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Shin ƙananan nanoparticles koyaushe sun fi kyau?Fahimtar tasirin nazarin halittu na tattara adadin nanoparticles na azurfa masu dogaro da girma a ƙarƙashin yanayin da suka dace da ilimin halitta
Marubuta: Bélteky P, Rónavári A, Zakupszky D, Boka E, Igaz N, Szerencsés B, Pfeiffer I, Vágvölgyi C, Kiricsi M, Kónya Z
Péter Bélteky,* Andrea Rónavári,1,* Dalma Zakupszky,1 Eszter Boka,1 Nóra Igaz,2 Bettina Szerencsés,3 Ilona Pfeiffer,3 Csaba Vágvölgyi,3 Mónika Kiricsi na Muhalli Chemistry, Hungary, Hungary a Faculty of Environmental Chemistry, Hungary, Hungary , Jami'ar Szeged;2 Sashen Kimiyyar Halittu da Kwayoyin Halitta, Sashen Kimiyya da Bayani, Jami'ar Szeged, Hungary;3 Ma'aikatar Microbiology, Faculty of Science and Information, Jami'ar Szeged, Hungary;4MTA-SZTE Reaction Kinetics da Surface Chemistry Research Group, Szeged, Hungary* Waɗannan marubutan sun ba da gudummawa daidai da wannan aikin.Sadarwa: 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 [Kariyar imel] Manufa: Silver nanoparticles (AgNPs) ne daya daga cikin nanomaterials da aka fi karantawa, musamman saboda aikace-aikacen su na biomedical.Duk da haka, saboda haɗuwa da nanoparticles, kyakkyawan cytotoxicity da aikin ƙwayoyin cuta sau da yawa ana lalata su a cikin kafofin watsa labaru na halitta.A cikin wannan aikin, an yi nazarin halayen haɗin kai da ayyukan nazarin halittu masu alaƙa na samfuran nanoparticle na azurfa daban-daban da aka ƙare da citrate tare da matsakaicin diamita na 10, 20, da 50 nm.Hanyar: Yi amfani da microscope na lantarki watsawa don haɗawa da siffanta nanoparticles, kimanta halayen haɗuwarsu a ma'auni daban-daban na pH, NaCl, glucose da glutamine maida hankali ta hanyar watsawar haske mai ƙarfi da ultraviolet-bayyan gani spectroscopy.Bugu da kari, a cikin al'adun tantanin halitta matsakaici sassa irin su Dulbecco yana inganta haɓaka halayen a cikin Matsakaicin Eagle Medium da Fetal Calf Serum.Sakamako: Sakamakon ya nuna cewa acidic pH da physiological electrolyte abun ciki gabaɗaya suna haifar da tarawar sikelin micron, wanda za'a iya shiga tsakani ta hanyar samuwar corona biomolecular.Yana da kyau a lura cewa manyan barbashi suna nuna juriya ga tasirin waje fiye da ƙananan takwarorinsu.An yi gwajin cytotoxicity na in vitro da gwaje-gwajen ƙwayoyin cuta ta hanyar kula da sel tare da tarin nanoparticle a matakan tarawa daban-daban.Kammalawa: Sakamakonmu yana nuna alaƙa mai zurfi tsakanin kwanciyar hankali na colloidal da guba na AgNPs, kamar yadda matsananciyar haɗuwa ke haifar da cikakkiyar asarar ayyukan nazarin halittu.Matsayi mafi girma na anti-aggregation da aka lura don ƙananan ƙwayoyin cuta yana da tasiri mai mahimmanci akan ƙwayar in vitro, saboda irin waɗannan samfurori suna riƙe da ƙarin aikin ƙwayoyin cuta da ƙwayoyin mammalian.Wadannan binciken sun kai ga ƙarshe cewa, duk da ra'ayi na gaba ɗaya a cikin wallafe-wallafen da suka dace, ƙaddamar da ƙananan ƙwayoyin nanoparticles bazai zama mafi kyawun aikin ba.Mahimman kalmomi: girma mai tsaka-tsakin iri, kwanciyar hankali na colloidal, halayen haɓaka mai dogara da girman girma, lalata lalata
Yayin da buƙatu da fitarwa na nanomaterials ke ci gaba da ƙaruwa, ana ƙara kulawa ga lafiyar halittu ko ayyukan halitta.Nanoparticles na Azurfa (AgNPs) ɗaya ne daga cikin waɗanda aka fi haɗawa, bincike da kuma amfani da wakilan wannan nau'in kayan saboda kyawawan kaddarorin su na haɓaka, gani da abubuwan halitta.1 An yi imani da cewa keɓaɓɓen halaye na nanomaterials (ciki har da AgNPs) an danganta su da babban yanki na musamman.Saboda haka, matsalar babu makawa ita ce duk wani tsari da ya shafi wannan mahimmin fasalin, kamar girman barbashi, shafi ko tari, ko zai lalata kaddarorin nanoparticles waɗanda ke da mahimmanci ga takamaiman aikace-aikace.
