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Iifotosensitizers ezisebenzayo zibaluleke kakhulu kusetyenziso olubanzi lwekliniki lwe-phototherapy.Nangona kunjalo, iifotosensitizers eziqhelekileyo zinengxaki yokufunxwa kobude obufutshane, ukungonelanga kwefoto, isivuno esisezantsi sohlobo lweoksijini esebenzayo (ROS), kunye nokucinywa kwe-ROS.Apha sichaza i-infrared (NIR) ye-supramolecular photosensitizer (RuDA) exutywe ngokuzimeleyo kwe-Ru (II) -arene organometallic complexes kwisisombululo samanzi.I-RuDA inokuvelisa kuphela i-oksijini ye-singlett (i-1O2) kwimeko edibeneyo, kwaye ibonisa ngokucacileyo ukuziphatha kwe-1O2 ye-aggregation-induced isizukulwana ngenxa yokwanda okukhulu kwenkqubo ye-crossover phakathi kwe-singlett-triplet system.Ngaphantsi kwesenzo sokukhanya kwe-laser ye-808 nm, i-RuDA ibonisa i-1O2 isivuno se-quantum ye-16.4% (i-FDA-evunyiweyo ye-indocyanine eluhlaza: ΦΔ = 0.2%) kunye nokusebenza kakuhle kwe-photothermal yokuguqulwa kwe-24.2% (i-nanorods zegolide zorhwebo) kunye ne-photostability egqwesileyo.: 21.0%, iinanoshell zegolide: 13.0%).Ukongezelela, i-RuDA-NPs ene-biocompatibility efanelekileyo inokuqokelela ngokukhethiweyo kwiindawo ze-tumor, ezibangela ukuhlehliswa kwe-tumor ngexesha lonyango lwe-photodynamic kunye ne-95.2% yokunciphisa umthamo we-tumor kwi-vivo.Olu nyango lokuphucula ukuhlanganiswa kwefotodynamic lubonelela ngesicwangciso sokuphuhlisa iifotosensitizers ezineempawu ezithandekayo zefotophysical kunye neefotochemical.
Xa kuthelekiswa nonyango oluqhelekileyo, unyango lwe-photodynamic (i-PDT) lunyango olukhangayo lomhlaza ngenxa yeenzuzo zalo ezibalulekileyo ezifana nolawulo oluchanekileyo lwe-spatiotemporal, ukungabikho kwe-invasiveness, ukunganyangeki kweziyobisi, kunye nokunciphisa imiphumo emibi ye-1,2,3.Ngaphantsi kokukhanya okukhanyayo, i-photosensitizers esetyenzisiweyo inokuthi isebenze ukuze yenze iintlobo ze-oksijini ezisebenzayo (ROS), ezikhokelela kwi-apoptosis / necrosis okanye iimpendulo ze-immune4,5. Nangona kunjalo, uninzi lweefotosensitizers eziqhelekileyo, ezinjengeklorin, i-porphyrins, kunye ne-anthraquinones, zinokufunxa amaza amaza amafutshane (i-frequency <680 nm), nto leyo ekhokelela ekungeneni kokukhanya okubi ngenxa yokufunxwa ngamandla kweemolekyuli zebhayoloji (umz., i-hemoglobin kunye ne-melanin) ummandla obonakalayo6,7. Nangona kunjalo, uninzi lweefotosensitizers eziqhelekileyo, ezinjengeklorin, i-porphyrins, kunye ne-anthraquinones, zinokufunxa amaza amaza amafutshane (i-frequency <680 nm), nto leyo ekhokelela ekungeneni kokukhanya okubi ngenxa yokufunxwa ngamandla kweemolekyuli zebhayoloji (umz., i-hemoglobin kunye ne-melanin) ummandla obonakalayo6,7. 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. Nangona kunjalo, uninzi lweefotosensitizers eziqhelekileyo ezifana neklorin, i-porphyrins kunye ne-anthraquinones zinexesha elifutshane lokufunxa kwamaza amaza (< 680 nm) okukhokelela ekungeneni okungahambi kakuhle kokukhanya ngenxa yokufunxa ngamandla kweemolekyuli zebhayoloji (umz. i-hemoglobin kunye ne-melanin) kwindawo ebonakalayo6,7.然而, 大多数 大多数 的 光敏剂, 如 氢 卟酚 卟酚, 卟啉 和 蒽醌, 具有 蒽醌 较 较 的 的 的 的 的 的 的 波长 的 波长导致光穿透性差.然而, 大多数大多数 传统 的, 二 氢 卟酚, 卟啉 蒽醌 蒽醌 较 较 短 短 的 波长 波长 波长 波长 波长吸收 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. Nangona kunjalo, uninzi lweefotosensitizers zemveli ezifana neklorin, iiporphyrins kunye ne-anthraquinones zinexesha elifutshane lokufunxa amaza amaza (i-frequency <680 nm) ngenxa yokufunxa ngamandla kwee-biomolecules ezifana ne-hemoglobin kunye ne-melanin ekhokelela ekungeneni kokukhanya okubi.Indawo ebonakalayo 6.7.Ngoko ke, i-infrared (NIR) efunxa i-photosensitizers esebenzayo kwi-700-900 nm "ifestile yonyango" ifaneleke kakuhle kwi-phototherapy.Ekubeni ukukhanya okukufutshane kwe-infrared yeyona nto incinci ifakwe kwizicubu zebhayoloji, kunokukhokelela ekungeneni okunzulu kunye ne-photodamage8,9.
Ngelishwa, i-NIR-absorbing photosensitizers ekhoyo ngokubanzi ine-photostability embi, i-oksijeni ephantsi ye-singlett (1O2) yokuvelisa amandla, kunye nokucima i-1O2 edibeneyo, ekhawulela ukusetyenziswa kweklinikhi10,11.Nangona iinzame ezinkulu zenziwe ukuphucula iipropathi zefotophysical kunye neefotochemical ze-photosensitizers eziqhelekileyo, ukuza kuthi ga ngoku iingxelo ezininzi ziye zaxela ukuba i-NIR-absorbing photosensitizers inokusombulula zonke ezi ngxaki.Ukongeza, ii-photosensitizers ezininzi zibonise isithembiso sokuvelisa ngokufanelekileyo i-1O212,13,14 xa i-irradiated ngokukhanya okungaphezulu kwe-800 nm, ekubeni amandla e-photon ehla ngokukhawuleza kwindawo ekufutshane ne-IR.I-Triphenylamine (TFA) njengomnikeli we-electron kunye [1,2,5] thiadiazole-[3,4-i] dipyrido [a, c] phenazine (TDP) njengeqela elamkela i-electron Donor-acceptor (DA) uhlobo lwedayi iklasi yedayi, ukufunxa kufuphi ne-infrared, eziye zafundwa ngokubanzi malunga ne-infrared bioimaging II kunye ne-photothermal therapy (PTT) ngenxa ye-bandgap yabo encinci.Ngaloo ndlela, iidayi zohlobo lwe-DA zingasetyenziselwa i-PDT nge-excitation ekufutshane ne-IR, nangona kunqabile ukuba zifundwe njenge-photosensitizers ye-PDT.
Kuyaziwa ukuba ukuphumelela okuphezulu kwe-intersystem crossing (ISC) ye-photosensitizers ikhuthaza ukubunjwa kwe-1O2.Isicwangciso esiqhelekileyo sokuqhubela phambili inkqubo ye-ISC kukuphucula i-spin-orbit coupling (SOC) yeefotosensitizers ngokuzisa ii-athomu ezinzima okanye i-organic moieties ekhethekileyo.Nangona kunjalo, le ndlela isenazo izinto ezingeloncedo kunye nezithintelo19,20.Kutshanje, i-supramolecular self-assembly ibonelele nge-bottom-up intelligent approach yokwenziwa kwezinto ezisebenzayo kwinqanaba le-molecular, i-21,22 kunye neenzuzo ezininzi kwi-phototherapy: (1) i-photosensitizers edibeneyo inokuba namandla okwenza izakhiwo zeribhoni.Ngokufana nezakhiwo ze-elektroniki ezinosasazo oluxineneyo lwamanqanaba amandla ngenxa yee-orbits ezidlulanayo phakathi kweebhloko zokwakha.Ngoko ke, umdlalo wamandla phakathi kwe-singlett ephantsi ye-excited state (S1) kunye ne-triplet engummelwane evuyayo (Tn) iya kuphuculwa, enenzuzo kwinkqubo ye-ISC 23, 24.(2) Indibano ye-Supramolecular iya kunciphisa i-relaxation engekho-radiative esekelwe kwindlela yokunciphisa i-intramolecular motion limitation (RIM), ekwakhuthaza inkqubo ye-ISC 25, 26.(3) Indibano ye-supramolecular inokukhusela iimolekyuli zangaphakathi ze-monomer kwi-oxidation kunye nokuthotywa, ngaloo ndlela iphucula kakhulu i-photosability ye-photosensitizer.Xa sinikwa ezi zibonelelo zingentla, sikholelwa ukuba iinkqubo ze-supramolecular photosensitizer zingayenye indlela ethembisayo ukoyisa iziphene zePDT.
