AgNP-III Yakawana redshift.6,29
Nezvekutanga colloidal kugadzikana kweAgNP system, DLS yakashandiswa kuyera avhareji hydrodynamic dhayamita uye zeta mukana wezvimedu papH 7.2.Mhedzisiro inoratidzwa muMufananidzo 2E inoratidza kuti AgNP-III ine colloidal kugadzikana kwepamusoro kupfuura AgNP-I kana AgNP-II, nekuti nhungamiro dzinozivikanwa dzinoratidza kuti zeta inogoneka ye30 mV zvachose inodiwa kuti igare kwenguva refu colloidal kugadzikana Izvi zvinotsigirwa zvakare kana. iyo Z yeavhareji kukosha (inowanikwa seavhareji hydrodynamic dhayamita yemahara uye akaunganidzwa zvimedu) inofananidzwa neiyo yekutanga particle saizi yakawanikwa neTEM, nekuti padyo neaya ma values ​​ari, hunyoro dhigirii Unganidza mumuenzaniso.Muchokwadi, avhareji yeZ yeAgNP-I neAgNP-II yakakwira zvine musoro pane yavo huru TEM-yakaongororwa saizi, saka kana ichienzaniswa neAgNP-III, aya masampuli anofungidzirwa kuti angangowedzera kuunganidza, uko kune yakanyanya kunaka zeta. inoperekedzwa nehukuru hwepedyo Iyo Z yepakati kukosha.
Tsananguro yechiitiko ichi inogona kuva mbiri.Kune rimwe divi, iyo citrate concentration inochengetwa padanho rakafanana mumatanho ese ekubatanidza, ichipa huwandu hwakanyanya hwemachaji epamusoro mapoka kudzivirira iyo chaiyo nzvimbo yezvimedu zvinokura kubva pakuderera.Nekudaro, maererano naLevak et al., mamorekuru madiki senge citrate anogona kuchinjika nyore nyore ne biomolecules pamusoro pe nanoparticles.Muchiitiko ichi, kugadzikana kwecolloidal kuchagadziriswa nekorona ye biomolecules inogadzirwa.31 Nekuti maitiro aya akaonekwawo muzviyero zvedu zvekuunganidza (zvakakurukurwa zvakadzama gare gare), citrate capping chete haigone kutsanangura chiitiko ichi.
Kune rimwe divi, saizi yechikamu inopesana nemaitiro ekuunganidza padanho re nanometer.Izvi zvinonyanya kutsigirwa nemaitiro echinyakare Derjaguin-Landau-Verwey-Overbeek (DLVO) nzira, uko kukwezva kwechidimbu kunotsanangurwa sehuwandu hwemasimba anokwezva uye anosemesa pakati pezvimedu.Maererano naHe et al., Kunyanya kukosha kweDLVO simba curve kunoderera nehukuru hwe nanoparticles mu hematite nanoparticles, zvichiita kuti zvive nyore kusvika kune zvishoma zvishoma simba rekutanga, nokudaro kukurudzira kusingagadziriswi aggregation (condensation).47 Zvisineyi, zvinofungidzirwa kuti kune zvimwe zvinhu zvinodarika miganhu yedzidziso yeDLVO.Kunyange zvazvo van der Waals giravhiti uye electrostatic double-layer repulsion zvakafanana nekuwedzera particle size, wongororo naHotze et al.inokurudzira kuti ine simba rakasimba pakuunganidza kupfuura DLVO inobvumira.14 Vanodavira kuti kukombama kwepamusoro kwenanoparticles hakuchagoni kufungidzirwa senzvimbo yakati sandara, zvichiita kuti fungidziro yemasvomhu isashande.Pamusoro pezvo, sezvo saizi yechikamu ichidzikira, chikamu chemaatomu aripo pamusoro chinova chakakwira, zvichitungamira kune yemagetsi chimiro uye pamusoro pechaji maitiro.Uye pamusoro reactivity shanduko, izvo zvinogona kutungamira mukudzikira kwechaji mumagetsi kaviri layer uye kukurudzira kuunganidza.