Tasirin girman barbashi da masu daidaitawa batutuwa ne da aka yi rubuce-rubuce sosai a cikin adabi.Misali, ra'ayin da aka yarda da shi shine cewa ƙananan nanoparticles sun fi guba fiye da manyan nanoparticles.2 Daidai da wallafe-wallafen gabaɗaya, karatunmu na baya sun nuna aikin dogaro da girman nanosilver akan ƙwayoyin mammalian da ƙwayoyin cuta.3- 5 Rufe saman wani sifa ce wacce ke da tasiri mai yawa akan kaddarorin nanomaterials.Kawai ta hanyar ƙara ko gyara masu daidaitawa a samansa, nau'in nanomaterial ɗaya na iya samun mabambanta na zahiri, sinadarai, da abubuwan halitta.Ana yin aikace-aikacen ma'aikatan capping galibi a matsayin ɓangare na haɗin nanoparticle.Alal misali, citrate-kare azurfa nanoparticles ne daya daga cikin mafi dacewa AgNPs a cikin bincike, wanda aka hada ta rage azurfa salts a zaba stabilizer bayani a matsayin dauki matsakaici.6 Citrate na iya samun sauƙin amfani da ƙarancin farashinsa, samuwa, haɓakawa, da ƙaƙƙarfan alaƙa ga azurfa, wanda za'a iya nunawa a cikin ma'amala daban-daban da aka ba da shawarar, daga adsorption mai jujjuyawar ƙasa zuwa hulɗar ionic.Kananan kwayoyin halitta da polyatomic ions kusa da 7,8, kamar su citrates, polymers, polyelectrolytes, da kwayoyin halitta suma ana amfani dasu don daidaita nano-azurfa da yin ayyuka na musamman akansa.9-12
Ko da yake yiwuwar canza aikin nanoparticles ta hanyar da gangan capping surface yanki ne mai ban sha'awa sosai, babban aikin wannan shafi mai mahimmanci ba shi da mahimmanci, yana samar da kwanciyar hankali na colloidal ga tsarin nanoparticle.Babban yanki na musamman na nanomaterials zai samar da babban makamashi na saman, wanda ke hana ikon thermodynamic na tsarin don isa mafi ƙarancin makamashi.13 Ba tare da daidaitawar da ta dace ba, wannan na iya haifar da ƙaranci na nanomaterials.Tari shine samuwar tarin barbashi na sifofi da girma dabam-dabam da ke faruwa a lokacin da barbashi da suka tarwatse suka hadu kuma hulɗar thermodynamic na yanzu suna ba da damar barbashi su manne da juna.Don haka, ana amfani da masu daidaitawa don hana tarawa ta hanyar gabatar da isasshe babban ƙarfi mai tsauri tsakanin ɓangarorin don magance sha'awar thermodynamic.14
Kodayake batun girman barbashi da ɗaukar hoto an binciko shi sosai a cikin mahallin tsarinsa na ayyukan nazarin halittu da ke haifar da nanoparticles, tara abubuwan da aka yi watsi da su.Babu kusan babu cikakken nazari don warware kwanciyar hankali na nanoparticles a ƙarƙashin yanayin da ya dace da ilimin halitta.10,15-17 Bugu da ƙari, wannan gudummawar ba ta da yawa, inda aka kuma yi nazarin gubar da ke tattare da tarawa, ko da zai iya haifar da mummunan halayen, kamar thrombosis na jini, ko asarar halayen da ake so, kamar gubarsa, kamar wanda aka nuna a cikin Hoto 1.18, 19 da aka nuna.A gaskiya ma, ɗaya daga cikin ƙananan hanyoyin da aka sani na juriya na nanoparticle na azurfa yana da alaƙa da tarawa, saboda an ba da rahoton wasu nau'in E. coli da Pseudomonas aeruginosa don rage girman nano-azurfa ta hanyar bayyana furotin flagellin, flagellin.Yana da babban kusanci ga azurfa, ta yadda zai haifar da tarawa.20
Akwai hanyoyi daban-daban masu alaƙa da guba na nanoparticles na azurfa, kuma tarawa yana rinjayar duk waɗannan hanyoyin.Hanyar da aka fi magana game da ayyukan nazarin halittu na AgNP, wani lokaci ana kiranta da tsarin "Trojan Horse", yana kula da AgNPs a matsayin Ag+ masu ɗaukar kaya.1,21 Tsarin doki na Trojan na iya tabbatar da karuwa mai yawa a cikin taro na Ag + na gida, wanda ke haifar da tsararrun ROS da membrane depolarization.22-24 Haɗin kai na iya rinjayar sakin Ag +, ta haka yana tasiri mai guba, saboda yana rage tasiri mai tasiri inda ions na azurfa za a iya oxidized da narkar da su.Koyaya, AgNPs ba kawai zasu nuna guba ta hanyar sakin ion ba.Dole ne a yi la'akari da girman girma da yawa da hulɗar da ke da alaƙa.Daga cikin su, girman da siffar nanoparticle surface ne ma'anar halaye.4,25 Tarin waɗannan hanyoyin ana iya rarraba su azaman “hanyoyin guba da aka jawo.”Akwai yuwuwar haɓakar halayen mitochondrial da saman membrane wanda zai iya lalata gabobin jiki kuma ya haifar da mutuwar tantanin halitta.25-27 Tun da samuwar tarukan a zahiri yana shafar girma da siffar abubuwa masu ɗauke da azurfa waɗanda tsarin rayuwa suka gane, waɗannan hulɗar kuma na iya shafar.
A cikin takardarmu da ta gabata game da tara nanoparticles na azurfa, mun nuna ingantaccen tsarin tantancewa wanda ya ƙunshi sinadarai da gwaje-gwajen halittu na in vitro don nazarin wannan matsala.19 Dynamic Light Scattering (DLS) ita ce dabarar da aka fi so don waɗannan nau'ikan dubawa saboda kayan na iya watsa photons a tsayin raƙuman ruwa kwatankwacin girman barbashi.Tun da saurin motsi na Brownian na barbashi a cikin matsakaicin ruwa yana da alaƙa da girman, ana iya amfani da canjin ƙarfin tarwatsewar haske don sanin matsakaicin diamita na hydrodynamic (Z-ma'ana) na samfurin ruwa.28 Bugu da ƙari, ta hanyar yin amfani da wutar lantarki zuwa samfurin, ana iya auna ƙarfin zeta (ζ m) na nanoparticle daidai da matsakaicin darajar Z.13,28 Idan madaidaicin ƙimar yuwuwar zeta ya isa sosai (bisa ga jagororin gabaɗaya> ± 30 mV), zai haifar da tsangwama mai ƙarfi tsakanin ɓangarorin don magance tari.Halayen bayyanar plasmon (SPR) wani lamari ne na musamman na gani, wanda aka fi danganta shi da nanoparticles na ƙarfe mai daraja (yafi Au da Ag).29 Dangane da motsin lantarki (plasmons na saman) na waɗannan kayan akan nanoscale, an san cewa AgNPs masu sassauƙa suna da ƙimar shayarwar UV-Vis kusa da 400 nm.30 Ana amfani da matsananciyar ƙarfi da tsayin raƙuman barbashi don ƙarin sakamako na DLS, saboda ana iya amfani da wannan hanyar don gano tari nanoparticle da tallan saman na biomolecules.