I-Ru (II) i-complexes esekelwe kwi-platform yezonyango ezithembisayo kwizicelo ezinokuthi zixilongwe kunye nonyango lwezifo ngenxa yeempawu zabo zebhayoloji ezikhethekileyo kunye ezikhangayo28,29,30,31,32,33,34.Ukongezelela, ubuninzi beemeko ezivuyisayo kunye neepropati ezithintekayo ze-photophysicochemical ze-Ru (II)-based complexes zibonelela ngeenzuzo ezinkulu zokuphuhliswa kwe-photosensitizers esekelwe kwi-Ru (II)35,36,37,38,39,40.Umzekelo ophawulekayo yi-ruthenium (II) i-polypyridyl complex TLD-1433, okwangoku kwiSigaba sesi-II solingo lwezonyango njenge-photosensitizer yonyango lwe-non-muscle invasive bladder cancer (NMIBC)41.Ukongezelela, i-ruthenium (II) i-arene organometallic complexes isetyenziswa ngokubanzi njengee-chemotherapeutic agents kunyango lomhlaza ngenxa yetyhefu ephantsi kunye nokulula kokuguqulwa42,43,44,45.Iimpawu ze-ionic ze-Ru (II) -arene organometallic complexes azikwazi nje ukuphucula i-solubility embi ye-DA chromophores kwii-solvents eziqhelekileyo, kodwa ziphucula indibano ye-chromophores ye-DA.Ukongeza, i-pseudooctahedral half-sandwich structure ye-organometallic complexes ye-Ru (II) -arenes inokuthintela ngokungqongqo i-H-aggregation ye-chromophores yohlobo lwe-DA, ngaloo ndlela iququzelela ukubunjwa kwe-J-aggregation kunye ne-redshifted absorption bands.Nangona kunjalo, izinto ezingalunganga ze-Ru (II) -arene complexes, ezifana nokuzinza okuphantsi kunye / okanye i-bioavailability embi, inokuchaphazela ukuphumelela konyango kunye nomsebenzi we-vivo we-arene-Ru (II) complexes.Nangona kunjalo, uphononongo lubonise ukuba ezi zinto zingeloncedo zinokoyiswa ngokufaka i-ruthenium complexes eneepolima ezihambelana ne-biocompatible ngokufaka i-encapsulation yomzimba okanye i-covalent conjugation.
Kulo msebenzi, sixela i-DA-conjugated complexes ye-Ru (II) -arene (RuDA) kunye ne-NIR trigger ngokusebenzisa ibhondi yokulungelelanisa phakathi kwe-chromophore ye-DAD kunye ne-Ru (II) -arene moiety.Ii-complexes ezibangelwayo zinokuzihlanganisa ngokwazo zibe yi-metalosupramolecular vesicles emanzini ngenxa yokunxibelelana okungahambelaniyo.Ngokucacileyo, indibano ye-supramolecular yanika i-RuDA nge-polymerization-induced intersystem crossing-over properties, eyandisa kakhulu ukusebenza kwe-ISC, eyayithandeka kakhulu kwi-PDT (Fig. 1A).Ukwandisa i-tumor accumulation kunye ne-vivo biocompatibility, i-FDA-evunyiweyo ye-Pluronic F127 (PEO-PPO-PEO) yayisetyenziselwa ukufaka i-RuDA47,48,49 ukudala i-RuDA-NP nanoparticles (Umfanekiso 1B) osebenza njenge-PDT / Dual- Imo yePTT yommeli .Kwi-phototherapy yomhlaza (Umfanekiso 1C), i-RuDA-NP yayisetyenziselwa ukuphatha iigundane ezinqunu kunye ne-MDA-MB-231 tumors ukufunda ukusebenza kwe-PDT kunye ne-PTT kwi-vivo.
Umzobo weSchematic we-photophysical mechanism ye-RuDA kwiifom ze-monomeric kunye neefom ezihlanganisiweyo ze-phototherapy yomhlaza, i-synthesis ye-B RuDA-NPs kunye ne-C RuDA-NPs ye-NIR-activated PDT kunye ne-PTT.
I-RuDA, equka ukusebenza kwe-TPA kunye ne-TDP, yalungiselelwa ngokwenkqubo eboniswe kwi-Supplementary Figure 1 (Figure 2A), kwaye i-RuDA ibonakaliswe yi-1H kunye ne-13C i-NMR spectra, i-electrospray ionization mass spectrometry, kunye nohlalutyo lwe-elemental (Amanani abongezelelweyo 2-4). ).Imephu ye-RuDA yoxinaniso lwe-electron ye-elektroni yenguqu ye-single ephantsi kakhulu yabalwa yi-theory exhomekeke kwi-dependent density functional (TD-DFT) ukufunda inkqubo yokudluliselwa kwentlawulo.Njengoko kubonisiwe kwi-Supplementary Figure 5, i-electron density drifts ikakhulu isuka kwi-triphenylamine ukuya kwiyunithi yokwamkela i-TDP emva kwe-photoexcitation, enokuthi ifakwe kwi-transition ye-intramolecular charge transfer (CT).
Ubume beMichiza yentsimbi ekrwada B Ukufunxa imbonakalo yentsimbi ekrwada kwimixube yomlinganiselo owahlukeneyo weDMF namanzi.C Amaxabiso aqhelekileyo okufunxa kwe-RuDA (800 nm) kunye ne-ICG (779 nm) ngokuchasene nexesha kwi-0.5 W cm-2 ye-808 nm laser ukukhanya.D I-photodegradation ye-ABDA ibonakaliswe yi-RuDA-induced formation ye-1O2 kwi-DMF / H2O imixube kunye nemixholo eyahlukeneyo yamanzi phantsi kwesenzo se-laser radiation kunye ne-wavelength ye-808 nm kunye namandla e-0.5 W / cm2.
I-Abstract-I-UV-ebonakalayo yokufunxa i-spectroscopy yasetyenziselwa ukufunda iipropati zokuzihlanganisa ze-Ore kwimixube ye-DMF kunye namanzi kwiiratio ezahlukeneyo.Njengoko kubonisiwe kwifig.I-2B, i-RuDA ibonisa iibhendi zokufunxa ukusuka kwi-600 ukuya kwi-900 nm kwi-DMF kunye nebhendi ephezulu yokufunxa kwi-729 nm.Ukunyusa umthamo wamanzi kukhokelele kutshintsho olubomvu ngokuthe ngcembe lokufunxa iOre ukuya kuma-800 nm, nto leyo ebonisa ukudityaniswa kwe-J ye-Ore kwinkqubo edityanisiweyo.I-photoluminescence spectra ye-RuDA kwii-solvents ezahlukeneyo ziboniswa kwi-Supplementary Figure 6. I-RuDA ibonakala ibonisa i-luminescence ye-NIR-II kunye ne-wavelength ephezulu yokukhutshwa kwe-ca.I-1050 nm kwi-CH2Cl2 kunye ne-CH3OH, ngokulandelanayo.I-Stokes shift enkulu (malunga ne-300 nm) ye-RuDA ibonisa utshintsho oluphawulekayo kwijometri yelizwe elinomdla kunye nokubunjwa kwamazwe aphantsi anomdla.Izivuno ze-luminescence quantum ze-Ore kwi-CH2Cl2 kunye ne-CH3OH zinqunywe ukuba zibe yi-3.3 kunye ne-0.6%, ngokulandelanayo.Nangona kunjalo, kumxube we-methanol kunye namanzi (5/95, v/v), i-redshift encinci yokukhutshwa kunye nokuncipha kwemveliso ye-quantum (0.22%) yabonwa, enokuthi ibe ngenxa yokuzihlanganisa kwe-Ore. .