Kana tichienzanisa mhedzisiro yeDLS yeAgNP-I, AgNP-II, uye AgNP-III muMufananidzo 3, takaona kuti ese matatu masampuli airatidza yakafanana pH inokurudzira kuunganidzwa.Nzvimbo ine acidic yakawanda (pH 3) inoshandura mukana we zeta wemuenzaniso kusvika ku0 mV, zvichiita kuti zvimedu zvigadzire ma micron-size aggregates, nepo alkaline pH inoshandura simba rayo rezeta kune yakakura yakaipa kukosha, apo zvikamu zvinoumba zviduku zvishoma (pH 5). )Uye 7.2)), kana kuramba isina kuunganidzwa zvachose (pH 9).Mimwe misiyano yakakosha pakati pemasampuli akasiyana akaonekwa zvakare.Munguva yose yekuedza, AgNP-I yakaratidza kuti ndiyo inonyanya kukoshesa pH-induced zeta inogona kuchinja, nokuti zeta inogona yezvikamu izvi yakaderedzwa pa pH 7.2 kana ichienzaniswa nepH 9, nepo AgNP-II neAgNP-III yakaratidza chete A. kuchinja kukuru mu ζ kwakapoteredza pH 3. Mukuwedzera, AgNP-II yakaratidza kuchinja kunononoka uye ine mwero zeta inogona, nepo AgNP-III yakaratidza hunyoro hutsika hwevatatu, nokuti hurongwa hwakaratidza huwandu hwepamusoro hwezeta kukosha uye kufamba zvishoma nezvishoma, zvichiratidza. AgNP-III Yakanyanya kupikisa pH-induced aggregation.Mhedzisiro iyi inoenderana neavhareji hydrodynamic diayamita yekuyera mhinduro.Tichifunga nezvehukuru hwemaprimers avo, AgNP-I yairatidza kuunganidzwa zvishoma nezvishoma papH yose, zvichida nekuda kwe10 mM NaCl kumashure, nepo AgNP-II neAgNP-III zvakangoratidza zvakakosha papH 3 yekuunganidza.Musiyano unonyanya kunakidza ndewekuti kunyangwe yakakura nanoparticle saizi, AgNP-III inoumba madiki akaunganidzwa papH 3 mumaawa makumi maviri nemana, ichiratidza hunhu hwayo hwekurwisa-aggregation.Nekukamura avhareji Z yeAgNPs papH 3 mushure memaawa makumi maviri nemana nekukosha kwemuenzaniso wakagadzirirwa, zvinogona kucherechedzwa kuti masaizi akaunganidzwa eAgNP-I neAgNP-II akawedzera ne50 times, 42 times, uye 22 times. , zvichiteerana.III.
Mufananidzo 3 Chiedza chine simba chinoparadzira mhedzisiro ye citrate-yakamiswa sirivheri nanoparticles sampu ine kuwedzera kukura (10 nm: AgNP-I, 20 nm: AgNP-II uye 50 nm: AgNP-III) inoratidzwa seavhareji hydrodynamic dhayamita (Z avhareji. ) (kurudyi) Pasi pemamiriro akasiyana epH, mukana we zeta (kuruboshwe) unoshanduka mukati meawa makumi maviri nemana.
Iyo yakacherechedzwa pH-inotsamira kuunganidzwa kwakakanganisawo hunhu hwepamusoro plasmon resonance (SPR) yemasampuri eAgNP, sezvinoratidzwa neiyo UV-Vis spectra.Zvinoenderana neSupplementary Figure S1, kuunganidzwa kweese matatu esirivha nanoparticle kumiswa kunoteverwa nekudzikiswa kwekusimba kwepamusoro pepamusoro peSPR uye shanduko tsvuku ine mwero.Hukuru hwekuchinja uku sebasa repH hunoenderana nehuwandu hwekuunganidza hwakafanotaurwa nemhedzisiro yeDLS, zvisinei, mamwe mafambiro anonakidza akaonekwa.Kupesana nekufungidzira, zvinozoitika kuti iyo yepakati-saizi AgNP-II ndiyo inonyanya kunzwa kune SPR shanduko, nepo mamwe maviri masampuli asinganyanye kunzwa.Mukutsvagisa kweSPR, 50 nm ndiyo theoretical particle size limit, iyo inoshandiswa kusiyanisa zvimedu zvichibva pane yavo dielectric