Dangane da bayanin da aka samu, ana yin amfani da kwayar halitta (MTT) da kuma gwajin ƙwayoyin cuta a cikin hanyar da aka kwatanta toxicity na AgNP a matsayin aiki na matakin haɗuwa, maimakon (mafi yawan amfani da factor) nanoparticle maida hankali.Wannan hanya ta musamman tana ba mu damar nuna mahimmancin mahimmancin matakin tarawa a cikin ayyukan nazarin halittu, saboda, alal misali, citrate-kashe AgNPs gaba daya sun rasa ayyukansu na halitta a cikin 'yan sa'o'i kadan saboda tarawa.19
A cikin aikin na yanzu, muna nufin fadada gudunmawarmu ta baya a cikin kwanciyar hankali na colloid masu alaka da kwayoyin halitta da tasirin su akan ayyukan nazarin halittu ta hanyar nazarin tasirin nanoparticle akan tarawar nanoparticle.Wannan babu shakka daya ne daga cikin nazarin nanoparticles.Hanya mafi girma da kuma 31 Don bincika wannan batu, an yi amfani da hanyar haɓakar iri don samar da AgNPs mai citrate da aka ƙare a cikin nau'i daban-daban guda uku (10, 20, da 50 nm).6,32 a matsayin daya daga cikin mafi yawan hanyoyin.Don nanomaterials waɗanda aka yadu kuma akai-akai amfani da su a cikin aikace-aikacen likita, an zaɓi citrate- ƙare AgNPs masu girma dabam dabam don nazarin yuwuwar dogaro girman girman abubuwan da ke da alaƙa da abubuwan halitta na nanosilver.Bayan haɗar AgNPs masu girma dabam dabam, mun siffata samfuran da aka samar ta hanyar microscopy na lantarki (TEM), sannan mu bincika barbashi ta amfani da hanyar nunawa da aka ambata.Bugu da ƙari, a gaban in vitro cell al'adun Dulbecco's Modified Eagle's Medium (DMEM) da Fetal Bovine Serum (FBS), an kimanta halayen haɗin gwiwar da ke dogara da girman girman da halayensa a wasu nau'o'in pH, NaCl, glucose, da glutamine.An ƙayyade halayen cytotoxicity a ƙarƙashin ingantattun yanayi.Ijma'in kimiyya ya nuna cewa gabaɗaya, ƙananan ƙwayoyin cuta sun fi dacewa;Bincikenmu yana samar da dandalin sinadarai da halittu don sanin ko haka lamarin yake.
Nanoparticles na azurfa guda uku tare da nau'ikan girman daban-daban an shirya su ta hanyar hanyar girma mai matsakaicin iri ta Wan et al., Tare da ƴan gyare-gyare.6 Wannan hanya ta dogara ne akan rage sinadarai, ta yin amfani da nitrate na azurfa (AgNO3) a matsayin tushen azurfa, sodium borohydride (NaBH4) a matsayin wakili mai ragewa, da sodium citrate a matsayin stabilizer.Na farko, shirya 75 ml na 9 mM citrate ruwa mai ruwa daga sodium citrate dihydrate (Na3C6H5O7 x 2H2O) da zafi zuwa 70 ° C.Sa'an nan, 2 ml na 1% w/v AgNO3 bayani an ƙara zuwa matsakaicin amsawa, sa'an nan kuma an zuba maganin sodium borohydride da aka shirya (2 mL 0.1% w / v) a cikin cakuda dropwise.Sakamakon dakatarwar rawaya-launin ruwan kasa an kiyaye shi a 70 ° C tare da motsawa mai ƙarfi na awa 1, sannan a sanyaya zuwa zafin jiki.Samfurin da aka samo (wanda ake kira AgNP-I daga yanzu) ana amfani dashi azaman tushen ci gaban iri-iri a cikin mataki na gaba.
Don haɗa matsakaiciyar dakatarwar barbashi (wanda aka nuna a matsayin AgNP-II), zafi 90 ml 7.6 mM citrate bayani zuwa 80 ° C, haɗa shi da 10 ml AgNP-I, sa'an nan kuma haxa 2 ml 1% w / v Maganin AgNO3 an ajiye shi a ƙarƙashin motsi mai ƙarfi na inji na awa 1, sa'an nan kuma an sanyaya samfurin zuwa zafin jiki.
Don mafi girman barbashi (AgNP-III), maimaita tsarin haɓaka iri ɗaya, amma a wannan yanayin, yi amfani da 10 ml na AgNP-II azaman dakatarwar iri.Bayan samfuran sun kai zafin daki, sun saita ƙimar su na AgNO3 dangane da jimlar abun ciki na AgNO3 zuwa 150 ppm ta ƙara ko fitar da ƙarin ƙarfi a 40 ° C, kuma a ƙarshe adana su a 4 ° C har sai an ƙara amfani.
Yi amfani da FEI Tecnai G2 20 X-Twin Transmission Electron Microscope (TEM) (FEI Hedkwatar Kamfanin, Hillsboro, Oregon, Amurka) tare da ƙarfin ƙarfin haɓakar kV 200 don bincika halayen ƙwayoyin nanoparticles da kama tsarin diffraction na lantarki (ED).Akalla hotunan wakilan 15 (~ 750 barbashi) an kimanta su ta amfani da fakitin software na ImageJ, kuma sakamakon histograms (da duk jadawali a cikin duka binciken) an ƙirƙira su a cikin OriginPro 2018 (OriginLab, Northampton, MA, Amurka) 33, 34.
Matsakaicin diamita na hydrodynamic (Z-matsakaici), yuwuwar zeta (ζ-yiwuwa) da kuma halayen haɓakar yanayin plasmon (SPR) na samfuran an auna su don nuna alamun farko na colloidal.Matsakaicin diamita na hydrodynamic da yuwuwar zeta na samfurin an auna su ta hanyar kayan aikin Malvern Zetasizer Nano ZS (Instruments Malvern, Malvern, UK) ta amfani da sel masu nannade capillary a 37 ± 0.1 ° C.Ocean Optics 355 DH-2000-BAL UV-Vis spectrophotometer (Halma PLC, Largo, FL, Amurka) da aka yi amfani da su don samun sifa ta SPR halaye daga UV-Vis bakan bakan na samfurori a cikin kewayon 250-800 nm.
A yayin duk gwajin, nau'ikan ma'auni daban-daban guda uku da suka danganci kwanciyar hankali na colloidal an gudanar da su a lokaci guda.Yi amfani da DLS don auna matsakaicin matsakaicin diamita na hydrodynamic (matsakaicin Z) da yuwuwar zeta (ζ yuwuwar) na barbashi, saboda matsakaicin Z yana da alaƙa da matsakaicin girman tarin nanoparticle, kuma yuwuwar zeta yana nuna ko ƙin electrostatic a cikin tsarin. yana da ƙarfi sosai don kashe sha'awar Van der Waals tsakanin nanoparticles.Ana yin ma'auni a cikin sau uku, kuma ana ƙididdige ma'auni na ma'anar Z da yuwuwar zeta ta software na Zetasizer.Siffar sifa ta SPR na barbashi ana kimanta su ta hanyar UV-Vis spectroscopy, saboda canje-canje a cikin ƙarfin kololuwa da tsayin raƙuman ruwa na iya nuna haɗuwa da hulɗar saman.29,35 A haƙiƙa, faɗakarwar plasmon sama a cikin karafa masu daraja yana da tasiri sosai wanda ya haifar da sabbin hanyoyin bincike na biomolecules.29,36,37 Ƙaddamarwar AgNPs a cikin cakuda gwaji shine game da 10 ppm, kuma manufar ita ce saita ƙarfin matsakaicin matsakaicin ƙaddamarwa na farko na SPR zuwa 1. An gudanar da gwajin a cikin hanyar dogara da lokaci a 0;1.5;3;6;12 da 24 hours a ƙarƙashin yanayi daban-daban masu dacewa da ilimin halitta.Ana iya ganin ƙarin cikakkun bayanai da ke kwatanta gwajin a cikin aikinmu na baya.19 A takaice, nau'o'in pH daban-daban (3; 5; 7.2 da 9), daban-daban sodium chloride (10 mM; 50 mM; 150 mM), glucose (3.9 mM; 6.7 mM) da glutamine (4 mM) maida hankali, da kuma Har ila yau, an shirya Dulbecco's Modified Eagle Medium (DMEM) da Fetal Bovine Serum (FBS) (a cikin ruwa da DMEM) a matsayin tsarin samfuri, kuma sun yi nazarin tasirin su akan haɓakar halayen nanoparticles na azurfa.pH Ana ƙididdige ƙimar, NaCl, glucose, da glutamine bisa ga ƙididdigar ilimin lissafi, yayin da adadin DMEM da FBS daidai suke da matakan da aka yi amfani da su a cikin duka gwajin in vitro.38-42 An yi duk ma'auni a pH 7.2 da 37 ° C tare da ƙaddamarwar gishiri na 10 mM NaCl na yau da kullum don kawar da duk wani hulɗar barbashi mai nisa (sai dai wasu gwaje-gwajen pH da NaCl, inda waɗannan halayen su ne masu canji a ƙarƙashin. karatu).28 An taƙaita jerin yanayi daban-daban a cikin Table 1. An yi amfani da gwajin da aka yiwa alama tare da † a matsayin ma'ana kuma ya dace da samfurin da ke dauke da 10 mM NaCl da pH 7.2.