Ukujonga i-self-assembly ye-ORE, sasebenzisa i-atomic force microscopy (AFM) ukuze sibone utshintsho lwe-morphological kwi-ORE kwisisombululo se-methanol emva kokongeza amanzi.Xa umthamo wamanzi wawungaphantsi kwe-80%, akukho kuhlanganiswa okucacileyo kwabonwa (umfanekiso owongezelelweyo wesi-7).Nangona kunjalo, ngokunyuka okungaphezulu kumxholo wamanzi ukuya kwi-90-95%, ii-nanoparticles ezincinci zavela, ezibonisa ukuzihlanganisa kwe-Ore.Ukongezwa, i-laser irradiation kunye ne-wavelength ye-808 nm ayizange ichaphazele ukunyanzeliswa kwe-RuDA emanzini. isisombululo (Umfanekiso 2C kunye neFig. 8).Ngokwahlukileyo, i-absorbection ye-indocyanine eluhlaza (ICG njengolawulo) yehla ngokukhawuleza kwi-779 nm, ebonisa i-photostability egqwesileyo ye-RuDA.Ukongezelela, ukuzinza kwe-RuDA-NPs kwi-PBS (pH = 5.4, 7.4 kunye ne-9.0), i-10% ye-FBS kunye ne-DMEM (i-glucose ephezulu) ihlolwe yi-UV-ebonakalayo ye-spectroscopy ye-spectroscopy ngamaxesha ahlukeneyo.Njengoko kuboniswe kwi-Supplementary Figure 9, utshintsho oluncinci kwiibhendi ze-RuDA-NP ze-absorption zabonwa kwi-PBS kwi-pH 7.4 / 9.0, i-FBS kunye ne-DMEM, ebonisa ukuzinza okuhle kakhulu kwe-RuDA-NP.Nangona kunjalo, kwi-acidic medium (рН = 5.4) i-hydrolysis ye-Ore ifunyenwe.Siphinde savavanya ngakumbi ukuzinza kwe-RuDA kunye ne-RuDA-NP usebenzisa iindlela eziphezulu ze-liquid chromatography (HPLC).Njengoko kuboniswe kwi-Supplementary Figure 10, i-RuDA yayizinzile kumxube we-methanol kunye namanzi (50/50, v / v) kwiyure yokuqala, kwaye i-hydrolysis yabonwa emva kweeyure ze-4.Nangona kunjalo, kuphela incopho ebanzi ye-concave-convex yabonwa kwi-RuDA NPs.Ngoko ke, i-gel permeation chromatography (GPC) yayisetyenziselwa ukuvavanya ukuzinza kwe-RuDA NPs kwi-PBS (pH = 7.4).Njengoko kuboniswe kwi-Supplementary Figure 11, emva kweeyure ze-8 zokufukamela phantsi kweemeko ezivavanyiweyo, ukuphakama kokuphakama, ububanzi obuphakamileyo kunye nommandla ophakamileyo weNP RuDA ayizange itshintshe kakhulu, ebonisa ukuzinza okuhle kakhulu kweNP RuDA.Ukongezelela, imifanekiso ye-TEM ibonise ukuba i-morphology ye-RuDA-NP nanoparticles yahlala ingatshintshi emva kweeyure ze-24 kwi-diluted PBS buffer (pH = 7.4, i-Supplementary Fig. 12).
Ngenxa yokuba i-self-assembly inokunika iimpawu ezahlukeneyo zokusebenza kunye neekhemikhali kwi-Ore, sabona ukukhululwa kwe-9,10-anthracenediylbis (methylene) dimalonic acid (ABDA, isalathisi se-1O2) kwimixube yamanzi e-methanol.Intsimbi ekrwada enomthamo wamanzi owahlukileyo50.Njengoko kubonisiwe kwi-Figure 2D kunye ne-Supplementary Figure 13, akukho ukuthotywa kwe-ABDA kwabonwa xa umthamo wamanzi ungaphantsi kwe-20%.Ngokunyuka komswakama ukuya kwi-40%, ukuthotywa kwe-ABDA kwenzeka, njengoko kubonakaliswe kukuncipha kokuqina kwe-ABDA fluorescence.Kwakhona kuye kwaphawulwa ukuba umxholo ophezulu wamanzi ubangela ukuthotywa ngokukhawuleza, ukuphakamisa ukuba i-RuDA self-assembly iyimfuneko kwaye inenzuzo ekuthotyweni kwe-ABDA.Le nto ihluke kakhulu kwi-ACQ yanamhlanje (i-aggregation-induced quenching) i-chromophores.Xa uhlanjululwa nge-laser nge-wavelength ye-808 nm, isivuno se-quantum se-1O2 RuDA kumxube we-98% H2O / 2% DMF yi-16.4%, amaxesha angama-82 aphezulu kune-ICG (ΦΔ = 0.2%)51, ebonisa isizukulwana esiphawulekayo sokusebenza kakuhle kwe-1O2 RuDA kwimeko yokudibanisa.
I-Electron spins isebenzisa i-2,2,6,6-tetramethyl-4-piperidinone (TEMP) kunye ne-5,5-dimethyl-1-pyrroline N-oxide (DMPO) njenge-spin traps I-resonance spectroscopy (ESR) yayisetyenziselwa ukuchonga iintlobo ezibangelwayo. AFK.nguRuDA.Njengoko kuboniswe kwi-Supplementary Figure 14, kuye kwaqinisekiswa ukuba i-1O2 yenziwe ngamaxesha ombane phakathi kwe-0 kunye ne-4 imizuzu.Ukongezelela, xa i-RuDA ifakwe kwi-DMPO phantsi kwe-irradiation, isibonakaliso se-EPR esine-line ye-1: 2: 2: 1 i-DMPO-OH · i-adduct yafunyanwa, ebonisa ukubunjwa kwe-hydroxyl radicals (OH ·).Ngokubanzi, ezi ziphumo zingentla zibonisa amandla e-RuDA ukuvuselela imveliso ye-ROS ngokusebenzisa uhlobo olubini lwenkqubo ye-photosensitization ye-I / II.
Ukuqonda kakuhle iimpawu ze-elektroniki ze-RuDA kwiifom ze-monomeric kunye neefom ezidibeneyo, i-frontier ye-molecular orbitals ye-RuDA kwiifom ze-monomeric kunye ne-dimeric zibalwe ngokusebenzisa indlela ye-DFT.Njengoko kubonisiwe kwifig.I-3A, i-molecular orbital ephezulu kakhulu (i-HOMO) ye-monomeric RuDA i-delocalized kunye ne-ligand backbone kunye ne-lowest unoccupied molecular orbital (LUMO) igxile kwiyunithi yokumkela i-TDP.Ngokuchasene noko, ubuninzi be-electron kwi-HOMO ye-dimeric igxininiswe kwi-ligand ye-molecule ye-RuDA, ngelixa i-electron density kwi-LUMO igxininiswe ngokukodwa kwiyunithi yokwamkela enye i-molecule ye-RuDA, ebonisa ukuba i-RuDA i-dimer.Iimpawu zeCT.
A I-HOMO kunye ne-LUMO ye-Ore zibalwa ngokohlobo lwe-monomeric kunye ne-dimeric.B Amanqanaba e-eneji enye kunye ne-triplet ye-Ore kwii-monomers kunye nee-dimers.C Amanqanaba aqikelelweyo e-RuDA kunye neendlela ezinokwenzeka ze-ISC njenge-monomeric C kunye ne-dimeric D. Iintolo zibonisa iziteshi ezinokwenzeka ze-ISC.
