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Ama-photosensitizer asebenzayo abaluleke kakhulu ekusetshenzisweni okubanzi komtholampilo kwe-phototherapy.Kodwa-ke, ama-photosensitizers avamile ngokuvamile ahlushwa ukumuncwa kwe-wavelength emifushane, ukungakwazi ukuthwebula izithombe okwanele, isivuno esiphansi se-oxygen esebenzayo (ROS), kanye nokucisha okubangelwa ukuhlanganisa kwe-ROS.Lapha sibika i-near-infrared (NIR) supramolecular photosensitizer (RuDA) mediated by self-assembly of Ru(II)-arene organometallic complexes in aqueous solution.I-RuDA ingakwazi ukukhiqiza kuphela i-singlett oxygen (1O2) esimweni esihlanganisiwe, futhi ibonisa ukuziphatha kwesizukulwane se-1O2 esibangelwa ukuhlanganisa okusobala ngenxa yokwanda okukhulu kwenqubo ye-crossover phakathi kwesistimu ye-singlett-triplet.Ngaphansi kwesenzo sokukhanya kwe-laser ye-808 nm, i-RuDA ibonisa isivuno se-quantum esingu-1O2 se-16.4% (i-indocyanine eluhlaza evunyelwe yi-FDA: ΦΔ=0.2%) kanye nokusebenza kahle kokuguqulwa kwe-photothermal kwe-24.2% (ama-nanorods egolide ezohwebo) nge-photostability enhle kakhulu.: 21.0%, ama-nanoshell egolide: 13.0%).Ukwengeza, ama-RuDA-NP ane-biocompatibility enhle angaqongelela ngokukhetha ezindaweni zesimila, okubangela ukwehla okukhulu kwesimila ngesikhathi sokwelashwa kwe-photodynamic ngokuncipha kwevolumu yesimila ku-vivo ngama-95.2%.Lokhu kwelashwa kwe-photodynamic okuthuthukisa ukuhlanganisa kunikeza isu lokuthuthukisa ama-photosensitizer anezakhiwo ezivumayo zokuthwebula izithombe namakhemikhali.
Uma kuqhathaniswa nokwelapha okuvamile, ukwelashwa kwe-photodynamic (i-PDT) kuyindlela yokwelapha ekhangayo yomdlavuza ngenxa yezinzuzo zayo ezibalulekile njengokulawula okunembile kwe-spatiotemporal, ukungahlaseli, ukumelana nezidakamizwa okungenamsebenzi, kanye nokunciphisa imiphumela emibi 1,2,3.Ngaphansi kwe-irradiation ekhanyayo, ama-photosensitizers asetshenzisiwe angasetshenziswa ukuze enze izinhlobo ze-oxygen esebenzayo kakhulu (ROS), okuholela ku-apoptosis/necrosis noma izimpendulo zokuzivikela komzimba4,5. Kodwa-ke, ama-photosensitizers amaningi avamile, njengama-chlorin, ama-porphyrins, nama-anthraquinones, anokumunca amaza amaza amafushane amaza (imvamisa engu-< 680 nm), okuholela ekungeneni kahle kokukhanya ngenxa yokumuncwa okukhulu kwama-molecule ebhayoloji (isb, i-hemoglobin ne-melanin) isifunda esibonakalayo6,7. Kodwa-ke, ama-photosensitizers amaningi avamile, njengama-chlorin, ama-porphyrins, nama-anthraquinones, anokumunca amaza amaza amafushane amaza (imvamisa engu-< 680 nm), okuholela ekungeneni kahle kokukhanya ngenxa yokumuncwa okukhulu kwama-molecule ebhayoloji (isb, i-hemoglobin ne-melanin) isifunda esibonakalayo6,7. 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. Kodwa-ke, ama-photosensitizers ajwayelekile njengama-chlorin, ama-porphyrins nama-anthraquinone anomunca amaza amaza amafushane amafushane uma kuqhathaniswa (< 680 nm) okuholela ekungeneni kokukhanya okungekuhle ngenxa yokumuncwa okukhulu kwama-molecule ebhayoloji (isb. i-hemoglobin ne-melanin) endaweni ebonakalayo6,7.然而, 大多数 传统 的 光敏剂, 如如 氢 氢 卟酚, 卟啉卟啉 蒽醌, 具有具有 较较 的的 的的 的的 波长的 (频率 <680 NM), 因此因此导致光穿透性差.然而, 大多数 传统 的 光敏剂, 二二 卟酚 卟酚, 卟啉卟啉, 具有具有 较相对 短的 的的 的的波长 (频率频率 频率 <680 nm) 因此 对 ,,,,,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. Kodwa-ke, ama-photosensitizers endabuko amaningi njengama-chlorin, ama-porphyrins nama-anthraquinones anomunca we-wavelength emfushane ngokuqhathaniswa (imvamisa engu-< 680 nm) ngenxa yokumuncwa okuqinile kwama-biomolecules njenge-hemoglobin ne-melanin okuholela ekungeneni kokukhanya okubi.Indawo ebonakalayo 6.7.Ngakho-ke, ama-photosensitizer amunca i-close-infrared (NIR) acushwe “ewindini lokwelapha” elingu-700–900 nm afaneleka kahle ku-phototherapy.Njengoba ukukhanya okuseduze kwe-infrared kukuncane kakhulu ukumuncwa izicubu zebhayoloji, kungaholela ekungeneni okujulile kanye nokulimala okuncane kwesithombe8,9.
Ngeshwa, izinzwa zezithombe ezimunca i-NIR ngokuvamile azikwazi ukuthwebula izithombe, amandla okukhiqiza i-singlett oxygen (1O2) aphansi, kanye nokucisha i-1O2 okubangwa ukuhlanganisa, okukhawulela ukusetshenziswa kwazo komtholampilo10,11.Nakuba kuye kwenziwa imizamo emikhulu yokuthuthukisa izakhiwo ze-photophysical kanye ne-photochemical yama-photosensitizers avamile, kuze kube manje imibiko eminingana ibike ukuthi ama-photosensitizers abamba i-NIR angakwazi ukuxazulula zonke lezi zinkinga.Ukwengeza, ama-photosensitizers ambalwa abonise isithembiso sokukhiqiza kahle kwe-1O212,13,14 uma ikhanya ngokukhanya okungaphezu kuka-800 nm, njengoba amandla e-photon ehla ngokushesha endaweni eseduze ne-IR.I-Triphenylamine (TFA) njengomnikezeli we-electron futhi [1,2,5]thiadiazole-[3,4-i]dipyrido[a,c]phenazine (TDP) njengeqembu lama-electron owamukelayo i-Donor-acceptor (DA) uhlobo lodayi ekilasini of odayi , abamunca eduze-infrared, eziye zacwaningwa kabanzi mayelana ne-infrared bioimaging II kanye ne-photothermal therapy (PTT) ngenxa ye-bandgap yabo encane.Ngakho-ke, odayi bohlobo lwe-DA bangasetshenziselwa i-PDT ngokuthakasela okuseduze kwe-IR, nakuba bengavamile ukufundwa njengezinzwa zezithombe ze-PDT.
Kuyaziwa ukuthi ukusebenza kahle okuphezulu kwe-intersystem crossing (ISC) yama-photosensitizers kukhuthaza ukwakheka kwe-1O2.Isu elivamile lokuthuthukisa inqubo ye-ISC ukuthuthukisa ukuhlangana kwe-spin-orbit (SOC) yama-photosensitizers ngokwethula ama-athomu asindayo noma ama-organic moieties akhethekile.Nokho, le ndlela isenobubi obuthile kanye nemikhawulo19,20.Muva nje, i-self-assembly ye-supramolecular inikeze indlela ehlakaniphile ephansi phezulu yokwenziwa kwezinto zokusebenza ezingeni lamangqamuzana, i-21,22 enezinzuzo eziningi ku-phototherapy: (1) ama-photosensitizers azihlanganisele angase abe namandla okwenza izakhiwo zeribhoni.Okufana nezakhiwo zikagesi ezinokusabalalisa okuminyene kwamaleveli wamandla ngenxa yezindlela ezidlulanayo phakathi kwamabhulokhi wokwakha.Ngakho-ke, ukufanisa amandla phakathi kwe-singlett ephansi yesimo esijabulisayo (S1) kanye nesimo esijabulisayo se-triplet (Tn) esiseduze sizothuthukiswa, okuzuzisa inqubo ye-ISC 23, 24.(2) Umhlangano we-Supramolecular uzonciphisa ukuphumula okungenawo umsakazo ngokusekelwe ku-intramolecular motion limitation mechanism (RIM), ebuye ikhuthaze inqubo ye-ISC 25, 26.(3) Umhlangano we-supramolecular ungavikela ama-molecule angaphakathi e-monomer ekugujwe yi-oxidation nasekuwohlokeni, ngaleyo ndlela ithuthukise kakhulu ukukwazi ukuthwebula izithombe kwe-photosensitizer.Uma kubhekwa lezi zinzuzo ezingenhla, sikholelwa ukuthi amasistimu e-supramolecular photosensitizer angaba enye indlela ethembisayo yokunqoba ukushiyeka kwe-PDT.
