Xov xwm

Ua tsaug rau koj tuaj xyuas Nature.com.Qhov browser version koj siv tau txwv CSS kev txhawb nqa.Rau qhov kev paub zoo tshaj plaws, peb xav kom koj siv qhov browser tshiab (lossis lov tes taw Compatibility Hom hauv Internet Explorer).Nyob rau lub sijhawm no, txhawm rau ua kom muaj kev txhawb nqa txuas ntxiv, peb yuav ua rau lub xaib tsis muaj qauv thiab JavaScript.
Cov tshuaj photosensitizers muaj txiaj ntsig tshwj xeeb yog qhov tseem ceeb rau kev siv tshuaj kho mob dav dav.Txawm li cas los xij, cov khoom siv photosensitizers feem ntau raug kev txom nyem los ntawm kev nqus lub wavelength luv, tsis txaus photostability, qis quantum yield ntawm reactive oxygen hom (ROS), thiab aggregation-induced quenching ntawm ROS.Ntawm no peb qhia txog qhov ze-infrared (NIR) supramolecular photosensitizer (RuDA) kho los ntawm tus kheej ntawm Ru(II)-arene organometallic complexes hauv cov tshuaj aqueous.RuDA tsuas tuaj yeem tsim cov pa oxygen (1O2) nyob rau hauv lub xeev sib sau ua ke, thiab nws pom tseeb kev sib sau ua ke vim yog 1O2 tiam tus cwj pwm vim muaj kev nce ntxiv hauv cov txheej txheem crossover ntawm singlet-triplet system.Raws li qhov kev txiav txim ntawm 808 nm laser teeb, RuDA nthuav tawm 1O2 quantum tawm los ntawm 16.4% (FDA-pom zoo indocyanine ntsuab: ΦΔ = 0.2%) thiab siab photothermal conversion efficiency ntawm 24.2% (kev lag luam kub nanorods) nrog zoo heev photostability.21.0%, kub nanoshells: 13.0%).Tsis tas li ntawd, RuDA-NPs nrog cov biocompatibility zoo tuaj yeem ua kom zoo dua ntawm cov qog nqaij hlav, ua rau cov qog nqaij hlav tseem ceeb thaum lub sij hawm kho photodynamic nrog 95.2% txo cov qog ntim hauv vivo.Qhov kev sib sau ua ke-txhim kho photodynamic txoj kev kho no muab ib lub tswv yim rau kev tsim cov photosensitizers nrog cov duab zoo nkauj thiab photochemical.
Piv nrog rau cov kev kho ib txwm siv, photodynamic therapy (PDT) yog ib qho kev kho mob zoo rau mob qog noj ntshav vim nws cov txiaj ntsig tseem ceeb xws li kev tswj hwm spatiotemporal, tsis muaj kev cuam tshuam, tsis muaj tshuaj tiv thaiv, thiab txo qis ntawm cov kev mob tshwm sim 1,2,3.Nyob rau hauv lub teeb irradiation, cov photosensitizers siv tau tuaj yeem qhib kom tsim cov pa oxygen ntau heev (ROS), ua rau apoptosis / necrosis lossis tiv thaiv kab mob 4,5. Txawm li cas los xij, feem ntau cov khoom siv hluav taws xob zoo li qub, xws li chlorins, porphyrins, thiab anthraquinones, muaj qhov nqus dej luv luv (frequency < 680 nm), yog li ua rau lub teeb pom kev tsis zoo vim yog kev nqus ntawm cov roj ntsha molecules (xws li hemoglobin thiab melanin). pom thaj tsam 6,7. Txawm li cas los xij, feem ntau cov khoom siv hluav taws xob zoo li qub, xws li chlorins, porphyrins, thiab anthraquinones, muaj qhov nqus dej luv luv (frequency < 680 nm), yog li ua rau lub teeb pom kev tsis zoo vim yog kev nqus ntawm cov roj ntsha molecules (xws li hemoglobin thiab melanin). pom thaj tsam 6,7. Однако большинство обычных фотосенсибилизаторов, таких как хлорины, порфирины и антрахиноны, обладают относительно коротковолновым поглощением (частота < 680 нм), что приводит к плохому проникновению света из-за интенсивного поглощения биологических молекул (например, гемоглобина и меланина) в видимая область6,7. Txawm li cas los xij, feem ntau cov photosensitizers xws li chlorins, porphyrins thiab anthraquinones muaj qhov luv wavelength absorption (< 680 nm) uas ua rau lub teeb pom kev tsis zoo vim yog kev nqus ntawm cov roj ntsha molecules (xws li hemoglobin thiab melanin) rau hauv thaj tsam pom 6,7.然而, 大多数 传统 光敏剂 光敏剂, 如如 氢, 卟啉卟啉 卟酚, 卟啉卟啉 蒽醌, 具有具有 较蒽醌 短蒽醌 的波长 的吸收 吸收吸收 吸收 (频率 <680 nm), 血红 蛋白 蛋白 黑色素) 的 强烈 黑色素) 的 强烈 吸收,导致光穿透性差.然而, 大多数 传统 光敏剂, 二二 卟酚, 卟啉 蒽醌, 卟啉, 具有 短蒽醌 短蒽醌 的蒽醌 的波长 吸收 (频率 频率 <对 分子 分子 分子) 的 黑色素 黑色素) 的 ,,,, 吸收 吸收吸收 吸收 吸收 吸收 HI导致光穿透性差. Однако большинство традиционных фотосенсибилизаторов, таких как хлорины, порфирины и антрахиноны, имеют относительно коротковолновое поглощение (частота < 680 нм) из-за сильного поглощения биомолекул, таких как гемоглобин и меланин, что приводит к плохому проникновению света. Txawm li cas los xij, feem ntau cov tshuaj photosensitizers xws li chlorins, porphyrins thiab anthraquinones muaj qhov luv wavelength absorption (frequency < 680 nm) vim muaj zog nqus ntawm biomolecules xws li hemoglobin thiab melanin ua rau lub teeb tsis zoo.Pom thaj tsam 6.7.Yog li ntawd, nyob ze-infrared (NIR) absorbing photosensitizers uas tau qhib rau hauv 700-900 nm "kho qhov rais" yog zoo suited rau kev kho mob phototherapy.Txij li thaum nyob ze lub teeb infrared yog qhov tsawg tshaj plaws absorbed los ntawm cov ntaub so ntswg lom, nws tuaj yeem ua rau nkag mus tob dua thiab tsawg dua photodamage8,9.
Hmoov tsis zoo, uas twb muaj lawm NIR-absorbing photosensitizers feem ntau muaj cov duab tsis zoo, tsis tshua muaj oxygen singlet (1O2) generating peev, thiab aggregation-induced 1O2 quenching, uas txwv lawv cov kev kho mob daim ntawv thov 10,11.Txawm hais tias muaj kev siv zog zoo los txhim kho cov duab hluav taws xob thiab cov khoom siv hluav taws xob ntawm cov pa photosensitizers, txog tam sim no ntau cov ntaub ntawv tau tshaj tawm tias NIR-absorbing photosensitizers tuaj yeem daws tag nrho cov teeb meem no.Tsis tas li ntawd, ntau cov photosensitizers tau pom cov lus cog tseg rau kev ua tau zoo ntawm 1O212,13,14 thaum irradiated nrog lub teeb siab tshaj 800 nm, txij li lub zog photon txo qis hauv thaj tsam ze-IR.Triphenylamine (TFA) raws li ib tug electron pub thiab [1,2,5]thiadiazole-[3,4-i]dipyrido[a,c]phenazine (TDP) raws li ib tug electron acceptor pab pawg Donor-acceptor (DA) hom dyes ib chav kawm ntawm dyes, absorbing ze-infrared, uas tau kawm ntau heev rau ze-infrared bioimaging II thiab photothermal therapy (PTT) vim lawv nqaim bandgap.Yog li, DA-type dyes tuaj yeem siv rau PDT nrog ze-IR excitation, txawm hais tias lawv tsis tshua muaj kev kawm ua cov duab rau PDT.
Nws paub zoo tias kev ua haujlwm siab ntawm intersystem hla (ISC) ntawm photosensitizers txhawb kev tsim ntawm 1O2.Ib lub tswv yim zoo rau kev ua kom cov txheej txheem ISC yog txhawm rau txhim kho qhov sib txuas-orbit coupling (SOC) ntawm photosensitizers los ntawm kev qhia hnyav atoms lossis tshwj xeeb cov organic moieties.Txawm li cas los xij, txoj hauv kev no tseem muaj qee qhov tsis zoo thiab kev txwv 19,20.Tsis ntev los no, supramolecular self-assembly tau muab cov kev txawj ntse hauv qab rau kev tsim cov ntaub ntawv ua haujlwm ntawm qib molecular, 21,22 nrog ntau qhov zoo hauv kev kho duab: (1) cov duab thaij duab tus kheej tuaj yeem muaj peev xwm tsim cov qauv ribbon.Zoo ib yam li cov qauv siv hluav taws xob nrog kev sib faib denser ntawm qib zog vim muaj kev sib tshooj ntawm lub tsev thaiv.Yog li ntawd, lub zog sib tw ntawm lub qis singlet zoo siab xeev (S1) thiab cov neeg nyob sib ze triplet zoo siab xeev (Tn) yuav raug txhim kho, uas muaj txiaj ntsig zoo rau cov txheej txheem ISC 23, 24.(2) Supramolecular los ua ke yuav txo qhov tsis muaj hluav taws xob so raws li cov txheej txheem intramolecular motion limitation mechanism (RIM), uas tseem txhawb nqa cov txheej txheem ISC 25, 26.(3) Lub rooj sib txoos supramolecular tuaj yeem tiv thaiv cov molecules sab hauv ntawm lub monomer los ntawm oxidation thiab degradation, yog li txhim kho cov duab zoo ntawm cov photosensitizer.Muab cov txiaj ntsig saum toj no, peb ntseeg tias cov tshuab supramolecular photosensitizer tuaj yeem yog lwm txoj hauv kev zoo los kov yeej qhov tsis txaus ntawm PDT.