Ukuhanjiswa kwee-electron kunye nemingxuma kwi-singlett ephantsi yamandla e-singlett evuyayo ye-RuDA kwiifom ze-monomeric kunye ne-dimeric zahlaziywa kusetyenziswa i-Multiwfn 3.852.53 software, eyabalwa ngokusebenzisa indlela ye-TD-DFT.Njengoko kubonisiwe kwileyibhile eyongezelelweyo.Njengoko kubonisiwe kwiMifanekiso 1-2, imingxuma ye-RDA ye-monomeric ininzi i-delocalized kunye ne-ligand backbone kula mazwe anomdla we-singlett, ngelixa i-electrons ininzi ifumaneka kwiqela le-TDP, ebonisa iimpawu ze-intramolecular ze-CT.Ukongeza, kwezi singlet imincili ithi, kukho ngaphezulu okanye ngaphantsi kokudibana phakathi kwemingxunya kunye nee-electron, ecebisa ukuba ezi singlet imincili amazwe ukwenza igalelo elithile excitation wendawo (LE).Kwi-dimers, ukongeza kwiimpawu ze-intramolecular CT kunye ne-LE, umlinganiselo othile weempawu ze-CT ze-intermolecular zabonwa kumazwe ahlukeneyo, ngakumbi i-S3, i-S4, i-S7, kunye ne-S8, ngokusekelwe kuhlalutyo lwe-intermolecular CT, kunye ne-CT intermolecular transitions njengezona ziphambili. (Itheyibhile eyoNgezelelweyo).3).
Ukuze siqonde ngcono iziphumo zovavanyo, siphinde sihlolisise iipropati ze-RuDA ezivuyisayo ukuba zihlolisise ukuhlukana phakathi kwe-monomers kunye ne-dimers (i-Supplementary Tables 4-5).Njengoko kubonisiwe kuMfanekiso 3B, amanqanaba amandla e-singlett kunye ne-triplet echulumancisayo amazwe e-dimer axinene kakhulu kunalawo e-monomer, enceda ukunciphisa umsantsa wamandla phakathi kwe-S1 kunye ne-Tn. Kuye kwaxelwa ukuba utshintsho lwe-ISC lunokuphunyezwa ngaphakathi kwe-gap encinci yamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 kunye ne-Tn54. Kuye kwaxelwa ukuba utshintsho lwe-ISC lunokuphunyezwa ngaphakathi kwe-gap encinci yamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 kunye ne-Tn54. Сообщалось, что переходы ISC inokwenza быть реализованы в пределах небольшой энергетической щели (ΔES1-Tn <0,3 эВ) между S1 и Tn Tn. Kuye kwaxelwa ukuba utshintsho lwe-ISC lunokuphunyezwa ngaphakathi kwesithuba esincinci samandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 kunye 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 может быть реализован в пределах небольшой энергетической щели (ΔES1-Tn < 0,3 эВ) между S1 и Tn54. Kuye kwaxelwa ukuba utshintsho lwe-ISC lunokuphunyezwa ngaphakathi kwe-gap encinci yamandla (ΔES1-Tn <0.3 eV) phakathi kwe-S1 kunye ne-Tn54.Ukongeza, inye kuphela i-orbital, ehleliyo okanye engasetyenziswanga, ekufuneka yohluke kwi-singlett ebotshelelweyo kunye ne-triplet state ukubonelela nge-non-zero ye-SOC edityanisiweyo.Ngaloo ndlela, ngokusekelwe kuhlalutyo lwamandla okuvuselela kunye nokuguqulwa kwe-orbital, zonke iziteshi ezinokwenzeka zenguqu ye-ISC ziboniswa kwiMifanekiso.3C,D.Ngokuphawulekayo, i-channel ye-ISC kuphela ekhoyo kwi-monomer, ngelixa ifom ye-dimeric ineendlela ezine ze-ISC ezinokuphucula utshintsho lwe-ISC.Ngoko ke, kunengqiqo ukucinga ukuba iamolekyu ezininzi ze-RuDA zidibene, zifikeleleke ngakumbi iziteshi ze-ISC.Ngoko ke, i-RuDA aggregates inokwenza i-electronic-band-band structures kwi-singlett kunye ne-triplet states, ukunciphisa i-gap yamandla phakathi kwe-S1 kunye ne-Tn ekhoyo, ngaloo ndlela ukwandisa ukusebenza kwe-ISC ukuququzelela i-1O2 isizukulwana.
Ukuqhubela phambili ukucacisa indlela engaphantsi, senze i-reference compound ye-arene-Ru (II) eyinkimbinkimbi (RuET) ngokutshintsha amaqela amabini e-ethyl kunye namaqela amabini e-phenyl e-triphenylamine kwi-RuDA (Umfanekiso we-4A, ukulinganisa okupheleleyo, jonga i-ESI, i-Supplementary 15 -21 ) Ukususela kumnikeli (diethylamine) ukuya kummkeli (TDF), i-RuET ineempawu ze-CT ze-intramolecular ezifana ne-RuDA.Njengoko kulindelwe, i-spectrum ye-absorption ye-RuET kwi-DMF ibonise ibhendi yokudlulisa intlawulo yamandla aphantsi kunye ne-absorption enamandla kwindawo ekufutshane ye-infrared kummandla we-600-1100 nm (Fig. 4B).Ukongezelela, i-RuET aggregation nayo yabonwa ngokunyuka kwamanzi, okubonakaliswe kwi-redshift ye-absorption maximum, eyaqinisekiswa ngakumbi nge-imaging ye-AFM yamanzi (i-Supplementary Fig. 22).Iziphumo zibonisa ukuba i-RuET, njenge-RuDA, inokwenza i-intramolecular states kunye nokuzihlanganisa kwizakhiwo ezidibeneyo.
Ubume beMichiza yeRuET.B I-spectra ye-Absorption ye-RuET kwimixube yee-ratio ezahlukeneyo ze-DMF kunye namanzi.Iiplati ze-C EIS Nyquist ze-RuDA kunye ne-RuET.Iimpendulo ze-Photocurrent D ze-RuDA kunye ne-RuET phantsi kwesenzo se-laser radiation kunye ne-wavelength ye-808 nm.
I-photodegradation ye-ABDA phambi kwe-RuET ihlolwe nge-irradiation nge-laser ene-wavelength ye-808 nm.Okumangalisa kukuba, akukho ukuthotywa kwe-ABDA kwabonwa kumacandelo ahlukeneyo amanzi (i-Supplementary Fig. 23).Isizathu esinokwenzeka kukuba i-RuET ayikwazi ukwenza ngokufanelekileyo i-electronic structure kuba ikhonkco ye-ethyl ayikhuthazi ukudluliselwa kwentlawulo ye-intermolecular.Ngoko ke, i-electrochemical impedance spectroscopy (EIS) kunye nemilinganiselo ye-photocurrent yesikhashana yenziwa ukuthelekisa iimpawu ze-photoelectrochemical ze-RuDA kunye ne-RuET.Ngokwecebo leNyquist (Umfanekiso 4C), i-RuDA ibonisa i-radius encinci kakhulu kune-RuET, oku kuthetha ukuba i-RuDA56 inothutho olukhawulezayo lwe-electron intermolecular kunye ne-conductivity engcono.Ukongezelela, i-photocurrent density ye-RuDA iphezulu kakhulu kune-RuET (Umfanekiso we-4D), eqinisekisa ukuhanjiswa kwentlawulo engcono ye-RuDA57.Ngaloo ndlela, iqela le-phenyl le-triphenylamine kwi-Ore lidlala indima ebalulekileyo ekuboneleleni ukuhanjiswa kwentlawulo ye-intermolecular kunye nokwakhiwa kwesakhiwo se-elektroniki esinebhanti.
