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Obere nanoparticles ọ dị mma mgbe niile?Ghọta nsonaazụ ndu nke nchịkọta nanoparticles ọlaọcha dabere nha n'okpuru ọnọdụ dabara adaba.
Ndị edemede: Bélteky P, Rónavári A, Zakupszky D, Boka E, Igaz N, Szerencsés B, Pfeiffer I, Vágvölgyi C, Kiricsi M, Kónya Z
Péter Bélteky,* Andrea Rónavári,* Dalma Zakupszky,1 Eszter Boka,1 Nóra Igaz,2 Bettina Szerencsés,3 Ilona Pfeiffer,3 Csaba Vágvölgyi,3 Mónika Kiricsi nke Chemistry Environmental, Hungary, Sayensị na Hungary , Mahadum Szeged;2 Ngalaba Biochemistry na Molecular Biology, Ngalaba Sayensị na Ozi, Mahadum Szeged, Hungary;3 Ngalaba Microbiology, Ngalaba Sayensị na Ozi, Mahadum Szeged, Hungary;4MTA-SZTE Reaction Kinetics na Surface Chemistry Research Group, Szeged, Hungary* Ndị ode akwụkwọ a nyere aka n'otu aka ahụ n'ọrụ a.Nkwukọrịta: 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] Nzube: Silver nanoparticles (AgNPs) bụ otu n'ime nanomaterials a na-amụkarị, ọkachasị n'ihi ngwa biomedical ha.Otú ọ dị, n'ihi nchịkọta nke nanoparticles, cytotoxicity ha magburu onwe ya na ọrụ antibacterial na-emebikarị na mgbasa ozi ndu.N'ime ọrụ a, a mụrụ omume nchịkọta na ihe omume ndị metụtara ndụ nke ihe nlele nanoparticle ọlaọcha atọ dị iche iche nke citrate kwụsịrị na nkezi dayameta nke 10, 20, na 50 nm.Usoro: Jiri microscope eletrọn nnyefe iji mepụta ma mara nanoparticles, nyochaa omume nchịkọta ha na ụkpụrụ pH dị iche iche, NaCl, glucose na glutamine n'ịgbasa ọkụ dị ike na ultraviolet-visible spectroscopy.Na mgbakwunye, n'ime ihe ndị na-ahụ maka omenaala cell dị ka Dulbecco na-eme ka omume nchịkọta dị na Eagle Medium na Serum nwa ebu n'afọ.Nsonaazụ: Nsonaazụ na-egosi na pH acidic na ọdịnaya electrolyte physiological na-ebute mkpokọta micron-scale, nke enwere ike ịgbazite ya site na nguzobe nke corona biomolecular.Ọ bụ ihe kwesịrị ịrịba ama na nnukwu irighiri ihe na-egosipụta nguzogide dị elu na mmetụta mpụga karịa obere ogbo ha.A na-eme cytotoxicity in vitro na nyocha nke nje bacteria site n'ịgwọ mkpụrụ ndụ nwere nchịkọta nanoparticle na ọkwa nchịkọta dị iche iche.Mkpebi: Nsonaazụ anyị na-ekpughe njikọ dị omimi n'etiti nkwụsi ike nke colloidal na nsi nke AgNP, ebe oke nchikota na-eduga na mfu zuru oke nke ọrụ ndu.Ọkwa dị elu nke mgbochi nchịkọta nke a na-ahụ maka ihe ndị buru ibu na-enwe mmetụta dị ukwuu na toxicity in vitro, n'ihi na ihe atụ ndị dị otú ahụ na-ejigide ọrụ antimicrobial na mammalian cell.Nchọpụta ndị a na-eduga ná nkwubi okwu na, n'agbanyeghị echiche n'ozuzu na akwụkwọ ndị dị mkpa, ịchọta nanoparticles kacha nta nwere ike ọ gaghị abụ ụzọ kachasị mma.Keywords: uto na-agbasa mkpụrụ, nkwụsi ike nke colloidal, omume nchịkọta na-adabere na nha, nchịkọta mmebi nsí
Ka ọchịchọ na mmepụta nke nanomaterials na-aga n'ihu na-abawanye, a na-akwụkwu nlebara anya na biosafety ma ọ bụ ọrụ ndu ha.Silver nanoparticles (AgNPs) bụ otu n'ime ndị nnọchi anya klaasị nke ihe a na-ejikarị ahazi, nyocha na nke ejiri n'ọrụ n'ihi ọmarịcha catalytic, ngwa anya na ihe ndu.1 A na-ekwenyekarị na njirimara pụrụ iche nke nanomaterials (gụnyere AgNPs) bụ nke a na-ekwukarị na nnukwu elu elu ha.Ya mere, nsogbu a na-apụghị izere ezere bụ usoro ọ bụla nke na-emetụta njirimara isi a, dị ka nha nha, mkpuchi elu ma ọ bụ nchịkọta, ma ọ ga-emebi ihe onwunwe nke nanoparticles nke dị oké mkpa maka ngwa ngwa.
