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ʻOi aku ka maikaʻi o nā nanoparticles liʻiliʻi?E hoʻomaopopo i nā hopena olaola o ka hōʻuluʻulu hilinaʻi nui o nā nanoparticles kālā ma lalo o nā kūlana pili olaola
Nā mea kākau: 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 o ka Environmental Chemistry, Hungari a me Hungary Informatics. , Ke Kulanui o Szeged;2 Keʻena o Biochemistry a me Molecular Biology, Faculty of Science and Information, University of Szeged, Hungary;3 Ke Keʻena ʻOihana Microbiology, Kumu ʻEpekema a me ka ʻIke, Ke Kulanui o Szeged, Hungary;4MTA-SZTE Reaction Kinetics and Surface Chemistry Research Group, Szeged, Hungary* Ua hāʻawi like kēia mau mea kākau i kēia hana.Kūkākūkā: 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] Ke kumu: Silver nanoparticles (AgNPs) are kekahi o nā nanomaterials i aʻo pinepine ʻia, ma muli o kā lākou noi biomedical.Eia nō naʻe, ma muli o ka hōʻuluʻulu ʻana o nā nanoparticles, ko lākou cytotoxicity maikaʻi loa a me ka hana antibacterial e hoʻohālikelike pinepine ʻia i ka media biological.Ma kēia hana, ua aʻo ʻia ka ʻano hoʻohui a me nā hana olaola e pili ana i ʻekolu mau ʻano nanoparticle kala i hoʻopau ʻia me ka awelika anawaena o 10, 20, a me 50 nm.Ke Kaʻina Hana: E hoʻohana i ka microscope electron transmission e hoʻohui a hōʻike i nā nanoparticles, e loiloi i kā lākou ʻano hoʻohui i nā waiwai pH like ʻole, NaCl, glucose a me ka glutamine ka hoʻopuʻi ʻana ma o ka hoʻopuʻi kukui uila a me ka spectroscopy ʻike ultraviolet.Eia kekahi, i loko o ka moʻomeheu cell moʻomeheu ʻāpana e like me Dulbecco e hoʻomaikaʻi i ka ʻano aggregation ma Eagle Medium a me Fetal Calf Serum.Nā hualoaʻa: Hōʻike nā hopena i ka pH acidic a me ka ʻike electrolyte physiological e hoʻoulu pinepine i ka micron-scale aggregation, hiki ke hoʻopili ʻia e ka hoʻokumu ʻana o ka corona biomolecular.He mea pono e hoʻomaopopo i ka hōʻike ʻana o nā ʻāpana nui i ke kūpaʻa kiʻekiʻe i nā mana o waho ma mua o ko lākou mau mea liʻiliʻi.Ua hana ʻia nā hoʻokolohua in vitro cytotoxicity a me antibacterial e ka mālama ʻana i nā cell me nā nanoparticle aggregates ma nā pae hoʻohui like ʻole.Ka Hopena: Hōʻike kā mākou mau hopena i ka pilina hohonu ma waena o ka colloidal stability a me ka ʻawaʻawa o AgNPs, no ka mea, ʻo ka hōʻuluʻulu nui e alakaʻi i ka nalowale o ka hana olaola.ʻO ke kiʻekiʻe kiʻekiʻe o ka anti-aggregation i ʻike ʻia no nā ʻāpana nui he hopena koʻikoʻi i ka in vitro toxicity, no ka mea, ʻoi aku ka nui o ka hana antimicrobial a me ka mammalian cell o ia mau laʻana.Ke alakaʻi nei kēia mau ʻike i ka hopena, ʻoiai ka manaʻo maʻamau i loko o nā palapala e pili ana, ʻo ka huli ʻana i nā nanoparticles liʻiliʻi loa ʻaʻole paha ʻo ia ka hana maikaʻi loa.Hua'ōlelo: ka ulu ʻana o ka hua, ka paʻa ʻana o ka colloidal, ka ʻano hoʻohui pili i ka nui, ka hoʻohui ʻana i nā mea ʻawaʻawa.
Ke hoʻomau nei ka noi a me ka hoʻopuka ʻana o nā nanomaterials, ʻoi aku ka nui o ka nānā ʻana i kā lākou biosafety a i ʻole ka hana biological.ʻO nā nanoparticles kālā (AgNPs) kekahi o nā mea maʻamau i hoʻohui ʻia, noiʻi ʻia a hoʻohana ʻia i nā ʻelele o kēia papa o nā mea no ka maikaʻi o kā lākou catalytic, optical a me nā waiwai biological.1 Ua manaʻoʻiʻo ʻia ʻo nā hiʻohiʻona kūʻokoʻa o nā nanomaterials (me nā AgNPs) ke kumu nui i ko lākou ʻāpana kikoʻī nui.No laila, ʻo ka pilikia inevitably ʻo kēlā me kēia kaʻina hana e pili ana i kēia hiʻohiʻona nui, e like me ka nui o ka particle, ka uhi ʻana o ka ʻili A i ʻole ka hōʻuluʻulu ʻana, inā paha e hōʻino nui ia i nā waiwai o nā nanoparticles i koʻikoʻi i nā noi kikoʻī.
ʻO nā hopena o ka nui o nā ʻāpana a me nā stabilizers nā kumuhana i kākau maikaʻi ʻia i ka palapala.No ka laʻana, ʻo ka manaʻo i ʻae ʻia he ʻoi aku ka ʻona o nā nanoparticles liʻiliʻi ma mua o nā nanoparticles nui.2 E like me ka palapala maʻamau, ua hōʻike kā mākou mau haʻawina mua i ka hana nui o ka nanosilver ma luna o nā pūnana mammalian a me nā microorganism.3- 5 ʻO ka uhi ʻana o ka ʻili he ʻano ʻē aʻe i loaʻa ka mana ākea i nā waiwai o nā mea nano.Ma ka hoʻohui ʻana a i ʻole ka hoʻololi ʻana i nā stabilizers ma luna o kona ʻili, hiki i ka nanomaterial like ke loaʻa i nā waiwai kino, kemika, a me nā waiwai olaola.Hoʻohana pinepine ʻia ka hoʻohana ʻana i nā mea capping ma ke ʻano he ʻāpana o ka nanoparticle synthesis.No ka laʻana, ʻo nā nanoparticles kālā i hoʻopau ʻia ka citrate kekahi o nā AgNP e pili ana i ka noiʻi, i hoʻohui ʻia e ka hoʻemi ʻana i nā paʻakai kālā i loko o kahi hopena stabilizer i koho ʻia e like me ke ʻano hana.6 Hiki iā Citrate ke hoʻohana maʻalahi i kona kumu kūʻai haʻahaʻa, loaʻa, biocompatibility, a me ka pilina ikaika no ke kālā, hiki ke ʻike ʻia i nā pilina like ʻole, mai ka hoʻohuli ʻana i ka ʻili a hiki i nā pilina ionic.ʻO nā mole liʻiliʻi a me nā iona polyatomic kokoke i 7,8, e like me nā citrates, polymers, polyelectrolytes, a me nā mea ola ola e hoʻohana mau ʻia e hoʻopaʻa i ka nano-kālā a hana i nā functionalizations kūʻokoʻa ma luna.9-12