Ukwandisa i-tumor accumulation kunye ne-vivo biocompatibility, siye safaka ngakumbi i-RuDA nge-F127.I-avareji ye-hydrodynamic diameter ye-RuDA-NPs inqunywe ukuba yi-123.1 nm kunye nokusabalalisa okuncinci (PDI = 0.089) usebenzisa indlela yokusabalalisa ukukhanya okuguquguqukayo (i-DLS) (Umfanekiso we-5A), owakhuthaza ukuqokelela kwe-tumor ngokunyusa ukugqithisa kunye nokugcinwa.EPR) isiphumo.Imifanekiso ye-TEM ibonise ukuba ii-Ore NP zinemilo engqukuva efanayo ne-avareji yedayamitha ye-86 nm.Ngokucacileyo, ubuninzi be-absorption ye-RuDA-NPs bubonakala kwi-800 nm (i-Supplementary Fig. 24), ebonisa ukuba i-RuDA-NPs inokugcina imisebenzi kunye neepropati zokuzihlanganisa i-RuDAs.Isivuno se-ROS esibaliweyo se-NP Ore yi-15.9%, efaniswa ne-Ore.Njengoko kubonisiwe kwifig.I-5B, C, iqela lokulawula (i-PBS kuphela) lafumana ukunyuka okuncinci kwiqondo lokushisa, ngelixa ukushisa kwesisombululo se-RuDA-NPs kwanda ngokukhawuleza ngokunyuka kweqondo lokushisa (ΔT) kwi-15.5, 26.1, kunye ne-43.0 ° C.Uxinzelelo oluphezulu lwaluyi-25, i-50, kunye ne-100 µM, ngokulandelanayo, ebonisa umphumo onamandla we-photothermal we-RuDA NPs.Ukongezelela, imilinganiselo yomjikelezo wokufudumeza / wokupholisa ithathwe ukuvavanya ukuzinza kwe-photothermal ye-RuDA-NP kunye nokuthelekisa ne-ICG.Ukushisa kwe-Ore NPs akuzange kunciphe emva kwemijikelezo emihlanu yokufudumeza / yokupholisa (Umfanekiso we-5D), obonisa ukuzinza okuhle kakhulu kwe-photothermal ye-Ore NPs.Ngokwahlukileyo, i-ICG ibonisa ukuzinza okuphantsi kwe-photothermal njengoko kubonwa ngokunyamalala okubonakalayo kwe-photothermal plateau yeqondo lokushisa phantsi kweemeko ezifanayo.Ngokwendlela yangaphambili58, i-photothermal conversion effective (PCE) ye-RuDA-NP ibalwa njenge-24.2%, ephezulu kunezinto ezikhoyo ze-photothermal ezifana ne-nanorods yegolide (21.0%) kunye ne-nanoshells yegolide (13.0%)59.Ngaloo ndlela, i-NP Ore ibonisa iimpawu ezibalaseleyo ze-photothermal, ezibenza bathembise i-PTT abameli.
Uhlalutyo lwe-DLS kunye nemifanekiso ye-TEM ye-RuDA NPs (inset).B Imifanekiso ye-Thermal yeentlobo ezahlukeneyo ze-RuDA NPs ezivezwe kwimitha ye-laser kwi-wavelength ye-808 nm (0.5 W cm-2).C Iijiko zoguqulo lwefotothermal yogxininiso olwahlukeneyo lwee-ore NPs, eziyidatha yobungakanani.B. D Ukunyuka kobushushu be-ORE NP kunye ne-ICG ngaphezu kwemijikelo emi-5 yokupholisa.
I-Photocytotoxicity ye-RuDA NPs ngokuchasene ne-MDA-MB-231 iiseli zomhlaza webele lomntu zavavanywa kwi-vitro.Njengoko kubonisiwe kwifig.I-6A, B, i-RuDA-NPs kunye ne-RuDA ibonise i-cytotoxicity engafanelekanga ngokungabikho kwe-irradiation, ebonisa ubuthi obuphantsi obumnyama be-RuDA-NPs kunye ne-RuDA.Nangona kunjalo, emva kokuvezwa kwimitha ye-laser kwi-wavelength ye-808 nm, i-RuDA kunye ne-RuDA NPs zibonise i-photocytotoxicity eyomeleleyo ngokuchasene ne-MDA-MB-231 iiseli zomhlaza ezinemilinganiselo ye-IC50 (isiqingatha esiphezulu soxinzelelo lwe-inhibitory) ye-5.4 kunye ne-9.4 μM, ngokulandelanayo, ebonisa i-IC50. ukuba i-RuDA-NP kunye ne-RuDA inamandla okwenza i-phototherapy yomhlaza.Ukongezelela, i-photocytotoxicity ye-RuDA-NP kunye ne-RuDA yaphandwa ngakumbi phambi kwe-vitamin C (Vc), i-ROS scavenger, ukucacisa indima ye-ROS kwi-cytotoxicity ebangelwa ukukhanya.Ngokucacileyo, ukusebenza kweeseli kunyukile emva kokongezwa kweVc, kwaye ixabiso le-IC50 ye-RuDA kunye ne-RuDA NPs yayiyi-25.7 kunye ne-40.0 μM, ngokulandelanayo, ebonisa indima ebalulekileyo ye-ROS kwi-photocytotoxicity ye-RuDA kunye ne-RuDA NPs.I-cytotoxicity eyenziwe ngokukhanya ye-RuDA-NPs kunye ne-RuDA kwi-MDA-MB-231 iiseli zomhlaza nge-cell staining ephilayo / efileyo usebenzisa i-calcein AM (i-fluorescence eluhlaza kwiiseli eziphilayo) kunye ne-propidium iodide (PI, i-fluorescence ebomvu yeeseli ezifileyo).kuqinisekiswa ziiseli) njengeprobes zefluorescent.Njengoko kuboniswe kwi-Figure 6C, iiseli eziphathwe nge-RuDA-NP okanye i-RuDA zahlala zisebenza ngaphandle kwe-irradiation, njengoko kubonakaliswe yi-fluorescence eluhlaza.Ngokuchasene noko, phantsi kwe-laser irradiation, kuphela i-fluorescence ebomvu yabonwa, eqinisekisa i-photocytotoxicity esebenzayo ye-RuDA okanye i-RuDA NPs.Kuyaphawuleka ukuba i-fluorescence eluhlaza ibonakala phezu kokongezwa kweVc, ebonisa ukuphulwa kwe-photocytotoxicity ye-RuDA kunye ne-RuDA NPs.Ezi ziphumo ziyahambelana ne-in vitro photocytotoxicity assays.
Ukusebenza okuxhomekeke kwi-dose ye-A RuDA- kunye ne-B RuDA-NP iiseli kwiiseli ze-MDA-MB-231 phambi okanye ukungabikho kwe-Vc (0.5 mM), ngokulandelanayo.Imivalo yemposiso, ithetha ± ukutenxa okusemgangathweni (n = 3). Iimvavanyo ezingabhaliswanga, ezinamacala amabini t *p <0.05, **p <0.01, kunye ne-**p <0.001. Iimvavanyo ezingabhaliswanga, ezinamacala amabini t *p <0.05, **p <0.01, kunye ne-**p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 kwaye ***p <0,001. Iimvavanyo ze-t ezinemisila emibini ezingadityaniswanga *p<0.05, **p<0.01, kunye 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 kwaye ***p <0,001. Iimvavanyo ze-t ezinemisila emibini ezingadityaniswanga *p<0.05, **p<0.01, kunye ne-**p<0.001.C Uhlalutyo lwebala leseli ephilayo/efileyo kusetyenziswa i-calcein AM kunye nepropidium iodide njenge-fluorescent probes.Isikali somgca: 30 µm.Imifanekiso emeleyo yophinda-phindo lwebhayoloji emithathu kwiqela ngalinye ibonisiwe.D Imifanekiso ye-Confocal fluorescence yokuveliswa kwe-ROS kwiiseli ze-MDA-MB-231 phantsi kweemeko zonyango ezahlukeneyo.I-fluorescence ye-DCF eluhlaza ibonisa ubukho be-ROS.I-Irradiate nge-laser ene-wavelength ye-808 nm ngamandla e-0.5 W / cm2 imizuzu eyi-10 (300 J / cm2).Isikali somgca: 30 µm.Imifanekiso emeleyo yophinda-phindo lwebhayoloji emithathu kwiqela ngalinye ibonisiwe.E Flow cytometry RuDA-NPs (50 µM) okanye i-RuDA (50 µM) yohlalutyo lonyango kunye okanye ngaphandle kwe-808 nm laser (0.5 W cm-2) phambi kobukho kunye nokungabikho kweVc (0.5 mM) kwi-10 min.Imifanekiso emeleyo yophinda-phindo lwebhayoloji emithathu kwiqela ngalinye ibonisiwe.F Nrf-2, HSP70 kunye ne-HO-1 ye-MDA-MB-231 iiseli eziphathwa nge-RuDA-NPs (50 µM) kunye okanye ngaphandle kwe-808 nm laser irradiation (0.5 W cm-2, 10 min, 300 J cm-2) , iiseli zivakalisa 2).Imifanekiso emeleyo yokuphinda kabini yebhayoloji kwiqela ngalinye ibonisiwe.