Mmetụta nke urughuru nha na stabilizers bụ isiokwu ndị e dịtụ mma akwụkwọ na akwụkwọ.Dịka ọmụmaatụ, echiche a na-anabatakarị bụ na obere nanoparticles na-egbu egbu karịa nanoparticles buru ibu.2 N'ikwekọ na akwụkwọ n'ozuzu, ọmụmụ ihe anyị gara aga egosipụtawo ọrụ nanosilver na-adabere na nha na mkpụrụ ndụ mammalian na microorganisms.3– 5 Mkpuchi elu bụ ihe ọzọ nwere mmetụta sara mbara na njirimara nke nanomaterials.Naanị site n'ịgbakwụnye ma ọ bụ gbanwee ihe nkwụsi ike n'elu ya, otu ihe nanomaterial ahụ nwere ike inwe ụdị anụ ahụ, kemịkalụ na nke ndụ dị iche iche.A na-emekarị ngwa nke ihe mkpuchi mkpuchi dị ka akụkụ nke njikọ nanoparticle.Dị ka ihe atụ, citrate-akwụsị silver nanoparticles bụ otu n'ime ndị kasị mkpa AgNPs na nnyocha, nke na-synthesized site mbenata ọlaọcha salts na a họrọ stabilizer ngwọta dị ka mmeghachi omume usoro.6 Citrate nwere ike iji ohere dị ọnụ ala ya dị ọnụ ala, nnweta, biocompatibility, na mmekọrịta siri ike maka ọlaọcha, nke nwere ike igosipụta na mmekọrịta dị iche iche a na-atụ aro, site na mgbasa ozi elu na-agbanwe agbanwe na mmekọrịta ionic.A na-ejikwa obere ụmụ irighiri ihe na ion polyatomic dị nso 7,8, dị ka citrates, polymers, polyelectrolytes, na ndị na-ahụ maka ihe ndị dị ndụ na-ejikarị eme ka nano-silver kwụsie ike ma rụọ ọrụ pụrụ iche na ya.9-12
Ọ bụ ezie na ohere nke ịgbanwe ọrụ nke nanoparticles site na ịkpachara anya n'elu capping bụ ebe na-adọrọ mmasị nke ukwuu, isi ọrụ nke mkpuchi elu a bụ ihe na-adịghị mma, na-enye nkwụsi ike colloidal maka usoro nanoparticle.Nnukwu ebe dị elu nke nanomaterials ga-emepụta ike dị elu, nke na-egbochi ike thermodynamic nke usoro ahụ iji ruo ike kacha nta.13 Enweghị nkwụsi ike nke ọma, nke a nwere ike iduga n'ịgbasa ihe nanomaterials.Mkpokọta bụ nguzobe nke aggregates nke irighiri nke dị iche iche shapes na nha na-eme mgbe achụsasịrị irighiri izute na ugbu a thermodynamic mmekọrịta ekwe ka ahụ na-rapara n'ahụ ibe ha.Ya mere, a na-eji stabilizers iji gbochie nchịkọta site na iwebata ike na-asọ oyi zuru oke n'etiti ihe ndị ahụ iji gbochie mmachi thermodynamic ha.14
Ọ bụ ezie na isiokwu nke urughuru nha na elu mkpuchi e nke ọma enyocha ya na onodu nke ya ụkpụrụ nke ndu omume triggered site nanoparticles, urughuru mkpokọta bụ a n'ụzọ na-eleghara anya na mpaghara.Ọ fọrọ nke nta ka ọ bụrụ ihe ọmụmụ nke ọma iji dozie nkwụsi ike colloidal nke nanoparticles n'okpuru ọnọdụ ndị metụtara ndụ.10,15-17 Tụkwasị na nke a, onyinye a na-adịkarị ụkọ, ebe a na-amụkwa ihe ndị na-egbu egbu na-ejikọta ya na nchịkọta, ọ bụrụgodị na ọ nwere ike ịkpata mmeghachi omume ọjọọ, dị ka thrombosis vaskụla, ma ọ bụ nkwụsị nke àgwà ndị a chọrọ, dị ka nsị ya, dị ka toxicity. egosiri na eserese 1.18, 19.N'ezie, otu n'ime usoro ole na ole a maara nke nguzogide nanoparticle ọlaọcha metụtara nchịkọta, n'ihi na a na-akọ na ụfọdụ E. coli na Pseudomonas aeruginosa strains na-ebelata mmetụta nano-ọlaọcha ha site n'igosipụta protein flagellin, flagellin.Ọ nwere mmekọrịta dị elu maka ọlaọcha, si otú a na-ebute mkpokọta.20
Enwere usoro dị iche iche dị iche iche metụtara nsị nke nanoparticles ọlaọcha, na nchịkọta na-emetụta usoro ndị a niile.Usoro AgNP kachasị atụle nke ọrụ ndu, mgbe ụfọdụ a na-akpọ usoro "Trojan Horse", na-ele AgNP dị ka ndị na-ebu Ag+.1,21 Usoro ịnyịnya nke Trojan nwere ike hụ na mmụba dị ukwuu na mpaghara Ag + mpaghara, nke na-eduga n'ọgbọ ROS na akpụkpọ anụ depolarization.22-24 Mkpokọta nwere ike imetụta ntọhapụ nke Ag +, si otú ahụ na-emetụta toxicity, n'ihi na ọ na-ebelata irè arụ ọrụ elu ebe ọlaọcha ion nwere ike oxidized na igbari.Agbanyeghị, AgNP agaghị egosipụta naanị nsi site na ntọhapụ ion.A ghaghị ịtụle ọtụtụ nha na mmekọrịta metụtara morphology.N'ime ha, nha na ọdịdị nke nanoparticle elu bụ njirimara njirimara.4,25 Enwere ike ịha mkpokọta usoro ndị a dị ka "usoro nsi na-akpata."Enwere ike inwe ọtụtụ mmeghachi omume mitochondrial na n'elu akpụkpọ ahụ nke nwere ike imebi akụkụ ahụ ma bute ọnwụ cell.25-27 Ebe ọ bụ na nguzobe nke mkpokọta na-emetụta nha na ọdịdị nke ihe ndị nwere ọlaọcha nke usoro ndụ ghọtara, mmekọrịta ndị a nwekwara ike imetụta.
N'akwụkwọ anyị gara aga na nchịkọta nke nanoparticles ọlaọcha, anyị gosipụtara usoro nyocha dị irè nke nwere kemịkal na in vitro biological tests iji mụọ nsogbu a.19 Dynamic Light Scattering (DLS) bụ usoro kachasị amasị maka ụdị nyocha ndị a n'ihi na ihe ahụ nwere ike ịgbasa foton n'ogo ogologo nke nha nha nha.Ebe ọ bụ na ngwa ngwa Brownian na-agba ọsọ nke ihe dị na mmiri mmiri na-ejikọta ya na nha, enwere ike iji mgbanwe na ike nke ọkụ gbasasịa mee ihe iji chọpụta nkezi hydrodynamic dayameta (Z- mean) nke ihe nlele mmiri.28 Tụkwasị na nke ahụ, site n'itinye voltaji na nlele ahụ, ike zeta (ζ nwere ike) nke nanoparticle nwere ike tụọ ya n'otu aka ahụ na uru Z.13,28 Ọ bụrụ na uru zuru oke nke ikike zeta dị elu (dị ka ntụziaka n'ozuzu> ± 30 mV), ọ ga-emepụta ihe mgbochi electrostatic siri ike n'etiti ihe ndị ahụ iji mebie nchịkọta ahụ.Njiri mara elu plasmon resonance (SPR) bụ ihe pụrụ iche ngwa anya, nke a na-ekwu na ọ bụ nanoparticles metal dị oké ọnụ ahịa (karịsịa Au na Ag).29 Dabere na oscillations eletrọnịkị (plasmons n'elu) nke ihe ndị a na nanoscale, a maara na AgNP dị okirikiri nwere njirimara UV-Vis absorption kacha nso 400 nm.30 A na-eji mgbanwe ike na ogologo ogologo nke ụmụ irighiri ihe iji gbakwunye nsonaazụ DLS, n'ihi na enwere ike iji usoro a chọpụta nchịkọta nanoparticle na mgbasa ozi elu nke biomolecules.