Layin kwayar cutar kansar ɗan adam (DU145) da keratinocytes ɗan adam mara mutuwa (HaCaT) an samo su daga ATCC (Manassas, VA, Amurka).Ana yin al'adar sel akai-akai a cikin mafi ƙarancin matsakaicin matsakaicin mikiya (DMEM) mai ɗauke da glucose 4.5 g/L (Sigma-Aldrich, Saint Louis, MO, Amurka), an ƙara su da 10% FBS, 2 mM L-glutamine, 0.01 % Streptomycin da 0.005% Penicillin (Sigma-Aldrich, St. Louis, Missouri, Amurka).Ana al'adar sel a cikin incubator 37°C ƙarƙashin 5% CO2 da 95% zafi.
Don bincika canje-canje a cikin cytotoxicity na AgNP da ke haifar da haɓakar ƙwayar cuta ta hanyar dogaro da lokaci, an yi gwajin MTT mai matakai biyu.Na farko, an auna yiwuwar nau'in tantanin halitta guda biyu bayan jiyya tare da AgNP-I, AgNP-II da AgNP-III.Don wannan karshen, an tsara nau'ikan kwayoyin halitta guda biyu cikin faranti 96-rijiya a yawan sel 10,000 / rijiya kuma ana bi da su tare da nau'ikan nau'ikan nau'ikan nau'ikan nanoparticles na azurfa daban-daban guda uku a cikin haɓaka ƙima a rana ta biyu.Bayan sa'o'i 24 na jiyya, an wanke sel tare da PBS kuma an sanya su tare da 0.5 mg / mL MTT reagent (SERVA, Heidelberg, Jamus) a cikin matsakaici na al'ada don 1 hour a 37 ° C.An narkar da lu'ulu'u na Formazan a cikin DMSO (Sigma-Aldrich, Saint Louis, MO, Amurka), kuma an auna sha a 570 nm ta amfani da mai karanta farantin HTX Synergy (BioTek-Hungary, Budapest, Hungary).Ana ɗaukar ƙimar sha na samfurin kulawa da ba a kula da shi a matsayin ƙimar rayuwa 100%.Yi aƙalla gwaje-gwaje 3 ta amfani da kwafin halittu masu zaman kansu guda huɗu.Ana ƙididdige IC50 daga madaidaicin amsawar kashi bisa sakamakon kuzari.
Bayan haka, a mataki na biyu, ta hanyar ƙaddamar da ƙwayoyin cuta tare da 150 mM NaCl na lokuta daban-daban (0, 1.5, 3, 6, 12, da 24 hours) kafin maganin tantanin halitta, an samar da nau'o'i daban-daban na tara nanoparticles na azurfa.Daga baya, an yi gwajin MTT iri ɗaya kamar yadda aka bayyana a baya don kimanta canje-canje a cikin yuwuwar tantanin halitta wanda ya shafa.Yi amfani da GraphPad Prism 7 don kimanta sakamako na ƙarshe, ƙididdige mahimmancin ƙididdiga na gwajin ta hanyar gwajin t-test mara ɗimbin yawa, kuma yi alama matakinsa kamar * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001). ) Kuma **** (p ≤ 0.0001).
Uku daban-daban masu girma dabam na azurfa nanoparticles (AgNP-I, AgNP-II da kuma AgNP-III) da aka yi amfani da antibacterial mai saukin kamuwa zuwa Cryptococcus neoformans IFM 5844 (IFM; Cibiyar Bincike don Pathogenic Fungi da Microbial Toxicology, Jami'ar Chiba) da Bacillus Test megaterium SZMC 603 (SZMC: Szeged Microbiology Collection) da E. coli SZMC 0582 a cikin RPMI 1640 matsakaici (Sigma-Aldrich Co.).Don yin la'akari da canje-canje a cikin ayyukan ƙwayoyin cuta da ke haifar da tarawar ƙwayoyin cuta, na farko, ƙananan ƙaddamarwar hanawa (MIC) an ƙaddara su ta hanyar microdilution a cikin 96-riji microtiter farantin.Zuwa 50 μL na daidaitaccen dakatarwar tantanin halitta (5 × 104 sel / ml a cikin RPMI 1640 matsakaici), ƙara 50 μL na dakatarwar nanoparticle na azurfa kuma a bi da su a hankali sau biyu na maida hankali (a cikin matsakaicin da aka ambata, kewayon shine 0 da 75 ppm, Wato, samfurin sarrafawa ya ƙunshi 50 μL na dakatarwar salula da 50 μL na matsakaici ba tare da nanoparticles ba).Bayan haka, an sanya farantin a 30 ° C na tsawon sa'o'i 48, kuma an auna ma'auni na al'ada a 620 nm ta amfani da SPECTROstar Nano plate reader (BMG LabTech, Offenburg, Jamus).An yi gwajin sau uku sau uku.