Ukuveliswa kwe-ROS ye-intracellular kwiiseli ze-MDA-MB-231 zavavanywa kusetyenziswa i-2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) indlela yokucoca.Njengoko kubonisiwe kwifig.I-6D, iiseli eziphathwe nge-RuDA-NPs okanye i-RuDA zibonise i-fluorescence eluhlaza eyahlukileyo xa i-radiated nge-808 nm laser, ebonisa ukuba i-RuDA-NPs kunye ne-RuDA inekhono elisebenzayo lokuvelisa i-ROS.Ngokuchasene noko, ngokungabikho kokukhanya okanye ubukho beVc, kuphela uphawu olubuthathaka lwe-fluorescent lweeseli lwabonwa, olubonisa ukubunjwa okuncinci kwe-ROS.Amanqanaba e-ROS e-Intracellular kwiiseli ze-RuDA-NP kunye neeseli ze-MDA-MB-231 eziphathwa nge-RuDA zaye zagqitywa ngakumbi nge-cytometry yokuhamba.Njengoko kuboniswe kwi-Supplementary Figure 25, ubukhulu be-fluorescence intensity (MFI) eyenziwa yi-RuDA-NPs kunye ne-RuDA phantsi kwe-808 nm laser irradiation yanda kakhulu malunga ne-5.1 kunye ne-4.8 amaxesha, ngokulandelanayo, xa kuthelekiswa neqela lokulawula, eliqinisekisa ukubunjwa kwabo okuhle kakhulu kwe-AFK.umthamo.Nangona kunjalo, amanqanaba e-ROS ye-intracellular kwi-RuDA-NP okanye i-MDA-MB-231 iiseli eziphathwe nge-RuDA zazifaniswa kuphela nokulawula ngaphandle kwe-laser irradiation okanye phambi kweVc, efana neziphumo zohlalutyo lwe-confocal fluorescence.
Kuye kwaboniswa ukuba i-mitochondria iyona nto ijoliswe kuyo ye-Ru (II) -arene complexes60.Ngoko ke, i-subcellular localization ye-RuDA kunye ne-RuDA-NPs yaphandwa.Njengoko kuboniswe kwi-Supplementary Figure 26, i-RuDA kunye ne-RuDA-NP ibonisa iiprofayili zokusabalalisa iiselula ezifanayo kunye nokuqokelela okuphezulu kwi-mitochondria (62.5 ± 4.3 kunye ne-60.4 ± 3.6 ng / mg iprotheni, ngokulandelanayo).Nangona kunjalo, kuphela inani elincinci le-Ru lifunyenwe kwiinqununu zenyukliya ze-Ore kunye ne-NP Ore (3.5 kunye ne-2.1%, ngokulandelanayo).I-cell fraction eseleyo iqulethe i-ruthenium eseleyo: 31.7% (30.6 ± 3.4 ng / mg protein) ye-RuDA kunye ne-42.9% (47.2 ± 4.5 ng / mg iprotheni) ye-RuDA-NPs.Ngokubanzi, iOre kunye neNP Ore ziqokelelwa ikakhulu kwimitochondria.Ukuvavanya ukungasebenzi kwe-mitochondrial, sasebenzisa i-JC-1 kunye ne-MitoSOX Red staining ukuvavanya i-membrane ye-mitochondrial enokubakho kunye nomthamo wemveliso ye-superoxide, ngokulandelanayo.Njengoko kuboniswe kwi-Supplementary Fig. 27, eluhlaza obunzima (JC-1) kunye nobomvu (i-MitoSOX Red) i-fluorescence yabonwa kwiiseli eziphathwa kunye ne-RuDA kunye ne-RuDA-NPs phantsi kwe-808 nm laser irradiation, ebonisa ukuba zombini i-RuDA kunye ne-RuDA-NPs i-fluorescent Inokwenza ngokufanelekileyo i-mitochondrial membrane depolarization kunye nemveliso ye-superoxide.Ukongezelela, indlela yokufa kweeseli yagqitywa ngokusebenzisa uhlalutyo lwe-cytometry esekelwe kwi-annexin V-FITC / propidium iodide (PI).Njengoko kuboniswe kwi-Figure 6E, xa i-irradiated nge-808 nm laser, i-RuDA kunye ne-RuDA-NP yenza ukuba kunyuke kakhulu izinga le-apoptosis yokuqala (i-quadrant esezantsi yasekunene) kwiiseli ze-MDA-MB-231 xa kuthelekiswa ne-PBS okanye i-PBS kunye ne-laser.iiseli ezigqityiweyo.Nangona kunjalo, xa i-Vc yongezwa, izinga le-apoptosis ye-RuDA kunye ne-RuDA-NP yehla kakhulu ukusuka kwi-50.9% kunye ne-52.0% ukuya kwi-15.8% kunye ne-17.8%, ngokulandelanayo, eqinisekisa indima ebalulekileyo ye-ROS kwi-photocytotoxicity ye-RuDA kunye ne-RuDA-NP..Ukongezelela, iiseli ezincinci ze-necrotic zabonwa kuwo onke amaqela avavanyiweyo (i-quadrant ephezulu ekhohlo), ebonisa ukuba i-apoptosis ingaba yindlela ephambili yokufa kweeseli eyenziwa yi-RuDA kunye ne-RuDA-NPs.
Ekubeni umonakalo wokuxinzezeleka kwe-oxidative iyona nto ibalulekileyo kwi-apoptosis, i-nuclear factor ehambelana ne-erythroid 2, i-2 (Nrf2) i-62, umlawuli oyintloko we-antioxidant system, iphandwa kwi-MDA-MB-231 ye-RuDA-NPs.Indlela yokusebenza ye-RuDA NPs ebangelwa yi-irradiation.Ngexesha elifanayo, ukubonakaliswa kweprotheyini esezantsi ye-heme oxygenase 1 (HO-1) nayo yafunyanwa.Njengoko kuboniswe kwi-Figure 6F kunye ne-Supplementary Figure 29, i-phototherapy ye-RuDA-NP-mediated yandisa i-Nrf2 kunye ne-HO-1 amanqanaba okuchaza xa kuthelekiswa neqela le-PBS, ebonisa ukuba i-RuDA-NPs inokuvuselela iindlela zokubonisa uxinzelelo lwe-oxidative.Ukongezelela, ukufunda umphumo we-photothermal we-RuDA-NPs63, ukubonakaliswa kweprotheyini ye-heat shock Hsp70 nayo yavavanywa.Kucacile ukuba iiseli eziphathwe nge-RuDA-NPs + 808 nm laser irradiation zibonise ukwanda kwe-Hsp70 xa kuthelekiswa namanye amaqela amabini, ebonisa impendulo yeselula kwi-hyperthermia.
Iziphumo eziphawulekayo ze-in vitro zibangele ukuba siphande ukusebenza kwe-vivo ye-RuDA-NP kwiigundane ezinqunu kunye ne-MDA-MB-231 tumors.Ukuhanjiswa kwezicubu ze-RuDA NPs kwafundwa ngokumisela umxholo we-ruthenium kwisibindi, intliziyo, i-spleen, izintso, imiphunga kunye nezicubu.Njengoko kubonisiwe kwifig.I-7A, umxholo ophezulu we-Ore NPs kwizitho eziqhelekileyo zibonakala ngexesha lokuqala lokujonga (i-4 h), ngelixa umxholo ophezulu unqunywe kwiisukisi ze-tumor kwiiyure ze-8 emva kwe-injection, mhlawumbi ngenxa ye-Ore NPs.Isiphumo se-EPR ye-LF.Ngokweziphumo zokusabalalisa, ixesha elifanelekileyo lonyango nge-NP ore lithathwe kwiiyure ze-8 emva kokulawulwa.Ukubonisa inkqubo yokuqokelela i-RuDA-NPs kwiindawo ze-tumor, iipropathi ze-photoacoustic (PA) ze-RuDA-NPs zibekwe iliso ngokurekhoda izibonakaliso ze-PA ze-RuDA-NPs ngamaxesha ahlukeneyo emva kwesitofu.Okokuqala, uphawu lwe-PA lwe-RuDA-NP kwi-vivo lwavavanywa ngokurekhoda imifanekiso ye-PA yesayithi ye-tumor emva kwe-injection ye-intratumoral ye-RuDA-NP.Njengoko kuboniswe kwi-Supplementary Figure 30, i-RuDA-NPs ibonise umqondiso oqinileyo we-PA, kwaye kukho ulungelelwaniso oluhle phakathi kwe-RuDA-NP yoxinaniso kunye ne-PA yesignali ye-PA (i-Supplementary Figure 30A).Emva koko, kwimifanekiso ye-PA ye-vivo yeendawo ze-tumor zarekhodwa emva kwenaliti ye-intravenous ye-RuDA kunye ne-RuDA-NP ngamaxesha ahlukeneyo emva kwesitofu.Njengoko kubonisiwe kwi-Figure 7B, isibonakaliso se-PA se-RuDA-NPs ukusuka kwindawo ye-tumor yanda ngokuthe ngcembe kunye nexesha kwaye yafikelela kwi-plateau kwiiyure ze-8 emva kwe-injection, ehambelana neziphumo zokusasazwa kwezicubu ezinqunywe nguhlalutyo lwe-ICP-MS.Ngokubhekiselele kwi-RuDA (i-Supplementary Fig. 30B), ubuninzi be-PA ubuninzi besignali bubonakala kwiiyure ze-4 emva kwe-injection, ebonisa izinga elikhawulezayo lokungena kwe-RuDA kwi-tumor.Ukongezelela, ukuziphatha kwe-excretory ye-RuDA kunye ne-RuDA-NPs yaphandwa ngokumisela inani le-ruthenium kumchamo kunye nendle usebenzisa i-ICP-MS.Indlela ephambili yokuphelisa i-RuDA (i-Supplementary Fig. 31) kunye ne-RuDA-NPs (i-Fig. 7C) ihamba ngendle, kunye nokukhutshwa okusebenzayo kwe-RuDA kunye ne-RuDA-NPs kwabonwa ngexesha le-8-day study, oku kuthetha ukuba i-RuDA kunye ne-RuDA-NPs inokupheliswa ngokufanelekileyo emzimbeni ngaphandle kobuthi bexesha elide.