Izakhiwo ezisekelwe ku-Ru(II) ziyinkundla yezokwelapha ethembisayo yezicelo ezingase zibe khona ekuxilongweni nasekwelapheni izifo ngenxa yezakhiwo zazo eziyingqayizivele nezikhangayo zebhayoloji28,29,30,31,32,33,34.Ngaphezu kwalokho, ukuchichima kwezimo ezijabulile kanye nezakhiwo ze-photophysicochemical eziguqulekayo ze-Ru(II)-based complexes kunikeza izinzuzo ezinhle zokuthuthukiswa kwe-photosensitizers esekelwe ku-Ru(II)35,36,37,38,39,40.Isibonelo esiphawulekayo i-ruthenium(II) polypyridyl complex TLD-1433, okwamanje esezivivinyweni zomtholampilo zeSigaba II njenge-photosensitizer yokwelapha umdlavuza wesinye ongahlaseli imisipha (NMIBC)41.Ukwengeza, i-ruthenium (II) i-arene organometallic complexes isetshenziswa kabanzi njengama-chemotherapeutic agents ekwelapheni umdlavuza ngenxa yobuthi bayo obuncane kanye nokulula kokuguqulwa42,43,44,45.Izakhiwo ze-ionic ze-Ru(II) -arene organometallic complexes azikwazi nje ukuthuthukisa ukuncibilika okungekuhle kwama-chromophore e-DA kuzincibilikisi ezivamile, kodwa futhi zithuthukise ukuhlangana kwamachromophore e-DA.Ngaphezu kwalokho, ukwakheka kwe-pseudooctahedral half-sandwich ye-organometallic complexes ye-Ru(II) -arenes ingavimbela ngokuqinile ukuhlanganiswa kwe-H kohlobo lwe-chromophores yohlobo lwe-DA, ngaleyo ndlela kube lula ukwakheka kwe-J-aggregation ngamabhande okumuncwa aphindeke kabili.Kodwa-ke, ububi bemvelo be-Ru(II) -arene complexes, njengokuzinza okuphansi kanye/noma ukungatholakali kahle kwe-bioavailability, kungathinta ukusebenza kahle kokwelapha kanye nomsebenzi we-vivo we-arene-Ru(II) complexes.Kodwa-ke, ucwaningo luye lwabonisa ukuthi lokhu kungalungile kunganqotshwa ngokuhlanganisa izakhiwo ze-ruthenium ezinama-polymers ahambisanayo nge-encapsulation ngokomzimba noma ukuhlangana okuhlangene.
Kulo msebenzi, sibika i-DA-conjugated complexes ye-Ru(II)-arene (RuDA) ene-NIR trigger nge-coordination bond phakathi kwe-chromophore ye-DAD kanye ne-Ru(II)-arene moiety.Izinkimbinkimbi eziwumphumela zingazihlanganisa ngokwazo zibe ama-metalosupramolecular vesicles emanzini ngenxa yokuxhumana okungahlangani.Ngokuphawulekayo, umhlangano we-supramolecular unikeze i-RuDA nge-polymerization-induced intersystem crossing-over properties, okwandise kakhulu ukusebenza kahle kwe-ISC, okwakuvuna kakhulu i-PDT (Fig. 1A).Ukwandisa ukuqoqwa kwesimila kanye ne-vivo biocompatibility, i-Pluronic F127 egunyazwe yi-FDA (PEO-PPO-PEO) yasetshenziselwa ukuhlanganisa i-RuDA47,48,49 ukuze kwakhiwe ama-nanoparticles e-RuDA-NP (Umfanekiso 1B) osebenza njenge-PDT/ Dual- Imodi ye-PTT yommeleli .Ku-phototherapy yomdlavuza (Umfanekiso 1C), i-RuDA-NP yayisetshenziselwa ukwelapha amagundane anqunu ngamathumba e-MDA-MB-231 ukuze kufundwe ukusebenza kahle kwe-PDT ne-PTT ku-vivo.
Umdwebo we-Schematic we-photophysical mechanism ye-RuDA kumafomu e-monomeric kanye ne-aggregated ye-phototherapy yomdlavuza, ukuhlanganiswa kwe-B RuDA-NPs kanye ne-C RuDA-NPs ye-NIR-activated PDT ne-PTT.
I-RuDA, ehlanganisa ukusebenza kwe-TPA ne-TDP, yalungiswa ngokuvumelana nenqubo ekhonjiswe ku-Supplementary Figure 1 (Figure 2A), futhi i-RuDA ibonakala nge-spectra ye-1H ne-13C NMR, i-electrospray ionization mass spectrometry, nokuhlaziywa kwe-elemental (Izibalo Ezingeziwe 2-4 ).Imephu yomehluko we-electron density ye-RuDA yenguquko ye-singlet ephansi kakhulu yenziwe ikhompyutha nge-time-dependent density functional theory (TD-DFT) ukuze kufundwe inqubo yokudlulisa imali.Njengoba kukhonjisiwe Emfanekisweni Owengeziwe wesi-5, ukuminyana kwama-electron akhukhuleka ikakhulukazi ukusuka ku-triphenylamine kuya kuyunithi yokwamukela i-TDP ngemva kwe-photoexcitation, okungabalelwa kushintsho oluvamile lwe-intramolecular charge transfer (CT).
Isakhiwo samakhemikhali se-Ore B I-spectra yokumuncwa kwe-Ore ezixubeni zezilinganiso ezihlukahlukene ze-DMF namanzi.C Amanani okumuncwa ajwayelekile e-RuDA (800 nm) ne-ICG (779 nm) uma kuqhathaniswa nesikhathi esingu-0.5 W cm-2 we-808 nm laser light.D I-photodegradation ye-ABDA iboniswa ukwakhiwa kwe-RuDA-induced ye-1O2 ku-DMF / H2O izingxube ezinokuqukethwe kwamanzi ahlukene ngaphansi kwesenzo semisebe ye-laser ene-wavelength ye-808 nm namandla angu-0.5 W / cm2.
I-Abstract-UV-visible absorption spectroscopy yasetshenziswa ukutadisha izici zokuzihlanganisa ze-Ore ezixubeni ze-DMF namanzi ngezilinganiso ezihlukahlukene.Njengoba kuboniswe emkhiwaneni.I-2B, i-RuDA ibonisa amabhendi okumunca kusuka ku-600 kuya ku-900 nm ku-DMF enebhendi yokumunca ephezulu ku-729 nm.Ukwenyusa inani lamanzi kuholele ekushintsheni okubomvu kancane kancane kokumuncwa kwe-Ore kufika ku-800 nm, okubonisa ukuhlanganiswa kuka-J kwe-Ore ohlelweni oluhlanganisiwe.I-spectra ye-photoluminescence ye-RuDA ku-solvents ehlukene ikhonjiswe ku-Supplementary Figure 6. I-RuDA ibonakala ibonisa ukukhanya kwe-NIR-II okujwayelekile nge-wavelength ephezulu yokukhishwa kwe-ca.1050 nm ku-CH2Cl2 naku-CH3OH, ngokulandelanayo.I-Stokes shift enkulu (mayelana ne-300 nm) ye-RuDA ibonisa ushintsho oluphawulekayo ku-geometry yesimo esijabulisayo kanye nokwakhiwa kwezizwe ezijabule eziphansi.Imikhiqizo ye-luminescence quantum ye-Ore ku-CH2Cl2 ne-CH3OH inqunywe ukuthi ibe ngu-3.3 no-0.6%, ngokulandelana.Kodwa-ke, kungxube ye-methanol namanzi (5/95, v/v), ukushintshwa okubomvu okuncane kokukhipha kanye nokuncipha kwesivuno se-quantum (0.22%) kubonwe, okungenzeka kube ngenxa yokuzihlanganisa kwe-Ore. .
Ukuze ubone ngeso lengqondo ukuzihlanganisa kwe-ORE, sisebenzise i-liquid atomic force microscopy (AFM) ukuze sibone ngeso lengqondo izinguquko ze-morphological ku-ORE kusixazululo se-methanol ngemva kokwengeza amanzi.Lapho okuqukethwe kwamanzi kwakungaphansi kwama-80%, akukho ukuhlanganisa okucacile okubonwe (I-Supplementary Fig. 7).Kodwa-ke, ngokunyuka okuqhubekayo kokuqukethwe kwamanzi ku-90-95%, kwavela ama-nanoparticles amancane, abonisa ukuzihlanganisa kwe-Ore. isixazululo (Fig. 2C kanye Supplementary Fig. 8).Ngokuphambene nalokho, ukumuncwa kwe-indocyanine eluhlaza (ICG njengokulawula) kwehle ngokushesha ku-779 nm, okubonisa ukuthi i-photostability enhle kakhulu ye-RuDA.Ukwengeza, ukuzinza kwe-RuDA-NPs ku-PBS (pH = 5.4, 7.4 no-9.0), i-10% ye-FBS ne-DMEM (i-glucose ephezulu) ihlolwe yi-spectroscopy yokumuncwa okubonakalayo kwe-UV ngesikhathi samaphuzu ahlukahlukene.Njengoba kuboniswe ku-Supplementary Figure 9, izinguquko ezincane kumabhendi we-RuDA-NP wokumunca zibonwe ku-PBS ku-pH 7.4 / 9.0, FBS kanye ne-DMEM, okubonisa ukuzinza okuhle kakhulu kwe-RuDA-NP.Kodwa-ke, ku-acidic medium (рН = 5.4) i-hydrolysis ye-Ore itholakele.Siphinde sahlola ukuzinza kwe-RuDA ne-RuDA-NP sisebenzisa izindlela ze-high performance liquid chromatography (HPLC).Njengoba kuboniswe ku-Supplementary Figure 10, i-RuDA yayizinzile engxube ye-methanol namanzi (50/50, v / v) ngehora lokuqala, futhi i-hydrolysis yabonwa ngemva kwamahora angu-4.Kodwa-ke, kuphela ukuphakama okubanzi kwe-concave-convex kwabonwa kuma-RuDA NPs.Ngakho-ke, i-gel permeation chromatography (GPC) yasetshenziselwa ukuhlola ukuzinza kwe-RuDA NPs ku-PBS (pH = 7.4).Njengoba kuboniswe ku-Supplementary Figure 11, ngemva kwamahora angu-8 okufukamela ngaphansi kwezimo ezihloliwe, ukuphakama okuphezulu, ububanzi obuphakeme kanye nendawo ephakeme ye-NP RuDA akuzange kushintshe kakhulu, okubonisa ukuzinza okuhle kakhulu kwe-NP RuDA.Ngaphezu kwalokho, izithombe ze-TEM zibonise ukuthi i-morphology ye-RuDA-NP nanoparticles yahlala ingashintshiwe ngemva kwamahora angu-24 ku-buffer ye-PBS ehlanjululwe (pH = 7.4, Fig. 12).