Dabere na ozi enwetara, a na-eme viability cell (MTT) na nyocha nje na-eme ihe n'ụzọ a na-akọwa toxicity AgNP dị ka ọrụ nke mkpokọta mkpokọta, karịa (ihe a na-ejikarị eme ihe) ntinye uche nanoparticle.Usoro a pụrụ iche na-enye anyị ohere igosi mkpa dị ukwuu nke nchịkọta nchịkọta na ọrụ ndu, n'ihi na, dịka ọmụmaatụ, citrate-treminated AgNPs kpamkpam na-efunahụ ọrụ ndụ ha n'ime awa ole na ole n'ihi nchịkọta.19
N'ime ọrụ ugbu a, anyị chọrọ iji gbasaa onyinye anyị gara aga na nkwụsi ike nke colloid metụtara bio na mmetụta ha na ọrụ ndu site n'ịmụ mmetụta nke nanoparticle nha na nchịkọta nanoparticle.Ihe ịrụ ụka adịghị ya na nke a bụ otu n'ime ọmụmụ nke nanoparticles.Nleba anya profaịlụ dị elu na 31 Iji nyochaa okwu a, a na-eji usoro uto na-agbasa mkpụrụ iji mepụta AgNP nke citrate kwụsịrị na nha atọ dị iche iche (10, 20, na 50 nm).6,32 dị ka otu n'ime ụzọ ndị a na-ahụkarị.Maka nanomaterials nke a na-ejikarị eme ihe na ngwa ahụike, a na-ahọrọ citrate-treminated AgNPs nke nha dị iche iche iji mụọ nkwekọ nha nke nanosilver metụtara nchịkọta.Mgbe emechara AgNP nke nha dị iche iche, anyị ji usoro nyocha eletrọnịkị eletrọnịkị (TEM) gosipụtara ihe ndị emepụtara, wee nyochaa ihe ndị ahụ site na iji usoro nyocha ahụ e kwuru n'elu.Na mgbakwunye, na ọnụnọ nke in vitro cell Culture Dulbecco's Modified Eagle's Medium (DMEM) na Fetal Bovine Serum (FBS), a na-enyocha omume nchịkọta na-adabere na nha na omume ya na ụkpụrụ pH dị iche iche, NaCl, glucose, na glutamine.A na-ekpebi njirimara nke cytotoxicity n'okpuru ọnọdụ zuru oke.Nkwekọrịta sayensị na-egosi na n'ozuzu, ihe ndị dị nta ka mma;nyocha anyị na-enye usoro kemịkalụ na usoro ndu iji chọpụta ma nke a ọ dị.
A kwadebere nanoparticles ọlaọcha atọ nwere oke nha dị iche iche site na usoro uto na-agbasa mkpụrụ nke Wan et al., na-enwe ntakịrị mgbanwe.6 Usoro a dabere na mbelata kemịkalụ, na-eji nitrate ọlaọcha (AgNO3) dị ka isi iyi ọla ọcha, sodium borohydride (NaBH4) dị ka onye na-ebelata mbelata, yana sodium citrate dị ka ihe nkwụsi ike.Nke mbụ, kwadebe 75 ml nke 9 mM citrate mmiri mmiri si na sodium citrate dihydrate (Na3C6H5O7 x 2H2O) na okpomọkụ ruo 70 ° C.Mgbe ahụ, 2 mL nke 1% w / v AgNO3 ngwọta na-agbakwunyere na mmeghachi omume mmeghachi omume, wee wụsa ngwọta sodium borohydride nke a kwadebere ọhụrụ (2 mL 0.1% w / v) n'ime ngwakọta dropwise.A na-edobe nkwusioru nke odo-aja aja na 70 Celsius C na-akpali akpali maka awa 1, wee mee ka ọ dị jụụ na ụlọ okpomọkụ.A na-eji ihe nlele (nke a na-akpọ AgNP-I site ugbu a gaa n'ihu) dị ka ihe ndabere maka uto na-agbasa mkpụrụ na usoro nhazi ọzọ.
Iji mepụta nkwusioru nke ọkara (nke a na-akpọ AgNP-II), kpoo 90 mL 7.6 mM citrate ngwọta na 80 ° C, jikọta ya na 10 ml AgNP-I, wee gwakọta 2 mL 1% w / v The AgNO3 ngwọta A na-edobe ya n'okpuru igwe siri ike na-akpali akpali maka awa 1, wee mee ka ihe nlele ahụ dị jụụ na ụlọ okpomọkụ.
Maka ihe kachasị ukwuu (AgNP-III), megharịa otu usoro uto ahụ, ma na nke a, jiri 10 ml nke AgNP-II dị ka nkwụsị nke mkpụrụ.Mgbe ihe nlele ahụ rutere n'ime ụlọ, ha na-edobe ntinye aha AgNO3 ha dabere na mkpokọta AgNO3 na 150 ppm site na ịgbakwunye ma ọ bụ na-ekpochapụ ihe mgbaze ọzọ na 40 Celsius, ma mesịa chekwaa ha na 4 Celsius C ruo mgbe ejiri ya ọzọ.
Jiri FEI Tecnai G2 20 X-Twin Transmission Electron Microscope (TEM) (FEI Corporate Headquarters, Hillsboro, Oregon, USA) na 200 kV acceleration voltaji iji nyochaa njirimara morphological nke nanoparticles wee weghara ụkpụrụ electron diffraction (ED) ha.Opekempe, a na-enyocha ihe oyiyi 15 nnọchiteanya (~ 750 particles) site na iji ngwugwu ngwanrọ ImageJ, na ihe ndekọ akụkọ ihe mere eme (na eserese niile dị na ọmụmụ ihe dum) ka emepụtara na OriginPro 2018 (OriginLab, Northampton, MA, USA) 33, 34.
Nkezi dayameta hydrodynamic (Z-nkezi), ikike zeta (ζ-potential) na njirimara elu plasmon resonance (SPR) nke ihe nlele ka a tụrụ iji gosi njirimara colloidal mbụ ha.A tụrụ nkezi dayameta hydrodynamic na zeta ikike nke ihe nlele ahụ site na ngwa Malvern Zetasizer Nano ZS (Malvern Instruments, Malvern, UK) na-eji mkpụrụ ndụ capillary apịaji apịaji na 37± 0.1°C.A na-eji Ocean Optics 355 DH-2000-BAL UV-Vis spectrophotometer (Halma PLC, Largo, FL, USA) nweta njirimara SPR dị iche iche site na UV-Vis absorption spectra nke samples na nso nke 250-800 nm.