A. Ukusabalalisa kwe-Ex vivo ye-RuDA-NP kwizicubu ze-mouse kunqunywe ngumxholo we-Ru (ipesenti ye-dose elawulwayo ye-Ru (ID) ngegram ye-tissue) ngamaxesha ahlukeneyo emva kwesitofu.Iinkcukacha zithetha ± ukutenxa okusemgangathweni (n = 3). Iimvavanyo ezingabhaliswanga, ezinamacala amabini t *p <0.05, **p <0.01, kunye ne-**p <0.001. Iimvavanyo ezingabhaliswanga, ezinamacala amabini t *p <0.05, **p <0.01, kunye ne-**p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 kwaye ***p <0,001. Iimvavanyo ze-t ezinemisila emibini ezingadityaniswanga *p<0.05, **p<0.01, kunye 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 kwaye ***p <0,001. Iimvavanyo ze-t ezinemisila emibini ezingadityaniswanga *p<0.05, **p<0.01, kunye ne-**p<0.001.Imifanekiso ye-B ye-PA ye-in vivo tumor sites kwi-808 nm excitation emva kokulawulwa kwe-intravenous ye-RuDA-NPs (10 µmol kg-1) ngamaxesha ahlukeneyo.Emva kokulawulwa kwe-intravenous ye-RuDA NPs (i-10 µmol kg-1), i-C Ru yakhutshwa kwiigundane ngomchamo kunye nendle ngamaxesha ahlukeneyo.Iinkcukacha zithetha ± ukutenxa okusemgangathweni (n = 3).
Umthamo wokufudumeza we-RuDA-NP kwi-vivo ufundwe kwiigundane ezinqunu kunye ne-MDA-MB-231 kunye ne-RuDA tumors ukuthelekisa.Njengoko kubonisiwe kwifig.I-8A kunye ne-Fig. 32, iqela lokulawula (i-saline) libonise utshintsho oluncinci lokushisa (ΔT ≈ 3 ° C) emva kwemizuzu ye-10 yokuvezwa okuqhubekayo.Nangona kunjalo, ukushisa kwe-RuDA-NPs kunye ne-RuDA kwanda ngokukhawuleza kunye nokushisa okuphezulu kwe-55.2 kunye ne-49.9 ° C, ngokulandelanayo, ukubonelela nge-hyperthermia eyaneleyo kwi-vivo yonyango lomhlaza.Ukunyuka okubonakalayo kwiqondo lokushisa eliphezulu kwi-RuDA NPs (ΔT ≈ 24 ° C) xa kuthelekiswa ne-RuDA (ΔT ≈ 19 ° C) ingaba ngenxa yokugqithiswa kwayo okungcono kunye nokuqokelela kwizicubu ze-tumor ngenxa yesiphumo se-EPR.
Imifanekiso ye-infrared thermal yeegundane kunye ne-MDA-MB-231 i-tumor i-radiated kunye ne-808 nm laser ngamaxesha ahlukeneyo kwiiyure ze-8 emva kwesitofu.Imifanekiso emeleyo yokuphindaphinda kwebhayoloji emine kwiqela ngalinye ibonisiwe.B Relative ithumba umthamo kunye C Umyinge thumba ubunzima amaqela ahlukeneyo iimpuku ngexesha unyango.D Iigophe zobunzima bomzimba bamaqela ahlukeneyo eempuku.I-Irradiate nge-laser ene-wavelength ye-808 nm ngamandla e-0.5 W / cm2 imizuzu eyi-10 (300 J / cm2).Imivalo yemposiso, ithetha ± ukutenxa okusemgangathweni (n = 3). Iimvavanyo ezingabhaliswanga, ezinamacala amabini t *p <0.05, **p <0.01, kunye ne-**p <0.001. Iimvavanyo ezingabhaliswanga, ezinamacala amabini t *p <0.05, **p <0.01, kunye ne-**p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 kwaye ***p <0,001. Iimvavanyo ze-t ezinemisila emibini ezingadityaniswanga *p<0.05, **p<0.01, kunye 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 kwaye ***p <0,001. Iimvavanyo ze-t ezinemisila emibini ezingadityaniswanga *p<0.05, **p<0.01, kunye ne-**p<0.001. I-E H & E ingcolisa imifanekiso yamalungu amakhulu kunye namathumba avela kumaqela ahlukeneyo onyango, kuquka iSaline, iSaline + Laser, iRuDA, iRuDA + Laser, iRuDA-NPs, kunye neRuDA-NPs + Laser amaqela. I-E H & E ingcolisa imifanekiso yamalungu amakhulu kunye namathumba avela kumaqela ahlukeneyo onyango, kuquka iSaline, iSaline + Laser, iRuDA, iRuDA + Laser, iRuDA-NPs, kunye neRuDA-NPs + Laser amaqela. 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. I-E H & E ingcolisa imifanekiso yamalungu amakhulu kunye namathumba avela kumaqela ahlukeneyo onyango, kuquka i-saline, i-saline + laser, i-RuDA, i-RuDA + iLaser, i-RuDA-NPs, kunye ne-RuDA-NPs + amaqela eLaser.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（激光、RuDA + 和激光NPs、RuDA + 和激光 NP來自不同治疗组的主要器官和肿瘤的E H&E Inkcazelo nge E H&E основных органов и опухолей из различных групп лечения, включая физиологический раствор, NPствор, NP-DAKDA, Rule, Rule I-E H & E staining yamalungu amakhulu kunye namathumba avela kumaqela ahlukeneyo onyango kuquka i-saline, i-saline + laser, i-RuDA, i-RuDA + laser, i-RuDA-NPs, kunye ne-RuDA-NPs + laser.Isikali sebar: 60 µm.