Ngenxa yokuthi ukuzihlanganisa kunganikeza izici ezihlukene zokusebenza namakhemikhali ku-Ore, sibone ukukhishwa kwe-9,10-anthracenediylbis(methylene)dimalonic acid (ABDA, inkomba 1O2) kuzingxube zamanzi e-methanol.Insimbi enamanzi ahlukene50.Njengoba kuboniswe kuMfanekiso 2D kanye Nomfanekiso Owengeziwe 13, akukho ukucekelwa phansi kwe-ABDA okubonwe lapho okuqukethwe kwamanzi kungaphansi kwama-20%.Ngokukhuphuka komswakama ukuya ku-40%, ukuwohloka kwe-ABDA kwenzeka, njengoba kufakazelwa ukwehla kokuqina kwe-ABDA fluorescence.Kuphinde kwaqashelwa ukuthi okuqukethwe kwamanzi aphezulu kubangela ukuwohloka okusheshayo, okuphakamisa ukuthi i-RuDA self-assembly iyadingeka futhi inenzuzo ekuwohlokeni kwe-ABDA.Lesi simo sihluke kakhulu kuma-chromophore esimanje (ACQ-induced quenching).Uma ukhanyisa nge-laser ene-wavelength ye-808 nm, isivuno se-quantum se-1O2 RuDA engxubeni ye-98% H2O/2% DMF yi-16.4%, okuyizikhathi ezingu-82 ezingaphezu kwe-ICG (ΦΔ = 0.2%)51, ekhombisa ukusebenza kahle kwesizukulwane esingu-1O2 RuDA esimweni sokuhlanganisa.
Ama-Electron spins asebenzisa i-2,2,6,6-tetramethyl-4-piperidinone (TEMP) kanye ne-5,5-dimethyl-1-pyrroline N-oxide (DMPO) njengama-spin trap Resonance spectroscopy (ESR) yasetshenziswa ukukhomba izinhlobo eziwumphumela. I-AFK.by RuDA.Njengoba kuboniswe ku-Supplementary Figure 14, kuqinisekisiwe ukuthi i-1O2 ikhiqizwa ngezikhathi zokukhipha imisebe phakathi kwe-0 ne-4 imizuzu.Ukwengeza, lapho i-RuDA ifakwe i-DMPO ngaphansi kwe-irradiation, isignali ye-EPR yemigqa emine ejwayelekile ye-1: 2: 2: i-1 DMPO-OH · i-adduct itholiwe, ebonisa ukwakheka kwe-hydroxyl radicals (OH ·).Sekukonke, imiphumela engenhla ibonisa ikhono le-RuDA lokugqugquzela ukukhiqizwa kwe-ROS ngokusebenzisa inqubo ye-photosensitivity yezinhlobo ezimbili ze-I/II.
Ukuze uqonde kangcono izakhiwo ze-elekthronikhi ze-RuDA kumafomu we-monomeric kanye ne-aggregated, i-frontier molecular orbitals ye-RuDA kumafomu e-monomeric kanye ne-dimeric ibalwe kusetshenziswa indlela ye-DFT.Njengoba kuboniswe emkhiwaneni.I-3A, i-molecular orbital (HOMO) ephakeme kakhulu ebanjwe kakhulu ye-monomeric RuDA ihlukaniswa eceleni kwe-ligand backbone kanye ne-lowest unoccupied molecular orbital (LUMO) igxile kuyunithi yokwamukela i-TDP.Ngokuphambene nalokho, ukuminyana kwe-electron ku-HOMO ye-dimeric kugxile ku-ligand ye-molecule eyodwa ye-RuDA, kuyilapho ukuminyana kwe-electron ku-LUMO kugxile kakhulu kuyunithi yokwamukela enye i-molecule ye-RuDA, ebonisa ukuthi i-RuDA iku-dimer.Izici ze-CT.
A I-HOMO ne-LUMO ye-Ore ibalwa ngamafomu e-monomeric kanye ne-dimeric.B Amazinga wamandla ama-Singlet kanye nama-triplet we-Ore kuma-monomers nama-dimers.C Amazinga alinganiselwe we-RuDA kanye namashaneli e-ISC angenzeka njenge-monomeric C kanye ne-dimeric D. Imicibisholo ibonisa iziteshi ze-ISC ezingaba khona.
Ukusatshalaliswa kwama-electron nezimbobo ku-singlett ye-low-energy ejabulisayo ye-RuDA kumafomu we-monomeric kanye ne-dimeric kwahlaziywa kusetshenziswa isofthiwe ye-Multiwfn 3.852.53, eyabalwa kusetshenziswa indlela ye-TD-DFT.Njengoba kukhonjisiwe kulebula eyengeziwe.Njengoba kukhonjisiwe ku-Figure 1-2, izimbobo ze-monomeric RDA zihlukaniswa kakhulu kumgogodla we-ligand kulezi zifundazwe ezijabulisayo ze-singlet, kuyilapho ama-electron atholakala kakhulu eqenjini le-TDP, abonisa izici ze-intramolecular ze-CT.Ukwengeza, kulezi zifundazwe ezijabulile ze-singlet, kunokugqagqana okuncane noma okuncane phakathi kwezimbobo nama-electron, okuphakamisa ukuthi lezi zifundazwe ezijabulile ze-singlet zenza umnikelo othile kusuka ekusaseni kwasendaweni (LE).Kuma-dimers, ngaphezu kwezici ze-intramolecular CT kanye ne-LE, ingxenye ethile yezici ze-CT ye-intermolecular yabonwa ezifundeni ezifanele, ikakhulukazi i-S3, i-S4, i-S7, ne-S8, ngokusekelwe ekuhlaziyweni kwe-CT ye-intermolecular, nokuguqulwa kwe-CT intermolecular njengokuyinhloko. (Ithebula Lokwengeza).3).
Ukuze siqonde kangcono imiphumela yokuhlola, siqhubeke sihlola izakhiwo ze-RuDA ezijabulile ukuze sihlole umehluko phakathi kwama-monomers nama-dimers (Amathebula Engeziwe 4-5).Njengoba kuboniswe kuMfanekiso 3B, amazinga wamandla we-singlet kanye nezimo ezijabulisayo ze-triplet ze-dimer aminyene kakhulu kunalawo e-monomer, okusiza ukwehlisa igebe lamandla phakathi kwe-S1 ne-Tn. Kubikwe ukuthi izinguquko ze-ISC zingenzeka phakathi kwegebe elincane lamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 ne-Tn54. Kubikwe ukuthi izinguquko ze-ISC zingenzeka phakathi kwegebe elincane lamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 ne-Tn54. I-Сообщалось, что переходы I-ISC ingakwazi ukuxhumanisa i-reализованы ku-пределах небольшой энергетической щели (ΔES1-Tn <0,3 эВ) между S1 kanye ne-Tn5. Kubikwe ukuthi ukuguqulwa kwe-ISC kungenziwa ngaphakathi kwegebe elincane lamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 ne-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 ingakwazi ukuxazulula inkinga nge-пределах небольшой энергетической щели (ΔES1-Tn < 0,3 эВ) между S1 и Tn54. Kubikwe ukuthi uguquko lwe-ISC lungenziwa phakathi kwegebe elincane lamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 ne-Tn54.Ukwengeza, i-orbital eyodwa kuphela, ematasatasa noma engahlali muntu, okufanele ihluke ezifundeni ze-singlet eziboshwe kanye nezintathu ukuze kuhlinzekwe okubalulekile kwe-SOC okungeyona iqanda.Ngakho-ke, ngokusekelwe ekuhlaziyweni kwamandla okuvusa kanye nokuguqulwa kwe-orbital, zonke iziteshi ezingenzeka zenguquko ye-ISC ziboniswa ku-Fig.3C,D.Ngokuphawulekayo, isiteshi se-ISC esisodwa kuphela esitholakala ku-monomer, kuyilapho ifomu le-dimeric lineziteshi ezine ze-ISC ezingathuthukisa uguquko lwe-ISC.Ngakho-ke, kunengqondo ukucabanga ukuthi ama-molecule e-RuDA engeziwe ahlanganisiwe, iziteshi ze-ISC zizofinyeleleka kakhulu.Ngakho-ke, i-RuDA aggregates ingakha izakhiwo ze-electronic-band ezimbili ku-singlet kanye ne-triplet states, ukunciphisa igebe lamandla phakathi kwe-S1 ne-Tn etholakalayo, ngaleyo ndlela ikhulise ukusebenza kahle kwe-ISC ukuze kube lula isizukulwane se-1O2.
Ukuze siqhubeke sicacisa indlela eyisisekelo, sihlanganise inhlanganisela yereferensi ye-arene-Ru(II) complex (RuET) ngokushintsha amaqembu amabili e-ethyl ngamaqembu amabili e-triphenylamine phenyl ku-RuDA (Fig. 4A, for characterization full, bheka i-ESI, Supplementary 15 -21 ) Kusuka kumnikeli (diethylamine) kuya kumamukeli (TDF), i-RuET inezici ze-CT ze-intramolecular ezifana ne-RuDA.Njengoba kulindelekile, i-spectrum yokumunca ye-RuET ku-DMF ibonise ibhande lokudlulisa icala eliphansi lamandla ngokumuncwa okuqinile endaweni eseduze ye-infrared esifundeni se-600-1100 nm (Fig. 4B).Ukwengeza, i-RuET aggregation iphinde yabonwa ngokuqukethwe kwamanzi okwandayo, okuboniswe ku-redshift ye-absorption maximum, eyaqinisekiswa ngokuqhubekayo yi-imaging ye-AFM ye-liquid (I-Supplementary Fig. 22).Imiphumela ibonisa ukuthi i-RuET, njenge-RuDA, ingakha izifunda ze-intramolecular futhi izihlanganisele ezakhiweni ezihlanganisiwe.
Isakhiwo samakhemikhali se-RuET.I-B Absorption spectra ye-RuET ngezingxube zezilinganiso ezihlukahlukene ze-DMF namanzi.I-Plots C EIS Nyquist ye-RuDA ne-RuET.Izimpendulo ze-Photocurrent D ze-RuDA ne-RuET ngaphansi kwesenzo semisebe ye-laser ene-wavelength ye-808 nm.