Sai dai an yi amfani da 50 μL na samfurori na nanoparticle guda ɗaya a wannan lokacin, irin wannan hanya kamar yadda aka bayyana a baya an yi amfani da shi don nazarin tasirin haɗuwa akan ayyukan ƙwayoyin cuta a kan nau'in da aka ambata.Jihohi daban-daban na tara nanoparticles na azurfa ana samar da su ta hanyar shigar da barbashi tare da 150 mM NaCl na lokuta daban-daban (0, 1.5, 3, 6, 12, da 24 hours) kafin sarrafa tantanin halitta.An yi amfani da dakatarwar da aka kara da 50 μL na RPMI 1640 matsakaici a matsayin kulawar girma, yayin da don sarrafa guba, an yi amfani da dakatarwa tare da nanoparticles marasa tarawa.An yi gwajin sau uku sau uku.Yi amfani da GraphPad Prism 7 don sake kimanta sakamakon ƙarshe, ta amfani da ƙididdigar ƙididdiga iri ɗaya kamar binciken MTT.
An kwatanta matakin haɗuwa na ƙananan ƙananan ƙwayoyin cuta (AgNP-I), kuma an buga sakamakon da aka buga a cikin aikinmu na baya, amma don kwatanta mafi kyau, duk barbashi an duba su sosai.Ana tattara bayanan gwaji kuma an tattauna su a cikin sassan masu zuwa.Girma uku na AgNP.19
Ma'aunai da TEM, UV-Vis da DLS suka yi sun tabbatar da nasarar haɓakar duk samfuran AgNP (Hoto 2A-D).Bisa ga jeri na farko na Hoto 2, mafi ƙanƙanta barbashi (AgNP-I) yana nuna nau'in nau'in nau'in halitta mai kama da matsakaicin diamita na kusan nm 10.Hanyar ci gaban iri-iri kuma tana ba da AgNP-II da AgNP-III tare da jeri daban-daban tare da matsakaicin diamita na barbashi na kusan 20 nm da 50 nm, bi da bi.Bisa ga daidaitattun daidaituwa na rarraba barbashi, masu girma na samfurori guda uku ba su zoba, wanda ke da mahimmanci don nazarin kwatancen su.Ta hanyar kwatanta matsakaicin ma'auni da rabon bakin ciki na tushen TEM na tsinkayar barbashi na 2D, ana ɗauka cewa ana kimanta girman ɓangarorin ta hanyar filogi mai tace siffar siffar ImageJ (Hoto 2E).43 Bisa ga bincike na siffar barbashi, su al'amari rabo (babban gefe / guntun gefen mafi karami bounding rectangle) ba ya shafar barbashi girma, kuma su thinness rabo (aunawa yanki na daidai da'irar / ka'idar yanki). ) sannu a hankali yana raguwa.Wannan yana haifar da ƙarin ƙwayoyin polyhedral, waɗanda suke daidai gwargwado a ka'idar, daidai da rabon bakin ciki na 1.
Hoto 2 Hoton microscope na lantarki (TEM) (A), ƙirar lantarki (ED) ƙirar (B), ƙididdigar girman girman girman (C), halayen ultraviolet-bayani (UV-Vis) bakan bakan haske (D), da matsakaicin ruwa Citrate - ƙare azurfa nanoparticles tare da inji diamita (Z-matsakaici), zeta m, al'amari rabo da kauri rabo (E) da uku daban-daban size jeri: AgNP-I ne 10 nm (saman jere), AgNP -II ne 20 nm (tsakiyar jere). ), AgNP-III (jere na kasa) shine 50 nm.
Ko da yake yanayin cyclic na hanyar girma ya shafi sifar barbashi zuwa wani ɗan lokaci, wanda ya haifar da ƙaramin girman girman AgNPs, duk samfuran ukun sun kasance masu ƙima-spherical.Bugu da kari, kamar yadda aka nuna a tsarin rarrabuwar wutar lantarki a cikin Hoto na 2B, nano Ba a shafan crystallinity na barbashi ba.Fitaccen zobe-wanda za a iya danganta shi da (111), (220), (200), da (311) Miller indices na azurfa-ya yi daidai da wallafe-wallafen kimiyya da gudummawar da muka bayar a baya.9, 19,44 Rarraba zobe na Debye-Scherrer na AgNP-II da AgNP-III saboda gaskiyar cewa an kama hoton ED a girman girman wannan, don haka girman ƙwayar ƙwayar cuta yana ƙaruwa, adadin ɓarnawar ɓarna a kowace. yankin naúrar yana ƙaruwa da raguwa .
Girma da siffar nanoparticles an san su da tasiri ga ayyukan nazarin halittu.3,45 Za'a iya yin bayanin abubuwan da suka dogara da sifofi da ayyukan ilimin halitta ta hanyar gaskiyar cewa siffofi daban-daban suna haɓaka wasu fuskokin crystal (suna da alamun Miller daban-daban), kuma waɗannan fuskokin crystal suna da ayyuka daban-daban.45,46 Tun da shirye barbashi samar da irin wannan ED sakamakon m sosai kama crystal halaye, za a iya zaci cewa a cikin m colloidal kwanciyar hankali da nazarin halittu gwaje-gwajen, duk wani lura bambance-bambancen da ya kamata a dangana ga Nanoparticle size, ba siffar da alaka Properties.
Sakamakon UV-Vis da aka taƙaita a cikin Hoto 2D yana ƙara jaddada girman yanayin yanayin haɗin AgNP, saboda kololuwar SPR na duk samfuran guda uku suna kusa da 400 nm, wanda shine ƙimar sifa ta nanoparticles na azurfa.29,30 Hotunan da aka kama sun kuma tabbatar da nasarar ci gaban nanosilver mai matsakaicin iri.Yayin da girman barbashi ya karu, tsayin tsayin da ya dace da matsakaicin ɗaukar haske na AgNP-II-mafi mahimmanci-Bisa ga wallafe-wallafen, AgNP-III ya ɗanɗana ja.6,29
Game da kwanciyar hankali na farko na tsarin AgNP, an yi amfani da DLS don auna matsakaicin diamita na hydrodynamic da yuwuwar zeta na barbashi a pH 7.2.Sakamakon da aka kwatanta a cikin Hoto 2E ya nuna cewa AgNP-III yana da kwanciyar hankali mafi girma fiye da AgNP-I ko AgNP-II, saboda jagororin gama gari sun nuna cewa yiwuwar zeta na 30 mV cikakke yana da mahimmanci don kwanciyar hankali na dogon lokaci Wannan binciken yana da goyon baya lokacin da Matsakaicin ƙimar Z (wanda aka samu azaman matsakaicin diamita na hydrodynamic na kyauta da tarawa) an kwatanta shi da girman barbashi na farko da TEM ta samu, saboda kusancin dabi'u biyu, mafi ƙarancin digiri Gather a cikin samfurin.A gaskiya ma, matsakaicin Z na AgNP-I da AgNP-II yana da ma'ana mafi girma fiye da girman girman su na TEM da aka kimanta, don haka idan aka kwatanta da AgNP-III, ana tsammanin waɗannan samfurori za su iya haɗuwa, inda mafi ƙarancin zeta. yana tare da girman kusanci Matsakaicin ƙimar Z.