N'oge nnwale ahụ dum, ụdị nha atọ dị iche iche metụtara nkwụsi ike colloidal ka emere n'otu oge.Jiri DLS tụọ nkezi hydrodynamic dayameta (Z nkezi) na ikike zeta (ζ nwere ike) nke ụmụ irighiri ihe, n'ihi na nkezi Z na-emetụta nha nha nke mkpokọta nanoparticle, ikike zeta na-egosi ma ọgbaghara electrostatic na sistemụ. siri ike nke ukwuu iji mebie mmasị Van der Waals n'etiti nanoparticles.A na-eme nha n'ime ugboro atọ, yana ngbanwe ọkọlọtọ nke Z pụtara na ikike zeta bụ nke Zetasizer na-agbakọ.A na-enyocha ihe ngosi SPR dị iche iche nke ihe ndị ahụ site na UV-Vis spectroscopy, n'ihi na mgbanwe nke oke ike na ogologo ogologo nwere ike igosi nchịkọta na mmekọrịta elu.29,35 N'ezie, n'elu plasmon resonance na oké ọnụ ahịa ọla na-enwe mmetụta nke na o butere ụzọ ọhụrụ nke nyocha nke biomolecules.29,36,37 Ntinye uche nke AgNP na ngwakọta nnwale bụ ihe dị ka 10 ppm, na ebumnuche bụ ịtọ ike nke mbido SPR mbụ kachasị na 1. A na-eme nnwale ahụ na oge dabere na 0;1.5;3;6;12 na 24 awa n'okpuru ọnọdụ dị iche iche metụtara ndu.Enwere ike ịhụ nkọwa ndị ọzọ na-akọwa nnwale ahụ na ọrụ anyị gara aga.19 Na nkenke, dị iche iche pH ụkpụrụ (3; 5; 7.2 na 9), dị iche iche sodium chloride (10 mM; 50 mM; 150 mM), glucose (3.9 mm; 6.7 mm) na glutamine (4 mM) ịta, na kwadokwara Dulbecco's Modified Eagle Medium (DMEM) na Fetal Bovine Serum (FBS) (n'ime mmiri na DMEM) dị ka usoro ihe atụ, wee mụọ mmetụta ha na omume nchịkọta nke nanoparticles silver synthesized.pH A na-enyocha ụkpụrụ nke, NaCl, glucose, na glutamine dabere na mkpokọta physiological, ebe ọnụọgụ DMEM na FBS bụ otu ọkwa ejiri na nnwale in vitro dum.38-42 A na-eme nha niile na pH 7.2 na 37 ° C na ntinye nnu nnu mgbe nile nke 10 mM NaCl iji kpochapụ mkparịta ụka ọ bụla dị anya (ma e wezụga ụfọdụ pH na NaCl, ebe àgwà ndị a bụ mgbanwe n'okpuru). ọmụmụ).28 A na-achikota ndepụta nke ọnọdụ dị iche iche na Tebụl 1. A na-eji nnwale ahụ akara na † mee ihe dị ka ihe atụ ma kwekọọ na ihe nlele nwere 10 mM NaCl na pH 7.2.
E nwetara ahịrị mkpụrụ ndụ cancer prostate mmadụ (DU145) na keratinocytes mmadụ anwụghị anwụ (HaCaT) sitere na ATCC (Manassas, VA, USA).A na-akụ mkpụrụ ndụ mgbe niile na Ugo kacha nta dị mkpa nke Dulbecco (DMEM) nwere 4.5 g/L glucose (Sigma-Aldrich, Saint Louis, MO, USA), gbakwunyere na 10% FBS, 2 mM L-glutamine, 0.01 % Streptomycin na 0.005% Penicillin (Sigma-Aldrich, St. Louis, Missouri, USA).A na-akụ mkpụrụ ndụ ahụ na incubator 37 Celsius C n'okpuru 5% CO2 na iru mmiri 95%.
Iji chọpụta mgbanwe ndị dị na cytotoxicity AgNP kpatara site na nchịkọta irighiri ihe na-adabere na oge, a na-eme nyocha MTT nzọụkwụ abụọ.Nke mbụ, a tụrụ ike ịdị ndụ nke ụdị cell abụọ ahụ mgbe a gwọchara ya na AgNP-I, AgNP-II na AgNP-III.Iji mezuo nke a, a na-akụ ụdị mkpụrụ ndụ abụọ ahụ n'ime efere 96-well na njupụta nke sel 10,000 / nke ọma ma mesoo ya na nha atọ dị iche iche nke nanoparticles ọlaọcha na-abawanye ụba n'ụbọchị nke abụọ.Mgbe awa 24 gasịrị, a na-asacha mkpụrụ ndụ ahụ na PBS ma tinye ya na 0.5 mg / mL MTT reagent (SERVA, Heidelberg, Germany) na-agbanye na omenala omenala maka 1 hour na 37 ° C.A gbazere kristal Formazan na DMSO (Sigma-Aldrich, Saint Louis, MO, USA), wee tụọ nnabata ahụ na 570 nm site na iji Synergy HTX plate reader (BioTek-Hungary, Budapest, Hungary).A na-ewere uru nnabata nke ihe nlele njikwa na-adịghị edozi ya dị ka ọnụọgụ nlanarị 100%.Mee ọ dịkarịa ala nnwale 3 site na iji ụdị mkpụrụ ndụ anọ nwere onwe ha.A na-agbakọ IC50 site na usoro nzaghachi dose dabere na nsonaazụ ike.
Mgbe nke ahụ gasịrị, na nzọụkwụ nke abụọ, site n'itinye ihe ndị ahụ na 150 mM NaCl maka oge dị iche iche (0, 1.5, 3, 6, 12, na 24 hours) tupu ọgwụgwọ cell, a na-emepụta ụdị nchịkọta dị iche iche nke ọlaọcha nanoparticles.Mgbe nke ahụ gasịrị, a na-eme otu nyocha MTT ahụ dị ka akọwara na mbụ iji nyochaa mgbanwe n'ime ndụ cell na-emetụta mkpokọta urughuru.Jiri GraphPad Prism 7 iji nyochaa nsonaazụ ikpeazụ, gbakọọ mkpa nyocha nke nnwale ahụ site na t-ule na-enweghị njikọ, ma kaa ọkwa ya dị ka * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001). ) Na **** (p ≤ 0.0001).
A na-eji nha atọ dị iche iche nke nanoparticles ọlaọcha (AgNP-I, AgNP-II na AgNP-III) maka ihe mgbochi nje na Cryptococcus neoformans IFM 5844 (IFM; Research Center for Pathogenic Fungi and Microbial Toxicology, Chiba University) na Bacillus Test megaterium SZMC 603 (SZMC: Szeged Microbiology Collection) na E. coli SZMC 0582 na RPMI 1640 ọkara (Sigma-Aldrich Co.).Iji nyochaa mgbanwe nke ọrụ nje bacteria kpatara site na nchịkọta nke ụmụ irighiri ihe, nke mbụ, a na-ekpebi itinye uche kacha nta ha (MIC) site na microdilution na efere 96-well microtiter.Na 50 μL nke nkwusioru cell ọkọlọtọ (5 × 104 sel / mL na RPMI 1640 ọkara), gbakwunye 50 μL nke nkwusioru nanoparticle ọlaọcha ma na-atụgharị ugboro abụọ na ntinye uche (na ọkara ahụ e kwuru n'elu, oke bụ 0 na 75 ppm, Ya bụ, ihe nlele njikwa nwere 50 μL nke nkwusioru cell na 50 μL nke ọkara na-enweghị nanoparticles).Mgbe nke ahụ gasịrị, a na-etinye efere ahụ na 30 ° C maka awa 48, a na-atụlekwa njupụta anya nke omenala na 620 nm site na iji SPECTROstar Nano plate reader (BMG LabTech, Offenburg, Germany).Emere nnwale ahụ ugboro atọ ugboro atọ.
Ewezuga na a na-eji 50 μL nke otu nchịkọta nanoparticle samples mee ihe n'oge a, otu usoro ahụ dịka akọwara na mbụ ka a na-eji nyochaa mmetụta nchịkọta na ọrụ antibacterial na ụdị ndị ahụ e kwuru n'elu.A na-emepụta steeti nchịkọta dị iche iche nke nanoparticles ọlaọcha site n'itinye ihe ndị ahụ na 150 mM NaCl maka oge dị iche iche (0, 1.5, 3, 6, 12, na 24) tupu nhazi cell.A na-eji nkwusioru agbakwunyere na 50 μL nke RPMI 1640 dị ka njikwa uto, ebe iji chịkwaa nsi, a na-eji nkwusioru na nanoparticles na-enweghị nchịkọta.Emere nnwale ahụ ugboro atọ ugboro atọ.Jiri GraphPad Prism 7 tulee nsonaazụ ikpeazụ ọzọ, na-eji otu nyocha ndekọ ọnụ ọgụgụ dị ka nyocha MTT.