I-photodegradation ye-ABDA phambi kwe-RuET ihlolwe nge-irradiation nge-laser ene-wavelength ye-808 nm.Ngokumangalisayo, akukho ukuwohloka kwe-ABDA okubonwe ezingxenyeni ezihlukahlukene zamanzi (Supplementary Fig. 23).Isizathu esingenzeka ukuthi i-RuET ayikwazi ukwenza kahle isakhiwo se-elekthronikhi esinebhande ngoba uchungechunge lwe-ethyl alukhuthazi ukudluliswa kwe-intermolecular charge.Ngakho-ke, i-electrochemical impedance spectroscopy (EIS) kanye nezilinganiso ze-photocurrent zesikhashana zenziwa ukuze kuqhathaniswe izakhiwo ze-photoelectrochemical ze-RuDA ne-RuET.Ngokusho kwesakhiwo seNyquist (Umfanekiso we-4C), i-RuDA ibonisa i-radius encane kakhulu kune-RuET, okusho ukuthi i-RuDA56 inokuthutha okusheshayo kwe-electron intermolecular kanye ne-conductivity engcono.Ukwengeza, i-photocurrent density ye-RuDA iphakeme kakhulu kune-RuET (Fig. 4D), eqinisekisa ukusebenza kahle kokudlulisa icala kwe-RuDA57.Ngakho, iqembu le-phenyl le-triphenylamine ku-Ore lidlala indima ebalulekile ekuhlinzekeni ukudluliswa kwe-intermolecular charge kanye nokwakhiwa kwesakhiwo se-elekthronikhi esinebhande.
Ukwandisa ukunqwabelana kwesimila kanye ne-vivo biocompatibility, siphinde sahlanganisa i-RuDA nge-F127.Isilinganiso se-hydrodynamic diameter ye-RuDA-NPs sinqunywe ukuba yi-123.1 nm ngokusatshalaliswa okuncane (PDI = 0.089) kusetshenziswa indlela yokusabalalisa ukukhanya okunamandla (i-DLS) (Umfanekiso we-5A), okukhuthaza ukuqoqwa kwe-tumor ngokwandisa ukuvuthwa nokugcinwa.EPR) umphumela.Izithombe ze-TEM zibonise ukuthi ama-Ore NP anomumo oyindilinga ofana nobubanzi obuyisilinganiso esingu-86 nm.Ngokuphawulekayo, ukumuncwa okuphezulu kwe-RuDA-NPs kubonakala ku-800 nm (I-Supplementary Fig. 24), ebonisa ukuthi i-RuDA-NPs ingase igcine imisebenzi kanye nezakhiwo zokuzihlanganisa i-RuDAs.Isivuno se-ROS quantum esibaliwe se-NP Ore yi-15.9%, esiqhathaniswa ne-Ore.Izakhiwo ze-photothermal ze-RuDA NPs zafundwa ngaphansi kwesenzo semisebe ye-laser ene-wavelength ye-808 nm usebenzisa ikhamera ye-infrared.Njengoba kuboniswe emkhiwaneni.I-5B, C, iqembu lokulawula (i-PBS kuphela) labhekana nokunyuka okuncane kwezinga lokushisa, kuyilapho izinga lokushisa lesisombululo se-RuDA-NPs landa ngokushesha ngokushisa okwandayo (ΔT) kuya ku-15.5, 26.1, no-43.0 ° C.Ukugxila okuphezulu kwakungama-25, 50, kanye ne-100 µM, ngokulandelana, okubonisa umphumela oqinile we-photothermal we-RuDA NPs.Ukwengeza, izilinganiso zomjikelezo wokushisa / wokupholisa zithathwe ukuze kuhlolwe ukuzinza kwe-photothermal ye-RuDA-NP futhi kuqhathaniswe ne-ICG.Ukushisa kwe-Ore NPs akuzange kunciphe ngemva kwemijikelezo emihlanu yokushisa / yokupholisa (Fig. 5D), ebonisa ukuzinza okuhle kakhulu kwe-photothermal ye-Ore NPs.Ngokuphambene, i-ICG ibonisa ukuzinza okuphansi kwe-photothermal njengoba kubonakala ngokunyamalala okubonakalayo kwethafa lokushisa kwe-photothermal ngaphansi kwezimo ezifanayo.Ngokwendlela yangaphambilini58, ukusebenza kahle kokuguqulwa kwe-photothermal (PCE) ye-RuDA-NP kubalwa njenge-24.2%, ephakeme kunezinto ezikhona ze-photothermal ezifana ne-nanorods yegolide (21.0%) nama-nanoshell egolide (13.0%)59.Ngakho-ke, i-NP Ore ibonisa izakhiwo ezinhle kakhulu ze-photothermal, ezibenza bathembise ama-PTT agents.
Ukuhlaziywa kwezithombe ze-DLS ne-TEM ze-RuDA NPs (inset).B Izithombe ezishisayo zokugxila okuhlukahlukene kwe-RuDA NPs ezivezwe emisebeni ye-laser ku-wavelength ye-808 nm (0.5 W cm-2).C Amajika okuguqulwa kwe-Photothermal okugxiliswa okuhlukahlukene kwe-ore NPs, okuyidatha yobuningi.B. D Ukukhuphuka kwezinga lokushisa kwe-ORE NP ne-ICG ngaphezu kwemijikelezo yokushisisa yokupholisa emi-5.
I-Photocytotoxicity ye-RuDA NPs ngokumelene ne-MDA-MB-231 yamangqamuzana omdlavuza webele womuntu yahlolwa ku-vitro.Njengoba kuboniswe emkhiwaneni.I-6A, B, i-RuDA-NPs ne-RuDA ibonise i-cytotoxicity enganaki lapho kungabikho i-irradiation, okusho ukuthi ubuthi obuphansi obumnyama be-RuDA-NPs ne-RuDA.Kodwa-ke, ngemuva kokuchayeka emisebeni ye-laser ebangeni elingu-808 nm, i-RuDA kanye ne-RuDA NPs babonise i-photocytotoxicity eqinile ngokumelene namaseli omdlavuza we-MDA-MB-231 anamanani e-IC50 (isigamu esiphezulu se-inhibitory concentration) se-5.4 kanye ne-9.4 μM, ngokulandelana, ekhombisa. ukuthi i-RuDA-NP ne-RuDA inamandla okwenza i-phototherapy yomdlavuza.Ukwengeza, i-photocytotoxicity ye-RuDA-NP ne-RuDA yaphinde yaphenywa phambi kwe-vitamin C (Vc), i-ROS scavenger, ukucacisa indima ye-ROS ku-cytotoxicity ebangelwa ukukhanya.Ngokusobala, ukusebenza kwamaseli kukhuphuke ngemuva kokwengezwa kwe-Vc, futhi amanani we-IC50 we-RuDA kanye ne-RuDA NPs ayengama-25.7 no-40.0 μM, ngokulandelana, okufakazela indima ebalulekile ye-ROS ku-photocytotoxicity ye-RuDA ne-RuDA NPs.I-cytotoxicity ebangelwa ukukhanya ye-RuDA-NPs kanye ne-RuDA kumaseli omdlavuza we-MDA-MB-231 ngokungcoliswa kwamaseli aphilayo/afile kusetshenziswa i-calcein AM (i-fluorescence eluhlaza yamaseli aphilayo) kanye ne-propidium iodide (PI, i-fluorescence ebomvu yamaseli afile).kuqinisekiswa amaseli) njengama-fluorescent probes.Njengoba kuboniswe ku-Figure 6C, amaseli aphathwa nge-RuDA-NP noma i-RuDA ahlala esebenza ngaphandle kwemisebe, njengoba kuboniswa yi-fluorescence eluhlaza kakhulu.Ngokuphambene nalokho, ngaphansi kwe-laser irradiation, i-fluorescence ebomvu kuphela yabonwa, eqinisekisa i-photocytotoxicity ephumelelayo ye-RuDA noma i-RuDA NPs.Kuyaphawuleka ukuthi i-fluorescence eluhlaza ivele phezu kokwengezwa kwe-Vc, okubonisa ukwephulwa kwe-photocytotoxicity ye-RuDA ne-RuDA NPs.Le miphumela ihambisana ne-in vitro photocytotoxicity assays.
Ukusebenza okuncike kumthamo wamaseli we-A RuDA- no-B RuDA-NP kumaseli e-MDA-MB-231 lapho kukhona noma ingekho i-Vc (0.5 mM), ngokulandelanayo.Amabha wephutha, asho ukuthi ± ukuchezuka okujwayelekile (n = 3). Ukuhlola okungu-t okungabhangqiwe, okunezinhlangothi ezimbili *p <0.05, **p <0.01, kanye ne-**p <0.001. Ukuhlola okungu-t okungabhangqiwe, okunezinhlangothi ezimbili *p <0.05, **p <0.01, kanye ne-**p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 futhi ***p <0,001. Ukuhlola kuka-t okunemisila emibili okungabhanqiwe *p<0.05, **p<0.01, kanye ne-**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 futhi ***p <0,001. Ukuhlola kuka-t okunemisila emibili okungabhanqiwe *p<0.05, **p<0.01, kanye ne-**p<0.001.C Ukuhlaziya amabala eseli aphilayo/afile kusetshenziswa i-calcein AM ne-propidium iodide njengama-fluorescent probe.Ibha yesikali: 30 µm.Izithombe ezimele izimpinda ezintathu zebhayoloji ezivela eqenjini ngalinye ziyaboniswa.D Izithombe ze-Confocal fluorescence zokukhiqizwa kwe-ROS kumaseli e-MDA-MB-231 ngaphansi kwezimo zokwelashwa ezihlukene.I-fluorescence ye-DCF eluhlaza ibonisa ukuba khona kwe-ROS.Irradiate nge-laser ene-wavelength engu-808 nm enamandla angu-0.5 W/cm2 imizuzu engu-10 (300 J/cm2).Ibha yesikali: 30 µm.Izithombe ezimele izimpinda ezintathu zebhayoloji ezivela eqenjini ngalinye ziyaboniswa.I-E Flow cytometry RuDA-NPs (50 µM) noma i-RuDA (50 µM) ukuhlaziywa kokwelashwa nge-laser engu-808 nm noma ngaphandle kwayo (0.5 W cm-2) lapho kukhona kanye nokungabikho kwe-Vc (0.5 mM) imizuzu engu-10.Izithombe ezimele izimpinda ezintathu zebhayoloji ezivela eqenjini ngalinye ziyaboniswa.I-F Nrf-2, HSP70 kanye ne-HO-1 yamaseli e-MDA-MB-231 aphathwe nge-RuDA-NPs (50 µM) noma ngaphandle kwawo 808 nm laser irradiation (0.5 W cm-2, 10 min, 300 J cm-2) , amaseli aveza 2).Izithombe ezimele izimpinda ezimbili zebhayoloji ezivela eqenjini ngalinye ziyaboniswa.