ʻOiai he wahi hoihoi loa ka hiki ke hoʻololi i ka hana o nā nanoparticles e ka hoʻopili ʻana i ka ʻili, ʻo ka hana nui o kēia uhi ʻili he mea ʻole, e hāʻawi ana i ka paʻa colloidal no ka ʻōnaehana nanoparticle.ʻO ka ʻāpana kikoʻī kikoʻī nui o nā nanomaterials e hoʻohua i ka ikehu ākea nui, kahi e pale ai i ka hiki ʻana o ka thermodynamic o ka ʻōnaehana e hiki i kona ikehu liʻiliʻi.13 Me ka hoʻopaʻa ʻole kūpono, hiki i kēia ke alakaʻi i ka agglomeration o nā nanomaterials.ʻO ka hōʻuluʻulu ʻana ka hoʻokumu ʻana o nā ʻāpana o nā ʻano like ʻole a me nā ʻano nui e kū ana i ka wā e hui ai nā ʻāpana i hoʻopuehu ʻia a me nā pilina thermodynamic o kēia manawa e ʻae i nā ʻāpana e pili kekahi i kekahi.No laila, hoʻohana ʻia nā stabilizers e pale i ka hōʻuluʻulu ʻana ma o ka hoʻokomo ʻana i kahi ikaika repulsive nui ma waena o nā ʻāpana e kūʻē i kā lākou hoʻoheheʻe thermodynamic.14
ʻOiai ua ʻimi pono ʻia ke kumuhana o ka nui o ka ʻāpana a me ka uhi o ka ʻili i loko o ka pōʻaiapili o kāna hoʻoponopono ʻana i nā hana olaola i hoʻāla ʻia e nā nanoparticles, ʻo ka hōʻuluʻulu ʻana he wahi i mālama ʻole ʻia.Aneane ʻaʻohe aʻo ʻana e hoʻoponopono i ke kūpaʻa colloidal o nā nanoparticles ma lalo o nā kūlana pili biologically.10,15-17 Eia kekahi, ʻokoʻa loa kēia hāʻawi, kahi i aʻo ʻia ai ka ʻona e pili ana i ka hoʻohui ʻana, ʻoiai inā hiki ke hoʻoulu i nā hopena ʻino, e like me ka thrombosis vascular, a i ʻole ka nalowale o nā ʻano i makemake ʻia, e like me kona ʻona, e like me hōʻike ʻia ma ke Kiʻi 1.18, 19 i hōʻike ʻia.ʻO kaʻoiaʻiʻo,ʻo kekahi o nāʻano hana liʻiliʻi iʻikeʻia o ke kū'ēʻana o ke kālā nanoparticle e pili ana i ka hōʻuluʻuluʻana, no ka mea, ua hōʻikeʻia kekahi mauʻano E. coli a me Pseudomonas aeruginosa e hoʻemi i ko lākou nano-kālā ma ka hōʻikeʻana i ka flagellin protein, flagellin.Loaʻa iā ia kahi pilina kiʻekiʻe no ke kālā, no laila e hoʻoulu ai i ka aggregation.20
Nui nā ʻano hana like ʻole e pili ana i ka ʻona o nā nanoparticles kālā, a ʻo ka hoʻohui ʻana e pili ana i kēia mau mīkini āpau.ʻO ke ʻano i kūkākūkā nui ʻia o ka hana biological AgNP, i kekahi manawa i kapa ʻia ʻo "Trojan Horse", manaʻo iā AgNPs ma ke ʻano he mea lawe Ag +.1,21 Hiki i ka mīkini lio Trojan ke hōʻoia i ka hoʻonui nui ʻana i ka neʻe ʻana o ka Ag + kūloko, kahi e alakaʻi ai i ka hanauna o ROS a me ka depolarization membrane.22-24 Hiki i ka hui ke hoʻopili i ka hoʻokuʻu ʻana o Ag+, no laila e hoʻopilikia ai i ka ʻawaʻawa, no ka mea e hōʻemi ana ia i ka ʻili hana pono kahi e hiki ai ke hoʻoheheʻe ʻia a hoʻoheheʻe ʻia nā iona kālā.Eia nō naʻe, ʻaʻole e hōʻike wale ʻo AgNP i ka ʻona ma o ka hoʻokuʻu ʻana i ka ion.Pono e noʻonoʻo ʻia ka nui a me ka morphology pili pili.I waena o lākou, ʻo ka nui a me ke ʻano o ka ʻili nanoparticle nā ​​hiʻohiʻona wehewehe.4,25 Hiki ke hoʻokaʻawale ʻia ka hōʻiliʻili ʻana o kēia mau ʻano hana ma ke ʻano he "mea hoʻohana ʻona."Nui paha nā ʻano mitochondrial a me ka membrane surface e hiki ai ke hōʻino i nā organelles a make i ka cell cell.25-27 No ka mea, pili maoli ka hana ʻana i nā mea hōʻuluʻulu i ka nui a me ke ʻano o nā mea kālā i ʻike ʻia e nā ʻōnaehana ola, hiki ke hoʻopili ʻia kēia mau pilina.
Ma kā mākou pepa mua e pili ana i ka hōʻuluʻulu ʻana o nā nanoparticles kālā, ua hōʻike mākou i kahi kaʻina hana hoʻomaʻamaʻa kūpono e pili ana i nā hoʻokolohua kemika a in vitro biological e aʻo i kēia pilikia.19 ʻO ka Dynamic Light Scattering (DLS) ka ʻenehana i makemake ʻia no kēia mau ʻano o ka nānā ʻana no ka mea hiki i ka mea ke hoʻopuehu i nā kiʻi ma kahi lōʻihi nalu e like me ka nui o kāna mau ʻāpana.No ka mea e pili ana ka wikiwiki o ka neʻe ʻana o Brownian o nā ʻāpana i loko o ka wai wai i ka nui, hiki ke hoʻohana ʻia ka hoʻololi ʻana i ka ikaika o ka mālamalama i hoʻopuehu ʻia e hoʻoholo ai i ka awelika hydrodynamic anawaena (Z-mean) o ka hāpana wai.28 Eia kekahi, ma ka hoʻohana ʻana i ka volta i ka hāpana, hiki ke ana like ka zeta hiki (ζ potential) o ka nanoparticle me ka waiwai awelika Z.13,28 Inā kiʻekiʻe ka waiwai paʻa o ka hiki zeta (e like me nā alakaʻi maʻamau> ± 30 mV), e hoʻoulu i ka hoʻokuʻu electrostatic ikaika ma waena o nā mea e kūʻē i ka hoʻohui.ʻO ke ʻano o ka plasmon resonance (SPR) kahi hiʻohiʻona ʻokoʻa kūʻokoʻa, pili nui ʻia i nā nanoparticles metala makamae (ʻo Au a me Ag).29 Ma muli o nā oscillations uila (surface plasmons) o kēia mau mea ma ka nanoscale, ua ʻike ʻia ʻo spherical AgNPs kahi hiʻohiʻona UV-Vis absorption peak kokoke i 400 nm.30 Hoʻohana ʻia ka ikaika a me ka lōʻihi o ka nalu o nā ʻāpana e hoʻohui i nā hopena DLS, no ka mea hiki ke hoʻohana ʻia kēia ʻano no ka ʻike ʻana i ka hōʻuluʻulu nanoparticle a me ka adsorption ʻili o nā biomolecules.