Ru(II)-raws li complexes yog ib qho kev cog lus kho mob platform rau cov kev siv hauv kev kuaj mob thiab kev kho mob ntawm cov kab mob vim lawv cov yam ntxwv tshwj xeeb thiab ntxim nyiam lom zem28,29,30,31,32,33,34.Tsis tas li ntawd, kev nplua nuj ntawm cov xeev zoo siab thiab cov khoom siv photophysicochemical tunable ntawm Ru(II)-based complexes muab cov txiaj ntsig zoo rau kev txhim kho ntawm Ru(II)-raws li photosensitizers35,36,37,38,39,40.Ib qho piv txwv tseem ceeb yog ruthenium(II) polypyridyl complex TLD-1433, uas yog tam sim no nyob rau hauv Phase II kev sim tshuaj raws li ib tug photosensitizer rau kev kho mob ntawm non-muscle invasive zais zis cancer (NMIBC)41.Tsis tas li ntawd, ruthenium (II) areene organometallic complexes tau dav siv los ua cov tshuaj chemotherapeutic rau kev kho mob qog noj ntshav vim lawv cov toxicity tsawg thiab yooj yim ntawm kev hloov kho42,43,44,45.Cov khoom ionic ntawm Ru(II)-arene organometallic complexes tsis tuaj yeem txhim kho cov neeg pluag solubility ntawm DA chromophores hauv cov kuab tshuaj, tab sis kuj txhim kho kev sib dhos ntawm DA chromophores.Tsis tas li ntawd, pseudooctahedral ib nrab-sandwich qauv ntawm organometallic complexes ntawm Ru (II)-arenes tuaj yeem tiv thaiv H-aggregation ntawm DA-hom chromophores, yog li ua kom yooj yim tsim J-aggregation nrog redshifted absorption bands.Txawm li cas los xij, qhov tsis zoo ntawm Ru(II)-arene complexes, xws li tsis tshua muaj kev ruaj ntseg thiab/lossis bioavailability tsis zoo, tuaj yeem cuam tshuam rau kev kho mob thiab kev ua haujlwm vivo ntawm arene-Ru(II) complexes.Txawm li cas los xij, cov kev tshawb fawb tau pom tias cov kev tsis zoo no tuaj yeem kov yeej los ntawm encapsulating ruthenium complexes nrog biocompatible polymers los ntawm lub cev encapsulation los yog covalent conjugation.
Hauv txoj haujlwm no, peb tshaj tawm DA-conjugated complexes ntawm Ru(II)-arene (RuDA) nrog NIR trigger los ntawm kev sib koom ua ke ntawm DAD chromophore thiab Ru(II)-arene moiety.Cov complexes uas ua rau nws tus kheej tuaj yeem sib sau ua ke rau hauv metalosupramolecular vesicles hauv dej vim tsis muaj kev sib cuam tshuam.Qhov tseem ceeb, lub rooj sib txoos supramolecular endowed RuDA nrog polymerization-induced intersystem hla-dhau cov khoom, uas ua rau ISC ua tau zoo, uas zoo heev rau PDT (Fig. 1A).Txhawm rau nce qog nqaij hlav thiab hauv vivo biocompatibility, FDA-pom zoo Pluronic F127 (PEO-PPO-PEO) tau siv los encapsulate RuDA47,48,49 los tsim RuDA-NP nanoparticles (Daim duab 1B) uas ua raws li PDT / Dual- hom PTT npe.Hauv kev kho mob qog noj ntshav (Daim duab 1C), RuDA-NP tau siv los kho cov nas liab qab nrog MDA-MB-231 qog los kawm txog kev ua tau zoo ntawm PDT thiab PTT hauv vivo.
Schematic illustration of the photophysical mechanism of RuDA in monomeric and aggregated forms for cancer phototherapy, synthesis of B RuDA-NPs and C RuDA-NPs for NIR-activated PDT and PTT.
RuDA, muaj xws li TPA thiab TDP functionality, tau npaj raws li cov txheej txheem qhia nyob rau hauv Ntxiv daim duab 1 (Daim duab 2A), thiab RuDA yog yus muaj los ntawm 1H thiab 13C NMR spectra, electrospray ionization mass spectrometry, thiab lub ntsiab tsom xam (Cov duab ntxiv 2-4. ).RuDA electron ceev sib txawv daim ntawv qhia ntawm qhov qis tshaj singlet hloov tau suav los ntawm lub sij hawm-dependent density functional theory (TD-DFT) los kawm txog tus nqi hloov pauv.Raws li pom nyob rau hauv Ntxiv daim duab 5, lub electron ceev drifts mas los ntawm triphenylamine mus rau lub TDP txais chav tsev tom qab photoexcitation, uas yuav raug ntaus nqi mus rau ib tug raug intramolecular them nqi hloov (CT) hloov.
Cov qauv tshuaj ntawm Ore B Absorption spectra ntawm Ore hauv kev sib xyaw ntawm ntau qhov sib piv ntawm DMF thiab dej.C Cov txiaj ntsig zoo nqus ntawm RuDA (800 nm) thiab ICG (779 nm) piv rau lub sijhawm ntawm 0.5 W cm-2 ntawm 808 nm laser teeb.D Lub photodegradation ntawm ABDA yog qhia los ntawm RuDA-induced tsim ntawm 1O2 hauv DMF / H2O sib tov nrog cov dej sib txawv nyob rau hauv qhov kev txiav txim ntawm laser hluav taws xob nrog lub wavelength ntawm 808 nm thiab lub zog ntawm 0.5 W / cm2.
Abstract-UV-pom pom kev nqus spectroscopy tau siv los kawm txog tus kheej cov khoom sib dhos ntawm Ore hauv kev sib xyaw ntawm DMF thiab dej hauv ntau qhov sib piv.Raws li qhia hauv daim duab.2B, RuDA nthuav tawm cov khoom nqus nqus ntawm 600 txog 900 nm hauv DMF nrog qhov siab tshaj plaws nqus ntawm 729 nm.Kev nce cov dej tau ua rau maj mam hloov pauv ntawm Ore nqus siab tshaj plaws mus rau 800 nm, uas qhia tias J-aggregation ntawm Ore hauv cov txheej txheem sib dhos.Lub photoluminescence spectra ntawm RuDA nyob rau hauv cov kuab tshuaj sib txawv yog qhia nyob rau hauv Ntxiv daim duab 6. RuDA zoo nkaus li nthuav tawm cov NIR-II luminescence nrog lub siab tshaj plaws emission wavelength ntawm ca.1050 nm hauv CH2Cl2 thiab CH3OH, feem.Qhov loj Stokes hloov pauv (txog 300 nm) ntawm RuDA qhia txog kev hloov pauv tseem ceeb hauv geometry ntawm lub xeev zoo siab thiab tsim cov xeev qis zog.Lub luminescence quantum yields ntawm Ore hauv CH2Cl2 thiab CH3OH tau txiav txim siab yog 3.3 thiab 0.6%, feem.Txawm li cas los xij, hauv kev sib xyaw ntawm methanol thiab dej (5/95, v / v), me ntsis redshift ntawm emission thiab txo qis hauv quantum yield (0.22%) tau pom, uas tej zaum yuav yog vim tus kheej ntawm Ore. .
Txhawm rau pom tus kheej ntawm ORE, peb siv cov kua atomic force microscopy (AFM) kom pom cov kev hloov pauv ntawm ORE hauv cov tshuaj methanol tom qab ntxiv dej.Thaum cov dej hauv qab no qis dua 80%, tsis pom muaj kev sib sau ua ke (Cov duab ntxiv 7).Txawm li cas los xij, nrog kev nce ntxiv hauv cov ntsiab lus dej mus rau 90-95%, me me nanoparticles tshwm sim, uas qhia tau hais tias tus kheej ntawm Ore. kev daws teeb meem (Fig. 2C thiab Supplementary Fig. 8).Nyob rau hauv sib piv, lub absorbance ntawm indocyanine ntsuab (ICG raws li kev tswj) poob sai sai ntawm 779 nm, qhia zoo heev photostability ntawm RuDA.Tsis tas li ntawd, kev ruaj ntseg ntawm RuDA-NPs hauv PBS (pH = 5.4, 7.4 thiab 9.0), 10% FBS thiab DMEM (siab qabzib) raug tshuaj xyuas los ntawm UV-pom pom kev nqus spectroscopy ntawm ntau lub sijhawm.Raws li pom nyob rau hauv Ntxiv Daim duab 9, kev hloov me ntsis ntawm RuDA-NP nqus bands tau pom nyob rau hauv PBS ntawm pH 7.4/9.0, FBS thiab DMEM, qhia tau hais tias zoo heev stability ntawm RuDA-NP.Txawm li cas los xij, hauv qhov nruab nrab acidic (рН = 5.4) hydrolysis ntawm Ore tau pom.Peb kuj tau soj ntsuam ntxiv kev ruaj ntseg ntawm RuDA thiab RuDA-NP siv cov kev ua haujlwm siab ua kua chromatography (HPLC).Raws li pom nyob rau hauv Ntxiv daim duab 10, RuDA yog ruaj khov nyob rau hauv ib tug sib tov ntawm methanol thiab dej (50/50, v / v) rau thawj teev, thiab hydrolysis tau soj ntsuam tom qab 4 teev.Txawm li cas los xij, tsuas yog qhov dav concave-convex ncov tau pom rau RuDA NPs.Yog li, gel permeation chromatography (GPC) tau siv los ntsuas qhov ruaj khov ntawm RuDA NPs hauv PBS (pH = 7.4).Raws li pom nyob rau hauv Ntxiv daim duab 11, tom qab 8 teev ntawm incubation nyob rau hauv lub xeem tej yam kev mob, lub ncov siab, ncov dav thiab ncov cheeb tsam ntawm NP RuDA tsis hloov loj, qhia tau hais tias zoo heev stability ntawm NP RuDA.Tsis tas li ntawd, TEM cov duab qhia tau hais tias lub morphology ntawm RuDA-NP nanoparticles tseem zoo li tsis hloov tom qab 24 teev nyob rau hauv diluted PBS tsis (pH = 7.4, Supplementary Fig. 12).