Ukukhiqizwa kwe-ROS ye-intracellular kumaseli e-MDA-MB-231 kwahlolwa kusetshenziswa indlela yokungcolisa i-2,7-dichlorodihydrofluorescein diacetate (DCFH-DA).Njengoba kuboniswe emkhiwaneni.I-6D, amaseli aphathwe nge-RuDA-NPs noma i-RuDA abonise i-fluorescence eluhlaza ehlukile lapho ekhishwa nge-laser engu-808 nm, ebonisa ukuthi i-RuDA-NPs ne-RuDA inekhono eliphumelelayo lokukhiqiza i-ROS.Ngokuphambene nalokho, ngokungabikho kokukhanya noma lapho kukhona i-Vc, kuphela isignali ye-fluorescent ebuthakathaka yamaseli yabonwa, ebonisa ukwakheka okuncane kwe-ROS.Amazinga e-ROS e-intracellular kumaseli e-RuDA-NP kanye namaseli e-MDA-MB-231 aphethwe yi-RuDA aphinde anqunywa ukugeleza kwe-cytometry.Njengoba kuboniswe ku-Supplementary Figure 25, isilinganiso se-fluorescence intensity (MFI) ekhiqizwa yi-RuDA-NPs kanye ne-RuDA ngaphansi kwe-808 nm laser irradiation yanda kakhulu cishe izikhathi ezingu-5.1 kanye ne-4.8, ngokulandelana, uma kuqhathaniswa neqembu lokulawula, eliqinisekisa ukwakheka kwabo okuhle kakhulu kwe-AFK.umthamo.Kodwa-ke, amazinga we-ROS we-intracellular kumaseli we-RuDA-NP noma we-MDA-MB-231 aphathwa nge-RuDA ayeqhathaniswa kuphela nezilawuli ngaphandle kwe-laser irradiation noma phambi kwe-Vc, efana nemiphumela yokuhlaziywa kwe-confocal fluorescence.
Kubonisiwe ukuthi i-mitochondria iyinkomba eyinhloko ye-Ru(II) -arene complexes60.Ngakho-ke, ukutholakala kwe-subcellular ye-RuDA ne-RuDA-NPs kuphenywe.Njengoba kuboniswe ku-Supplementary Figure 26, i-RuDA ne-RuDA-NP ibonisa amaphrofayili afanayo okusabalalisa amaselula ngokuqoqwa okuphezulu kakhulu ku-mitochondria (62.5 ± 4.3 kanye ne-60.4 ± 3.6 ng/mg amaprotheni, ngokulandelana).Kodwa-ke, inani elincane kuphela le-Ru elitholakala ezingxenyeni zenuzi ze-Ore ne-NP Ore (3.5 no-2.1%, ngokulandelana).Ingxenye esele yeseli iqukethe i-ruthenium esele: 31.7% (30.6 ± 3.4 ng / mg amaprotheni) we-RuDA kanye ne-42.9% (47.2 ± 4.5 ng / mg amaprotheni) we-RuDA-NPs.Ngokuvamile, i-Ore kanye ne-NP Ore ikakhulukazi inqwabelana ku-mitochondria.Ukuhlola ukungasebenzi kahle kwe-mitochondrial, sisebenzise i-JC-1 ne-MitoSOX Red staining ukuze sihlole amandla olwelwesi lwe-mitochondrial kanye namandla okukhiqiza e-superoxide, ngokulandelana.Njengoba kuboniswe ku-Supplementary Fig. 27, i-fluorescence eluhlaza kakhulu (JC-1) nobomvu (MitoSOX Red) yabonwa kumaseli aphathwe kokubili i-RuDA ne-RuDA-NPs ngaphansi kwe-808 nm laser irradiation, okubonisa ukuthi kokubili i-RuDA ne-RuDA-NPs i-fluorescent kakhulu. Ingenza ngempumelelo i-mitochondrial membrane depolarization kanye nokukhiqizwa kwe-superoxide.Ngaphezu kwalokho, indlela yokufa kweseli yanqunywa kusetshenziswa ukuhlaziya okusekelwe ku-flow cytometry ye-annexin V-FITC/propidium iodide (PI).Njengoba kuboniswe ku-Figure 6E, lapho i-radiated nge-808 nm laser, i-RuDA ne-RuDA-NP yenze izinga le-apoptosis elanda kakhulu (i-quadrant engezansi kwesokudla) kumaseli e-MDA-MB-231 uma kuqhathaniswa ne-PBS noma i-PBS plus laser.amaseli acutshunguliwe.Kodwa-ke, lapho i-Vc ingeziwe, izinga le-apoptosis le-RuDA ne-RuDA-NP lehle kakhulu ukusuka ku-50.9% no-52.0% kuya ku-15.8% no-17.8%, ngokulandelana, okuqinisekisa indima ebalulekile ye-ROS ku-photocytotoxicity ye-RuDA ne-RuDA-NP..Ukwengeza, amangqamuzana amancane e-necrotic abonwa kuwo wonke amaqembu ahlolwe (i-quadrant ephezulu kwesokunxele), ephakamisa ukuthi i-apoptosis ingase ibe uhlobo oluvelele lokufa kwamaseli okubangelwa i-RuDA ne-RuDA-NPs.
Njengoba ukulimala kokucindezeleka kwe-oxidative kuyisici esiyinhloko se-apoptosis, isici se-nyukliya esihlotshaniswa ne-erythroid 2, i-factor 2 (Nrf2) 62, umlawuli oyinhloko wesistimu ye-antioxidant, yaphenywa ku-MDA-MB-231 ye-RuDA-NPs.Indlela yokwenza ye-RuDA NPs ebangelwa i-irradiation.Ngesikhathi esifanayo, ukubonakaliswa kwephrotheni engezansi ye-heme oxygenase 1 (HO-1) nayo yatholwa.Njengoba kuboniswe ku-Figure 6F kanye ne-Supplementary Figure 29, i-phototherapy ye-RuDA-NP-mediated yanda amazinga we-Nrf2 kanye ne-HO-1 uma kuqhathaniswa neqembu le-PBS, okubonisa ukuthi i-RuDA-NPs ingase ikhuthaze izindlela zokubonisa ukucindezeleka kwe-oxidative.Ukwengeza, ukutadisha umphumela we-photothermal we-RuDA-NPs63, ukuvezwa kwe-heat shock protein Hsp70 nakho kwahlolwa.Kuyacaca ukuthi amaseli aphathwe nge-RuDA-NPs + 808 nm laser irradiation abonise ukwanda kwe-Hsp70 uma kuqhathaniswa namanye amaqembu amabili, okubonisa impendulo yeselula ku-hyperthermia.
Imiphumela ye-in vitro ephawulekayo isishukumisele ukuthi siphenye ukusebenza kwe-in vivo kwe-RuDA-NP kumagundane anqunu anamathumba e-MDA-MB-231.Ukusatshalaliswa kwezicubu ze-RuDA NPs kwacwaningwa ngokunquma okuqukethwe kwe-ruthenium esibindini, inhliziyo, ubende, izinso, amaphaphu, namathumba.Njengoba kuboniswe emkhiwaneni.I-7A, okuqukethwe okuphezulu kwama-Ore NPs ezithweni ezijwayelekile kuvele ngesikhathi sokuqala sokubhekwa (4 h), kuyilapho okuqukethwe okuphezulu kunqunywa ezicutshini zesimila amahora angu-8 ngemva komjovo, okungenzeka ngenxa ye-Ore NPs.Umphumela we-EPR we-LF.Ngokwemiphumela yokusabalalisa, isikhathi esifanele sokwelashwa nge-NP ore sithathwe amahora angu-8 ngemva kokuphathwa.Ukubonisa inqubo yokuqoqwa kwe-RuDA-NPs ezindaweni ze-tumor, izakhiwo ze-photoacoustic (PA) ze-RuDA-NPs ziqashwe ngokurekhoda izimpawu ze-PA ze-RuDA-NPs ngezikhathi ezahlukene ngemva kokujova.Okokuqala, isignali ye-PA ye-RuDA-NP ku-vivo yahlolwa ngokurekhoda izithombe ze-PA zesayithi lesimila ngemuva komjovo we-intratumoral we-RuDA-NP.Njengoba kuboniswe ku-Supplementary Figure 30, i-RuDA-NPs ibonise isignali eqinile ye-PA, futhi kwakukhona ukuhlobana okuhle phakathi kokuhlushwa kwe-RuDA-NP kanye nokuqina kwesignali ye-PA (I-Supplementary Figure 30A).Khona-ke, ku-vivo PA izithombe zezindawo zesimila zarekhodwa ngemva komjovo womjovo we-RuDA ne-RuDA-NP ngezikhathi ezihlukene ngemva komjovo.Njengoba kuboniswe ku-Figure 7B, isignali ye-PA ye-RuDA-NPs evela endaweni ye-tumor yanda kancane kancane ngokuhamba kwesikhathi futhi yafinyelela ithafa emahoreni angu-8 ngemva kokujova, ngokuhambisana nemiphumela yokusabalalisa izicubu ezinqunywe ukuhlaziywa kwe-ICP-MS.Ngokuphathelene ne-RuDA (I-Supplementary Fig. 30B), ubukhulu besignali ye-PA buvele amahora angu-4 ngemva kokujova, okubonisa izinga elisheshayo lokungena kwe-RuDA ku-tumor.Ukwengeza, ukuziphatha kwe-excretory ye-RuDA ne-RuDA-NPs kwaphenywa ngokunquma inani le-ruthenium emchameni nasendle usebenzisa i-ICP-MS.Umzila oyinhloko wokuqedwa kwe-RuDA (I-Supplementary Fig. 31) kanye ne-RuDA-NPs (Fig. 7C) ihamba ngendle, futhi ukukhishwa okuphumelelayo kwe-RuDA kanye ne-RuDA-NPs kwabonwa phakathi nenkathi yokufunda yezinsuku ezingu-8, okusho ukuthi i-RuDA kanye ne-RuDA-NPs ingase isuswe ngokuphumelelayo emzimbeni ngaphandle kobuthi besikhathi eside.