Ma muli o ka ʻike i loaʻa, hana ʻia ka cell viability (MTT) a me ka antibacterial assays ma ke ʻano i wehewehe ʻia ai ka mea ʻawaʻawa AgNP ma ke ʻano he hana o ka pae hōʻuluʻulu, ma mua o (ka mea i hoʻohana pinepine ʻia) nanoparticle concentration.ʻO kēia ala kūikawā e hiki ai iā mākou ke hōʻike i ke koʻikoʻi hohonu o ka pae hoʻohui i loko o ka hana olaola, no ka mea, no ka laʻana, nalowale loa nā AgNPs citrate-terminated i kā lākou hana olaola i loko o kekahi mau hola ma muli o ka hoʻohui.19
Ma ka hana o kēia manawa, manaʻo mākou e hoʻonui nui i kā mākou mau haʻawina mua i ka paʻa o nā colloids pili bio a me kā lākou hopena i ka hana olaola ma ke aʻo ʻana i ka hopena o ka nui nanoparticle ma ka nanoparticle aggregation.ʻO kēia kekahi o nā haʻawina o nā nanoparticles.ʻO kahi hiʻohiʻona kiʻekiʻe a me 31 No ka noiʻi ʻana i kēia pilikia, ua hoʻohana ʻia kahi ala ulu i hoʻopaʻa ʻia i nā hua no ka hana ʻana i nā AgNP i hoʻopau ʻia i ka citrate i ʻekolu mau pae nui like ʻole (10, 20, a me 50 nm).6,32 e like me kekahi o nā ala maʻamau.No nā nanomaterials i hoʻohana nui ʻia a hoʻohana mau ʻia i nā noi lapaʻau, koho ʻia nā AgNP i hoʻopau ʻia i ka citrate o nā nui like ʻole e aʻo i ka nui o ka hilinaʻi ʻana o nā waiwai kūlohelohe pili i ka nanosilver.Ma hope o ka synthesizing AgNPs o nā ʻano nui like ʻole, hōʻike mākou i nā laʻana i hana ʻia e ka lawe ʻana i ka microscopy electron (TEM), a laila nānā i nā ʻāpana me ka hoʻohana ʻana i ke kaʻina hana screening.Eia kekahi, ma ke alo o nā moʻomeheu cell in vitro ʻO Dulbecco's Modified Eagle's Medium (DMEM) a me Fetal Bovine Serum (FBS), ua loiloi ʻia ka ʻano hoʻohui i hilinaʻi nui ʻia a me kāna ʻano ma nā ʻano pH like ʻole, NaCl, glucose, a me ka glutamine.Hoʻoholo ʻia nā ʻano o ka cytotoxicity ma lalo o nā kūlana ākea.Hōʻike ka manaʻo ʻepekema ma ke ʻano laulā, ʻoi aku ka maikaʻi o nā ʻāpana liʻiliʻi;Hāʻawi kā mākou noiʻi i kahi kahua kemika a me ke olaola e hoʻoholo ai inā ʻo ia ka hihia.
Ua hoʻomākaukau ʻia ʻekolu nanoparticles kālā me nā pae nui like ʻole e ke ʻano o ka ulu ʻana o nā hua i manaʻo ʻia e Wan et al., me nā hoʻololi liʻiliʻi.6 Hoʻokumu ʻia kēia ʻano hana i ka hoʻemi kemika, me ka hoʻohana ʻana i ka nitrate kālā (AgNO3) ma ke kumu kālā, ka sodium borohydride (NaBH4) ma ke ʻano he mea hoʻemi, a me ka sodium citrate ma ke ʻano he stabilizer.E hoomakaukau mua i 75 mL o 9 mM citrate aqueous solution mai ka sodium citrate dihydrate (Na3C6H5O7 x 2H2O) a me ka wela a hiki i 70°C.A laila, hoʻohui ʻia ka 2 mL o 1% w/v AgNO3 solution i ka mea hoʻonā, a laila ninini ʻia ka wai hou sodium borohydride (2 mL 0.1% w/v) i loko o ka hui ʻana.ʻO ka hopena o ka hoʻokuʻu ʻana i ka melemele-ʻeleʻele i mālama ʻia ma 70 ° C me ka hoʻoulu ʻana ikaika no 1 hola, a laila hoʻomaha ʻia i ka mahana wela.Hoʻohana ʻia ka laʻana i loaʻa (i kapa ʻia ʻo AgNP-I mai kēia manawa) ma ke kumu no ka ulu ʻana o ka hua i hoʻopaʻa ʻia i ka hana synthesis e hiki mai ana.
No ka hoʻohui ʻana i kahi hoʻokuʻu ʻia ʻana o ka mea liʻiliʻi liʻiliʻi (i hōʻike ʻia ʻo AgNP-II), wela 90 mL 7.6 mM citrate solution i 80°C, hui pū me 10 mL AgNP-I, a laila hui pū me 2 mL 1% w/v ʻO ka hopena AgNO3 mālama ʻia ma lalo o ka hoʻoulu ʻana i ka mīkini ikaika no 1 hola, a laila hoʻomaʻalili ʻia ka hāpana i ka lumi wela.
No ka ʻāpana nui loa (AgNP-III), e hana hou i ke kaʻina ulu like, akā i kēia hihia, e hoʻohana i 10 mL o AgNP-II e like me ka hoʻokuʻu ʻana i nā hua.Ma hope o ka hōʻea ʻana o nā laʻana i ka lumi wela, hoʻonoho lākou i kā lākou helu Ag nominal ma muli o ka nui o ka AgNO3 maʻiʻo i 150 ppm ma ka hoʻohui ʻana a i ʻole ka hoʻohemo ʻana i ka solvent hou ma 40 ° C, a hoʻopaʻa hope iā lākou ma 4 ° C a hiki i ka hoʻohana hou ʻana.
E hoʻohana i ka FEI Tecnai G2 20 X-Twin Transmission Electron Microscope (TEM) (FEI Corporate Headquarters, Hillsboro, Oregon, USA) me 200 kV acceleration voltage e nānā i nā hiʻohiʻona morphological o nā nanoparticles a hopu i kā lākou electron diffraction (ED).Ma ka liʻiliʻi he 15 mau kiʻi ʻelele (~ 750 mau ʻāpana) i loiloi ʻia me ka hoʻohana ʻana i ka pūʻolo polokalamu ImageJ, a ua hana ʻia nā histograms (a me nā kiʻi āpau i ka noiʻi holoʻokoʻa) ma OriginPro 2018 (OriginLab, Northampton, MA, USA) 33, 34.
ʻO ka awelika hydrodynamic anawaena (Z-average), hiki i ka zeta (ζ-potential) a me ke ʻano o ka plasmon plasmon resonance (SPR) o nā laʻana i ana ʻia e hōʻike i ko lākou mau waiwai colloidal mua.Ua ana ka awelika hydrodynamic anawaena a me ka zeta hiki o ka hāpana e ka Malvern Zetasizer Nano ZS mea kani (Malvern Instruments, Malvern, UK) me ka hoʻohana ʻana i nā pūnana capillary i hoʻopili ʻia ma 37±0.1°C.Ua hoʻohana ʻia ʻo Ocean Optics 355 DH-2000-BAL UV-Vis spectrophotometer (Halma PLC, Largo, FL, USA) no ka loaʻa ʻana o nā hiʻohiʻona SPR mai ka UV-Vis absorption spectra o nā laʻana i ka laulā o 250-800 nm.
I loko o ka hoʻokolohua holoʻokoʻa, ua lawe ʻia ʻekolu ʻano ana ʻokoʻa e pili ana i ka paʻa colloidal i ka manawa like.E hoʻohana i ka DLS no ke ana i ka awelika hydrodynamic anawaena (Z awelika) a me ka zeta hiki (ζ hiki) o na particles, no ka mea, ua pili ka awelika Z i ka awelika nui o ka nanoparticle aggregates, a me ka zeta hiki ke hoike ina ka electrostatic repulsion i loko o ka pūnaewele. Ua lawa ka ikaika e hoʻopau iā Van der Waals i ka huki ʻana ma waena o nā nanoparticles.Hana ʻia nā ana ʻekolu, a ʻo ka hoʻokaʻawale maʻamau o Z mean a me zeta hiki ke helu ʻia e ka polokalamu Zetasizer.ʻIke ʻia ka hiʻohiʻona SPR spectra o nā ʻāpana e ka UV-Vis spectroscopy, no ka mea hiki ke hoʻololi i ka ikaika kiʻekiʻe a me ka lōʻihi o ka nalu e hōʻike i ka hoʻohui ʻana a me ka pilina ma luna.29,35 ʻO ka ʻoiaʻiʻo, ʻoi aku ka mana o ka resonance plasmon surface i nā metala makamae i alakaʻi ai i nā ala hou o ka nānā ʻana i nā biomolecules.29,36,37 ʻO ka manaʻo o AgNPs i loko o ka hui hoʻokolohua e pili ana i 10 ppm, aʻo ka manaʻo e hoʻonohonoho i ka ikaika o ka nui loa o ka SPR absorption mua i ka 1. Ua hanaʻia ka hoʻokolohua ma keʻano o ka manawa ma ka 0;1.5;3;6;12 a me 24 mau hola ma lalo o nā kūlana pili olaola like ʻole.Hiki ke ʻike ʻia nā kikoʻī hou aku e wehewehe ana i ka hoʻokolohua ma kā mākou hana mua.19 I ka pōkole, nā helu pH like ʻole (3; 5; 7.2 a me 9), ʻokoʻa ka sodium chloride (10 mM; 50 mM; 150 mM), glucose (3.9 mM; 6.7 mM) a me ka glutamine (4 mM), a Ua hoʻomākaukau pū ʻia ʻo Dulbecco's Modified Eagle Medium (DMEM) a me Fetal Bovine Serum (FBS) (i ka wai a me DMEM) ma ke ʻano he kumu hoʻohālike, a ua aʻo i kā lākou hopena i ka ʻano hoʻohui o nā nanoparticles kālā synthesized.pH ʻIke ʻia nā waiwai o, NaCl, glucose, a me ka glutamine e pili ana i ka physiological concentrations, aʻo ka nui o DMEM a me FBS e like me nā pae i hoʻohana ʻia i ka hoʻokolohua in vitro holoʻokoʻa.38-42 Ua hana ʻia nā ana a pau ma ka pH 7.2 a me 37°C me ka paʻakai paʻakai paʻa mau o 10 mM NaCl e hoʻopau i nā pānaʻi ʻāpana lōʻihi (koe naʻe kekahi mau hoʻokolohua pili i ka pH a me ka NaCl, kahi o kēia mau ʻano nā mea hoʻololi ma lalo. haʻawina).28 Ua hōʻuluʻulu ʻia ka papa inoa o nā kūlana like ʻole ma ka Papa 1. Hoʻohana ʻia ka hoʻokolohua i kaha ʻia me † ma ke ʻano he kuhikuhi a pili i kahi laʻana i loaʻa ka 10 mM NaCl a me ka pH 7.2.