Vim tias nws tus kheej tuaj yeem sib tham txog kev ua haujlwm thiab tshuaj lom neeg sib txawv ntawm Ore, peb tau pom qhov tso tawm ntawm 9,10-anthracenediylbis (methylene) dimalonic acid (ABDA, qhov taw qhia 1O2) hauv methanol-dej sib tov.Ore nrog dej sib txawv50.Raws li pom nyob rau hauv daim duab 2D thiab ntxiv daim duab 13, tsis muaj degradation ntawm ABDA tau pom thaum cov dej ntsiab lus qis dua 20%.Nrog nce av noo mus rau 40%, ABDA degradation tshwm sim, raws li pov thawj los ntawm kev txo qis ntawm ABDA fluorescence.Nws kuj tau pom tias cov ntsiab lus dej siab dua ua rau muaj kev puas tsuaj sai dua, qhia tias RuDA kev sib dhos tus kheej yog qhov tsim nyog thiab muaj txiaj ntsig zoo rau ABDA degradation.Qhov tshwm sim no txawv heev ntawm ACQ niaj hnub (kev sib sau ua ke-vim quenching) chromophores.Thaum irradiated nrog ib tug laser nrog ib tug wavelength ntawm 808 nm, lub quantum tawm los ntawm 1O2 RuDA nyob rau hauv ib tug sib tov ntawm 98% H2O / 2% DMF yog 16.4%, uas yog 82 lub sij hawm siab tshaj ntawm ICG (ΦΔ = 0.2%)51, Ua kom pom qhov ua tau zoo tshaj plaws tiam 1O2 RuDA hauv lub xeev ntawm kev sib sau.
Electron spinning siv 2,2,6,6-tetramethyl-4-piperidinone (TEMP) thiab 5,5-dimethyl-1-pyrroline N-oxide (DMPO) raws li spin traps Resonance spectroscopy (ESR) tau siv los txheeb xyuas cov txiaj ntsig ntawm hom AFK.los ntawm RuDA.Raws li pom nyob rau hauv Ntxiv Daim duab 14, nws tau raug lees paub tias 1O2 yog tsim los ntawm irradiation lub sij hawm ntawm 0 thiab 4 feeb.Tsis tas li ntawd, thaum RuDA tau incubated nrog DMPO nyob rau hauv irradiation, ib tug raug plaub-kab EPR teeb liab ntawm 1: 2: 2: 1 DMPO-OH · adduct raug kuaj pom, qhia qhov tsim ntawm hydroxyl radicals (OH·).Zuag qhia tag nrho, cov txiaj ntsig saum toj no qhia tau tias muaj peev xwm ntawm RuDA los txhawb ROS ntau lawm los ntawm cov txheej txheem dual hom I / II photosensitization.
Txhawm rau kom nkag siab zoo dua cov khoom siv hluav taws xob ntawm RuDA hauv monomeric thiab cov ntaub ntawv sib sau ua ke, frontier molecular orbitals ntawm RuDA hauv monomeric thiab dimeric cov ntaub ntawv raug suav siv txoj kev DFT.Raws li qhia hauv daim duab.3A, qhov siab tshaj plaws nyob hauv molecular orbital (HOMO) ntawm monomeric RuDA yog delocalized raws li ligand caj qaum thiab qhov qis tshaj plaws uas tsis muaj nyob hauv molecular orbital (LUMO) yog nyob rau ntawm TDP txais chav tsev.Ntawm qhov tsis sib xws, electron ceev hauv dimeric HOMO yog concentrated nyob rau hauv ligand ntawm ib tug RuDA molecule, thaum lub electron ceev nyob rau hauv lub LUMO yog mas concentrated nyob rau hauv lub receiver unit ntawm lwm RuDA molecule, uas qhia tias RuDA nyob rau hauv lub dimer.Feature ntawm CT.
Lub HOMO thiab LUMO ntawm Ore yog xam nyob rau hauv monomeric thiab dimeric daim ntawv.B Singlet thiab triplet zog theem ntawm Ore hauv monomers thiab dimers.C kwv yees theem ntawm RuDA thiab ua tau ISC raws li monomeric C thiab dimeric D. Xub qhia tau ISC raws.
Kev faib tawm ntawm cov hluav taws xob thiab qhov nyob rau hauv qhov qis-zog singlet zoo siab xeev ntawm RuDA nyob rau hauv cov ntaub ntawv monomeric thiab dimeric tau soj ntsuam siv Multiwfn 3.852.53 software, uas tau suav nrog txoj kev TD-DFT.Raws li tau teev nyob rau ntawm daim ntawv lo ntxiv.Raws li pom nyob rau hauv daim duab 1-2, monomeric RDA qhov feem ntau delocalized raws li ligand caj qaum nyob rau hauv cov singlet zoo siab lub xeev, thaum electrons feem ntau nyob rau hauv pawg TDP, qhia lub intramolecular yam ntxwv ntawm CT.Tsis tas li ntawd, rau cov xeev singlet zoo siab, muaj ntau dua los yog tsawg dua sib tshooj ntawm lub qhov thiab cov hluav taws xob, qhia tias cov singlet zoo siab lub xeev ua rau qee qhov kev koom tes los ntawm lub zos excitation (LE).Rau cov dimers, ntxiv rau cov yam ntxwv ntawm intramolecular CT thiab LE, qee qhov kev faib ua feem ntawm intermolecular CT nta tau pom nyob rau hauv cov xeev, tshwj xeeb tshaj yog S3, S4, S7, thiab S8, raws li kev tsom xam CT intermolecular, nrog CT intermolecular hloov pauv raws li cov tseem ceeb. (Cov lus ntxiv).3).
Txhawm rau kom nkag siab zoo dua cov txiaj ntsig kev sim, peb tau tshawb xyuas ntxiv cov khoom ntawm RuDA zoo siab lub xeev los tshawb txog qhov sib txawv ntawm monomers thiab dimers (Cov Lus Ntxiv 4–5).Raws li pom nyob rau hauv daim duab 3B, lub zog theem ntawm singlet thiab triplet zoo siab lub xeev ntawm lub dimer yog ntau denser tshaj cov monomer, uas yuav pab txo tau lub zog sib txawv ntawm S1 thiab Tn. Nws tau raug tshaj tawm tias qhov kev hloov pauv ntawm ISC tuaj yeem pom tau hauv qhov sib txawv me me (ΔES1-Tn <0.3 eV) ntawm S1 thiab Tn54. Nws tau tshaj tawm tias qhov kev hloov pauv ntawm ISC tuaj yeem pom tau hauv qhov sib txawv me me ntawm lub zog (ΔES1-Tn <0.3 eV) ntawm S1 thiab Tn54. Сообщалось, что переходы ISC могут быть реализованы в пределах небольшой энергетической щели (5 Вус, 1-Tn) Nws tau raug tshaj tawm tias ISC kev hloov pauv tuaj yeem pom tau hauv qhov sib txawv me me ntawm lub zog (ΔES1-Tn <0.3 eV) ntawm S1 thiab Tn54.据报道，ISC 跃迁可以在S1 和Tn54 之间的小能隙（ΔES1-Tn < 0.3 eV）内实现.据报道，ISC 跃迁可以在S1 和Tn54 之间的小能隙（ΔES1-Tn < 0.3 eV）内实现. Сообщалось, что переход ISC может быть реализован в пределах небольшой энергетической щели (ΔэES1.3Tn) <0 Nws tau raug tshaj tawm tias qhov kev hloov pauv ntawm ISC tuaj yeem pom tau hauv qhov sib txawv me me ntawm lub zog (ΔES1-Tn <0.3 eV) ntawm S1 thiab Tn54.Tsis tas li ntawd xwb, tsuas yog ib qho orbital, nyob lossis tsis nyob, yuav tsum sib txawv hauv kev khi ib leeg thiab triplet lub xeev los muab qhov tsis yog xoom SOC qhov tseem ceeb.Yog li, raws li kev txheeb xyuas ntawm lub zog excitation thiab kev hloov pauv ntawm lub orbital, tag nrho cov kev hloov pauv tau ntawm ISC tau qhia hauv daim duab.3 C, ib d.Qhov tseem ceeb, tsuas yog ib qho ISC channel muaj nyob rau hauv monomer, thaum daim ntawv dimeric muaj plaub ISC raws uas tuaj yeem txhim kho ISC kev hloov pauv.Yog li, nws tsim nyog xav tias ntau dua RuDA molecules sib sau ua ke, qhov nkag tau yooj yim dua ISC channels yuav yog.Yog li, RuDA aggregates tuaj yeem tsim ob-band hluav taws xob cov qauv hauv singlet thiab triplet xeev, txo qhov sib txawv ntawm lub zog ntawm S1 thiab muaj Tn, yog li ua kom cov kev ua tau zoo ntawm ISC los pab txhawb 1O2 tiam.
Txhawm rau kom paub meej ntxiv txog cov txheej txheem hauv qab no, peb tau tsim cov khoom siv sib txuas ntawm arene-Ru(II) complex (RuET) los ntawm kev hloov ob pawg ethyl nrog ob pawg triphenylamine phenyl hauv RuDA (Fig. 4A, rau tag nrho cov yam ntxwv, saib ESI, Ntxiv 15. -21) Los ntawm pub (diethylamine) mus rau tus neeg txais (TDF), RuET muaj tib yam intramolecular CT yam ntxwv li RuDA.Raws li qhov xav tau, qhov nqus spectrum ntawm RuET hauv DMF tau pom tias muaj lub zog qis zog hloov pauv nrog lub zog nqus tau zoo nyob rau thaj tsam ze ntawm infrared thaj tsam ntawm 600-1100 nm (Fig. 4B).Tsis tas li ntawd, RuET aggregation kuj tau soj ntsuam nrog cov ntsiab lus dej nce ntxiv, uas tau pom nyob rau hauv redshift ntawm qhov nqus siab tshaj plaws, uas tau lees paub ntxiv los ntawm cov kua AFM imaging (Supplementary Fig. 22).Cov txiaj ntsig tau pom tias RuET, zoo li RuDA, tuaj yeem tsim cov xeev intramolecular thiab nws tus kheej sib sau ua ke hauv cov qauv sib sau.
Cov qauv tshuaj ntawm RuET.B Absorption spectra ntawm RuET hauv kev sib xyaw ntawm ntau qhov sib piv ntawm DMF thiab dej.Cov phiaj xwm C EIS Nyquist rau RuDA thiab RuET.Cov lus teb Photocurrent D ntawm RuDA thiab RuET nyob rau hauv qhov kev txiav txim ntawm laser hluav taws xob nrog lub wavelength ntawm 808 nm.