A. Ukusatshalaliswa kwe-Ex vivo kwe-RuDA-NP kumathishu egundane kwanqunywa okuqukethwe kwe-Ru (iphesenti lomthamo olawulwayo we-Ru (ID) ngegremu ngalinye lesithishu) ngezikhathi ezihlukile ngemuva komjovo.Idatha isho ukuchezuka okujwayelekile ± (n = 3). Ukuhlola okungu-t okungabhangqiwe, okunezinhlangothi ezimbili *p <0.05, **p <0.01, kanye ne-**p <0.001. Ukuhlola okungu-t okungabhangqiwe, okunezinhlangothi ezimbili *p <0.05, **p <0.01, kanye ne-**p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 futhi ***p <0,001. Ukuhlola kuka-t okunemisila emibili okungabhanqiwe *p<0.05, **p<0.01, kanye ne-**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 futhi ***p <0,001. Ukuhlola kuka-t okunemisila emibili okungabhanqiwe *p<0.05, **p<0.01, kanye ne-**p<0.001.Izithombe ze-B PA zezindawo zesimila ku-vivo ku-808 nm excitation ngemva kokulawulwa komthambo we-RuDA-NPs (10 µmol kg-1) ngezikhathi ezahlukene.Ngemuva kokuphathwa ngomthambo i-RuDA NPs (10 µmol kg-1), i-C Ru yakhishwa emagundwaneni anomchamo nendle ngezikhathi ezahlukene.Idatha isho ukuchezuka okujwayelekile ± (n = 3).
Amandla okushisa we-RuDA-NP ku-vivo afundwe kumagundane anqunu ane-MDA-MB-231 kanye ne-RuDA tumors ukuze kuqhathaniswe.Njengoba kuboniswe emkhiwaneni.I-8A kanye ne-Fig. 32 eyengeziwe, iqembu lokulawula (i-saline) libonise ushintsho oluncane lokushisa (ΔT ≈ 3 ° C) ngemva kwemizuzu ye-10 yokuchayeka okuqhubekayo.Kodwa-ke, izinga lokushisa le-RuDA-NPs ne-RuDA lenyuka ngokushesha ngamazinga okushisa aphezulu angu-55.2 no-49.9 ° C, ngokulandelana, ukuhlinzeka nge-hyperthermia eyanele yokwelashwa komdlavuza we-vivo.Ukwanda okuphawuliwe kokushisa okuphezulu kwe-RuDA NPs (ΔT ≈ 24 ° C) uma kuqhathaniswa ne-RuDA (ΔT ≈ 19 ° C) kungase kube ngenxa yokungena kwayo okungcono nokuqoqwa kwezicubu ze-tumor ngenxa yomphumela we-EPR.
Izithombe ezishisayo ze-infrared zamagundane anamathumba e-MDA-MB-231 afakwe i-laser engu-808 nm ngezikhathi ezihlukene amahora angu-8 ngemva komjovo.Izithombe ezimele izimpinda ezine zebhayoloji ezivela eqenjini ngalinye ziyaboniswa.B Ivolumu yesimila esihlobene kanye no-C Isilinganiso sesimila esimaphakathi samaqembu ahlukene amagundane ngesikhathi sokwelashwa.D Amajika esisindo somzimba amaqembu ahlukene amagundane.Irradiate nge-laser ene-wavelength engu-808 nm enamandla angu-0.5 W/cm2 imizuzu engu-10 (300 J/cm2).Amabha wephutha, asho ukuthi ± ukuchezuka okujwayelekile (n = 3). Ukuhlola okungu-t okungabhangqiwe, okunezinhlangothi ezimbili *p <0.05, **p <0.01, kanye ne-**p <0.001. Ukuhlola okungu-t okungabhangqiwe, okunezinhlangothi ezimbili *p <0.05, **p <0.01, kanye ne-**p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 futhi ***p <0,001. Ukuhlola kuka-t okunemisila emibili okungabhanqiwe *p<0.05, **p<0.01, kanye ne-**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 futhi ***p <0,001. Ukuhlola kuka-t okunemisila emibili okungabhanqiwe *p<0.05, **p<0.01, kanye ne-**p<0.001. Izithombe ze-E H&E ezingcolisa izitho ezinkulu nezimila ezivela emaqenjini ahlukene okwelapha, okuhlanganisa i-Saline, i-Saline + Laser, i-RuDA, i-RuDA + Laser, i-RuDA-NPs, ne-RuDA-NPs + Laser amaqembu. Izithombe ze-E H&E ezingcolisa izitho ezinkulu nezimila ezivela emaqenjini ahlukene okwelapha, okuhlanganisa i-Saline, i-Saline + Laser, i-RuDA, i-RuDA + Laser, i-RuDA-NPs, ne-RuDA-NPs + Laser amaqembu. 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. Izithombe ze-E H&E ezingcolisa izitho ezinkulu nezimila ezivela emaqenjini ahlukene okwelapha, okuhlanganisa i-saline, i-saline + laser, i-RuDA, i-RuDA + Laser, i-RuDA-NPs, kanye namaqembu e-RuDA-NPs + Laser.來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 +和激光（RuDA-NPs、RuDA + 和激光 NPs、RuDA + 激光）來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, пухолей из различных групп лечения, включая физиологический раствор, физир DA, Rule, Rule, NP-DA, Rule, +DA, Rule, +DA I-E H&E staining yezitho ezinkulu namathumba avela emaqenjini ahlukene okwelapha ahlanganisa i-saline, i-saline + laser, i-RuDA, i-RuDA + laser, i-RuDA-NPs, ne-RuDA-NPs + laser.Ibha yesikali: 60 µm.
Umphumela we-phototherapy ku-vivo nge-RuDA kanye ne-RuDA NPs wahlolwa lapho amagundane anqunu ane-MDA-MB-231 tumors ajovwe nge-intravenously nge-RuDA noma i-RuDA NPs ngomthamo owodwa we-10.0 µmol kg-1 nge-vein yomsila, bese i-8 amahora ngemuva kokujova.i-laser irradiation ene-wavelength engu-808 nm.Njengoba kuboniswe kuMfanekiso 8B, amavolumu e-tumor anyuswe kakhulu ku-saline kanye namaqembu e-laser, okubonisa ukuthi i-saline noma i-laser 808 irradiation ibe nomthelela omncane ekukhuleni kwesimila.Njengaseqenjini le-saline, ukukhula kwe-tumor okusheshayo kuphinde kwabonwa kumagundane aphathwa nge-RuDA-NPs noma i-RuDA ngokungabikho kwe-laser irradiation, okubonisa ubuthi bawo obumnyama obuphansi.Ngokuphambene, ngemva kokushiswa kwe-laser, kokubili ukwelashwa kwe-RuDA-NP kanye ne-RuDA kubangele ukuhlehla kwe-tumor okuphawulekayo ngokunciphisa umthamo we-tumor ka-95.2% no-84.3%, ngokulandelana, uma kuqhathaniswa neqembu eliphathwa ngosawoti, okubonisa i-PDT ye-synergistic enhle kakhulu., ilamule umphumela we-RuDA/CHTV.- I-NP noma i-Ore. Uma kuqhathaniswa ne-RuDA, i-RuDA NPs ibonise umphumela ongcono we-phototherapeutic, owawubangelwa ikakhulukazi umphumela we-EPR we-RuDA NPs.Imiphumela yokuvimbela ukukhula kwe-tumor yabuye yahlolwa ngesisindo se-tumor ekhishwe ngosuku lwe-15 yokwelashwa (Fig. 8C kanye ne-Supplementary Fig. 33).Isilinganiso se-tumor mass ku-RuDA-NP amagundane aphathwayo kanye namagundane aphathwe i-RuDA kwakuyi-0.08 kanye ne-0.27 g, ngokulandelanayo, eyayilula kakhulu kuneqembu lokulawula (1.43 g).