Bayanin wannan lamari na iya zama kashi biyu.A daya hannun, da citrate taro da aka kiyaye a irin wannan matakin a duk kira matakai, samar da wani in mun gwada da babban adadin caje kungiyoyin don hana takamaiman surface yankin na girma barbashi daga raguwa.Koyaya, a cewar Levak et al., ƙananan ƙwayoyin cuta kamar citrate ana iya samun sauƙin musanya ta biomolecules akan saman nanoparticles.A wannan yanayin, za a ƙayyade kwanciyar hankali ta hanyar korona na biomolecules da aka samar.31 Domin an kuma lura da wannan hali a cikin ma'aunin mu na tarawa (an tattauna dalla-dalla daga baya), citrate capping kadai ba zai iya bayyana wannan lamari ba.
A daya hannun, girman barbashi yana da inversely gwargwado zuwa tara hali a matakin nanometer.Wannan galibi yana samun goyan bayan hanyar gargajiya ta Derjaguin-Landau-Verwey-Overbeek (DLVO), inda aka kwatanta jan hankalin barbashi a matsayin jimlar karfi mai ban sha'awa da kyama tsakanin barbashi.A cewar He et al., Matsakaicin ƙimar ƙarfin ƙarfin DLVO yana raguwa tare da girman nanoparticles a cikin nanoparticles na hematite, yana sauƙaƙa don isa mafi ƙarancin makamashi na farko, ta haka yana haɓaka haɓakar da ba za a iya jurewa ba (condensation).47 Duk da haka, ana hasashe cewa akwai wasu fannonin da suka wuce iyakokin ka'idar DLVO.Ko da yake van der Waals nauyi da electrostatic biyu-Layer repulsion suna kama da ƙara barbashi size, wani bita ta Hotze et al.yana ba da shawarar cewa yana da tasiri mai ƙarfi akan tarawa fiye da yadda DLVO ya ƙyale.14 Sun yi imanin cewa ba za a iya ƙididdige lanƙwan saman nanoparticles a matsayin ƙasa mai lebur ba, wanda ba za a iya amfani da kimar lissafi ba.Bugu da kari, yayin da girman barbashi ya ragu, adadin atom da ke kan saman ya zama mafi girma, wanda ke haifar da tsarin lantarki da halayen cajin saman.Kuma sake kunnawa saman yana canzawa, wanda zai iya haifar da raguwar cajin a cikin layin lantarki biyu da haɓaka haɓakawa.
Lokacin kwatanta sakamakon DLS na AgNP-I, AgNP-II, da AgNP-III a cikin Hoto 3, mun lura cewa duk samfurori guda uku sun nuna irin wannan haɗin pH.Yanayin acidic mai nauyi (pH 3) yana canza yuwuwar zeta na samfurin zuwa 0 mV, yana haifar da barbashi don samar da adadin micron-sized aggregates, yayin da alkaline pH ya canza yuwuwar zeta zuwa mafi girman ƙimar mara kyau, inda barbashi ke samar da ƙarami tara (pH 5). ).Kuma 7.2)), ko zama gaba ɗaya ba tare da tarawa ba (pH 9).An kuma lura da wasu mahimman bambance-bambance tsakanin samfurori daban-daban.A cikin gwajin, AgNP-I ya tabbatar da cewa ya zama mafi mahimmanci ga pH-induced yuwuwar canje-canjen zeta, saboda an rage yiwuwar zeta na waɗannan ƙwayoyin a pH 7.2 idan aka kwatanta da pH 9, yayin da AgNP-II da AgNP-III kawai sun nuna A A. babban canji a cikin ζ yana kusa da pH 3. Bugu da ƙari, AgNP-II ya nuna sauye-sauye a hankali da matsakaicin matsakaicin zeta, yayin da AgNP-III ya nuna hali mafi sauƙi na uku, saboda tsarin ya nuna mafi girman darajar zeta da jinkirin motsi, yana nuna AgNP-III Mafi juriya ga tarawar pH.Waɗannan sakamakon sun yi daidai da matsakaicin sakamakon auna diamita na hydrodynamic.Idan aka yi la'akari da girman ɓangarorin abubuwan da suka fi dacewa, AgNP-I ya nuna tari a hankali akai-akai a duk ƙimar pH, mai yuwuwa saboda 10 mM NaCl baya, yayin da AgNP-II da AgNP-III kawai sun nuna mahimmanci a pH 3 Na taro.Bambanci mafi ban sha'awa shi ne cewa duk da girman girman nanoparticle, AgNP-III yana samar da mafi ƙanƙanta a cikin pH 3 a cikin sa'o'i 24, yana nuna kaddarorin anti-aggregation.Ta hanyar rarraba matsakaicin Z na AgNPs a pH 3 bayan sa'o'i 24 ta darajar samfurin da aka shirya, ana iya lura da cewa girman girman girman AgNP-I da AgNP-II sun karu da sau 50, sau 42, da sau 22. , bi da bi.III.
Hoto 3 Sakamakon watsawar haske mai ƙarfi na samfurin citrate- ƙarewar azurfa nanoparticles samfurin tare da ƙara girman (10 nm: AgNP-I, 20 nm: AgNP-II da 50 nm: AgNP-III) an bayyana shi azaman matsakaicin diamita na hydrodynamic (matsakaicin Z). ) (dama) A ƙarƙashin yanayin pH daban-daban, yuwuwar zeta (hagu) yana canzawa cikin sa'o'i 24.