Lub photodegradation ntawm ABDA nyob rau hauv lub xub ntiag ntawm RuET tau soj ntsuam los ntawm irradiation nrog ib tug laser nrog ib tug wavelength ntawm 808 nm.Kuj ceeb tias, tsis muaj degradation ntawm ABDA tau pom nyob rau hauv ntau yam dej feem (Supplementary Fig. 23).Ib qho laj thawj yog vim li cas RuET tsis tuaj yeem tsim cov qauv siv hluav taws xob zoo vim tias cov saw hlau ethyl tsis txhawb kev hloov pauv hloov pauv hloov pauv hloov pauv tau zoo.Yog li ntawd, electrochemical impedance spectroscopy (EIS) thiab kev ntsuas photocurrent tam sim no tau ua los sib piv cov duab hluav taws xob ntawm RuDA thiab RuET.Raws li Nyquist zajlus (Daim duab 4C), RuDA qhia ib lub vojvoog me me dua li RuET, uas txhais tau hais tias RuDA56 muaj kev thauj mus los ntawm cov hluav taws xob hauv nruab nrab sai thiab zoo dua conductivity.Tsis tas li ntawd, qhov photocurrent ntom ntom ntawm RuDA yog ntau dua li ntawm RuET (Fig. 4D), lees paub qhov zoo dua tus nqi hloov pauv ntawm RuDA57.Yog li, pawg phenyl ntawm triphenylamine hauv Ore ua lub luag haujlwm tseem ceeb hauv kev muab cov nqi hloov pauv hloov pauv thiab tsim cov qauv hluav taws xob banded.
Txhawm rau ua kom cov qog nqaij hlav thiab nyob rau hauv vivo biocompatibility, peb ntxiv encapsulated RuDA nrog F127.Qhov nruab nrab hydrodynamic txoj kab uas hla ntawm RuDA-NPs tau txiav txim siab yog 123.1 nm nrog ib qho nqaim faib (PDI = 0.089) siv cov dynamic light scattering (DLS) txoj kev (Daim duab 5A), uas txhawb cov qog tsub zuj zuj los ntawm kev nce permeability thiab tuav.EPR) cuam tshuam.Cov duab TEM tau pom tias Ore NPs muaj cov duab kheej kheej kheej kheej nrog qhov nruab nrab txoj kab uas hla ntawm 86 nm.Qhov tseem ceeb, qhov nqus siab tshaj plaws ntawm RuDA-NPs tau tshwm sim ntawm 800 nm (Ntxiv daim duab 24), qhia tias RuDA-NPs tuaj yeem khaws cov haujlwm thiab cov khoom ntawm tus kheej sib sau RuDAs.Kev suav ROS quantum yield rau NP Ore yog 15.9%, uas yog piv rau Ore. Cov duab hluav taws xob ntawm RuDA NPs tau kawm nyob rau hauv qhov kev txiav txim ntawm laser hluav taws xob nrog lub wavelength ntawm 808 nm siv lub koob yees duab infrared.Raws li qhia hauv daim duab.5B, C, pawg tswj hwm (PBS nkaus xwb) tau ntsib qhov kub thiab txias me ntsis, thaum qhov kub ntawm cov tshuaj RuDA-NPs tau nce sai nrog qhov kub thiab txias (ΔT) mus rau 15.5, 26.1, thiab 43.0 ° C.Cov concentrations siab yog 25, 50, thiab 100 µM, feem, uas qhia tau hais tias muaj zog photothermal nyhuv ntawm RuDA NPs.Tsis tas li ntawd, kev ntsuas cua sov / cua txias tau raug coj los ntsuas qhov kev ruaj ntseg photothermal ntawm RuDA-NP thiab sib piv nrog ICG.Qhov kub ntawm Ore NPs tsis txo qis tom qab tsib lub cua sov / cua txias (Daim duab 5D), uas qhia tau hais tias zoo heev photothermal stability ntawm Ore NPs.Hauv qhov sib piv, ICG nthuav tawm qis dua photothermal stability raws li pom los ntawm qhov pom tseeb ploj ntawm photothermal kub toj siab nyob rau hauv tib yam mob.Raws li txoj kev dhau los 58, qhov kev hloov pauv photothermal (PCE) ntawm RuDA-NP tau suav tias yog 24.2%, uas yog siab dua li cov khoom siv photothermal uas twb muaj lawm xws li kub nanorods (21.0%) thiab kub nanoshells (13.0%) 59.Yog li, NP Ore nthuav tawm cov khoom zoo nkauj photothermal, uas ua rau lawv cog lus PTT cov neeg sawv cev.
Kev soj ntsuam ntawm DLS thiab TEM cov duab ntawm RuDA NPs (inset).B Cov duab tshav kub ntawm ntau qhov ntau ntawm RuDA NPs raug rau laser hluav taws xob ntawm lub wavelength ntawm 808 nm (0.5 W cm-2).C Photothermal hloov dua siab tshiab nkhaus ntawm ntau qhov ntau ntawm ore NPs, uas yog cov ntaub ntawv ntau.B. D Kub nce ntawm ORE NP thiab ICG tshaj 5 lub voj voog cua sov-txias.
Photocytotoxicity ntawm RuDA NPs tiv thaiv MDA-MB-231 tib neeg cov qog nqaij hlav cancer mis raug soj ntsuam hauv vitro.Raws li qhia hauv daim duab.6A, B, RuDA-NPs thiab RuDA nthuav tawm cytotoxicity tsis txaus ntseeg thaum tsis muaj irradiation, implying qis toxicity ntawm RuDA-NPs thiab RuDA.Txawm li cas los xij, tom qab raug laser hluav taws xob ntawm lub wavelength ntawm 808 nm, RuDA thiab RuDA NPs tau pom muaj zog photocytotoxicity tiv thaiv MDA-MB-231 cov qog nqaij hlav qog noj ntshav nrog IC50 qhov tseem ceeb (ib nrab qhov siab tshaj plaws inhibitory concentration) ntawm 5.4 thiab 9.4 μM, ntsig txog, ua qauv qhia. tias RuDA-NP thiab RuDA muaj peev xwm kho mob qog noj ntshav.Tsis tas li ntawd, photocytotoxicity ntawm RuDA-NP thiab RuDA tau tshawb xyuas ntxiv nyob rau hauv lub xub ntiag ntawm vitamin C (Vc), ib tug ROS scavenger, kom elucidate lub luag hauj lwm ntawm ROS nyob rau hauv lub teeb-induced cytotoxicity.Obviously, cell viability nce tom qab qhov sib ntxiv ntawm Vc, thiab IC50 qhov tseem ceeb ntawm RuDA thiab RuDA NPs yog 25.7 thiab 40.0 μM, feem, uas ua pov thawj lub luag haujlwm tseem ceeb ntawm ROS hauv photocytotoxicity ntawm RuDA thiab RuDA NPs.Lub teeb-induced cytotoxicity ntawm RuDA-NPs thiab RuDA hauv MDA-MB-231 cov qog nqaij hlav qog noj ntshav los ntawm kev nyob / tuag cell staining siv calcein AM (ntsuab fluorescence rau cov hlwb nyob) thiab propidium iodide (PI, liab fluorescence rau cov hlwb tuag).lees paub los ntawm cov hlwb) raws li kev sojntsuam fluorescent.Raws li pom nyob rau hauv daim duab 6C, cov hlwb kho nrog RuDA-NP lossis RuDA tseem siv tau yam tsis muaj irradiation, raws li muaj pov thawj los ntawm cov xim ntsuab fluorescence.Ntawm qhov tsis sib xws, nyob rau hauv laser irradiation, tsuas yog liab fluorescence tau pom, uas lees paub qhov zoo photocytotoxicity ntawm RuDA lossis RuDA NPs.Nws yog ib qho tseem ceeb uas cov fluorescence ntsuab tshwm nyob rau ntawm qhov sib ntxiv ntawm Vc, uas qhia txog kev ua txhaum ntawm photocytotoxicity ntawm RuDA thiab RuDA NPs.Cov txiaj ntsig no zoo ib yam nrog hauv vitro photocytotoxicity assay.
Dose-dependent viability ntawm A RuDA- thiab B RuDA-NP hlwb hauv MDA-MB-231 hlwb nyob rau hauv muaj lossis tsis muaj Vc (0.5 mM), feem.Cov khoom yuam kev, txhais tau tias ± tus qauv sib txawv (n = 3). Unpaired, ob-sided t tests *p <0.05, **p <0.01, thiab ***p <0.001. Unpaired, ob-sided t tests *p <0.05, **p <0.01, thiab ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 и ***p <0,001. Unpaired ob-tailed t-tests *p<0.05, **p<0.01, thiab ***p<0.001.未配对的双边t 检验*p <0.05、**p <0.01 和***p <0.001.未配对的双边t 检验*p <0.05、**p <0.01 和***p <0.001. Непарные двусторонние t-тесты *p <0,05, **p <0,01 thiab ***p <0,001. Unpaired ob-tailed t-tests *p<0.05, **p<0.01, thiab ***p<0.001.C Live/dead cell staining analysis siv calcein AM thiab propidium iodide li fluorescent probes.Scale bar: 30 µm.Cov duab sawv cev ntawm peb lub cev rov ua dua los ntawm txhua pab pawg tau pom.D Confocal fluorescence dluab ntawm ROS ntau lawm nyob rau hauv MDA-MB-231 hlwb nyob rau hauv ntau yam kev kho mob.Ntsuab DCF fluorescence qhia tias muaj ROS.Irradiate nrog lub laser nrog lub wavelength ntawm 808 nm nrog lub zog ntawm 0.5 W / cm2 rau 10 feeb (300 J / cm2).Scale bar: 30 µm.Cov duab sawv cev ntawm peb lub cev rov ua dua los ntawm txhua pab pawg tau pom.E Flow cytometry RuDA-NPs (50 µM) lossis RuDA (50 µM) kev kho mob tsom xam nrog lossis tsis muaj 808 nm laser (0.5 W cm-2) nyob rau hauv lub xub ntiag thiab tsis muaj Vc (0.5 mM) rau 10 min.Cov duab sawv cev ntawm peb lub cev rov ua dua los ntawm txhua pab pawg tau pom.F Nrf-2, HSP70 thiab HO-1 ntawm MDA-MB-231 hlwb kho nrog RuDA-NPs (50 µM) nrog lossis tsis muaj 808 nm laser irradiation (0.5 W cm-2, 10 min, 300 J cm-2), cell nthuav qhia 2).Cov duab sawv cev ntawm ob qho kev rov ua dua tshiab los ntawm txhua pab pawg tau pom.