Ua loaʻa mai ka laina cell cancer prostate kanaka (DU145) a me nā keratinocytes kanaka ola ʻole (HaCaT) mai ATCC (Manassas, VA, USA).Hoʻomaʻamaʻa mau ʻia nā kelepona ma Dulbecco's minimum essential medium Eagle (DMEM) loaʻa ka 4.5 g/L glucose (Sigma-Aldrich, Saint Louis, MO, USA), hoʻohui ʻia me 10% FBS, 2 mM L-glutamine, 0.01 % Streptomycin a me 0.005% Penicillin (Sigma-Aldrich, St. Louis, Missouri, USA).Hoʻomaʻamaʻa ʻia nā cell i kahi incubator 37 ° C ma lalo o 5% CO2 a me 95% humidity.
No ka ʻimi ʻana i nā hoʻololi o ka cytotoxicity AgNP i hoʻokumu ʻia e ka hoʻohui ʻana i ka ʻāpana i ke ʻano o ka manawa, ua hana ʻia kahi hoʻokolohua MTT ʻelua.ʻO ka mea mua, ua ana ʻia ka viability o nā ʻano cell ʻelua ma hope o ka mālama ʻana me AgNP-I, AgNP-II a me AgNP-III.No kēia hopena, ua kanu ʻia nā ʻano cell ʻelua i loko o nā papa 96-well ma kahi kikoʻī o 10,000 cell / well a mālama ʻia me ʻekolu mau nui like ʻole o nā nanoparticles kālā i ka hoʻonui ʻana i nā manaʻo i ka lua o ka lā.Ma hope o 24 mau hola o ka mālama ʻana, holoi ʻia nā cell me ka PBS a hoʻokomo ʻia me 0.5 mg/mL MTT reagent (SERVA, Heidelberg, Kelemānia) i hoʻoheheʻe ʻia i loko o ka moʻomeheu moʻomeheu no 1 hola ma 37 ° C.Ua hoʻoheheʻe ʻia nā kristal Formazan i DMSO (Sigma-Aldrich, Saint Louis, MO, USA), a ua ana ʻia ka absorption ma 570 nm me ka hoʻohana ʻana i kahi mea heluhelu papa Synergy HTX (BioTek-Hungary, Budapest, Hungary).Manaʻo ʻia ka waiwai absorption o ka hāpana hoʻomalu i mālama ʻole ʻia he 100% ola ola.Hana i 3 mau hoʻokolohua ma ka hoʻohana ʻana i ʻehā mau hoʻopiʻi kūlohelohe kūʻokoʻa.Ua helu ʻia ʻo IC50 mai kahi ʻāpana pane pane ma muli o nā hopena ola.
Ma hope iho, ma ka lua o ka ʻanuʻu, ma ka hoʻoulu ʻana i nā ʻāpana me 150 mM NaCl no nā manawa like ʻole (0, 1.5, 3, 6, 12, a me 24 mau hola) ma mua o ka mālama ʻana i ke kelepona, ua hana ʻia nā mokuʻāina aggregation o nā nanoparticles kālā.Ma hope mai, ua hana ʻia ka MTT assay e like me ka mea i wehewehe mua ʻia no ka loiloi ʻana i nā loli i ka hiki ʻana o ke kelepona i hoʻopili ʻia e ka ʻāpana ʻāpana.E hoʻohana i ka GraphPad Prism 7 no ka loiloi i ka hopena hope loa, e helu i ke koʻikoʻi helu helu o ka hoʻokolohua ma ka hoʻāʻo t hoʻohui ʻole, a e kaha i kona pae me * (p ≤ 0.05), ** (p ≤ 0.01), *** (p ≤ 0.001 ) A me **** (p ≤ 0.0001).
ʻEkolu ʻano nui o nā nanoparticles kālā (AgNP-I, AgNP-II a me AgNP-III) i hoʻohana ʻia no ka antibacterial susceptibility iā Cryptococcus neoformans IFM 5844 (IFM; Research Center for Pathogenic Fungi and Microbial Toxicology, Chiba University) a me Bacillus Test megaterium SZMC 6031 (SZMC: Szeged Microbiology Collection) a me E. coli SZMC 0582 ma RPMI 1640 medium (Sigma-Aldrich Co.).I mea e loiloi ai i nā hoʻololi o ka hana antibacterial i hoʻokumu ʻia e ka hōʻuluʻulu ʻana o nā ʻāpana, ʻo ka mea mua, ua hoʻoholo ʻia ko lākou haʻahaʻa haʻahaʻa haʻahaʻa (MIC) e ka microdilution i loko o kahi pā microtiter 96-well.I ka 50 μL o ka standardized cell suspension (5 × 104 cell / mL in RPMI 1640 medium), e hoʻohui i 50 μL o ke kālā nanoparticle suspension a me ka serially dilute ʻelua i ka ʻike (i ka ʻōlelo i ʻōlelo ʻia, ʻo 0 a me 75 ppm ka laulā, ʻo ia hoʻi, ʻO ka hāpana hoʻomalu he 50 μL o ka hoʻokuʻu ʻana o ke kelepona a me 50 μL o ka medium me ka ʻole o nā nanoparticles).Ma hope iho, ua hoʻokomo ʻia ka pā ma 30 ° C no nā hola 48, a ua ana ʻia ka nui o ka moʻomeheu i ka 620 nm me ka hoʻohana ʻana i kahi mea heluhelu papa SPECTROstar Nano (BMG LabTech, Offenburg, Kelemānia).Ua hana ʻia ka hoʻokolohua ʻekolu manawa ma ka ʻekolu.
Ma waho aʻe o ka 50 μL o nā mea hoʻohālike nanoparticle hoʻokahi i hoʻohanaʻia i kēia manawa, ua hoʻohanaʻia ke kaʻina hana like e like me ka mea i ho'ākāka muaʻia e nānā i ka hopena o ka hōʻuluʻuluʻana i ka hana antibacterial ma nā mea i'ōlelo muaʻia.Hoʻokumu ʻia nā mokuʻāina hoʻohui like ʻole o nā nanoparticles kālā ma ka hoʻoulu ʻana i nā ʻāpana me 150 mM NaCl no nā manawa like ʻole (0, 1.5, 3, 6, 12, a me 24 mau hola) ma mua o ka hana ʻana o ke kelepona.Ua hoʻohana ʻia kahi hoʻokuʻu i hoʻohui ʻia me 50 μL o RPMI 1640 medium ma ke ʻano he mana ulu, ʻoiai i mea e hoʻomalu ai i ka ʻawaʻawa, ua hoʻohana ʻia kahi hoʻokuʻu me nā nanoparticles non-aggregated.Ua hana ʻia ka hoʻokolohua ʻekolu manawa ma ka ʻekolu.E hoʻohana i ka GraphPad Prism 7 no ka loiloi hou ʻana i ka hopena hope, me ka hoʻohana ʻana i ka ʻikepili helu like me ka nānā ʻana MTT.