properties.Zvimedu zvidiki pane 50 nm (AgNP-I uye AgNP-II) zvinogona kutsanangurwa sezviri nyore dielectric dipoles, nepo zvimedu zvinosvika kana kudarika muganho uyu (AgNP-III) zvine zvakanyanya kuomarara dielectric properties, uye resonance yavo Iyo bhendi inotsemuka kuita multimodal shanduko. .Panyaya yemasampula maviri madiki madiki, maAgNP anogona kutorwa seakareruka dipoles, uye iyo plasma inogona kupindirana zviri nyore.Sezvo saizi yechikamu ichiwedzera, kubatanidza uku kunoburitsa plasma yakakura, iyo inogona kutsanangura kunzwisiswa kwepamusoro kunoonekwa.29 Zvisineyi, kune zvimedu zvakakura, fungidziro yakareruka ye dipole haishande kana dzimwe nyika dzekubatanidza dzingaitika zvakare, izvo zvinogona kutsanangura kudzikira kwemaitiro eAgNP-III kuratidza shanduko yekuona.29
Pasi pemamiriro edu ekuedza, zvinoratidzwa kuti pH kukosha ine mhedzisiro yakakura pakugadzikana kwecolloidal ye citrate-yakavharwa sirivheri nanoparticles ehukuru hwakasiyana.Mune aya masisitimu, kugadzikana kunopihwa neakaipa -COO- mapoka ari pamusoro peAgNPs.Iyo carboxylate inoshanda boka reiyo citrate ion inoburitswa muhuwandu hukuru hweH + ion, saka iyo yakagadzirwa carboxyl boka haichakwanise kupa electrostatic repulsion pakati pezvimedu, sezvakaratidzwa mumutsara wepamusoro weMufananidzo 4. Maererano neLe Chatelier's musimboti, AgNP masampuli anokurumidza kuunganidza pa pH 3, asi zvishoma nezvishoma anowedzera kugadzikana sezvo pH inowedzera.
Mufananidzo 4 Schematic mechanism yekudyidzana kwepasi inotsanangurwa nekuunganidza pasi pe pH yakasiyana (yepamusoro mutsara), NaCl concentration (yepakati mutsara), uye biomolecules (yezasi mutsara).
Zvinoenderana neFigure 5, iyo colloidal kugadzikana muAgNP kumiswa kwehukuru hwakasiyana kwakaongororwawo pasi pekuwedzera munyu.Kubva pane zeta inogona, iyo yakawedzera nanoparticle saizi mune idzi citrate-yakamiswa AgNP masisitimu zvakare inopa kukwidziridzwa kuramba kune ekunze pesvedzero kubva kuNaCl.MuAgNP-I, 10 mM NaCl inokwana kukurudzira kuunganidzwa kwakapfava, uye munyu we50 mM unopa mhedzisiro yakafanana.MuAgNP-II uye AgNP-III, 10 mM NaCl haina zvakanyanya kukanganisa zeta inogona nekuti maitiro avo anoramba ari pa (AgNP-II) kana pazasi (AgNP-III) -30 mV.Kuwedzera huwandu hweNaCl kusvika ku50 mM uye pakupedzisira kusvika ku150 mM NaCl yakakwana kuti ideredze zvakanyanya kukosha kwezeta inogona mumasampuli ose, kunyange zvazvo zvikamu zvakakura zvinochengetedza zvakanyanya kuipa.Mhedzisiro iyi inowirirana neavhareji inotarisirwa hydrodynamic dhayamita yeAgNPs;iyo Z avhareji mitsetse yakayerwa pa10, 50, uye 150 mM NaCl inoratidza zvakasiyana, zvishoma nezvishoma ichiwedzera kukosha.Pakupedzisira, ma micron-saizi aggregates akaonekwa mune ese matatu 150 mM miedzo.
Mufananidzo 5 Chiedza chine simba chinoparadzira mhedzisiro ye citrate-yakapera sirivha nanoparticles sampu ine kuwedzera kukura (10 nm: AgNP-I, 20 nm: AgNP-II uye 50 nm: AgNP-III) inoratidzwa seavhareji hydrodynamic dhayamita (Z avhareji. ) (kurudyi) uye zeta zvinogona (kuruboshwe) kuchinja mukati meawa 24 pasi pezvikamu zvakasiyana zveNaCl.