Intracellular ROS ntau lawm hauv MDA-MB-231 hlwb raug tshuaj xyuas siv 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) staining txoj kev.Raws li qhia hauv daim duab.6D, hlwb kho nrog RuDA-NPs lossis RuDA nthuav tawm txawv ntsuab fluorescence thaum irradiated nrog 808 nm laser, qhia tias RuDA-NPs thiab RuDA muaj peev xwm tsim tau ROS.Ntawm qhov tsis sib xws, nyob rau hauv qhov tsis muaj lub teeb lossis nyob rau hauv lub xub ntiag ntawm Vc, tsuas yog ib qho tsis muaj zog fluorescent teeb liab ntawm lub hlwb tau pom, uas qhia tau hais tias me ntsis tsim ntawm ROS.Intracellular ROS theem hauv RuDA-NP hlwb thiab RuDA-kho MDA-MB-231 hlwb tau txiav txim siab ntxiv los ntawm kev ntws cytometry.Raws li pom nyob rau hauv Ntxiv daim duab 25, qhov nruab nrab fluorescence siv (MFI) generated los ntawm RuDA-NPs thiab RuDA nyob rau hauv 808 nm laser irradiation yog ho nce li ntawm 5.1 thiab 4.8 zaug, feem, piv rau pawg tswj, paub meej tias lawv zoo heev tsim AFK.muaj peev xwm.Txawm li cas los xij, intracellular ROS theem hauv RuDA-NP lossis MDA-MB-231 hlwb kho nrog RuDA tsuas yog piv rau kev tswj tsis muaj laser irradiation lossis nyob rau hauv lub xub ntiag ntawm Vc, zoo ib yam li cov txiaj ntsig ntawm confocal fluorescence tsom xam.
Nws tau pom tias mitochondria yog lub hom phiaj tseem ceeb ntawm Ru(II)-arene complexes60.Yog li ntawd, lub subcellular localization ntawm RuDA thiab RuDA-NPs tau tshawb xyuas.Raws li pom nyob rau hauv Ntxiv daim duab 26, RuDA thiab RuDA-NP qhia zoo sib xws cellular faib profiles nrog lub siab tshaj plaws tsub zuj zuj nyob rau hauv mitochondria (62.5 ± 4.3 thiab 60.4 ± 3.6 ng / mg protein, feem).Txawm li cas los xij, tsuas yog ib qho me me ntawm Ru tau pom nyob rau hauv cov khoom siv nuclear ntawm Ore thiab NP Ore (3.5 thiab 2.1%, feem).Cov seem ntawm tes muaj cov seem ruthenium: 31.7% (30.6 ± 3.4 ng / mg protein) rau RuDA thiab 42.9% (47.2 ± 4.5 ng / mg protein) rau RuDA-NPs.Feem ntau, Ore thiab NP Ore yog tsim nyob rau hauv mitochondria.Txhawm rau ntsuas mitochondrial tsis ua haujlwm, peb siv JC-1 thiab MitoSOX Liab staining los ntsuas mitochondrial membrane peev xwm thiab superoxide ntau lawm, raws li.Raws li pom nyob rau hauv Ntxiv Fig. 27, khaus ntsuab (JC-1) thiab liab (MitoSOX Liab) fluorescence tau pom nyob rau hauv cov hlwb kho nrog RuDA thiab RuDA-NPs nyob rau hauv 808 nm laser irradiation, qhia tias RuDA thiab RuDA-NPs heev fluorescent. Nws tuaj yeem ua tau zoo induce mitochondrial membrane depolarization thiab superoxide ntau lawm.Tsis tas li ntawd, cov txheej txheem ntawm kev tuag ntawm tes tau txiav txim siab siv cov dej cytometry raws kev tsom xam ntawm annexin V-FITC / propidium iodide (PI).Raws li pom nyob rau hauv daim duab 6E, thaum irradiated nrog 808 nm laser, RuDA thiab RuDA-NP induced ib tug tseem ceeb nce apoptosis thaum ntxov (txo txoj cai quadrant) nyob rau hauv MDA-MB-231 hlwb piv rau PBS los yog PBS ntxiv laser.ua cov hlwb.Txawm li cas los xij, thaum Vc tau ntxiv, apoptosis tus nqi ntawm RuDA thiab RuDA-NP txo qis los ntawm 50.9% thiab 52.0% mus rau 15.8% thiab 17.8%, feem, uas lees paub lub luag haujlwm tseem ceeb ntawm ROS hauv photocytotoxicity ntawm RuDA thiab RuDA-NP..Tsis tas li ntawd, me ntsis necrotic hlwb tau pom nyob rau hauv tag nrho cov pab pawg kuaj (sab laug quadrant), qhia tias apoptosis tej zaum yuav yog hom tseem ceeb ntawm cell tuag vim yog RuDA thiab RuDA-NPs.
Txij li thaum oxidative kev nyuaj siab puas tsuaj yog ib qho tseem ceeb ntawm kev txiav txim siab ntawm apoptosis, qhov cuam tshuam nuclear cuam tshuam nrog erythroid 2, yam 2 (Nrf2) 62, tus tswj hwm tseem ceeb ntawm cov kab mob antioxidant, tau tshawb xyuas hauv RuDA-NPs-kho MDA-MB-231.Mechanism of action of RuDA NPs induced by irradiation.Nyob rau tib lub sijhawm, kev qhia ntawm cov dej ntws qis ntawm cov protein heme oxygenase 1 (HO-1) kuj tau kuaj pom.Raws li pom nyob rau hauv daim duab 6F thiab ntxiv daim duab 29, RuDA-NP-mediated phototherapy nce Nrf2 thiab HO-1 qhia qib piv rau pawg PBS, qhia tias RuDA-NPs tuaj yeem ua rau oxidative stress signaling pathways.Tsis tas li ntawd, txhawm rau kawm txog cov nyhuv photothermal ntawm RuDA-NPs63, qhov kev qhia ntawm lub tshav kub poob siab protein Hsp70 kuj tau soj ntsuam.Nws yog qhov tseeb tias cov hlwb kho nrog RuDA-NPs + 808 nm laser irradiation tau nthuav tawm ntau ntxiv ntawm Hsp70 piv rau lwm ob pawg, qhia txog cov lus teb ntawm tes rau hyperthermia.
Cov txiaj ntsig zoo kawg hauv vitro tau ua rau peb tshawb xyuas qhov kev ua tau zoo hauv vivo ntawm RuDA-NP hauv cov nas liab liab nrog MDA-MB-231 cov qog.Cov ntaub so ntswg faib ntawm RuDA NPs tau kawm los ntawm kev txiav txim siab cov ntsiab lus ntawm ruthenium hauv daim siab, lub plawv, tus po, ob lub raum, lub ntsws, thiab cov qog.Raws li qhia hauv daim duab.7A, cov ntsiab lus siab tshaj plaws ntawm Ore NPs hauv lub cev ib txwm muaj tshwm sim thaum thawj zaug soj ntsuam (4 h), thaum cov ntsiab lus siab tshaj plaws tau txiav txim siab hauv cov qog nqaij hlav 8 teev tom qab txhaj tshuaj, tej zaum yog vim Ore NPs.EPR cuam tshuam ntawm LF.Raws li cov txiaj ntsig kev faib tawm, lub sijhawm zoo ntawm kev kho mob nrog NP ore tau noj 8 teev tom qab kev tswj hwm.Txhawm rau piav qhia txog cov txheej txheem ntawm kev sib sau ntawm RuDA-NPs hauv cov chaw qog, cov khoom siv photoacoustic (PA) ntawm RuDA-NPs tau raug saib xyuas los ntawm kev sau cov cim PA ntawm RuDA-NPs ntawm lub sijhawm sib txawv tom qab txhaj tshuaj.Ua ntej, PA teeb liab ntawm RuDA-NP hauv vivo raug soj ntsuam los ntawm kev kaw cov duab PA ntawm qhov chaw qog tom qab txhaj tshuaj ntawm RuDA-NP.Raws li pom nyob rau hauv Ntxiv Daim duab 30, RuDA-NPs tau pom lub teeb liab PA muaj zog, thiab muaj kev sib raug zoo ntawm RuDA-NP concentration thiab PA teeb liab siv zog (Cov duab ntxiv 30A).Tom qab ntawd, hauv vivo PA cov duab ntawm cov chaw qog tau sau tseg tom qab txhaj tshuaj ntawm RuDA thiab RuDA-NP ntawm lub sijhawm sib txawv tom qab txhaj tshuaj.Raws li pom nyob rau hauv daim duab 7B, PA teeb liab ntawm RuDA-NPs los ntawm cov qog nqaij hlav maj mam nce nrog lub sij hawm thiab mus txog lub toj siab ntawm 8 teev tom qab txhaj tshuaj, raws li cov ntaub so ntswg faib tau txiav txim los ntawm ICP-MS tsom xam.Nrog rau RuDA (Cov Lus Qhia Ntxiv 30B), qhov siab tshaj plaws PA teeb liab siv tau tshwm sim 4 teev tom qab txhaj tshuaj, qhia txog qhov ceev ntawm RuDA nkag mus rau hauv cov qog.Tsis tas li ntawd, tus cwj pwm excretory ntawm RuDA thiab RuDA-NPs tau tshawb xyuas los ntawm kev txiav txim siab tus nqi ntawm ruthenium hauv cov zis thiab faeces siv ICP-MS.Txoj hauv kev tseem ceeb ntawm kev tshem tawm rau RuDA (Cov Lus Qhia Ntxiv 31) thiab RuDA-NPs (Daim duab 7C) yog los ntawm faeces, thiab kev tshem tawm ntawm RuDA thiab RuDA-NPs tau pom zoo thaum lub sijhawm kawm 8-hnub, uas txhais tau hais tias RuDA. thiab RuDA-NPs tuaj yeem tshem tawm ntawm lub cev zoo yam tsis muaj tshuaj lom mus ntev.