A kọwapụtala ọkwa nchịkọta nke obere ntakịrị (AgNP-I), na ebipụtala nsonaazụ ya na ọrụ anyị gara aga, mana maka ntụnyere ka mma, a na-enyocha ihe niile.A na-anakọta ma tụlee data nnwale ahụ na ngalaba ndị a.Nha atọ nke AgNP.19
Ntụle ndị TEM, UV-Vis na DLS mere gosipụtara na njikọ na-aga nke ọma nke ihe atụ AgNP niile (Ọnụ ọgụgụ 2A-D).Dị ka ahịrị mbụ nke eserese 2 si dị, ihe kacha nta (AgNP-I) na-egosi ụdị ọdịdị dị iche iche nwere otu dayameta nke ihe dịka 10 nm.Usoro uto na-agbasa mkpụrụ na-enyekwa AgNP-II na AgNP-III nwere oke nha dị iche iche yana dayameta irighiri ihe ruru 20 nm na 50 nm, n'otu n'otu.Dị ka ọkọlọtọ ọkọlọtọ nke nkesa urughuru, nha nke atọ sample adịghị overlack, nke dị mkpa maka ha comparative analysis.Site n'ịtụle nkezi akụkụ na nha ịdị gịrịgịrị nke ntule 2D nke dabere na TEM, a na-eche na a na-enyocha nha nke akụkụ ahụ site na nkwụnye ihe nzacha nke ImageJ (Figure 2E).43 Dị ka nyocha nke udi nke irighiri ihe, akụkụ ha (nnukwu akụkụ / mkpirisi akụkụ nke kacha nta bounding rektangulu) adịghị emetụta urughuru ibu, na ha thinness ruru (tụtara ebe kwekọrọ zuru okè okirikiri / theoretical mpaghara. ) ji nwayọọ nwayọọ na-ebelata.Nke a na-ebute ọtụtụ irighiri ihe polyhedral, nke zuru oke na tiori, kwekọrọ na nha dị gịrịgịrị nke 1.
Onyonyo 2 Mfefe elektrọn microscope (TEM) oyiyi (A), eletrọn diffraction (ED) ụkpụrụ (B), size nkesa histogram (C), àgwà ultraviolet-visible (UV-Vis) ìhè absorption ụdịdị dị iche iche (D), na nkezi mmiri Citrate. -kwụsị ọlaọcha nanoparticles na n'ibu dayameta (Z-nkezi), zeta ikike, akụkụ ruru na ọkpụrụkpụ ruru (E) nwere atọ dị iche iche size range: AgNP-I bụ 10 nm (n'elu ahịrị), AgNP -II bụ 20 nm (n'etiti ahịrị). ), AgNP-III (ahịrị ala) bụ 50 nm.
Ọ bụ ezie na ọdịdị cyclic nke usoro uto na-emetụta ọdịdị urughuru ahụ ruo n'ókè ụfọdụ, na-ebute ntanarị nke nnukwu AgNPs, ihe nlele atọ ahụ nọgidere na-adị okirikiri.Na mgbakwunye, dị ka e gosiri na eletrọn diffraction ụkpụrụ na Figure 2B, nano The crystallinity nke ahụ adịghị emetụta.Mgbanaka mgbanaka ahụ ama ama-nke enwere ike ijikọ ya na (111), (220), (200), na (311) Miller indices of silver- dabara na akwụkwọ sayensị yana onyinye anyị gara aga.9, 19,44 The fragmentation nke Debye-Scherrer mgbanaka nke AgNP-II na AgNP-III bụ n'ihi na eziokwu na ED image weghaara na otu ihe na-ebuwanye ibu, otú ahụ urughuru size na-abawanye, ọnụ ọgụgụ nke diffracted irighiri kwa. mpaghara otu na-abawanye ma na-ebelata .
A maara nha na ọdịdị nke nanoparticles na-emetụta ọrụ ndu.Enwere ike ịkọwa 3,45 catalytic na usoro ndu nke na-adabere na ọdịdị ya site n'eziokwu ahụ bụ na ụdị dị iche iche na-agbasawanye ụfọdụ ihu kristal (nwere ụdị Miller dị iche iche), na ihu kristal ndị a nwere ọrụ dị iche iche.45,46 Ebe ọ bụ na ihe ndị a kwadebere na-enye nsonaazụ ED yiri nke dabara na njirimara kristal yiri nke ahụ, enwere ike iche na na nkwụsi ike colloidal anyị na-esote na nyocha ọrụ ndu, a ga-ekwupụta ọdịiche ọ bụla hụrụ na nha Nanoparticle, ọ bụghị ihe metụtara ọdịdị.
Nsonaazụ UV-Vis a chịkọtara na eserese 2D na-emesi ike na ọdịdị okirikiri dị egwu nke AgNP agbakwunyere, n'ihi na ọnụ ọgụgụ SPR nke ihe nlele atọ niile dị gburugburu 400 nm, nke bụ njirimara njirimara nke nanoparticles ọlaọcha okirikiri.29,30 Ihe onyonyo a weghaara gosikwara na uto nke nanosilver sitere na mkpụrụ na-aga nke ọma.Dị ka urughuru nha na-abawanye, ogologo wavelength kwekọrọ na kacha ìhè absorption nke AgNP-II-karịa pụtara ìhè-Dịka akwụkwọ, AgNP-III nwetara a redshift.6,29
Banyere nkwụsi ike colloidal mbụ nke sistemụ AgNP, a na-eji DLS tụọ nkezi hydrodynamic dayameta na ikike zeta nke ụmụ irighiri ihe na pH 7.2.Nsonaazụ e gosipụtara na eserese 2E na-egosi na AgNP-III nwere nkwụsi ike colloidal dị elu karịa AgNP-I ma ọ bụ AgNP-II, n'ihi na ntuziaka ndị a na-ahụkarị na-egosi na ikike zeta nke 30 mV zuru oke dị mkpa maka nkwụsi ike colloidal ogologo oge a na-akwado nchoputa a mgbe. A na-atụnyere uru nkezi Z (enwetara dị ka nkezi hydrodynamic dayameta nke free na aggregated particles) tụnyere nke mbụ urughuru size nwetara site TEM, n'ihi na nso nso abụọ ụkpụrụ bụ, milder ogo Kpọkọtanụ na sample.N'ezie, nkezi Z nke AgNP-I na AgNP-II nwere ezi uche dị elu karịa isi ihe ha na-enyocha TEM, ya mere e jiri ya tụnyere AgNP-III, a na-ebu amụma na ihe atụ ndị a ga-enwe ike ikpokọta, ebe ikike zeta na-adịghị mma. na-esonyere nha nso Nkezi uru Z.