Ngaphezu kwalokho, isisindo somzimba wamagundane sabhalwa njalo ezinsukwini ezintathu ukuze kufundwe ubuthi obumnyama be-RuDA-NPs noma i-RuDA ku-vivo.Njengoba kuboniswe ku-Figure 8D, akukho mehluko omkhulu wesisindo somzimba owabonwa kuwo wonke amaqembu okwelapha. Ngaphezu kwalokho, ukungcoliswa kwe-hematoxylin ne-eosin (H&E) kwezitho ezinkulu (inhliziyo, isibindi, ubende, amaphaphu, nezinso) kwenziwa emaqenjini ahlukene okwelapha. Ngaphezu kwalokho, ukungcoliswa kwe-hematoxylin ne-eosin (H&E) kwezitho ezinkulu (inhliziyo, isibindi, ubende, amaphaphu, nezinso) kwenziwa emaqenjini ahlukene okwelapha. Кроме того, было проведено окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селезенки, легких и порганов). Ukwengeza, ukungcoliswa kwe-hematoxylin ne-eosin (H&E) kwezitho ezinkulu (inhliziyo, isibindi, ubende, amaphaphu, nezinso) ezivela emaqenjini ahlukene okwelapha kwenziwa.此外，对不同治疗组的主要器官（心脏、肝脏、脾脏、肺和肾脏）进苏木精和伊红(H&E) (H&E) Кроме того, проводили окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селезенки, легких и почерук) в разлихучуные Ngaphezu kwalokho, ukungcoliswa kwe-hematoxylin ne-eosin (H&E) kwezitho ezinkulu (inhliziyo, isibindi, ubende, amaphaphu nezinso) kwenziwa emaqenjini ahlukene okwelapha.Njengoba kuboniswe ku-Fig.I-8E, izithombe ezingcolile ze-H&E zezitho ezinhlanu ezinkulu ezivela kumaqembu e-RuDA-NPs kanye ne-RuDA azibonisi ukungahambi kahle okusobala noma ukulimala kwezitho. I-8E, izithombe ezingcolile ze-H&E zezitho ezinhlanu ezinkulu ezivela kumaqembu e-RuDA-NPs kanye ne-RuDA azibonisi ukungahambi kahle okusobala noma ukulimala kwezitho.Njengoba kuboniswe emkhiwaneni.8E, изображения окрашивания I-H&E ithuthukisa ukusebenza kwe-RuDA-NPs kanye ne-RuDA ayikaze ibe nemiphumela emibi kakhulu. I-8E, izithombe ze-H&E ezingcolisa izitho ezinhlanu ezinkulu ezivela kumaqembu e-RuDA-NPs kanye ne-RuDA azibonisi ukungahambi kahle kwesitho esisobala noma izilonda.如图8E 所示，來自RuDA-NPs 和RuDA 组的五个主要器官的H&E 染色图像沒有显示出明显的异常或器官。如图8E 所示，來自RuDA-NPs 和RuDA 组的五个主要器官的H&E Njengoba i-рисунке 8E iqala, futhi i-H&E ikhokha kakhulu i-RuDA-NPs kanye ne-RuDA ayikaze isebenze ngokuzenzakalelayo. Njengoba kuboniswe ku-Figure 8E, izithombe ezingcolile ze-H & E zezitho ezinhlanu ezinkulu ezivela ku-RuDA-NPs kanye namaqembu e-RuDA abonisanga ukungahambi kahle okusobala noma ukulimala kwesitho.Le miphumela ibonise ukuthi i-RuDA-NP noma i-RuDA ayizange ibonise izimpawu zobuthi ku-vivo. Ngaphezu kwalokho, izithombe ezingcolile ze-H & E zezimila zibonise ukuthi kokubili amaqembu e-RuDA + Laser kanye ne-RuDA-NPs + Laser angabangela ukubhujiswa okukhulu kwamangqamuzana omdlavuza, okubonisa ukusebenza kahle kwe-vivo phototherapeutic ye-RuDA ne-RuDA-NPs. Ngaphezu kwalokho, izithombe ezingcolile ze-H & E zezimila zibonise ukuthi kokubili amaqembu e-RuDA + Laser kanye ne-RuDA-NPs + Laser angabangela ukubhujiswa okukhulu kwamangqamuzana omdlavuza, okubonisa ukusebenza kahle kwe-vivo phototherapeutic ye-RuDA ne-RuDA-NPs.Ngaphezu kwalokho, izithombe ze-tumor ezinamabala e-hematoxylin-eosin zibonise ukuthi womabili amaqembu e-RuDA+Laser kanye ne-RuDA-NPs+Laser angabangela ukubhujiswa okukhulu kwamangqamuzana omdlavuza, okubonisa ukusebenza kahle kwe-phototherapeutic kwe-RuDA kanye ne-RuDA-NPs ku-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坏，请入明的时公细坏，话公给坏，话公导坏细公平坏。此外 , 肿瘤 的 & e 染色 显示 , ruda + laser kanye ruda-nps + laser 组均 导致 的 癌细胞 破坏 , 证明 组组均。 ...Ngaphezu kwalokho, i-hematoxylin ne-eosin enezithombe ze-tumor izithombe zabonisa ukuthi kokubili amaqembu e-RuDA+Laser kanye ne-RuDA-NPs+Laser abangele ukubhujiswa okukhulu kwamangqamuzana omdlavuza, okubonisa ukusebenza kahle kwe-phototherapeutic kwe-RuDA kanye ne-RuDA-NPs ku-vivo.
Ekuphetheni, i-Ru (II) -arene (RuDA) i-organometallic complex ene-DA-type ligands yenzelwe ukwenza lula inqubo ye-ISC isebenzisa indlela yokuhlanganisa.I-RuDA ehlanganisiwe ingakwazi ukuzihlanganisa ngokusebenzisa ukuxhumana okungeyona i-covalent ukuze yenze izinhlelo ze-supramolecular ezitholakala ku-RuDA, ngaleyo ndlela yenze kube lula ukwakheka kwe-1O2 kanye nokuguqulwa kwe-photothermal okuphumelelayo kokwelashwa komdlavuza obangelwa ukukhanya.Kuyaphawuleka ukuthi i-monomeric RuDA ayizange ikhiqize i-1O2 ngaphansi kwe-laser irradiation ku-808 nm, kodwa ingakhiqiza inani elikhulu le-1O2 esimweni esihlanganisiwe, ebonisa ukuqonda nokusebenza kahle komklamo wethu.Ucwaningo olwalandela luye lwabonisa ukuthi umhlangano we-supramolecular unikeza i-RuDA ngezakhiwo ezithuthukisiwe ze-photophysical kanye ne-photochemical, njengokumuncwa kwe-redshift kanye nokumelana ne-photobleaching, okufiseleka kakhulu ekucubunguleni i-PDT ne-PTT.Kokubili ukuhlolwa kwe-in vitro kanye ne-vivo kubonise ukuthi ama-RuDA NP ane-biocompatibility enhle kanye nokuqoqwa okuhle kwesimila abonisa umsebenzi omuhle kakhulu wokulwa nomdlavuza obangelwa ukukhanya phezu kwe-laser irradiation ebangeni elingu-808 nm.Ngakho-ke, ama-RuDA NPs njengama-reagents e-bimodal supramolecular PDT/PTW azocebisa isethi yama-photosensitizers acushwe kumaza wamaza angaphezu kuka-800 nm.Idizayini yomqondo yesistimu ye-supramolecular inikeza umzila osebenzayo wama-photosensitizers acushwe yi-NIR anemiphumela emihle kakhulu ye-photosensitizing.
Wonke amakhemikhali nezincibilikisi kutholwe kubahlinzeki bezentengiselwano futhi kwasetshenziswa ngaphandle kokuhlanzwa okwengeziwe.I-RuCl3 ithengwe kwa-Boren Precious Metals Co., Ltd. (Kunming, China).[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione) kanye ne-4,7-bis[4-(N,N-diphenylamino)phenyl]-5 , I-6-Diamino-2,1,3-benzothiadiazole yahlanganiswa ngokuvumelana nezifundo zangaphambilini64,65.Izibukeli ze-NMR zirekhodwe ku-spectrometer ye-Bruker Avance III-HD 600 MHz e-Southeastern University Analytical Test Center kusetshenziswa i-d6-DMSO noma i-CDCl3 njengesinyibilikisi.Ukushintsha kwamakhemikhali δ kunikezwa nge-ppm.ngokuphathelene ne-tetramethylsilane, kanye nokusetshenziswana okungaguquki J kunikezwa ngamavelu aphelele ku-hertz.I-High resolution mass spectrometry (HRMS) yenziwe ngethuluzi le-Agilent 6224 ESI/TOF MS.Ukuhlaziywa okuyisisekelo kwe-C, H, kanye no-N kwenziwa ku-Vario MICROCHNOS elemental analyzer (Elementar).I-spectra ebonakalayo ye-UV ikalwe nge-spectrophotometer ye-Shimadzu UV3600.I-spectra ye-fluorescence irekhodwe ku-spectrofluorimeter ye-Shimadzu RF-6000.Izibukeli ze-EPR zarekhodwa ethuluzini le-Bruker EMXmicro-6/1.I-morphology nesakhiwo samasampuli alungisiwe kwacwaningwa kumathuluzi e-FEI Tecnai G20 (TEM) kanye ne-Bruker Icon (AFM) asebenza ngevolthi engu-200 kV.I-Dynamic light scattering (DLS) yenziwe ku-Nanobrook Omni analyzer (Brookhaven).Izakhiwo ze-Photoelectrochemical zikalwa ngokusethwa kwe-electrochemical (CHI-660, China).Izithombe ze-Photoacoustic zitholwe kusetshenziswa uhlelo lwe-FUJIFILM VisualSonics Vevo® LAZR.Izithombe ze-Confocal zitholwe kusetshenziswa i-Olympus FV3000 confocal microscope.Ukuhlaziywa kwe-FACS kwenziwa nge-cytometer ye-BD Calibur flow.Ukuhlolwa kokusebenza okuphezulu kwe-liquid chromatography (HPLC) kwenziwa kusistimu ye-Waters Alliance e2695 kusetshenziswa umtshina ongu-2489 UV/Vis.Ukuhlolwa kwe-Gel Permeation Chromatography (GPC) kwarekhodwa ensimbini ye-Thermo ULTIMATE 3000 kusetshenziswa umtshina we-ERC RefratoMax520 refractive index.
[(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, I-N-diphenylamino)phenyl]-5,6-diamino-2,1,3-benzothiadiazole 65 (652.0 mg, 1.0 mmol) kanye ne-glacial acetic acid (30 mL) yashukunyiswa esiqandisini se-reflux amahora angu-12.I-solvent yabe ikhishwa ku-vacuo kusetshenziswa i-evaporator ejikelezayo.Izinsalela ezibangelwayo zahlanjululwa nge-flash column chromatography (i-silica gel, CH2Cl2: MeOH = 20: 1) ukuthola i-RuDA njengempushana eluhlaza (isivuno: 877.5 mg, 80%).indunu.Ibalwe ku-C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.Kutholwe: 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).I-13C NMR (150 MHz, D6-DMSO), Δ (PPM) 158.03, 152.81, 147.31, 147.20, 145.51, 147.68, 138.8, 135.51, 124.68, 138.41, 123.49, 103.49 , 103. , 86.52, 84.75, 63.29, 30.90, 22.29, 18.83.ESI-MS: m/z [M-Cl]+ = 1097.25.
Ukuhlanganiswa kwe-4,7-bis[4-(N,N-diethylamino)phenyl-5,6-diamino-2,1,3-benzothiadiazole (L2): I-L2 yahlanganiswa ngezinyathelo ezimbili.I-Pd(PPh3)4 (46 mg, 0.040 mmol) yengezwe ku-N,N-diethyl-4-(tributylstannyl)aniline (1.05 g, 2.4 mmol) kanye nesixazululo esingu-4,7-dibromo-5,6-dinitro - 2, I-1,3-benzothiadiazole (0.38 g, 1.0 mmol) ku-toluene eyomile (100 ml).Ingxube yashukunyiswa ku-100 ° C amahora angu-24.Ngemva kokukhipha i-toluene ku-vacuo, okuqinile okwaba khona kwagezwa nge-petroleum ether.Khona-ke ingxube yalesi sakhi (234.0 mg, 0.45 mmol) kanye ne-iron powder (0.30 g, 5.4 mmol) ku-acetic acid (20 ml) yanyakaziswa ku-80 ° C. amahora angu-4.Ingxube yokusabela yathululelwa emanzini futhi okuqinile okunsundu okwavela kwaqoqwa ngokuhlunga.Umkhiqizo wahlanzwa kabili nge-vacuum sublimation ukuze unikeze okuqinile okuluhlaza (126.2 mg, 57% isivuno).indunu.Ibalwe ku-C26H32N6S: C 67.79, H 7.00, N 18.24.Kutholwe: 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).13С 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.