Ua hōʻike ʻia ka pae hōʻuluʻulu o nā mea liʻiliʻi loa (AgNP-I), a ua paʻi ʻia nā hopena i kā mākou hana mua, akā no ka hoʻohālikelike maikaʻi ʻana, ua nānā pono ʻia nā ʻāpana āpau.ʻOhi ʻia nā ʻikepili hoʻokolohua a kūkākūkā ʻia ma nā ʻāpana aʻe.ʻEkolu nui o AgNP.19
ʻO nā ana i hana ʻia e TEM, UV-Vis a me DLS i hōʻoia i ka hoʻohui maikaʻi ʻana o nā hōʻailona AgNP āpau (Figure 2A-D).Wahi a ka lālani mua o ke Kiʻi 2, hōʻike ka ʻāpana liʻiliʻi loa (AgNP-I) i kahi morphology spherical like me ka awelika anawaena o kahi 10 nm.Hāʻawi pū ke ʻano o ka ulu ʻana o ka hua i ka AgNP-II a me AgNP-III me nā pae nui like ʻole me ka awelika o nā ʻāpana ʻāpana ma kahi o 20 nm a me 50 nm.E like me ka hoʻokaʻawale maʻamau o ka puʻunaue ʻāpana, ʻaʻole i uhi ka nui o nā laʻana ʻekolu, he mea nui ia no kā lākou hoʻohālikelike hoʻohālikelike.Ma ka hoʻohālikelike ʻana i ka lākiō hiʻohiʻona awelika a me ka lākiō lahilahi o nā projections 2D ma muli o TEM, ua manaʻo ʻia ua loiloi ʻia ka sphericity o nā ʻāpana e ka hoʻopili kānana kinona o ImageJ (Figure 2E).43 E like me ka nānā ʻana i ke ʻano o nā ʻāpana, ʻaʻole i pili ko lākou ʻano lākiō (ʻaoʻao nui / ʻaoʻao pōkole o ka ʻāpana liʻiliʻi liʻiliʻi) i ka ulu ʻana o nā ʻāpana, a me kā lākou ratio lahilahi (ana ʻia o ka pōʻai kūpono kūpono / ʻāpana theoretical. ) emi mālie.Loaʻa kēia i nā ʻāpana polyhedral hou aʻe, he poepoe kūpono i ke kumumanaʻo, e pili ana i ka ratio thinness o 1.
Kiʻi 2 Kiʻi kiʻi ʻoniʻoni electron kiʻi (TEM) (A), ʻano hoʻohālike electron diffraction (ED) (B), histogram puʻunaue nui (C), hiʻohiʻona ultraviolet-ʻike ʻia (UV-Vis) kukui absorption spectrum (D), a me ka wai maʻamau Citrate ʻO nā nanoparticles kālā i hoʻopau ʻia me ke anawaena mechanical (Z-average), hiki i ka zeta, ka lākiō hiʻohiʻona a me ka mānoanoa o ka mānoanoa (E) i ʻekolu mau ʻano nui like ʻole: AgNP-I ʻo 10 nm (laina luna), ʻo AgNP -II ʻo 20 nm (laina waena. ), ʻo AgNP-III (lālani lalo) he 50 nm.
ʻOiai ʻo ke ʻano pōʻaiapuni o ke ʻano ulu i hoʻopili ʻia i ke ʻano o ka ʻāpana i kekahi ʻano, e hopena i ka sphericity liʻiliʻi o nā AgNP nui, ua mau nā mea ʻekolu a pau he quasi-spherical.Eia kekahi, e like me ka mea i hōʻike ʻia ma ke ʻano hoʻohālikelike electron ma ke Kiʻi 2B, nano ʻAʻole pili ʻia ka crystallinity o nā ʻāpana.ʻO ke apo ʻokoʻa koʻikoʻi-hiki ke hoʻopili ʻia me ka (111), (220), (200), a me (311) Miller mau helu kālā-ua kūlike loa me ka palapala ʻepekema a me kā mākou mau haʻawina mua.9, 19,44 ʻO ka ʻāpana o ke apo Debye-Scherrer o AgNP-II a me AgNP-III ma muli o ka paʻa ʻana o ke kiʻi ED i ka hoʻonui like, no laila e piʻi aʻe ka nui o ka nui, ka helu o nā ʻāpana ʻokoʻa i kēlā me kēia. piʻi a emi iho ka ʻāpana ʻāpana .
ʻIke ʻia ka nui a me ke ʻano o nā nanoparticles e pili ana i ka hana biological.3,45 Hiki ke ho'ākāka 'ia ka hana catalytic a me ka hana biological e pili ana i ke kino ma muli o ka ho'onui 'ana o nā kinona i kekahi mau maka aniani (he mau helu Miller 'oko'a), a he mau hana 'oko'a ko kēia mau maka aniani.45,46 Ma muli o ka hāʻawi ʻana o nā ʻāpana i hoʻomākaukau ʻia i nā hopena ED e pili ana i nā hiʻohiʻona kristal like loa, hiki ke manaʻo ʻia i loko o kā mākou mau hoʻokolohua colloidal paʻa a me nā hana biological, pono e hoʻopili ʻia nā ʻokoʻa i ʻike ʻia i ka nui Nanoparticle, ʻaʻole nā ​​​​mea pili i ke ʻano.
ʻO nā hualoaʻa UV-Vis i hōʻuluʻulu ʻia ma ka Figure 2D e hoʻokūpaʻa hou i ke ʻano spherical nui o ka AgNP synthesized, no ka mea, ʻo nā piko SPR o nā laʻana ʻekolu ma kahi o 400 nm, ʻo ia ka waiwai o nā nanoparticles kālā spherical.29,30 Ua hoʻokūpaʻa pū ka spectra i hopu ʻia i ka ulu kūleʻa o ka nanosilver.Ke piʻi aʻe ka nui o ka ʻāpana, ʻo ka lōʻihi o ka nalu e pili ana i ka hoʻomaʻamaʻa māmā loa o AgNP-II-ʻoi aku ka nui-E like me ka palapala, AgNP-III Ua ʻike i kahi ʻulaʻula.6,29
E pili ana i ke kūpaʻa colloidal mua o ka ʻōnaehana AgNP, ua hoʻohana ʻia ʻo DLS e ana i ke anawaena hydrodynamic awelika a me ka hiki ke zeta o nā ʻāpana ma ka pH 7.2.Hōʻike nā hopena i hōʻike ʻia ma ka Figure 2E he ʻoi aku ka nui o ka colloidal paʻa o AgNP-III ma mua o AgNP-I a i ʻole AgNP-II, no ka mea, hōʻike nā alakaʻi maʻamau he pono ka zeta o 30 mV pono no ka paʻa colloidal lōʻihi. ua hoʻohālikelike ʻia ka waiwai awelika Z (loaʻa e like me ka awelika hydrodynamic anawaena o nā ʻāpana manuahi a hoʻohui ʻia) me ka nui o ka ʻāpana nui i loaʻa e TEM, no ka mea, ʻoi aku ka kokoke o nā waiwai ʻelua, ʻoi aku ka māmā o ke degere E hōʻiliʻili i ka hāpana.ʻO ka ʻoiaʻiʻo, ʻoi aku ka kiʻekiʻe o ka awelika Z o AgNP-I a me AgNP-II ma mua o kā lākou nui TEM-evaluated particle size, no laila ke hoʻohālikelike ʻia me AgNP-III, ua wānana ʻia kēia mau laʻana e ʻoi aku ka nui o ka hōʻuluʻulu ʻana, kahi o ka zeta maikaʻi loa. hele pū ʻia me ka nui kokoke ʻO ka waiwai awelika Z.