Nkọwa maka ihe omume a nwere ike ịbụ okpukpu abụọ.N'otu aka ahụ, a na-edobe ịta citrate n'otu ọkwa ahụ na usoro nhazi niile, na-enye ọnụ ọgụgụ dị elu nke ebubo elu dị iche iche iji gbochie ebe dị elu nke ihe ndị na-eto eto ka ọ ghara ibelata.Otú ọ dị, dị ka Levak et al., obere ụmụ irighiri dị ka citrate nwere ike ịgbanwe ngwa ngwa site biomolecules n'elu nke nanoparticles.N'okwu a, a ga-ekpebi nkwụsi ike nke colloidal site na corona nke biomolecules emepụtara.31 N'ihi na a hụkwara omume nke a na nchikota anyị (a tụlere ya n'ụzọ zuru ezu ma emechaa), citrate capping naanị enweghị ike ịkọwa ihe a.
N'aka nke ọzọ, urughuru nha bụ inversely hà nhata na mkpokọta ọchịchọ na nanometer larịị.Nke a na-akwado nke ọma site na usoro omenala Derjaguin-Landau-Verwey-Overbeek (DLVO), ebe a na-akọwa mmachi ihe dị ka nchikota nke ihe na-adọrọ adọrọ na nke na-asọ oyi n'etiti ụmụ irighiri ihe.Dị ka He et al., Uru kachasị elu nke DLVO ume ike na-ebelata na nha nke nanoparticles na hematite nanoparticles, na-eme ka ọ dịkwuo mfe iru ike kachasị nta, si otú ahụ na-akwalite nchịkọta na-enweghị ike ịgbanwe (condensation).47 Otú ọ dị, a na-eche na e nwere akụkụ ndị ọzọ gafere oke nke ozizi DLVO.Ọ bụ ezie na van der Waals ike ndọda na electrostatic okpukpu abụọ-oyi repulsion yiri na-amụba urughuru size, a nyochaa site Hotze et al.na-atụ aro na ọ nwere mmetụta siri ike na nchịkọta karịa DLVO na-enye ohere.14 Ha kwenyere na a pụghị ịtụle ngọgọ elu nke nanoparticles dị ka ala dị larịị, na-eme ka atụmatụ mgbakọ na mwepụ ghara ịdị irè.Na mgbakwunye, dị ka urughuru size ebelata, pasent nke atọm dị n'elu na-aghọ elu, na-eduga eletrọnịkị Ọdịdị na elu ụgwọ omume.Na mgbanwe mmeghachi omume elu, nke nwere ike iduga mbelata ụgwọ na oyi akwa eletrik abụọ ma kwalite mkpokọta.
Mgbe anyị tụlere nsonaazụ DLS nke AgNP-I, AgNP-II, na AgNP-III na eserese 3, anyị hụrụ na nlele atọ ahụ gosipụtara nchịkọta pH yiri nke ahụ.Mpaghara acidic siri ike (pH 3) na-agbanwe ikike zeta nke ihe nlele ahụ na 0 mV, na-eme ka ụmụ irighiri ihe na-emepụta mkpokọta micron-sized, ebe pH alkaline na-agbanwe ikike zeta ya na uru na-adịghị mma, ebe ihe ndị ahụ na-etolite obere mkpokọta (pH 5). ).Na 7.2)), ma ọ bụ nọgide na-enweghị mkpokọta (pH 9).A hụkwara ụfọdụ ọdịiche dị mkpa n'etiti ụdị dị iche iche.N'ime nnwale ahụ dum, AgNP-I gosipụtara na ọ bụ onye kachasị emetụ n'ahụ na pH na-eme ka zeta nwere ike gbanwee, n'ihi na ike zeta nke ihe ndị a ebelatala na pH 7.2 ma e jiri ya tụnyere pH 9, ebe AgNP-II na AgNP-III gosipụtara naanị A. mgbanwe dị ukwuu na ζ dị gburugburu pH 3. Tụkwasị na nke ahụ, AgNP-II gosipụtara mgbanwe nwayọọ nwayọọ na ikike zeta na-agafeghị oke, ebe AgNP-III gosipụtara omume dị nro nke atọ ahụ, n'ihi na usoro ahụ gosipụtara uru zeta zuru oke na ngwa ngwa na-emekarị, na-egosi. AgNP-III Kacha eguzogide mkpokọta pH butere.Nsonaazụ ndị a dabara na nkezi nsonaazụ nha dayameta hydrodynamic.N'iburu n'uche nha nha nke primers ha, AgNP-I gosipụtara nchịkọta nwayọọ nwayọọ na-adịgide adịgide na ụkpụrụ pH niile, o yikarịrị ka ọ bụ n'ihi 10 mM NaCl ndabere, ebe AgNP-II na AgNP-III gosipụtara dị ịrịba ama na pH 3 nke mkpokọta.Ihe dị iche na-adọrọ mmasị bụ na n'agbanyeghị nnukwu nanoparticle size, AgNP-III na-emepụta nchịkọta kachasị nta na pH 3 n'ime awa 24, na-egosipụta njirimara mgbochi nchịkọta ya.Site n'ikewa nkezi Z nke AgNP na pH 3 mgbe awa 24 gachara site na uru nke ihe nlele a kwadebere, enwere ike ịhụ na nha nha nha AgNP-I na AgNP-II abawanyela ugboro 50, ugboro 42 na ugboro 22. , n'otu n'otu.III.
Ọnụọgụ 3 A na-egosipụta nsonaazụ ọkụ dị ike nke nanoparticles ọlaọcha citrate kwụsịrị na-abawanye nha (10 nm: AgNP-I, 20 nm: AgNP-II na 50 nm: AgNP-III) dị ka nkezi hydrodynamic dayameta (Z nkezi ) (n'aka nri) N'okpuru ọnọdụ pH dị iche iche, ikike zeta (aka ekpe) gbanwere n'ime awa 24.