Iyo UV-Vis inoburitsa muSupplementary Figure S2 inoratidza kuti iyo SPR ye50 uye 150 mM NaCl mumasamples ese matatu ine nekukasira uye kuderera kukuru.Izvi zvinogona kutsanangurwa neDLS, nekuti NaCl-based aggregation inoitika nekukurumidza kupfuura pH-inotsamira kuedza, iyo inotsanangurwa nemusiyano mukuru pakati pekutanga (0, 1.5, uye 3 maawa) kuyerwa.Pamusoro pezvo, kuwedzera kusanganiswa kwemunyu kuchawedzerawo kubvumirwa kwechiyedzo chesvikiro, icho chichava nemhedzisiro yakakura pamusoro peplasmon resonance.29
Mhedzisiro yeNaCl inopfupikiswa mumutsara wepakati weMufananidzo 4. Kazhinji, zvinogona kugumiswa kuti kuwedzera kusanganiswa kwesodium chloride kune simba rakafanana nekuwedzera acidity, nokuti Na + ions ine tsika yekubatanidza kumativi e-carboxylate mapoka. kudzvinyirira zvakaipa zeta zvinogona AgNPs.Pamusoro pezvo, 150 mM NaCl yakagadzira micron-size aggregates mumasampuli ese matatu, zvichiratidza kuti physiological electrolyte concentration inokanganisa kugadzikana kwecolloidal kwecitrate-terinated AgNPs.Nekutarisisa iyo yakakosha condensing concentration (CCC) yeNaCl pane yakafanana AgNP masisitimu, izvi mhedzisiro zvinogona kuiswa nehungwaru mumabhuku akakodzera.Huynh et al.yakaverenga kuti CCC yeNaCl ye citrate-terinated silver nanoparticles ine avhareji dhayamita ye71 nm yaive 47.6 mM, nepo El Badawy et al.akacherechedza kuti CCC ye10 nm AgNPs ine citrate coating yaiva 70 mM.10,16 Mukuwedzera, iyo CCC yakakwirira zvikuru yeinenge 300 mM yakayerwa naHe et al., izvo zvakaita kuti nzira yavo yekubatanidza ive yakasiyana kubva pakadhindwa yakambotaurwa.48 Kunyange zvazvo mupiro wemazuva ano usina kunangana nekuongorora kwakadzama kwezvinokosha izvi, nokuti mamiriro edu ekuedza ari kuwedzera mukuoma kwechidzidzo chese, biologically yakakosha NaCl concentration ye 50 mM, kunyanya 150 mM NaCl, inoratidzika kunge yakakwirira.Induced coagulation, ichitsanangura shanduko dzakasimba dzakaonekwa.
Nhanho inotevera muyedzo yepolymerization ndeye kushandisa mamorekuru akareruka asi ane biologically anoenderana nekuita nanoparticle-biomolecule.Kubva paDLS (Figure 6 ne7) uye UV-Vis zvawanikwa (Supplementary Figures S3 uye S4), dzimwe mhedziso dzakajairwa dzinogona kusimbiswa.Pasi pemamiriro edu ekuyedza, mamorekuru akadzidzwa glucose uye glutamine haazokonzeresa kuunganidzwa mune chero AgNP system, nekuti iyo Z-kureva maitiro ane hukama nehukoshi hunoenderana nekuyera kukosha.Kunyangwe kuvepo kwavo kusingakanganise kuunganidzwa, mhedzisiro yekuyedza inoratidza kuti mamorekuru aya adsorbed zvishoma pamusoro peAgNPs.Mhedzisiro inonyanya kutsigira maonero aya ndeyekuonekwa shanduko mukutorwa kwechiedza.Kunyangwe zvazvo AgNP-I isingaratidze hurefu hune chinangwa kana shanduko yehusimba, inogona kucherechedzwa zvakajeka nekuyera zvimedu zvakakura, izvo zvinowanzoitika nekuda kwekunzwa kukuru kwemaziso kwambotaurwa.Pasinei nekusungwa, glucose inogona kukonzera kuchinja kukuru kutsvuku mushure memaawa 1.5 kana ichienzaniswa nechiyero chekutonga, iyo inenge 40 nm muAgNP-II uye inenge 10 nm muAgNP-III, iyo inoratidza kuitika kwekubatana kwepamusoro.Glutamine yakaratidza maitiro akafanana, asi shanduko yakanga isiri pachena.Mukuwedzera, zvakakoshawo kutaura kuti glutamine inogona kuderedza zvachose zeta inogona yepakati uye yakakura zvidimbu.Zvisinei, sezvo idzi shanduko dzezeta dzisingaite sedzinokanganisa chiyero chekubatanidza, zvinogona kufungidzirwa kuti kunyange madiki biomolecules se glutamine anogona kupa imwe dhigirii yekuramba kwepakati pakati pezvikamu.
Mufananidzo 6 Chiedza chine simba chinoparadzira mhedzisiro ye citrate-yakamiswa sirivheri nanoparticle samples ine kuwedzera kukura (10 nm: AgNP-I, 20 nm: AgNP-II uye 50 nm: AgNP-III) inoratidzwa seavhareji hydrodynamic dhayamita (Z avhareji) (kurudyi) Pasi pemamiriro ekunze ezvakasiyana-siyana zveglucose, iyo zeta inogona (kuruboshwe) inoshanduka mukati meawa 24.