ʻO ka wehewehe ʻana no kēia hanana hiki ke ʻelua.Ma ka ʻaoʻao hoʻokahi, mālama ʻia ka manaʻo citrate ma kahi pae like i nā ʻanuʻu synthesis āpau, e hāʻawi ana i kahi kiʻekiʻe o nā pūʻulu ʻili i hoʻopaʻa ʻia e pale aku i ka ʻāpana kikoʻī o nā ʻāpana ulu mai ka emi ʻana.Eia naʻe, e like me ka Levak et al., hiki ke hoʻololi maʻalahi nā molekole liʻiliʻi e like me ka citrate e nā biomolecules ma ka ʻili o nā nanoparticles.I kēia hihia, e hoʻoholo ʻia ka paʻa colloidal e ke corona o nā biomolecules i hana ʻia.31 No ka mea, ua ʻike ʻia kēia ʻano i loko o kā mākou ana hui ʻana (e kūkākūkā hou ʻia ma hope), ʻaʻole hiki i ka citrate capping wale nō ke wehewehe i kēia ʻano.
Ma ka ʻaoʻao ʻē aʻe, ʻokoʻa ka nui o ka ʻāpana me ke ʻano hoʻohui i ka pae nanometer.Kākoʻo nui ʻia kēia e ke ʻano kuʻuna Derjaguin-Landau-Verwey-Overbeek (DLVO), kahi i wehewehe ʻia ai ka huki ʻana o nā ʻāpana like me ka huina o nā mana hoihoi a hoʻopailua ma waena o nā ʻāpana.Wahi a He et al., ke emi nei ka nui o ka ikehu DLVO me ka nui o ka nanoparticles i loko o ka hematite nanoparticles, e maʻalahi ka hiki i ka ikehu kumu haʻahaʻa loa, a laila e hoʻoikaika i ka hoʻohui ʻana (condensation).47 Eia naʻe, ua manaʻo ʻia aia kekahi mau mea ʻē aʻe ma waho o nā palena o ka manaʻo DLVO.ʻOiai ʻo van der Waals ua like ka gravity a me ka electrostatic double-layer repulsion me ka hoʻonui ʻana i ka nui o nā ʻāpana, kahi loiloi a Hotze et al.ke manaʻo nei he ʻoi aku ka ikaika o ka hoʻohui ʻana ma mua o ka DLVO e ʻae ai.14 Manaʻo lākou ʻaʻole hiki ke koho hou ʻia ka curvature o ka ʻili o nā nanoparticles ma ke ʻano he ʻili palahalaha, ʻaʻole hiki ke hoʻohana ʻia ka manaʻo makemakika.Eia kekahi, i ka emi ʻana o ka nui o nā ʻāpana, ʻoi aku ka kiʻekiʻe o ka pākēneka o nā mana i ka ʻili, e alakaʻi ana i ke ʻano uila a me ke ʻano o ka uku o ka ʻili.A hoʻololi i ka hoʻoulu ʻana o ka ʻili, hiki ke alakaʻi i ka hoʻemi ʻana i ka uku ma ka papa pālua uila a hoʻoikaika i ka hoʻohui.
I ka hoʻohālikelike ʻana i nā hualoaʻa DLS o AgNP-I, AgNP-II, a me AgNP-III ma ke Kiʻi 3, ʻike mākou ua hōʻike ʻia nā hōʻailona ʻekolu a pau i ka pH hoʻouluulu ʻana.Hoʻololi ke kaiapuni ʻakika nui (pH 3) i ka hiki o ka zeta o ka hāpana i 0 mV, e hana ana nā ʻāpana i nā ʻāpana micron-nui, aʻo ka pH alkaline e hoʻololi i kona hiki zeta i kahi waiwai maikaʻi ʻole nui, kahi e hana ai nā ʻāpana i nā hui liʻiliʻi (pH 5. ).A me 7.2) ), a i ʻole e noho ʻole ʻia (pH 9).Ua ʻike pū ʻia kekahi mau ʻokoʻa koʻikoʻi ma waena o nā laʻana like ʻole.I loko o ka hoʻokolohua holoʻokoʻa, ua hōʻike ʻo AgNP-I i ka mea maʻalahi loa i ka pH-induced zeta hiki ke hoʻololi ʻia, no ka mea, ua hoʻemi ʻia ka hiki o ka zeta o kēia mau ʻāpana i ka pH 7.2 i hoʻohālikelike ʻia me ka pH 9, ʻoiai ʻo AgNP-II a me AgNP-III wale nō i hōʻike iā A. ʻO ka hoʻololi nui o ka ζ ma kahi o ka pH 3. Eia kekahi, ua hōʻike ʻo AgNP-II i nā loli lohi a me ka hiki ke zeta maʻalahi, ʻoiai ʻo AgNP-III i hōʻike i ka ʻano haʻahaʻa o nā ʻekolu, no ka mea, ua hōʻike ka ʻōnaehana i ka waiwai zeta kiʻekiʻe loa a me ka neʻe lohi, e hōʻike ana. ʻO AgNP-III ʻoi loa ka pale ʻana i ka pH-induced aggregation.Kūlike kēia mau hualoaʻa me ka awelika hydrodynamic anawaena hopena.Ma muli o ka nui o ka nui o kā lākou mau kumu, ua hōʻike ʻo AgNP-I i ka hōʻuluʻulu mālie mau i nā waiwai pH āpau, ʻoi aku paha ma muli o ka 10 mM NaCl hope, ʻoiai ʻo AgNP-II a me AgNP-III wale nō i hōʻike nui i ka pH 3 o ka hōʻiliʻili ʻana.ʻO ka ʻokoʻa hoihoi loa ʻoiai ʻo kona nui nanoparticle nui, ʻo AgNP-III e hana i nā mea liʻiliʻi loa i ka pH 3 i nā hola 24, e hōʻike ana i kāna mau waiwai anti-aggregation.Ma ka puunaue ana i ka awelika Z o AgNPs ma ka pH 3 ma hope o 24 mau hola e ka waiwai o ka laana i hoomakaukauia, hiki ke ikeia ua pii ka nui o na huina pili o AgNP-I a me AgNP-II i 50 manawa, 42 manawa, a me 22 manawa. , pakahi.III.
Kiʻi 3 ʻO nā hopena hoʻopuehu kukui ikaika o ka citrate-terminated silver nanoparticles sample me ka hoʻonui ʻana i ka nui (10 nm: AgNP-I, 20 nm: AgNP-II a me 50 nm: AgNP-III) i hōʻike ʻia e like me ka awelika hydrodynamic anawaena (Z awelika. ) ('ākau) Ma lalo o nā kūlana pH like'ole, ho'ololi ka zeta (hema) i loko o 24 hola.