Izinhlanganisela zalungiswa futhi zahlanzwa ngokulandela izinqubo ezifana ne-RuDA.indunu.Ibalwe ku-C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.Kutholakele: 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).I-13C NMR (151 MHz, CDCL3), Δ (PPM) 158.20, 153.56, 148.40, 135.0, 1313, 115.0, 87.0, 88.7, 87.0, 84.4, 107.0, 84.4, 107.0, 84.4., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m/z [M-Cl]+ = 905.24.
I-RuDA yahlakazwa ku-MeOH/H2O (5/95, v/v) ekuhlanganiseni kwe-10 μM.I-spectrum yokumuncwa ye-RuDA ikalwa njalo emizuzwini engu-5 ku-spectrophotometer ye-Shimadzu UV-3600 ngaphansi kwemisebe ene-laser light ene-wavelength engu-808 nm (0.5 W/cm2).I-spectra ye-ICG irekhodwe ngaphansi kwezimo ezifanayo nezinga.
I-spectra ye-EPR irekhodwe ku-spectrometer ye-Bruker EMXmicro-6/1 enamandla e-microwave angu-20 mW, uhla lokuskena lwe-100 G, kanye nokushintshwa kwenkundla kwe-1 G. 2,2,6,6-tetramethyl-4-piperidone (TEMP) kanye ne-5,5-dimethyl-1-pyrroline N-oxide (DMPO) zisetshenziswe njengezicupho ezijikelezayo.I-electron spin resonance spectra yarekhodelwa izixazululo ezixubile ze-RuDA (50 µM) ne-TEMF (20 mM) noma i-DMPO (20 mM) ngaphansi kwesenzo semisebe ye-laser enobude obungamaza obungu-808 nm (0.5 W/cm2).
Izibalo ze-DFT ne-TD-DFT ze-RuDA zenziwa emazingeni e-PBE1PBE/6–31 G*//LanL2DZ ngesisombululo samanzi kusetshenziswa uhlelo lwe-Gaussian 1666,67,68.I-HOMO-LUMO, imbobo kanye nokusatshalaliswa kwe-electron ye-RuDA yesimo esijabulisayo se-singlet esine-low-energy yahlelwa kusetshenziswa uhlelo lwe-GaussView (inguqulo 5.0).
Siqale sazama ukukala ukusebenza kahle kwesizukulwane se-1O2 RuDA sisebenzisa i-spectroscopy evamile ye-UV-ebonakalayo nge-ICG (ΦΔ = 0.002) njengento evamile, kodwa ukuchithwa kwezithombe ze-ICG kuthinte kakhulu imiphumela.Ngakho-ke, isivuno se-quantum se-1O2 RuDA silinganiswe ngokuthola ushintsho ekuqineni kwe-ABDA fluorescence cishe ku-428 nm lapho i-radiated nge-laser ene-wavelength engu-808 nm (0.5 W / cm2).Ukuhlolwa kwenziwa ku-RuDA ne-RuDA NPs (20 μM) emanzini/DMF (98/2, v/v) equkethe i-ABDA (50 μM).Isivuno se-quantum se-1O2 sibalwa kusetshenziswa ifomula elandelayo: ΦΔ (PS) = ΦΔ (ICG) × (rFS/APS)/(rICG/AICG).I-rPS kanye ne-rICG izinga lokuphendula le-ABDA ne-1O2 etholwe ku-photosensitizer ne-ICG, ngokulandelanayo.I-APS ne-AICG ukumuncwa kwe-photosensitizer ne-ICG ku-808 nm, ngokulandelana.
Izilinganiso ze-AFM zenziwa ngaphansi kwezimo eziwuketshezi kusetshenziswa imodi yokuskena kusistimu ye-AFM ye-Bruker Dimension Icon.Ngokusebenzisa isakhiwo esivulekile esinamaseli awuketshezi, amangqamuzana ahlanzwa kabili nge-ethanol futhi omiswa ngomfudlana we-nitrogen.Faka amaseli omisiwe ekhanda le-optical le-microscope.Ngokushesha faka iconsi lesampula echibini loketshezi bese ulibeka ku-cantilever usebenzisa isirinji yepulasitiki elahlwayo eyinyumba kanye nenaliti eyinyumba.Okunye ukudonsa kufakwa ngokuqondile kusampula, futhi lapho ikhanda le-optical lehliswa, amaconsi amabili ayahlangana, enze i-meniscus phakathi kwesampula kanye ne-liquid reservoir.Izilinganiso ze-AFM zenziwe kusetshenziswa i-nitride cantilever emise okwe-SCANASYST-FLUID V (Bruker, hardness k = 0.7 N m-1, f0 = 120–180 kHz).
Amachromatogram e-HPLC atholwe kusistimu ye-Waters e2695 efakwe ikholomu ye-phoenix C18 (250×4.6 mm, 5 µm) kusetshenziswa umtshina ongu-2489 UV/Vis.Ubude begagasi bomtshina bungama-650 nm.Izigaba zamaselula A no-B kwakungamanzi ne-methanol, ngokulandelana, futhi izinga lokugeleza lesigaba seselula lalingu-1.0 ml·min-1.I-gradient (i-solvent B) ibimi kanje: 100% isuka ku-0 iye kumizuzu emi-4, isuka ku-100% iye ku-50% isuka emizuzwini emi-5 kuye kwengama-30, iphinde isethelwe ku-100% isuka ku-31 iye kumizuzu engama-40.I-ore yancibilika kusixazululo esixubile se-methanol namanzi (50/50, ngevolumu) ekuhlanganiseni kuka-50 μM.Umthamo womjovo wawungama-20 μl.
Izivivinyo ze-GPC zirekhodwe kuthuluzi le-Thermo ULTIMATE 3000 elifakwe amakholomu amabili e-PL aquagel-OH MIXED-H (2×300×7.5 mm, 8 µm) kanye nomtshina wenkomba we-ERC RefratoMax520.Ikholomu ye-GPC ikhishwe ngamanzi ngesilinganiso sokugeleza esingu-1 ml/min ku-30°C.Ama-NP ore ahlakazwa kusixazululo se-PBS (pH = 7.4, 50 μM), umthamo womjovo wawungu-20 μL.
Ama-photocurrents akalwe ngokusetha kwe-electrochemical (CHI-660B, China).Izimpendulo ze-optoelectronic lapho i-laser ivuliwe futhi ivaliwe (808 nm, 0.5 W/cm2) ikalwe ngevoltheji engu-0.5 V ebhokisini elimnyama, ngokulandelanayo.Iseli elijwayelekile elinama-electrode amathathu lasetshenziswa ne-glassy carbon electrode (GCE) emise okwe-L njenge-electrode esebenzayo, i-calomel electrode evamile (SCE) njenge-reference electrode, kanye nediski yeplatinamu njenge-electrode ephikisayo.Isixazululo esingu-0.1 M Na2SO4 sisetshenziswe njenge-electrolyte.
Ulayini wamangqamuzana omdlavuza webele womuntu we-MDA-MB-231 wathengwa kwa-KeyGEN Biotec Co., LTD (Nanjing, China, inombolo yekhathalogi: KG033).Amaseli atshalwa kuma-monolayers ku-Dulbecco's Modified Eagle's Medium (DMEM, i-glucose ephezulu) ehlanganiswe nesisombululo se-fetal bovine serum (FBS), i-penicillin (100 μg/ml) ne-streptomycin (100 μg/ml).Wonke amaseli akhuliswe ku-37°C endaweni enomswakama equkethe u-5% CO2.
Ukuhlolwa kwe-MTT kwasetshenziselwa ukucacisa i-cytotoxicity ye-RuDA ne-RuDA-NPs lapho kukhona khona kanye nokungabikho kwe-irradiation ekhanyayo, noma ngaphandle kwe-Vc (0.5 mM).Amaseli omdlavuza we-MDA-MB-231 akhuliswe kumapuleti anemithombo engama-96 kubuningi beseli cishe obuyi-1 x 105 amaseli/ml/umthombo futhi afakwa amahora angu-12 ku-37.0°C endaweni engu-5% CO2 kanye no-95% womoya.I-RuDA ne-RuDA NPs ehlakazekile emanzini yengezwa kumaseli.Ngemva kwamahora angu-12 okufukamela, amangqamuzana avezwa emisebeni ye-laser engu-0.5 W cm -2 ebangeni elingamaza angu-808 nm imizuzu engu-10 (300 J cm -2) bese efakwa ebumnyameni amahora angu-24.Amaseli abe esefakwa i-MTT (5 mg/ml) amanye amahora ama-5.Okokugcina, shintsha okumaphakathi kuye ku-DMSO (200 µl) ukuze uncibilikise umphumela wamakristalu we-formazan onsomi.Amanani e-OD akalwa kusetshenziswa isifundi se-microplate esinobude begagasi obungu-570/630 nm.Inani le-IC50 lesampula ngayinye libalwe kusetshenziswa isofthiwe ye-SPSS kusukela kumajika okuphendula umthamo atholwe okungenani ekuhlolweni okuthathu okuzimele.
Amaseli e-MDA-MB-231 aphathwe nge-RuDA kanye ne-RuDA-NP ekuhlanganiseni kwe-50 μM.Ngemva kwamahora angu-12 okufukamela, amangqamuzana akhanyiselwe nge-laser ene-wavelength engu-808 nm namandla angu-0.5 W/cm2 amaminithi angu-10 (300 J/cm2).Eqenjini likavithamini C (Vc), amaseli alashwa nge-0.5 mM Vc ngaphambi kokufakwa kwemisebe ye-laser.Amaseli abe esefukanyelwa ebumnyameni amahora angu-24 engeziwe, abese egcotshwa i-calcein AM ne-propidium iodide (20 μg/ml, 5 μl) imizuzu engu-30, bese egezwa nge-PBS (10 μl, pH 7.4).izithombe zamaseli anebala.
Isikhathi sokuthumela: Sep-23-2022