Mkpokọta pH a hụrụ na-emetụtakwa njirimara elu plasmon resonance (SPR) nke ihe nlele AgNP, dịka ihe ngosi UV-Vis ha gosipụtara.Dị ka ihe atụ S1 mgbakwunye si kwuo, nchikota nke nkwusioru ọlaọcha atọ nanoparticle na-esochi mbelata n'ike nke elu SPR ha yana ngbanwe na-acha uhie uhie na-agafeghị oke.Ogologo mgbanwe ndị a dị ka ọrụ nke pH kwekọrọ n'ogo nchịkọta nke nsonaazụ DLS buru amụma, agbanyeghị, a hụla ụfọdụ usoro na-adọrọ mmasị.N'adịghị ka nghọta, ọ na-apụta na AgNP-II nke na-ajụ ase bụ ihe kachasị emetụ n'ahụ maka mgbanwe SPR, ebe ihe atụ abụọ ndị ọzọ na-enwechaghị mmetụta.N'ime nyocha SPR, 50 nm bụ oke nha nha nha, nke a na-eji amata ihe dị iche iche dabere na akụrụngwa dielectric ha.Enwere ike ịkọwa ihe ndị dị ntakịrị karịa 50 nm (AgNP-I na AgNP-II) dị ka dielectric dipoles dị mfe, ebe ụmụ irighiri ihe ruru ma ọ bụ gafere oke a (AgNP-III) nwere ihe dielectric dị mgbagwoju anya karị, na resonance The band kewara n'ime mgbanwe multimodal. .N'ihe gbasara ihe nlele abụọ dị ntakịrị, AgNP nwere ike ịre dị ka dipoles dị mfe, na plasma nwere ike ịgbakọta ngwa ngwa.Ka oke urughuru na-abawanye, njikọ a na-emepụta plasma ka ukwuu, nke nwere ike ịkọwa mmetụta dị elu nke a hụrụ.29 Otú ọ dị, maka akụkụ kachasị ukwuu, nleba anya dipole dị mfe abaghị uru mgbe steeti njikọta ndị ọzọ nwekwara ike ime, nke nwere ike ịkọwa mbelata nke AgNP-III iji gosi mgbanwe mgbanwe.29
N'okpuru ọnọdụ nnwale anyị, e gosipụtara na uru pH nwere mmetụta dị ukwuu na nkwụsi ike colloidal nke nanoparticles ọlaọcha na-ekpuchi citrate nke nha dị iche iche.N'ime usoro ndị a, a na-enye nkwụsi ike site n'aka ndị ebubo na-adịghị mma -COO n'elu AgNP.The carboxylate functional group nke citrate ion na-protonated na a nnukwu ọnụ ọgụgụ nke H + ion, otú ahụ emepụtara carboxyl otu enweghị ike inye electrostatic repulsion n'etiti ahụ, dị ka e gosiri na n'elu ahịrị nke Figure 4. Dị ka Le Chatelier si ụkpụrụ, AgNP. samples na-agbakọta ngwa ngwa na pH 3, mana nke nta nke nta na-abawanyewanye ka pH na-abawanye.
Ọgụgụ 4 Usoro nhazi nke mmekọrịta dị n'elu nke akọwapụtara site na nchịkọta n'okpuru pH dị iche iche (ahịrị elu), ntinye uche NaCl (ahịrị etiti), na biomolecules (ahịrị ala).
Dị ka eserese 5 si kwuo, a na-enyocha nkwụsi ike colloidal na nkwụsị AgNP nke nha dị iche iche n'okpuru ụbara nnu.Dabere na ikike zeta, mmụba nanoparticle nha na sistemu AgNP citrate kwụsịrị ọzọ na-enye nkwalite nkwalite na mmetụta mpụga sitere na NaCl.Na AgNP-I, 10 mM NaCl zuru ezu iji weta nchịkọta dị nro, na ntinye nnu nke 50 mM na-enye nsonaazụ yiri nke ahụ.Na AgNP-II na AgNP-III, 10 mM NaCl anaghị emetụta ikike zeta n'ihi na ụkpụrụ ha dị na (AgNP-II) ma ọ bụ n'okpuru (AgNP-III) -30 mV.Ịbawanye ntinye uche NaCl na 50 mM na n'ikpeazụ ruo 150 mM NaCl zuru ezu iji belata uru zuru oke nke ikike zeta na ihe nlele niile, ọ bụ ezie na akụkụ buru ibu na-ejigide ụgwọ na-adịghị mma.Nsonaazụ ndị a kwekọrọ na dayameta hydrodynamic nke AgNP na-atụ anya ya;nkezi usoro usoro Z tụrụ na 10, 50, na 150 mM NaCl na-egosi dị iche iche, jiri nwayọọ nwayọọ na-abawanye ụkpụrụ.N'ikpeazụ, ahụrụ mkpokọta micron na nnwale 150 mm atọ niile.
Ọnụọgụ 5 A na-egosipụta nsonaazụ ọkụ na-agbanwe agbanwe nke citrate-akwụsị silver nanoparticles sample na-abawanye nha (10 nm: AgNP-I, 20 nm: AgNP-II na 50 nm: AgNP-III) dị ka nkezi hydrodynamic dayameta (Z nkezi. ) (n'aka nri) na zeta ikike (aka ekpe) gbanwere n'ime awa 24 n'okpuru nchịkọta NaCl dị iche iche.
Nsonaazụ UV-Vis na eserese S2 mgbakwunye na-egosi na SPR nke 50 na 150 mM NaCl na nlele atọ ahụ nwere mbelata ozugbo na nke ukwuu.DLS nwere ike ịkọwa nke a, n'ihi na nchịkọta nke NaCl na-eme ngwa ngwa karịa nyocha ndị dabere na pH, nke a kọwara site na nnukwu ọdịiche dị n'etiti mbido (0, 1.5, na 3 awa).Na mgbakwunye, ịba ụba ntinye nnu ga-abawanye ikike ikike nke usoro nnwale, nke ga-enwe mmetụta dị ukwuu na resonance plasmon.29
A na-achikota mmetụta nke NaCl n'ahịrị etiti nke Figure 4. N'ozuzu, enwere ike ikwubi na ịba ụba nke sodium chloride nwere mmetụta yiri nke ahụ dị ka ịba ụba acidity, n'ihi na Na + ions nwere ọchịchọ ịhazi gburugburu otu carboxylate, na-egbochi zeta na-adịghị mma AgNPs.Na mgbakwunye, 150 mM NaCl mepụtara mkpokọta micron-sized n'ime ihe atụ atọ ahụ, na-egosi na ntinye uche nke electrolyte physiological na-emebi nkwụsi ike nke colloidal nke citrate-akwụsị AgNP.Site n'ịtụle ntinye uche dị oke egwu (CCC) nke NaCl na usoro AgNP yiri ya, enwere ike itinye nsonaazụ ndị a n'ụzọ aghụghọ na akwụkwọ ndị dị mkpa.Huynh et al.gbakọrọ na CCC nke NaCl maka nanoparticles ọlaọcha citrate kwụsịrị na nkezi dayameta nke 71 nm bụ 47.6 mM, ebe El Badawy et al.hụrụ na CCC nke 10 nm AgNP nwere mkpuchi citrate bụ 70 mM.10,16 Tụkwasị na nke ahụ, a na-atụle CCC dị elu nke dị ihe dị ka 300 mM site na He et al., Nke mere ka usoro nhazi ha dị iche na mbipụta ahụ a kpọtụrụ aha na mbụ.48 Ọ bụ ezie na onyinye dị ugbu a abụghị maka nyocha zuru oke nke ụkpụrụ ndị a, n'ihi na ọnọdụ nnwale anyị na-abawanye na mgbagwoju anya nke ọmụmụ ihe dum, ihe ọmụmụ NaCl dị mkpa nke 50 mM, karịsịa 150 mM NaCl, yiri ka ọ dị elu.Na-akpata coagulation, na-akọwa mgbanwe siri ike achọpụtara.