Ua hoʻopili pū ʻia ka hōʻuluʻulu ʻana o ka pH i ka ʻano o ka plasmon resonance (SPR) o nā mea hoʻohālike AgNP, e like me ka hōʻike ʻia e kā lākou UV-Vis spectra.Wahi a Supplementary Figure S1, ʻo ka hōʻuluʻulu ʻana o nā mea hoʻokuʻu nanoparticle kālā ʻekolu e ukali ʻia e ka hoʻemi ʻana i ka ikaika o ko lākou mau piko SPR a me kahi hoʻololi ʻulaʻula haʻahaʻa.ʻO ka nui o kēia mau hoʻololi ma ke ʻano he hana o ka pH e kūlike me ke kiʻekiʻe o ka hōʻuluʻulu ʻana i wānana ʻia e nā hopena DLS, akā naʻe, ua ʻike ʻia kekahi mau ʻano hoihoi.Kū'ē i ka intuition, ua hoʻololiʻiaʻo ka AgNP-II nui loa ka mea maʻalahi i nā hoʻololi SPR, aʻo nā mea'ē aʻeʻelua i emi iho ka maʻalahi.Ma ka noiʻi SPR, ʻo 50 nm ka palena nui o ka nui o ka nui, i hoʻohana ʻia e hoʻokaʻawale i nā ʻāpana e pili ana i kā lākou mau waiwai dielectric.Hiki ke wehewehe ʻia nā ʻāpana liʻiliʻi ma mua o 50 nm (AgNP-I a me AgNP-II) ma ke ʻano he dipoles dielectric maʻalahi, ʻoiai ʻo nā ʻāpana i hiki a ʻoi aku paha i kēia palena (AgNP-III) i ʻoi aku ka paʻakikī o nā waiwai dielectric, a me kā lākou resonance Ua māhele ʻia ka band i nā loli multimodal. .I ka hihia o ʻelua mau ʻāpana liʻiliʻi liʻiliʻi, hiki ke manaʻo ʻia nā AgNP he dipoles maʻalahi, a hiki ke hoʻopili maʻalahi ka plasma.Ke piʻi aʻe nei ka nui o ka ʻāpana, hoʻopuka maoli kēia hui ʻana i ka plasma nui, hiki ke wehewehe i ka ʻike kiʻekiʻe i ʻike ʻia.29 Eia naʻe, no nā ʻāpana nui loa, ʻaʻole kūpono ka manaʻo dipole maʻalahi i ka wā e hiki mai ai nā mokuʻāina hoʻohui ʻē aʻe, hiki ke wehewehe i ka manaʻo emi o AgNP-III e hōʻike i nā loli spectral.29
Ma lalo o kā mākou mau kūlana hoʻokolohua, ua hōʻike ʻia he hopena koʻikoʻi ka pH waiwai i ka paʻa colloidal o nā nanoparticles kālā citrate-coated o nā ʻano nui.I loko o kēia mau ʻōnaehana, hāʻawi ʻia ka paʻa e nā hui -COO- i hoʻopiʻi maikaʻi ʻia ma ka ʻili o AgNPs.ʻO ka hui hana carboxylate o ka citrate ion ua protonated i ka nui o nā ion H+, no laila ʻaʻole hiki i ka hui carboxyl i hana ʻia ke hāʻawi hou i ka hoʻihoʻi electrostatic ma waena o nā ʻāpana, e like me ka mea i hōʻike ʻia ma ka lālani kiʻekiʻe o ke Kiʻi 4. E like me ke kumu o Le Chatelier, AgNP hōʻuluʻulu koke nā laʻana i ka pH 3, akā e lilo mālie a paʻa i ka piʻi ʻana o ka pH.
Kiʻi 4 ʻO ke ʻano hoʻolālā o ka launa ʻana o ka ʻili i wehewehe ʻia e ka hōʻuluʻulu ʻana ma lalo o ka pH like ʻole (laina luna), ka ʻike NaCl (laina waena), a me nā biomolecules (laina lalo).
Wahi a ke Kiʻi 5, ua nānā pū ʻia ke kūpaʻa colloidal i nā hoʻokuʻu ʻana o AgNP o nā nui like ʻole ma lalo o ka hoʻonui ʻana i ka paʻakai.Ma muli o ka hiki zeta, hoʻonui hou ka nui nanoparticle i kēia mau ʻōnaehana AgNP i hoʻopau ʻia i ka citrate e hoʻonui i ke kū'ē i nā mana o waho mai NaCl.Ma AgNP-I, lawa ka 10 mM NaCl i mea e hoʻoulu ai i ka hoʻohui ʻoluʻolu, a ʻo ka paʻakai paʻakai o 50 mM hāʻawi i nā hopena like loa.Ma AgNP-II a me AgNP-III, 10 mM NaCl ʻaʻole i pili nui i ka hiki zeta no ka mea e mau ana ko lākou mau waiwai ma (AgNP-II) a i lalo (AgNP-III) -30 mV.ʻO ka hoʻonui ʻana i ka manaʻo o ka NaCl i 50 mM a i ka 150 mM NaCl ua lawa ia e hōʻemi nui i ka waiwai paʻa o ka hiki i ka zeta i nā laʻana a pau, ʻoiai ʻo nā ʻāpana nui e paʻa i ka uku maikaʻi ʻole.Kūlike kēia mau hopena me ke anawaena hydrodynamic awelika i manaʻo ʻia o nā AgNP;ʻO nā laina maʻamau Z i ana ʻia ma 10, 50, a me 150 mM NaCl hōʻike ʻokoʻa, e hoʻonui mālie ana i nā waiwai.ʻO ka hope, ua ʻike ʻia nā hōʻuluʻulu micron-nui ma nā hoʻokolohua 150 mM ʻekolu.
Kiʻi 5 ʻO nā hopena hoʻopuehu kukui ikaika o ka citrate-terminated silver nanoparticles sample me ka hoʻonui ʻana i ka nui (10 nm: AgNP-I, 20 nm: AgNP-II a me 50 nm: AgNP-III) i hōʻike ʻia e like me ka awelika hydrodynamic anawaena (Z awelika. ) ('ākau) a me ka hiki zeta (hema) hoʻololi i loko o 24 hola ma lalo o nā ʻokoʻa NaCl.
Hōʻike nā hopena UV-Vis i ke Kiʻi S2 hou i ka SPR o 50 a me 150 mM NaCl i nā laʻana ʻekolu he emi koke a nui.Hiki ke wehewehe ʻia kēia e DLS, no ka mea, ʻoi aku ka wikiwiki o ka hoʻohui ʻana o NaCl ma mua o nā hoʻokolohua pili i ka pH, i wehewehe ʻia e ka ʻokoʻa nui ma waena o nā ana mua (0, 1.5, a me 3 mau hola).Eia hou, e hoʻonui ka hoʻonui ʻana i ka paʻakai e hoʻonui i ka permittivity pili o ka mea hoʻokolohua, kahi e loaʻa ai kahi hopena nui i ka resonance plasmon surface.29
Ua hōʻuluʻulu ʻia ka hopena o ka NaCl i ka lālani waena o ke Kiʻi 4. Ma ke ʻano laulā, hiki ke hoʻoholo ʻia ʻo ka hoʻonui ʻana i ka sodium chloride ka hopena like e like me ka hoʻonui ʻana i ka acidity, no ka mea, loaʻa i nā ion Na + ka hoʻonohonoho ʻana a puni nā hui carboxylate. ke kāohi nei i nā AgNP hiki ʻole i ka zeta.Eia kekahi, 150 mM NaCl i hoʻopuka i ka micron-sized aggregates i loko o nā hōʻailona ʻekolu, e hōʻike ana he mea hōʻino ka physiological electrolyte i ka paʻa colloidal o nā AgNPs citrate-terminated.Ma ka noʻonoʻo ʻana i ka manaʻo condensing koʻikoʻi (CCC) o NaCl ma nā ʻōnaehana AgNP like, hiki ke kau ʻia kēia mau hopena i loko o nā palapala pili.Huynh et al.helu 'ia ka CCC o NaCl no ka citrate-hoopau kala nanoparticles me ka awelika anawaena o 71 nm he 47.6 mM, oiai El Badawy et al.ʻike ʻia ka CCC o 10 nm AgNPs me ka citrate coating he 70 mM.10,16 Eia kekahi, ʻo ka CCC kiʻekiʻe kiʻekiʻe ma kahi o 300 mM ua ana ʻia e He et al., ʻo ia ka mea i ʻokoʻa ai kā lākou ʻano hana synthesis mai ka paʻi i haʻi ʻia ma mua.48 ʻOiai ʻaʻole i manaʻo ʻia ka hāʻawi ʻana i kēia manawa i ka loiloi piha ʻana o kēia mau waiwai, no ka mea ke piʻi nei kā mākou mau hoʻokolohua i ka paʻakikī o ka haʻawina holoʻokoʻa, ʻike ʻia ke kiʻekiʻe o ka naʻau NaCl pili i ka biologically o 50 mM, ʻoi aku ka 150 mM NaCl.Hoʻokomo ʻia ka coagulation, e wehewehe ana i nā loli ikaika i ʻike ʻia.