Mufananidzo 7 Chiedza chine simba chinoparadzira mhedzisiro ye citrate-yakamiswa sirivheri nanoparticles sampu ine kuwedzera kukura (10 nm: AgNP-I, 20 nm: AgNP-II uye 50 nm: AgNP-III) inoratidzwa seavhareji hydrodynamic dhayamita (Z avhareji. ) (kurudyi) Pamberi pe glutamine, mukana we zeta (kuruboshwe) unoshanduka mukati meawa makumi maviri nemana.
Muchidimbu, madiki biomolecules senge glucose uye glutamine haakonzere kugadzikana kwecolloidal pakuyerwa kwekuyerwa: kunyangwe achikanganisa mukana we zeta uye UV-Vis mhedzisiro kune akasiyana madhigirii, iyo Z avhareji mhedzisiro haina kuenderana.Izvi zvinoratidza kuti kunyura kwepamusoro kwemamorekuru kunotadzisa kudzorwa nemagetsi, asi panguva imwe chete kunopa kugadzikana kwedimensional.
Kuti tibatanidze zvakamboitika nezvakange zvabuda kare uye titevedzere mamiriro ebhayoloji zvine hunyanzvi, takasarudza mamwe anonyanya kushandiswa maseru etsika uye akaashandisa semamiriro ekuyedza ekudzidza kugadzikana kweAgNP colloids.Mune yese in vitro kuyedza, rimwe remabasa akanyanya kukosha eDMEM sepakati ndiko kumisikidza zvinodiwa osmotic mamiriro, asi kubva kune kemikari yekuona, iyo yakaoma munyu mhinduro ine yakazara ionic simba yakafanana ne150 mM NaCl. .40 Kana iri FBS, isanganiswa yakaoma yebiomolecules-kunyanya mapuroteni-kubva pakuona kwepamusoro adsorption, ine zvimwe zvakafanana nemigumisiro yekuedza yeglucose uye glutamine, pasinei nekugadzirwa kwemakemikari uye zvakasiyana-siyana Bonde rinonyanya kuoma.19 DLS uye UV-Migumisiro inooneka inoratidzwa muMufananidzo 8 uye Supplementary Figure S5, maererano, inogona kutsanangurwa nekuongorora makemikari ezvinyorwa izvi uye kuzvibatanidza nezviyero muchikamu chekare.
Mufananidzo 8 Chiedza chine simba chinoparadzira mhedzisiro ye citrate-yakamiswa sirivheri nanoparticles sampu ine kuwedzera kukura (10 nm: AgNP-I, 20 nm: AgNP-II uye 50 nm: AgNP-III) inoratidzwa seavhareji hydrodynamic dhayamita (Z avhareji. ) (kurudyi) Muhupo hwesero tsika zvikamu DMEM neFBS, zeta zvinogona (kuruboshwe) zvinoshanduka mukati meawa 24.
Dilution yeAgNPs yehukuru hwakasiyana muDMEM ine mhedzisiro yakafanana pakugadzikana kwecolloidal kune iyo inocherechedzwa muhupo hwehuwandu hweNaCl.Kupararira kweAgNP mu50 v / v% DMEM yakaratidza kuti kuunganidza kukuru kwakaonekwa nekuwedzera kwezeta inogona uye Z-avhareji kukosha uye kuderera kwakanyanya kweSPR yakasimba.Izvo zvakakosha kuti ticherechedze kuti iyo yakakura aggregate saizi yakakonzerwa neDMEM mushure memaawa makumi maviri nemana inopesana nehukuru hweprimer nanoparticles.
Kudyidzana pakati peFBS neAgNP kwakafanana neinoonekwa pamberi pemamorekuru madiki akadai seglucose uye glutamine, asi mhedzisiro yacho yakasimba.Avhareji yeZ yezvimedu inoramba isina kubatwa, nepo kuwedzera kwezeta kugona kunoonekwa.Iyo SPR peak yakaratidza kutsvuka kudiki kuchinja, asi pamwe zvinonyanya kufadza, iyo SPR kusimba haina kuderera zvakanyanya senge muyero yekudzora.Iyi mhedzisiro inogona kutsanangurwa neiyo innate adsorption ye macromolecules pamusoro pe nanoparticles (yezasi mutsara muMufananidzo 4), iyo ikozvino inonzwisiswa sekuumbwa kweiyo biomolecular corona mumuviri.49