Nzọụkwụ na-esote na nnwale polymerization bụ iji ụmụ irighiri ihe dị mfe mana nke dabara n'ụzọ ndụ iji mee ka mmekọrịta nanoparticle-biomolecule.Dabere na DLS (Figure 6 na 7) yana nsonaazụ UV-Vis (Nkọwa mgbakwunye S3 na S4), enwere ike ikwupụta nkwubi okwu ụfọdụ.N'okpuru ọnọdụ nnwale anyị, glucose na glutamine molecules ndị a na-amụ agaghị ebute mkpokọta na sistemụ AgNP ọ bụla, n'ihi na usoro Z-pụtara nwere njikọ chiri anya na nha nha ntụaka kwekọrọ.Ọ bụ ezie na ọnụnọ ha anaghị emetụta mkpokọta, nsonaazụ nnwale na-egosi na a na-etinye akụkụ nke mkpụrụ ndụ ndị a n'elu AgNP.Nsonaazụ kachasị pụta ìhè na-akwado echiche a bụ mgbanwe a hụrụ na ntinye ọkụ.Ọ bụ ezie na AgNP-I anaghị egosipụta mgbanwe ntụgharị ma ọ bụ mgbanwe siri ike bara uru, enwere ike ịhụ ya nke ọma site n'ịtụ irighiri ihe buru ibu, nke nwere ike ịbụ n'ihi nhụta anya dị ukwuu a kpọtụrụ aha na mbụ.N'agbanyeghị itinye uche, glucose nwere ike ime mgbanwe na-acha ọbara ọbara ka awa 1.5 gachara ma e jiri ya tụnyere njikwa njikwa, nke dị ihe dị ka 40 nm na AgNP-II na ihe dịka 10 nm na AgNP-III, nke na-egosi na mmekọrịta dị n'elu.Glutamine gosipụtara omume yiri nke ahụ, mana mgbanwe ahụ edoghị anya.Na mgbakwunye, ọ dịkwa mma ịkọ na glutamine nwere ike belata ikike zeta zuru oke nke ọkara na nnukwu ihe.Otú ọ dị, ebe ọ bụ na mgbanwe zeta ndị a adịghị ka ọ na-emetụta ọkwa nchịkọta, ọ nwere ike ịkọ na ọbụna obere biomolecules dị ka glutamine nwere ike inye ụfọdụ nkwụsị nke oghere n'etiti ihe ndị ahụ.
Ọnụọgụ 6 A na-egosipụta nsonaazụ ọkụ na-ekpo ọkụ nke citrate kwụsịrị ọlaọcha nanoparticle sample na-abawanye nha (10 nm: AgNP-I, 20 nm: AgNP-II na 50 nm: AgNP-III) dị ka nkezi hydrodynamic dayameta (Z nkezi) (n'aka nri) N'okpuru ọnọdụ mpụga nke mkpokọta glucose dị iche iche, ikike zeta (aka ekpe) na-agbanwe n'ime awa 24.
Ọnụọgụ 7 A na-egosipụta nsonaazụ ọkụ dị ike nke nanoparticles ọlaọcha citrate kwụsịrị na-abawanye nha (10 nm: AgNP-I, 20 nm: AgNP-II na 50 nm: AgNP-III) dị ka nkezi hydrodynamic dayameta (nkezi Z). ) (n'aka nri) N'ihu glutamine, ikike zeta (aka ekpe) gbanwere n'ime awa 24.
Na nkenke, obere biomolecules dị ka glucose na glutamine adịghị emetụta nkwụsi ike nke colloidal na ntinye a tụrụ atụ: ọ bụ ezie na ha na-emetụta ikike zeta na nsonaazụ UV-Vis na ọkwa dịgasị iche iche, nkezi nke Z adịghị agbanwe agbanwe.Nke a na-egosi na mgbasa ozi dị n'elu nke ụmụ irighiri ihe na-egbochi mmechi electrostatic, ma n'otu oge ahụ na-enye nkwụsi ike akụkụ.
Iji jikọta nsonaazụ ndị gara aga na nsonaazụ gara aga ma mee ka ọnọdụ ndụ dị mma nke ọma, anyị ahọpụtara ụfọdụ akụkụ omenala cell a na-ejikarị eme ihe ma jiri ya dị ka ọnọdụ nnwale maka ịmụ nkwụsi ike nke AgNP colloid.N'ime nnwale in vitro dum, otu n'ime ọrụ kachasị mkpa nke DMEM dị ka onye na-ajụ ase bụ ịmepụta ọnọdụ osmotic dị mkpa, ma site na echiche nke kemịkalụ, ọ bụ ngwọta nnu dị mgbagwoju anya nke nwere ike ionic zuru oke yiri 150 mM NaCl. .40 Maka FBS, ọ bụ ngwakọta dị mgbagwoju anya nke biomolecules-karịsịa protein-site n'echiche nke mgbasa ozi elu, ọ nwere ụfọdụ myirịta na nsonaazụ nnwale nke glucose na glutamine, n'agbanyeghị ihe mejupụtara kemịkalụ na ụdị dị iche iche Mmekọahụ dị mgbagwoju anya karị.19 DLS na UV-Nsonaazụ a na-ahụ anya nke egosiri na eserese 8 na S5 mgbakwunye, nwere ike ịkọwa site n'inyocha ihe mejupụtara kemịkalụ nke ihe ndị a ma jikọta ha na nha ndị dị na ngalaba gara aga.
Ọnụọgụ 8 A na-egosipụta nsonaazụ ọkụ dị ike nke nanoparticles ọlaọcha citrate kwụsịrị na-abawanye nha (10 nm: AgNP-I, 20 nm: AgNP-II na 50 nm: AgNP-III) dị ka nkezi hydrodynamic dayameta (nkezi Z). ) (n'aka nri) N'ihu nke akụkụ omenala cell DMEM na FBS, ikike zeta (aka ekpe) gbanwere n'ime awa 24.
Ntughari nke AgNP nke nha dị iche iche na DMEM nwere mmetụta yiri nke ahụ na nkwụsi ike nke colloidal na nke a hụrụ na ọnụnọ nke nnukwu NaCl.Mgbasa nke AgNP na 50 v / v% DMEM gosiri na a chọtara nchịkọta buru ibu na mmụba nke zeta nwere ike na uru Z-nkezi na mbelata nke ukwuu nke ike SPR.Ọ dị mma ịmara na oke mkpokọta DMEM kpalitere mgbe awa 24 gachara dabara na nha nke nanoparticles primer.
Mmekọrịta dị n'etiti FBS na AgNP yiri nke ahụ a hụrụ na ọnụnọ nke obere ụmụ irighiri dị ka glucose na glutamine, mana mmetụta dị ike karị.Nkezi Z nke ụmụ irighiri ihe ndị ahụ ka emetụtaghị ya, ebe a na-achọpụta mmụba nke ikike zeta.Ọnụ ọgụgụ kasị elu SPR gosipụtara ntakịrị mgbanwe na-acha uhie uhie, ma eleghị anya na-adọrọ mmasị karị, ike SPR ebelataghị ka ọ dị na njikwa njikwa.Enwere ike ịkọwa nsonaazụ ndị a site na mgbasa ozi ebumpụta ụwa nke macromolecules n'elu nanoparticles (ahịrị ala na eserese 4), nke a ghọtara ugbu a dị ka nguzobe nke corona biomolecular n'ime ahụ.49