ʻO ka hana hou aʻe i ka hoʻokolohua polymerization ʻo ia ka hoʻohana ʻana i nā molekole maʻalahi akā pili i ka biologically e hoʻohālikelike i nā pilina nanoparticle-biomolecule.Ma muli o ka DLS (Nā Kiʻi 6 a me 7) a me nā hopena o UV-Vis (Nā Kiʻi Pākuʻi S3 a me S4), hiki ke hōʻoia i kekahi mau hopena maʻamau.Ma lalo o kā mākou mau kūlana hoʻokolohua, ʻaʻole e hoʻoulu ʻia nā molekele i aʻo ʻia ka glucose a me ka glutamine i ka hoʻohui ʻana i kekahi ʻōnaehana AgNP, no ka mea, pili loa ke ʻano Z-mean i ka waiwai ana o ke ana.ʻOiai ʻaʻole pili ko lākou hele ʻana i ka hōʻuluʻulu ʻana, hōʻike nā hopena hoʻokolohua e hoʻopili ʻia kēia mau molekala ma ka ʻili o AgNPs.ʻO ka hopena koʻikoʻi e kākoʻo ana i kēia manaʻo, ʻo ia ka loli i ʻike ʻia i ka lawe māmā ʻana.ʻOiai ʻaʻole hōʻike ʻo AgNP-I i ka lōʻihi o ka nalu a i ʻole ka hoʻololi ʻana i ka ikaika, hiki ke ʻike maopopo ʻia ma ke ana ʻana i nā ʻāpana nui, ʻo ia ka mea ma muli o ka ʻoi aku o ka ʻike optical i ʻōlelo ʻia ma mua.Ma waho aʻe o ka manaʻo, hiki i ka glucose ke hoʻololi i ka ʻulaʻula nui aʻe ma hope o 1.5 mau hola i hoʻohālikelike ʻia me ke ana mana, ʻo ia ka 40 nm ma AgNP-II a ma kahi o 10 nm i AgNP-III, e hōʻike ana i ka hiki ʻana mai o nā pilina ma luna.Ua hōʻike ʻo Glutamine i kahi ʻano like, akā ʻaʻole maopopo ka loli.Eia kekahi, pono e haʻi ʻia e hiki i ka glutamine ke hōʻemi i ka hiki ʻana o ka zeta o nā ʻāpana waena a me nā mea nui.Eia naʻe, no ka mea ʻaʻole pili kēia mau hoʻololi zeta i ka pae hōʻuluʻulu, hiki ke manaʻo ʻia ʻo nā biomolecules liʻiliʻi e like me ka glutamine hiki ke hāʻawi i kahi kiʻekiʻe o ka repulsion spatial ma waena o nā ʻāpana.
Kiʻi 6 ʻO nā hopena hoʻopuehu kukui ikaika o nā mea nanoparticle kālā i hoʻopau ʻia i ka citrate me ka hoʻonui nui ʻana (10 nm: AgNP-I, 20 nm: AgNP-II a me 50 nm: AgNP-III) i hōʻike ʻia e like me ka awelika hydrodynamic anawaena (Z awelika) ('ākau) Ma lalo o nā kūlana kūwaho o nā ʻokoʻa glucose ʻokoʻa, hoʻololi ka hiki zeta (hema) i loko o 24 mau hola.
Kiʻi 7 ʻO nā hopena hoʻopuehu kukui ikaika o ka citrate-terminated silver nanoparticles sample me ka hoʻonui ʻana i ka nui (10 nm: AgNP-I, 20 nm: AgNP-II a me 50 nm: AgNP-III) i hōʻike ʻia e like me ka awelika hydrodynamic anawaena (Z awelika ) ('ākau) Ma ke alo o ka glutamine, hiki ke loli ka zeta (hema) i loko o 24 mau hola.
I ka pōkole, ʻaʻole pili nā biomolecules liʻiliʻi e like me ka glucose a me ka glutamine i ke kūpaʻa colloidal i ke ana ʻana: ʻoiai e pili ana lākou i ka hiki o ka zeta a me nā hopena UV-Vis i nā pae like ʻole, ʻaʻole kūlike nā hopena awelika Z.Hōʻike kēia i ka adsorption o ka ʻili o nā molekole e kāohi i ka repulsion electrostatic, akā i ka manawa like e hāʻawi i ke kūpaʻa dimensional.
I mea e hoʻopili ai i nā hualoaʻa mua me nā hualoaʻa mua a hoʻohālikelike i nā kūlana olaola me ka mākaukau, ua koho mākou i kekahi o nā mea hoʻohana maʻamau o ka moʻomeheu cell a hoʻohana iā lākou i mau kūlana hoʻokolohua no ke aʻo ʻana i ke kūpaʻa o nā colloids AgNP.I loko o ka hoʻokolohua in vitro holoʻokoʻa, ʻo kekahi o nā hana koʻikoʻi o DMEM ma ke ʻano he ʻano mea e hoʻokumu i nā kūlana osmotic e pono ai, akā mai kahi ʻano kemika, he paʻakai paʻakai paʻakikī me ka ikaika ionic āpau e like me 150 mM NaCl. .40 No ka FBS, he hui paʻakikī ia o nā biomolecules-ʻoi aku ka nui o nā protein-mai ka ʻike o ka adsorption o ka ʻili, aia kekahi mau mea like me nā hopena hoʻokolohua o ka glucose a me ka glutamine, ʻoiai ʻoi aku ka paʻakikī o ka hui ʻana a me ka ʻokoʻa.19 DLS a me UV- Hiki ke wehewehe ʻia nā hopena ʻike ʻia i hōʻike ʻia ma ke Kiʻi 8 a me ke Kiʻi S5 ma ka nānā ʻana i ka haku mele ʻana o kēia mau mea a me ka hoʻopili ʻana iā lākou me nā ana ma ka pauku mua.
Kiʻi 8 ʻO nā hopena hoʻopuehu kukui ikaika o ka citrate-terminated silver nanoparticles sample me ka hoʻonui ʻana i ka nui (10 nm: AgNP-I, 20 nm: AgNP-II a me 50 nm: AgNP-III) i hōʻike ʻia e like me ka awelika hydrodynamic anawaena (Z awelika ) ('ākau) Ma ke alo o nā mea moʻomeheu cell DMEM a me FBS, hiki ke hoʻololi i ka zeta (hema) i loko o 24 mau hola.
ʻO ka hoʻoheheʻe ʻana o nā AgNP o nā ʻano nui like ʻole i DMEM he hopena like ia i ka paʻa colloidal i ka mea i ʻike ʻia ma ke alo o nā kiʻekiʻe NaCl.Ua hōʻike ʻia ka hoʻopuehu ʻana o AgNP ma 50 v/v% DMEM ua ʻike ʻia ka hōʻuluʻulu nui me ka piʻi ʻana o ka mana zeta a me ka waiwai Z-average a me ka emi ʻana o ka ikaika SPR.Pono e hoʻomaopopo ʻia ʻo ka nui aggregate nui i hoʻokomo ʻia e DMEM ma hope o 24 mau hola he inversely proportional i ka nui o nā nanoparticles primer.
ʻO ka pilina ma waena o FBS a me AgNP e like me ka mea iʻikeʻia ma ke alo o nā mole liʻiliʻi e like me ka glucose a me ka glutamine, akāʻoi aku ka ikaika o ka hopena.ʻAʻole paʻa ka awelika Z o nā ʻāpana, ʻoiai ke ʻike ʻia ka piʻi ʻana o ka hiki ke zeta.Hōʻike ka piko SPR i kahi ʻulaʻula liʻiliʻi, akā ʻoi aku ka hoihoi, ʻaʻole i emi iki ka ikaika o ka SPR e like me ke ana mana.Hiki ke wehewehe ʻia kēia mau hualoaʻa e ka adsorption innate o nā macromolecules ma ka ʻili o nā nanoparticles (laina lalo i ka Figure 4), i kēia manawa i hoʻomaopopo ʻia ʻo ke ʻano o ka biomolecular corona i loko o ke kino.49