A. Ex vivo faib ntawm RuDA-NP hauv cov ntaub so ntswg nas tau txiav txim siab los ntawm Ru cov ntsiab lus (feem pua ​​​​ntawm cov koob tshuaj Ru (ID) ib gram ntawm cov ntaub so ntswg) nyob rau lub sijhawm sib txawv tom qab txhaj tshuaj.Cov ntaub ntawv yog txhais tau tias ± tus qauv sib txawv (n = 3). Unpaired, ob-sided t tests *p <0.05, **p <0.01, thiab ***p <0.001. Unpaired, ob-sided t tests *p <0.05, **p <0.01, thiab ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 и ***p <0,001. Unpaired ob-tailed t-tests *p<0.05, **p<0.01, thiab ***p<0.001.未配对的双边t 检验*p <0.05、**p <0.01 和***p <0.001.未配对的双边t 检验*p <0.05、**p <0.01 和***p <0.001. Непарные двусторонние t-тесты *p <0,05, **p <0,01 thiab ***p <0,001. Unpaired ob-tailed t-tests *p<0.05, **p<0.01, thiab ***p<0.001.B PA cov duab ntawm hauv vivo qog qhov chaw ntawm 808 nm excitation tom qab txhaj tshuaj ntawm RuDA-NPs (10 µmol kg-1) ntawm lub sijhawm sib txawv.Tom qab txhaj tshuaj ntawm RuDA NPs (10 µmol kg-1), C Ru raug tso tawm los ntawm nas nrog zis thiab quav nyob rau lub sijhawm sib txawv.Cov ntaub ntawv yog txhais tau tias ± tus qauv sib txawv (n = 3).
Lub peev xwm cua sov ntawm RuDA-NP hauv vivo tau kawm hauv cov nas liab liab nrog MDA-MB-231 thiab RuDA qog rau kev sib piv.Raws li qhia hauv daim duab.8A thiab ntxiv Fig. 32, pawg tswj hwm (saline) tau pom qhov ntsuas kub tsawg dua (ΔT ≈ 3 ° C) tom qab 10 feeb ntawm kev sib kis tas mus li.Txawm li cas los xij, qhov kub ntawm RuDA-NPs thiab RuDA tau nce sai nrog qhov kub siab tshaj plaws ntawm 55.2 thiab 49.9 ° C, raws li, muab cov hyperthermia txaus rau hauv vivo kev kho mob qog noj ntshav.Qhov pom kev nce siab hauv qhov kub thiab txias rau RuDA NPs (ΔT ≈ 24 ° C) piv rau RuDA (ΔT ≈ 19 ° C) yuav yog vim nws qhov permeability zoo dua thiab tsub zuj zuj hauv cov qog nqaij hlav vim EPR cov nyhuv.
Infrared thermal dluab ntawm nas nrog MDA-MB-231 qog irradiated nrog 808 nm laser ntawm txawv sij hawm 8 teev tom qab txhaj tshuaj.Cov duab sawv cev ntawm plaub qhov kev rov ua dua tshiab los ntawm txhua pab pawg tau pom.B txheeb ze qog ntim thiab C nruab nrab qog loj ntawm ntau pawg nas thaum kho.D Curves ntawm lub cev hnyav ntawm ntau pawg nas.Irradiate nrog lub laser nrog lub wavelength ntawm 808 nm nrog lub zog ntawm 0.5 W / cm2 rau 10 feeb (300 J / cm2).Cov khoom yuam kev, txhais tau tias ± tus qauv sib txawv (n = 3). Unpaired, ob-sided t tests *p <0.05, **p <0.01, thiab ***p <0.001. Unpaired, ob-sided t tests *p <0.05, **p <0.01, thiab ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 и ***p <0,001. Unpaired ob-tailed t-tests *p<0.05, **p<0.01, thiab ***p<0.001.未配对的双边t 检验*p <0.05、**p <0.01 和***p <0.001.未配对的双边t 检验*p <0.05、**p <0.01 和***p <0.001. Непарные двусторонние t-тесты *p <0,05, **p <0,01 thiab ***p <0,001. Unpaired ob-tailed t-tests *p<0.05, **p<0.01, thiab ***p<0.001. E H&E staining dluab ntawm cov kabmob loj thiab cov qog los ntawm ntau pawg kho mob, suav nrog Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, thiab RuDA-NPs + Laser pawg. E H&E staining dluab ntawm cov kabmob loj thiab cov qog los ntawm ntau pawg kho mob, suav nrog Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, thiab RuDA-NPs + Laser pawg. Изображения окрашивания E H&E основных органов и опухолей из разных групп лечения, включая группы физиологического раствора, физиологического раствора + лазера, RuDA, RuDA + Laser, RuDA-NPs и RuDA-NPs + Laser. E H&E staining dluab ntawm cov kabmob loj thiab cov qog los ntawm ntau pawg kho mob, suav nrog saline, saline + laser, RuDA, RuDA + Laser, RuDA-NPs, thiab RuDA-NPs + Laser pawg.来自不同治疗组的主要器官和肿瘤的E H&E 染色图像，包括盐水、盐水+ 激光、RuDA、RuDA、RuDA、RuDA、RuDA、RuDA、RuDA来自不同治疗组的主要器官和肿瘤的E H&E Окрашивание E H&E основных органов и опухолей из различных групп лечения, включая физиологический раствор, физиологический раствор + лазер, RuDA, RuDA + лазер, RuDA-NPs и RuDA-NPs + лазер. E H&E staining ntawm lub cev loj thiab cov qog los ntawm ntau pawg kho mob suav nrog saline, saline + laser, RuDA, RuDA + laser, RuDA-NPs, thiab RuDA-NPs + laser.Scale bar: 60 µm.
Cov nyhuv ntawm phototherapy nyob rau hauv vivo nrog RuDA thiab RuDA NPs tau soj ntsuam nyob rau hauv cov nas liab qab nrog MDA-MB-231 cov qog tau txhaj rau hauv cov hlab ntsha nrog RuDA lossis RuDA NPs ntawm ib koob ntawm 10.0 µmol kg-1 ntawm tus Tsov tus tw leeg, thiab tom qab ntawd 8. teev tom qab txhaj tshuaj.laser irradiation nrog lub wavelength ntawm 808 nm.Raws li pom nyob rau hauv daim duab 8B, cov qog nqaij hlav tau nce ntau hauv cov saline thiab laser pawg, qhia tias saline los yog laser 808 irradiation tsis tshua muaj kev cuam tshuam rau cov qog loj hlob.Raws li nyob rau hauv pab pawg neeg saline, cov qog loj hlob sai kuj tau pom nyob rau hauv cov nas kho nrog RuDA-NPs lossis RuDA thaum tsis muaj laser irradiation, ua rau pom lawv qhov tsis tshua muaj toxicity.Nyob rau hauv sib piv, tom qab laser irradiation, ob qho tib si RuDA-NP thiab RuDA kev kho mob induced loj hlav regression nrog qog ntim txo ntawm 95.2% thiab 84.3%, feem, piv rau cov saline kho pab pawg neeg, qhia tau hais tias zoo heev synergistic PDT., kho los ntawm RuDA/CHTV effect.Piv nrog RuDA, RuDA NPs tau pom tias muaj cov nyhuv phototherapeutic zoo dua, uas yog vim EPR nyhuv ntawm RuDA NPs.Cov qog loj hlob inhibition tau raug soj ntsuam ntxiv los ntawm qog qhov hnyav excised rau hnub 15 ntawm kev kho mob (Fig. 8C thiab Supplementary Fig. 33).Qhov nruab nrab qog loj hauv RuDA-NP kho cov nas thiab RuDA kho cov nas yog 0.08 thiab 0.27 g, raws li, uas yog sib zog dua hauv pawg tswj hwm (1.43 g).
Tsis tas li ntawd, lub cev hnyav ntawm nas tau sau tseg txhua peb hnub los kawm txog qhov tsaus ntuj toxicity ntawm RuDA-NPs lossis RuDA hauv vivo.Raws li pom hauv daim duab 8D, tsis muaj qhov sib txawv tseem ceeb hauv lub cev hnyav tau pom rau txhua pab pawg kho mob. Tsis tas li ntawd, hematoxylin thiab eosin (H&E) staining ntawm lub cev loj (lub plawv, siab, tus po, ntsws, thiab lub raum) tau ua los ntawm ntau pab pawg kho mob. Tsis tas li ntawd, hematoxylin thiab eosin (H&E) staining ntawm lub cev loj (lub plawv, siab, tus po, ntsws, thiab lub raum) tau ua los ntawm ntau pab pawg kho mob. Кроме того, было проведено окрашивание гематоксилином и эозином (H&E) основных органов (серашивание зердца, печилени , Tsis tas li ntawd, hematoxylin thiab eosin (H&E) staining ntawm lub cev loj (lub plawv, siab, tus po, ntsws, thiab ob lub raum) tau ua los ntawm ntau pab pawg kho mob.此外，对不同治疗组的主要器官（心脏、肝脏、脾脏、肺和肾脏）进行苏期精和伊粓。 (H&E) Кроме того, проводили окрашивание гематоксилином и эозином (H&E) основных органов (сердца, пердца, печекни, сепкел, сердца, печекни, сепкелеле, сердца, печекни, сепкеле сердца Tsis tas li ntawd, hematoxylin thiab eosin (H&E) staining ntawm lub cev loj (lub plawv, siab, tus po, ntsws, thiab lub raum) tau ua nyob rau hauv ntau pab pawg kho mob.Raws li qhia hauv Fig.8E, H&E staining dluab ntawm tsib lub cev tseem ceeb los ntawm pawg RuDA-NPs thiab RuDA tsis pom muaj qhov txawv txav lossis kev puas tsuaj ntawm lub cev. 8E, H&E staining dluab ntawm tsib lub cev tseem ceeb los ntawm pawg RuDA-NPs thiab RuDA tsis pom muaj qhov txawv txav lossis kev puas tsuaj ntawm lub cev.Raws li qhia hauv daim duab.8E, изображения окрашивания H&E пяти основных органов из групп RuDA-NPs и RuDA не демонстрируют явовных анинила 8E, H&E staining dluab ntawm tsib lub hauv nruab nrog cev loj los ntawm RuDA-NPs thiab RuDA pawg qhia tsis pom qhov txawv txav ntawm lub cev lossis qhov txhab.如图8E 所示，来自RuDA-NPs 和RuDA 组的五个主要器官的H&E 染色图像没有显示出明帾的异如图8E 所示，来自RuDA-NPs 和RuDA 组的五个主要器官的H&E Как показано на рисунке 8E, изображения окрашивания H&E пяти основных органов из групп RuDA-NPs thiab RuDA не и лижазанов Raws li pom nyob rau hauv daim duab 8E, H&E staining dluab ntawm tsib lub cev tseem ceeb los ntawm RuDA-NPs thiab RuDA pawg tsis pom muaj qhov txawv txav los yog kev puas tsuaj ntawm lub cev.Cov txiaj ntsig no tau pom tias tsis yog RuDA-NP lossis RuDA tsis pom cov tsos mob toxicity hauv vivo. Tsis tas li ntawd, H&E staining dluab ntawm cov qog pom tau tias ob qho tib si RuDA + Laser thiab RuDA-NPs + Laser pawg tuaj yeem ua rau mob qog noj ntshav loj heev, ua kom pom qhov zoo tshaj plaws hauv vivo phototherapeutic efficiency ntawm RuDA thiab RuDA-NPs. Tsis tas li ntawd, H&E staining dluab ntawm cov qog pom tau tias ob qho tib si RuDA + Laser thiab RuDA-NPs + Laser pawg tuaj yeem ua rau mob qog noj ntshav loj heev, ua kom pom qhov zoo tshaj plaws hauv vivo phototherapeutic efficiency ntawm RuDA thiab RuDA-NPs.Tsis tas li ntawd, hematoxylin-eosin stained qog cov duab qhia tau hais tias ob qho tib si RuDA + Laser thiab RuDA-NPs + Laser pawg tuaj yeem ua rau muaj kev puas tsuaj loj ntawm cov qog nqaij hlav cancer, ua kom pom qhov ua tau zoo tshaj ntawm kev kho mob ntawm RuDA thiab RuDA-NPs hauv vivo.此外，肿瘤的H&E 染色图像显示，RuDA + Laser 和RuDA-NPs + Laser 组均可导致严重的癌细胞破嘎式。此外 , 肿瘤的 & e 染色 显示 , ruda + laser 和 ruda-nps + laser 组均导致的癌的细胞。 。 破坏 ， 证 ruda-nps ...Tsis tas li ntawd, hematoxylin thiab eosin stained qog cov duab qhia tau hais tias ob qho tib si RuDA + Laser thiab RuDA-NPs + Laser pawg ua rau muaj kev puas tsuaj loj ntawm cov qog nqaij hlav cancer, ua kom pom qhov zoo tshaj ntawm kev kho mob zoo ntawm RuDA thiab RuDA-NPs hauv vivo.