Abubuwan da aka lura da pH ɗin da aka lura kuma sun shafi halayen haɓakar plasmon (SPR) na samfuran AgNP, kamar yadda aka nuna ta UV-Vis spectra.Dangane da Ƙarin Hoto S1, tara duk dakatarwar nanoparticle na azurfa guda uku yana biye da raguwa a cikin ƙarfin kololuwar SPR ɗin su da matsakaita ja.Girman waɗannan canje-canje a matsayin aikin pH ya yi daidai da matakin haɗuwa da sakamakon DLS ya annabta, duk da haka, an lura da wasu abubuwa masu ban sha'awa.Sabanin fahimta, ya bayyana cewa AgNP-II mai matsakaici shine mafi mahimmanci ga canje-canjen SPR, yayin da sauran samfurori guda biyu ba su da hankali.A cikin bincike na SPR, 50 nm shine ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙayyadaddun, wanda aka yi amfani da shi don bambanta barbashi bisa ga kaddarorin dielectric.Barbashi ƙasa da 50 nm (AgNP-I da AgNP-II) za a iya bayyana su azaman dielectric dipoles masu sauƙi, yayin da barbashi da suka kai ko suka wuce wannan iyaka (AgNP-III) suna da ƙarin hadaddun kaddarorin dielectric, kuma rawarsu Ƙungiyar ta rabu cikin canje-canjen multimodal. .A cikin yanayin ƙananan samfurori guda biyu, ana iya ɗaukar AgNPs azaman dipoles masu sauƙi, kuma plasma na iya haɗuwa cikin sauƙi.Yayin da girman barbashi ya ƙaru, wannan haɗin kai da gaske yana samar da plasma mafi girma, wanda zai iya yin bayanin mafi girman hankali da aka gani.29 Duk da haka, don mafi girma ɓangarorin, ƙididdigar dipole mai sauƙi ba ta da inganci lokacin da sauran jihohin haɗin gwiwa na iya faruwa, wanda zai iya bayyana ragi na AgNP-III don nuna canje-canje na gani.29
A ƙarƙashin yanayin gwajin mu, an tabbatar da cewa ƙimar pH yana da tasiri mai zurfi akan kwanciyar hankali na colloidal na nanoparticles na azurfa da aka rufe da citrate na masu girma dabam.A cikin waɗannan tsarin, ana ba da kwanciyar hankali ta hanyar da aka caje-ƙungiyoyin -COO akan saman AgNPs.Ƙungiyar aikin carboxylate na citrate ion ana yin protonated a cikin adadi mai yawa na H+ ions, don haka ƙungiyar carboxyl da aka haifar ba za ta iya samar da wutar lantarki tsakanin barbashi ba, kamar yadda aka nuna a saman jere na Hoto 4. Bisa ga ka'idar Le Chatelier, AgNP samfurori da sauri suna tarawa a pH 3, amma a hankali suna ƙara karuwa yayin da pH ke ƙaruwa.
Hoto 4 Tsarin tsari na hulɗar saman da aka bayyana ta hanyar tarawa a ƙarƙashin pH daban-daban (jere na sama), NaCl maida hankali (jere na tsakiya), da biomolecules (jere na ƙasa).
Dangane da Hoto na 5, an kuma bincika kwanciyar hankali na colloidal a cikin dakatarwar AgNP masu girma dabam dabam a ƙarƙashin ƙara yawan gishiri.Dangane da yuwuwar zeta, ƙãra girman nanoparticle a cikin waɗannan tsarin AgNP da aka ƙare citrate kuma yana ba da ingantaccen juriya ga tasirin waje daga NaCl.A cikin AgNP-I, 10 mM NaCl ya isa ya haifar da tarawa mai laushi, kuma ƙwayar gishiri na 50 mM yana samar da sakamako iri ɗaya.A cikin AgNP-II da AgNP-III, 10 mM NaCl ba zai tasiri tasirin zeta sosai ba saboda ƙimar su ta kasance a (AgNP-II) ko ƙasa (AgNP-III) -30 mV.Ƙara yawan ƙaddamarwar NaCl zuwa 50 mM kuma a ƙarshe zuwa 150 mM NaCl ya isa ya rage mahimmancin ƙimar yuwuwar zeta a cikin duk samfurori, kodayake manyan ƙwayoyin cuta suna riƙe da ƙarin cajin mara kyau.Waɗannan sakamakon sun yi daidai da matsakaicin matsakaicin diamita na hydrodynamic na AgNPs;Matsakaicin layukan yanayin Z da aka auna akan 10, 50, da 150 mM NaCl suna nuna daban-daban, a hankali suna haɓaka ƙima.A ƙarshe, an gano tari mai girman micron a duk gwaje-gwajen 150 mM guda uku.
Hoto 5 Sakamakon watsawar haske mai ƙarfi na samfurin nanoparticles na azurfa citrate da aka ƙare tare da haɓaka girma (10 nm: AgNP-I, 20 nm: AgNP-II da 50 nm: AgNP-III) an bayyana su azaman matsakaicin diamita na hydrodynamic (matsakaicin Z). ) (dama) da yuwuwar zeta (hagu) suna canzawa a cikin sa'o'i 24 a ƙarƙashin ƙididdigar NaCl daban-daban.
Sakamakon UV-Vis a Ƙarin Hoto S2 ya nuna cewa SPR na 50 da 150 mM NaCl a cikin dukkanin samfurori guda uku suna da raguwa mai sauri da mahimmanci.Wannan za a iya bayyana ta DLS, saboda haɗin tushen NaCl yana faruwa da sauri fiye da gwaje-gwaje masu dogara da pH, wanda aka bayyana ta babban bambanci tsakanin farkon (0, 1.5, da 3 hours) ma'auni.Bugu da ƙari, haɓaka ƙaddamarwar gishiri kuma zai ƙara yawan izinin dangi na matsakaicin gwaji, wanda zai yi tasiri mai zurfi a kan tasirin plasmon.29
An taƙaita tasirin NaCl a cikin layi na tsakiya na Hoto 4. Gabaɗaya, ana iya ƙaddamar da cewa ƙara yawan ƙwayar sodium chloride yana da irin wannan tasiri kamar ƙara yawan acidity, saboda Na + ions suna da hali don daidaitawa a kusa da ƙungiyoyin carboxylate, danne mummunan zeta m AgNPs.Bugu da ƙari, 150 mM NaCl ya samar da nau'i-nau'i na micron a cikin dukkanin samfurori guda uku, yana nuna cewa ƙaddamarwar electrolyte na ilimin lissafi yana da lahani ga zaman lafiyar colloidal na citrate-terminated AgNPs.Ta hanyar la'akari da mahimmancin ƙaddamarwa (CCC) na NaCl akan tsarin AgNP irin wannan, waɗannan sakamakon za a iya sanya su cikin basira a cikin wallafe-wallafen da suka dace.Huyn et al.an ƙididdige cewa CCC na NaCl don citrate- ƙare azurfa nanoparticles tare da matsakaicin diamita na 71 nm shine 47.6 mM, yayin da El Badawy et al.lura cewa CCC na 10 nm AgNPs tare da citrate shafi shine 70 mM.10,16 Bugu da ƙari, babban CCC mai girma na kimanin 300 mM an auna ta He et al., wanda ya sa hanyar haɗin su ya bambanta da littafin da aka ambata a baya.48 Ko da yake gudummawar da ake bayarwa na yanzu ba ta nufin cikakken bincike na waɗannan dabi'un ba, saboda yanayin gwajin mu yana karuwa a cikin rikitarwa na dukan binciken, ƙaddamarwar NaCl na ilimin halitta na 50 mM, musamman 150 mM NaCl, yana da alama yana da girma sosai.Ƙunƙarar coagulation, yana bayanin sauye-sauye masu ƙarfi da aka gano.