Hauv kev xaus, Ru(II)-arene (RuDA) organometallic complex nrog DA-type ligands tau tsim los pab txhawb cov txheej txheem ISC siv txoj kev sib sau ua ke.Synthesized RuDA tuaj yeem sib sau ua tus kheej los ntawm kev sib cuam tshuam uas tsis yog-covalent los tsim RuDA-derived supramolecular systems, yog li pab txhawb 1O2 tsim thiab muaj txiaj ntsig photothermal hloov dua siab tshiab rau kev kho mob qog noj ntshav.Nws yog ib qho tseem ceeb uas monomeric RuDA tsis tau tsim 1O2 nyob rau hauv laser irradiation ntawm 808 nm, tab sis yuav tsim ib tug loj npaum li cas ntawm 1O2 nyob rau hauv lub aggregated lub xeev, qhia lub rationality thiab efficiency ntawm peb tsim.Cov kev tshawb fawb tom qab tau pom tias lub rooj sib tham supramolecular endows RuDA nrog kev txhim kho cov duab ntawm lub cev thiab cov tshuaj photochemical, xws li redshift absorption thiab photobleaching tsis kam, uas yog qhov xav tau rau PDT thiab PTT ua.Ob qho tib si hauv vitro thiab hauv vivo thwmsim tau pom tias RuDA NPs nrog biocompatibility zoo thiab zoo tsub zuj zuj hauv cov qog ua rau pom cov teeb pom kev zoo los tiv thaiv qog noj ntshav thaum laser irradiation ntawm lub wavelength ntawm 808 nm.Yog li, RuDA NPs ua tau zoo bimodal supramolecular PDT / PTW reagents yuav txhawb nqa cov txheej txheem ntawm photosensitizers qhib ntawm wavelengths saum 800 nm.Lub tswv yim tsim ntawm supramolecular system muab txoj hauv kev zoo rau NIR-activated photosensitizers nrog cov teebmeem zoo heev.
Tag nrho cov tshuaj thiab cov kuab tshuaj tau txais los ntawm cov chaw muag khoom lag luam thiab siv yam tsis muaj kev lim dej ntxiv.RuCl3 tau yuav los ntawm Boren Precious Metals Co., Ltd. (Kunming, Suav).[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione) thiab 4,7-bis[4-(N,N-diphenylamino)phenyl]-5 ,6-Diamino-2,1,3-benzothiadiazole tau ua ke raws li kev tshawb fawb yav dhau los64,65.NMR spectra raug kaw rau ntawm Bruker Avance III-HD 600 MHz spectrometer ntawm Southeastern University Analytical Test Center siv d6-DMSO lossis CDCl3 ua hnyav.Cov tshuaj hloov pauv δ yog muab rau hauv ppm.nrog rau kev hwm rau tetramethylsilane, thiab kev sib cuam tshuam tas li J tau muab rau hauv qhov tseem ceeb hauv hertz.Kev daws teeb meem loj spectrometry (HRMS) tau ua tiav ntawm Agilent 6224 ESI / TOF MS ntsuas.Elemental tsom xam ntawm C, H, thiab N tau ua tiav ntawm Vario MICRCHNOS elemental analyzer (Elementar).UV-pom spectra tau ntsuas ntawm Shimadzu UV3600 spectrophotometer.Fluorescence spectra raug kaw rau ntawm Shimadzu RF-6000 spectrofluorimeter.EPR spectra tau kaw rau ntawm Bruker EMXmicro-6/1 ntsuas.Cov morphology thiab cov qauv ntawm cov qauv npaj tau kawm ntawm FEI Tecnai G20 (TEM) thiab Bruker Icon (AFM) cov cuab yeej ua haujlwm ntawm qhov hluav taws xob ntawm 200 kV.Dynamic light scattering (DLS) tau ua ntawm Nanobrook Omni analyzer (Brookhaven).Photoelectrochemical zog tau ntsuas los ntawm kev teeb tsa electrochemical (CHI-660, Suav).Cov duab photoacoustic tau txais los ntawm FUJIFILM VisualSonics Vevo® LAZR system.Confocal duab tau txais los ntawm Olympus FV3000 confocal microscope.FACS tsom xam tau ua tiav ntawm BD Calibur flow cytometer.Kev ua haujlwm siab ua kua chromatography (HPLC) kev sim tau ua tiav ntawm Waters Alliance e2695 system siv 2489 UV / Vis ntes.Gel Permeation Chromatography (GPC) cov kev ntsuam xyuas tau kaw rau ntawm Thermo ULTIMATE 3000 ntsuas siv ERC RefratoMax520 refractive index detector.
[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione)64 (481.0 mg, 1.0 mmol), 4,7-bis[4 -(N, N-diphenylamino)phenyl]-5,6-diamino-2,1,3-benzothiadiazole 65 (652.0 mg, 1.0 mmol) thiab glacial acetic acid (30 mL) tau nplawm ntawm reflux tub yees rau 12 teev.Tom qab ntawd cov kuab tshuaj tau muab tshem tawm hauv lub tshuab nqus tsev siv lub tshuab evaporator teb.Qhov tshwm sim residue tau lim los ntawm flash kem chromatography (silica gel, CH2Cl2: MeOH = 20: 1) kom tau RuDA raws li cov hmoov ntsuab (yield: 877.5 mg, 80%).qhov quav.Muab xam rau C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.Pom tau: C 67.92, H 4.26, N 9.82.1H NMR (600 MHz, d6-DMSO) δ 10.04 (s, 2H), 8.98 (s, 2H), 8.15 (s, 2H), 7.79 (s, 4H), 7.44 (s, 8H), 7.21 (d, J = 31.2 Hz, 16H), 6.47 (s, 2H), 6.24 (s, 2H), 2.69 (s, 1H), 2 .25 (s, 3H), 0.99 (s, 6H).13C NMR (150 MHz, D6-DMOPS), δ (PPM) 158.0,5.5.5.5,5,51, 120.8.49, 103.49 , 103., 86.52, 84.75, 63.29, 30.90, 22.29, 18.83.ESI-MS: m/z [M-Cl]+ = 1097.25.
Synthesis ntawm 4,7-bis [4-(N, N-diethylamino) phenyl-5,6-diamino-2,1,3-benzothiadiazole (L2): L2 yog synthesized nyob rau hauv ob kauj ruam.Pd (PPh3)4 (46 mg, 0.040 mmol) tau ntxiv rau N, N-diethyl-4-(tributylstannyl) aniline (1.05 g, 2.4 mmol) thiab 4,7-dibromo-5,6-dinitro tov - 2, 1,3-benzothiadiazole (0.38 g, 1.0 mmol) nyob rau hauv qhuav toluene (100 ml).Qhov sib tov yog nplawm ntawm 100 ° C rau 24 teev.Tom qab tshem tawm cov toluene hauv lub tshuab nqus tsev, cov khoom tsim tau raug ntxuav nrog roj av ether.Tom qab ntawd sib tov ntawm no compound (234.0 mg, 0.45 mmol) thiab hlau hmoov (0.30 g, 5.4 mmol) nyob rau hauv acetic acid (20 ml) yog stirred ntawm 80 ° C. rau 4 teev.Cov tshuaj tiv thaiv sib xyaw yog nchuav rau hauv dej thiab cov khoom xim av tau sau los ntawm kev pom.Cov khoom raug ntxuav ob zaug los ntawm lub tshuab nqus tsev sublimation kom muab cov khoom ntsuab (126.2 mg, 57% tawm los).qhov quav.Muab xam rau C26H32N6S: C 67.79, H 7.00, N 18.24.Pom tau: C 67.84, H 6.95, H 18.16.1H NMR (600 MHz, CDCl3), δ (ppm) 7.42 (d, 4H), 6.84 (d, 4H), 4.09 (s, 4H), 3.42 (d, 8H), 1.22 (s, 12H).13C NMR (150 MHz, CDCl3), δ (ppm) 151.77, 147.39, 138.07, 131.20, 121.09, 113.84, 111.90, 44.34, 12.77.ESI-MS: m/z [M+H]+ = 461.24.