Mataki na gaba a gwajin polymerization shine a yi amfani da sauƙi amma masu dacewa da kwayoyin halitta don daidaita hulɗar nanoparticle-biomolecule.Dangane da DLS (Hoto na 6 da 7) da sakamakon UV-Vis (Ƙarin Figures S3 da S4), za'a iya tabbatar da wasu ƙarshe na gaba ɗaya.A ƙarƙashin yanayin gwajin mu, ƙwayoyin ƙwayoyin glucose da glutamine da aka yi karatu ba za su haifar da tarawa a cikin kowane tsarin AgNP ba, saboda yanayin Z-ma'ana yana da alaƙa da ma'aunin ma'aunin ma'auni daidai.Ko da yake kasancewar su ba ya shafar tarawa, sakamakon gwaji ya nuna cewa waɗannan kwayoyin halitta an ɗanɗana su a saman AgNPs.Babban sakamakon da ke goyan bayan wannan ra'ayi shine canjin da aka gani a cikin ɗaukar haske.Kodayake AgNP-I baya nuna ma'ana mai ma'ana ko canje-canje masu ƙarfi, ana iya lura da shi sosai ta hanyar auna ɓangarorin da suka fi girma, wanda ya fi dacewa saboda ƙwarewar gani da aka ambata a baya.Ba tare da la'akari da maida hankali ba, glucose na iya haifar da babban canjin ja bayan sa'o'i 1.5 idan aka kwatanta da ma'aunin sarrafawa, wanda shine kusan 40 nm a cikin AgNP-II da kusan 10 nm a cikin AgNP-III, wanda ke tabbatar da abin da ya faru na hulɗar saman.Glutamine ya nuna irin wannan yanayin, amma canjin bai fito fili ba.Bugu da ƙari, yana da mahimmanci a ambaci cewa glutamine na iya rage cikakkiyar damar zeta na matsakaici da manyan barbashi.Duk da haka, tun da waɗannan canje-canjen zeta ba su da alama sun shafi matakin tarawa, ana iya yin la'akari da cewa ko da ƙananan kwayoyin halitta kamar glutamine na iya samar da wani nau'i na ƙyamar sararin samaniya tsakanin barbashi.
Hoto 6 Sakamakon watsawar haske mai ƙarfi na citrate- ƙarewar samfuran nanoparticle na azurfa tare da haɓaka girma (10 nm: AgNP-I, 20 nm: AgNP-II da 50 nm: AgNP-III) an bayyana su azaman matsakaicin diamita na hydrodynamic (Z matsakaicin) (dama) A ƙarƙashin yanayin waje na ƙididdigar glucose daban-daban, yuwuwar zeta (hagu) yana canzawa cikin sa'o'i 24.
Hoto 7 Sakamakon watsawar haske mai ƙarfi na samfurin nanoparticles na azurfa citrate da aka ƙare tare da haɓaka girma (10 nm: AgNP-I, 20 nm: AgNP-II da 50 nm: AgNP-III) an bayyana su azaman matsakaicin diamita na hydrodynamic (matsakaicin Z). ) (dama) A gaban glutamine, yuwuwar zeta (hagu) yana canzawa cikin sa'o'i 24.
A takaice dai, ƙananan ƙwayoyin halitta kamar glucose da glutamine ba su shafar zaman lafiyar colloidal a ma'auni mai mahimmanci: ko da yake suna rinjayar yiwuwar zeta da sakamakon UV-Vis zuwa digiri daban-daban, matsakaicin sakamakon Z ba daidai ba ne.Wannan yana nuna cewa tallan da ke sama na ƙwayoyin cuta yana hana haɓakar electrostatic, amma a lokaci guda yana samar da kwanciyar hankali.
Domin danganta sakamakon da ya gabata tare da sakamakon da ya gabata da kuma kwatanta yanayin ilimin halitta da fasaha, mun zaɓi wasu abubuwan da aka fi amfani da su na al'adun tantanin halitta kuma mun yi amfani da su azaman yanayin gwaji don nazarin kwanciyar hankali na AgNP colloid.A cikin duka gwaje-gwajen in vitro, ɗayan mahimman ayyukan DMEM a matsayin matsakaici shine kafa yanayin osmotic da ake buƙata, amma daga mahangar sinadarai, maganin gishiri ne mai rikitarwa tare da jimlar ionic ƙarfin kama da 150 mM NaCl. .40 Amma ga FBS, shi ne hadadden cakuda biomolecules-yafi sunadaran gina jiki-daga ra'ayi na surface adsorption, yana da wasu kamance da sakamakon gwaji na glucose da glutamine, duk da sinadaran abun da ke ciki da kuma bambancin Jima'i ya fi rikitarwa.19 DLS da UV-Sakamakon bayyane da aka nuna a cikin Hoto na 8 da Ƙarin Hoto S5, bi da bi, ana iya bayyana su ta hanyar nazarin sinadarai na waɗannan kayan da kuma daidaita su da ma'auni a cikin sashin da ya gabata.
Hoto 8 Sakamakon watsawar haske mai ƙarfi na samfurin nanoparticles na azurfa citrate da aka ƙare tare da haɓaka girma (10 nm: AgNP-I, 20 nm: AgNP-II da 50 nm: AgNP-III) an bayyana su azaman matsakaicin diamita na hydrodynamic (matsakaicin Z). ) (dama) A gaban sassan al'adun tantanin halitta DMEM da FBS, yiwuwar zeta (hagu) yana canzawa a cikin sa'o'i 24.
Dilution na AgNPs na daban-daban masu girma dabam a cikin DMEM yana da irin wannan tasiri akan kwanciyar hankali na colloidal zuwa abin da aka gani a gaban babban adadin NaCl.Watsawa na AgNP a cikin 50 v / v% DMEM ya nuna cewa an gano babban taro tare da karuwar yiwuwar zeta da ƙimar Z-matsakaici da raguwa mai tsanani na SPR.Yana da kyau a lura cewa matsakaicin girman jimlar da DMEM ta jawo bayan sa'o'i 24 ya yi daidai da girman nanoparticles na farko.
Ma'amala tsakanin FBS da AgNP yayi kama da wanda aka gani a gaban ƙananan ƙwayoyin cuta kamar glucose da glutamine, amma tasirin ya fi ƙarfi.Matsakaicin Z na barbashi ya kasance ba a shafa ba, yayin da aka gano haɓakar yuwuwar zeta.Ƙimar SPR ta nuna ɗan ƙaramin ja, amma watakila mafi ban sha'awa, ƙarfin SPR bai ragu ba kamar yadda yake a cikin ma'aunin sarrafawa.Ana iya bayyana waɗannan sakamakon ta hanyar haɗaɗɗun macromolecules a saman nanoparticles (jere na ƙasa a cikin hoto na 4), wanda yanzu aka fahimta a matsayin samuwar corona biomolecular a cikin jiki.49