Cov tshuaj tau npaj thiab ua kom huv raws li cov txheej txheem zoo ib yam li RuDA.qhov quav.Muab xam rau C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.Pom tau: C, 61.32, H, 5.12, N, 11.81,1H NMR (600 MHz, d6-DMSO), δ (ppm) 10.19 (s, 2H), 9.28 (s, 2H), 8.09 (s, 2H), 7.95 (s, 4H), 6.93 (s, 4H), 6.48 (d, 2H), 6.34 (s, 2H), 3.54 (t, 8H), 2.80 (m, 1H), 2.33 (s, 3H), 1.31 (t, 12H), 1.07 (s, 6H).13C NMR (151 MHZ, CDCL3), δ (PPM) 158.20, 148.7,5,5,8.77, 104.23, 87.0., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m/z [M-Cl]+ = 905.24.
RuDA tau yaj hauv MeOH / H2O (5/95, v / v) ntawm qhov concentration ntawm 10 μM.Kev nqus spectrum ntawm RuDA tau ntsuas txhua 5 feeb ntawm Shimadzu UV-3600 spectrophotometer nyob rau hauv irradiation nrog lub teeb laser nrog lub wavelength ntawm 808 nm (0.5 W / cm2).Lub ICG spectra tau kaw nyob rau hauv tib yam kev mob raws li tus qauv.
EPR spectra tau kaw rau ntawm Bruker EMXmicro-6/1 spectrometer nrog lub microwave zog ntawm 20 mW, scanning ntau ntawm 100 G, thiab ib qho kev hloov pauv ntawm 1 G. 2,2,6,6-tetramethyl-4-piperidone (TEMP) thiab 5,5-dimethyl-1-pyrroline N-oxide (DMPO) tau siv los ua cov ntxiab ntxiab.Electron spin resonance spectra tau sau tseg rau cov kev daws teeb meem sib xyaw ntawm RuDA (50 µM) thiab TEMF (20 mM) lossis DMPO (20 mM) nyob rau hauv qhov kev txiav txim ntawm laser hluav taws xob nrog lub wavelength ntawm 808 nm (0.5 W / cm2).
Kev suav DFT thiab TD-DFT rau RuDA tau ua tiav ntawm PBE1PBE / 6–31 G*//LanL2DZ qib hauv cov kua dej uas siv Gaussian program 1666,67,68.HOMO-LUMO, qhov thiab hluav taws xob faib tawm ntawm lub zog qis zog singlet zoo siab lub xeev RuDA tau npaj siv GaussView program (version 5.0).
Peb thawj zaug sim ntsuas qhov ua tau zoo ntawm 1O2 RuDA siv cov pa UV-pom spectroscopy nrog ICG (ΦΔ = 0.002) raws li tus qauv, tab sis photodegradation ntawm ICG cuam tshuam rau cov txiaj ntsig.Yog li, quantum yield ntawm 1O2 RuDA tau ntsuas los ntawm kev ntsuas qhov kev hloov pauv ntawm ABDA fluorescence ntawm li 428 nm thaum irradiated nrog lub laser nrog lub wavelength ntawm 808 nm (0.5 W / cm2).Kev sim tau ua rau RuDA thiab RuDA NPs (20 μM) hauv dej / DMF (98/2, v / v) uas muaj ABDA (50 μM).Lub quantum yield ntawm 1O2 tau suav nrog cov qauv hauv qab no: ΦΔ (PS) = ΦΔ (ICG) × (rFS/APS)/(rICG/AICG).rPS thiab rICG yog cov tshuaj tiv thaiv ntawm ABDA nrog 1O2 tau los ntawm photosensitizer thiab ICG, feem.APS thiab AICG yog qhov nqus ntawm photosensitizer thiab ICG ntawm 808 nm, feem.
Kev ntsuas AFM tau ua tiav hauv cov kua dej siv hom scan ntawm Bruker Dimension Icon AFM system.Siv cov qauv qhib nrog cov kua hlwb, cov hlwb raug ntxuav ob zaug nrog ethanol thiab qhuav nrog cov kwj ntawm nitrogen.Ntxig cov hlwb qhuav rau hauv lub taub hau kho qhov muag ntawm lub tshuab tsom.Tam sim ntawd tso ib tug poob ntawm cov qauv mus rau hauv lub pas dej ua ke ntawm cov kua thiab muab tso rau ntawm lub cantilever siv ib tug sterile pov tseg yas syringe thiab ib tug sterile koob.Lwm qhov poob yog muab tso ncaj qha rau ntawm tus qauv, thiab thaum lub taub hau kho qhov muag txo qis, ob lub tee sib koom ua ke, ua ib qho meniscus ntawm cov qauv thiab cov dej tso dej.Kev ntsuas AFM tau ua tiav siv SCANASYST-FLUID V-shaped nitride cantilever (Bruker, hardness k = 0.7 N m-1, f0 = 120-180 kHz).
HPLC chromatograms tau txais ntawm Waters e2695 system nruab nrog phoenix C18 kem (250 × 4.6 mm, 5 µm) siv 2489 UV / Vis ntes.Lub wavelength ntawm lub ntes yog 650 nm.Txawb theem A thiab B yog dej thiab methanol, feem, thiab cov theem mobile ntws yog 1.0 ml·min-1.Cov gradient (cov kuab tshuaj B) yog raws li nram no: 100% ntawm 0 mus rau 4 feeb, 100% mus rau 50% ntawm 5 mus rau 30 feeb, thiab rov qab mus rau 100% ntawm 31 mus rau 40 feeb.Ore yog yaj nyob rau hauv ib tug tov tov ntawm methanol thiab dej (50/50, los ntawm ntim) ntawm ib tug concentration ntawm 50 μM.Kev txhaj tshuaj yog 20 μl.
GPC kev soj ntsuam tau kaw rau ntawm Thermo ULTIMATE 3000 ntsuas nruab nrog ob lub PL aquagel-OH MIXED-H txhua kab (2 × 300 × 7.5 mm, 8 µm) thiab ERC RefratoMax520 refractive index detector.GPC kem tau eluted nrog dej ntawm tus nqi ntws ntawm 1 ml / min ntawm 30 ° C.Ore NPs tau yaj hauv PBS tov (pH = 7.4, 50 μM), txhaj tshuaj ntim yog 20 μL.
Photocurrents tau ntsuas ntawm kev teeb tsa hluav taws xob (CHI-660B, Tuam Tshoj).Cov lus teb optoelectronic thaum lub laser tau qhib thiab tawm (808 nm, 0.5 W / cm2) tau ntsuas ntawm qhov hluav taws xob ntawm 0.5 V hauv lub thawv dub, raws li.Tus qauv peb-electrode cell tau siv nrog L-shaped glassy carbon electrode (GCE) ua cov khoom siv hluav taws xob ua haujlwm, tus qauv calomel electrode (SCE) raws li cov khoom siv electrode, thiab platinum disk ua lub txee electrode.A 0.1 M Na2SO4 tov yog siv los ua ib qho electrolyte.
Tib neeg txoj kab mob qog noj ntshav MDA-MB-231 tau yuav los ntawm KeyGEN Biotec Co., LTD (Nanjing, Suav teb, tus lej xov tooj: KG033).Cells tau loj hlob hauv monolayers hauv Dulbecco's Modified Eagle's Medium (DMEM, siab qabzib) ntxiv nrog kev daws ntawm 10% fetal bovine serum (FBS), penicillin (100 μg / ml) thiab streptomycin (100 μg / ml).Tag nrho cov hlwb raug coj los ntawm 37 ° C nyob rau hauv qhov chaw ntub dej uas muaj 5% CO2.
Kev soj ntsuam MTT tau siv los txiav txim siab cytotoxicity ntawm RuDA thiab RuDA-NPs nyob rau hauv lub xub ntiag thiab tsis muaj lub teeb irradiation, nrog los yog tsis muaj Vc (0.5 mM).MDA-MB-231 cov qog nqaij hlav qog noj ntshav tau loj hlob hauv 96-zoo daim hlau ntawm lub cell ceev ntawm kwv yees li 1 x 105 hlwb / ml / zoo thiab incubated rau 12 teev ntawm 37.0 ° C nyob rau hauv ib qho chaw ntawm 5% CO2 thiab 95% cua.RuDA thiab RuDA NPs yaj hauv dej tau ntxiv rau lub hlwb.Tom qab 12 teev ntawm incubation, lub hlwb raug rau 0.5 W cm -2 laser hluav taws xob ntawm ib tug wavelength ntawm 808 nm rau 10 feeb (300 J cm -2) thiab ces incubated nyob rau hauv qhov tsaus ntuj rau 24 teev.Tom qab ntawd cov hlwb raug tsim nrog MTT (5 mg / ml) rau lwm 5 teev.Thaum kawg, hloov qhov nruab nrab mus rau DMSO (200 µl) kom yaj cov xim liab formalan crystals.OD qhov tseem ceeb tau ntsuas siv lub microplate nyeem ntawv nrog lub wavelength ntawm 570/630 nm.Tus nqi IC50 rau txhua tus qauv tau suav nrog siv SPSS software los ntawm cov koob tshuaj tiv thaiv kab mob uas tau txais los ntawm tsawg kawg peb qhov kev sim ywj pheej.
MDA-MB-231 hlwb raug kho nrog RuDA thiab RuDA-NP ntawm qhov concentration ntawm 50 μM.Tom qab 12 teev ntawm incubation, lub hlwb tau irradiated nrog ib tug laser nrog ib tug wavelength ntawm 808 nm thiab lub zog ntawm 0.5 W / cm2 rau 10 min (300 J / cm2).Hauv pawg vitamin C (Vc), cov hlwb raug kho nrog 0.5 mM Vc ua ntej laser irradiation.Tom qab ntawd cov cell raug muab tso rau hauv qhov tsaus ntuj rau 24 teev ntxiv, tom qab ntawd pleev xim nrog calcein AM thiab propidium iodide (20 μg / ml, 5 μl) rau 30 feeb, tom qab ntawd ntxuav nrog PBS (10 μl, pH 7.4).cov duab ntawm stained cells.