Umphumo we-phototherapy kwi-vivo kunye ne-RuDA kunye ne-RuDA NPs yavandlakanywa apho iigundane ze-MDA-MB-231 zifakwe nge-intravenously kunye ne-RuDA okanye i-RuDA NPs ngedosi enye ye-10.0 µmol kg-1 nge-vein yomsila, emva koko i-8 iiyure emva kokutofa.I-laser irradiation ene-wavelength ye-808 nm.Njengoko kubonisiwe kwi-Figure 8B, imiqulu ye-tumor yanda kakhulu kwiqela le-saline kunye ne-laser, ebonisa ukuba i-saline okanye i-laser 808 irradiation yayinempembelelo encinane ekukhuleni kwethumba.Njengoko kwiqela le-saline, ukukhula kwe-tumor okukhawulezayo kwabonwa kwiigundane eziphathwe nge-RuDA-NPs okanye i-RuDA ngokungabikho kwe-laser irradiation, ebonisa ubuthi babo obuphantsi obumnyama.Ngokwahlukileyo, emva kwe-laser irradiation, zombini i-RuDA-NP kunye ne-RuDA unyango luye lwabangela ukuguqulwa kwe-tumor okuphawulekayo kunye nokunciphisa umthamo we-tumor ye-95.2% kunye ne-84.3%, ngokulandelanayo, xa kuthelekiswa neqela eliphathwe nge-saline, ebonisa i-PDT ebalaseleyo ye-synergistic., idibene ne-RuDA / CHTV effect.- I-NP okanye i-Ore Xa kuthelekiswa ne-RuDA, i-RuDA NPs ibonise umphumo ongcono we-phototherapeutic, eyona nto yayibangelwa yimpembelelo ye-EPR ye-RuDA NPs.Iziphumo zokuthintela ukukhula kwe-tumor ziye zavavanywa ngakumbi nge-tumor weight excised ngosuku lwe-15 yonyango (umzobo we-8C kunye ne-Supplementary Fig. 33).Ubuninzi be-tumor mass in RuDA-NP iphathwe iigundane kunye ne-RuDA yokuphatha iigundane yi-0.08 kunye ne-0.27 g, ngokulandelanayo, eyayilula kakhulu kuneqela lokulawula (1.43 g).
Ukongezelela, ubunzima bomzimba weegundane zabhalwa rhoqo ngeentsuku ezintathu ukufunda ubuthi obumnyama be-RuDA-NPs okanye i-RuDA kwi-vivo.Njengoko kuboniswe kwi-Figure 8D, akukho ntlukwano ebalulekileyo kubunzima bomzimba obuqatshelwe kuwo onke amaqela onyango. Ngapha koko, ukungcoliswa kwe-hematoxylin kunye ne-eosin (H&E) kwamalungu amakhulu (intliziyo, isibindi, udakada, imiphunga, kunye nezintso) kumaqela ahlukeneyo onyango lwenziwa. Ngaphezu koko, i-hematoxylin kunye ne-eosin (H & E) ukungcoliswa kwamalungu amakhulu (intliziyo, isibindi, i-spleen, imiphunga kunye nezintso) ezivela kumaqela ahlukeneyo onyango lwenziwa. Кроме того, было проведено окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селезенки, легких и позкупы) и основных органов. Ukongezelela, i-hematoxylin kunye ne-eosin (H & E) i-staining yamalungu amakhulu (intliziyo, isibindi, i-spleen, imiphunga kunye nezintso) ezivela kumaqela ahlukeneyo onyango lwenziwa.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 разлипуные Ukongezelela, i-hematoxylin kunye ne-eosin (H & E) i-staining yamalungu amakhulu (intliziyo, isibindi, i-spleen, imiphunga kunye nezintso) yenziwa kumaqela ahlukeneyo onyango.Njengoko kubonisiwe kwiFig.I-8E, imifanekiso ye-H & E ye-staining yamalungu amahlanu amakhulu avela kwi-RuDA-NPs kunye namaqela e-RuDA abonisa ukungahambi kakuhle okucacileyo okanye umonakalo wamalungu. I-8E, imifanekiso ye-H & E ye-staining yamalungu amahlanu amakhulu avela kwi-RuDA-NPs kunye namaqela e-RuDA abonisa ukungahambi kakuhle okucacileyo okanye umonakalo wamalungu.Njengoko kubonisiwe kwifig.8E, изображения окрашивания H&E пяти основных органов из групп I-RuDA-NPs kunye ne-RuDA ayisayi kuphinda isebenze. I-8E, imifanekiso ye-H & E ye-staining yamalungu amahlanu amakhulu avela kwi-RuDA-NPs kunye namaqela e-RuDA abonisa ukungahambi kakuhle kwamalungu okanye izilonda ezicacileyo.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 Как показано на рисунке 8E, изображения окрашивания H&E ihlawulela i-RuDA-NPs kunye ne-RuDA ayisayi kuphinda isebenze. Njengoko kuboniswe kwi-Figure 8E, imifanekiso ye-H & E ye-staining yamalungu amahlanu amakhulu avela kwi-RuDA-NPs kunye namaqela e-RuDA abonisa ukungahambi kakuhle okucacileyo okanye umonakalo womzimba.Ezi ziphumo zibonise ukuba akukho RuDA-NP okanye i-RuDA ebonisa iimpawu zetyhefu kwi-vivo. Ngaphezu koko, imifanekiso eqingqiweyo ye-H & E ye-tumor ibonise ukuba zombini i-RuDA + Laser kunye ne-RuDA-NPs + amaqela e-Laser anokubangela ukutshabalalisa i-cell cell cell, ebonisa ukugqwesa kwi-vivo phototherapeutic efficacy ye-RuDA kunye ne-RuDA-NPs. Ngaphezu koko, imifanekiso eqingqiweyo ye-H & E ye-tumor ibonise ukuba zombini i-RuDA + Laser kunye ne-RuDA-NPs + amaqela e-Laser anokubangela ukutshabalalisa i-cell cell cell, ebonisa ukugqwesa kwi-vivo phototherapeutic efficacy ye-RuDA kunye ne-RuDA-NPs.Ukongezelela, imifanekiso ye-tumor ene-hematoxylin-eosin ibonise ukuba zombini i-RuDA + Laser kunye ne-RuDA-NPs + i-Laser amaqela anokubangela ukutshatyalaliswa okukhulu kweeseli zomhlaza, ebonisa ukusebenza kwe-phototherapeutic ephezulu ye-RuDA kunye ne-RuDA-NPs kwi-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，话公義的智慧。此外 , 肿瘤 的 & e 染色 显示 , ruda + laser kunye neruda-nps + laser 组均 致 的 癌细胞 破坏 , 证明 组入 ruda . ...Ukongeza, i-hematoxylin kunye ne-eosin enemibala ye-tumor imifanekiso ibonise ukuba zombini i-RuDA + Laser kunye ne-RuDA-NPs + i-Laser amaqela abangele ukutshatyalaliswa okukhulu kweeseli zomhlaza, ebonisa ukusebenza kwe-phototherapeutic ephezulu ye-RuDA kunye ne-RuDA-NPs kwi-vivo.
Ekugqibeleni, i-Ru (II) -arene (RuDA) i-organometallic complex kunye ne-DA-type ligands yenzelwe ukuququzelela inkqubo ye-ISC ngokusebenzisa indlela yokudibanisa.I-RuDA edibeneyo inokuzihlanganisa ngokusebenzisa i-non-covalent interactions ukwenza i-RuDA-derived supramolecular systems, ngaloo ndlela iququzelela ukubunjwa kwe-1O2 kunye nokuguqulwa okusebenzayo kwe-photothermal kunyango lomhlaza olubangelwa ukukhanya.Kuyaphawuleka ukuba i-RuDA ye-monomeric ayizange ivelise i-1O2 phantsi kwe-laser irradiation kwi-808 nm, kodwa inokuvelisa inani elikhulu le-1O2 kwisimo esihlanganisiweyo, ebonisa ingqiqo kunye nokusebenza kakuhle koyilo lwethu.Uphononongo olulandelayo lubonise ukuba indibano ye-supramolecular inika i-RuDA ngokuphuculweyo kwe-photophysical kunye ne-photochemical properties, ezifana ne-redshift absorption kunye ne-photobleaching resistance, ezinqweneleka kakhulu kwi-PDT kunye ne-PTT processing.Zombini i-in vitro kunye ne-vivo experiments zibonise ukuba i-RuDA NPs kunye ne-biocompatibility efanelekileyo kunye nokuqokelela okulungileyo kwi-tumor ibonisa umsebenzi obalaseleyo we-anticancer owenziwe ukukhanya kwi-laser irradiation kwi-wavelength ye-808 nm.Ngaloo ndlela, i-RuDA NPs njengee-reagents ze-bimodal supramolecular PDT / PTW ziya kutyebisa isethi ye-photosensitizers esebenzayo kwi-wavelengths ngaphezu kwe-800 nm.Uyilo lwengqikelelo yenkqubo ye-supramolecular ibonelela ngendlela esebenzayo yeefotosensitizers esebenzayo ye-NIR eneziphumo ezibalaseleyo zefotosensitizing.
Zonke iikhemikhali kunye ne-solvents zifunyenwe kubanikezeli bezorhwebo kwaye zisetyenziswe ngaphandle kokucocwa ngakumbi.I-RuCl3 yathengwa kwi-Boren Precious Metals Co., Ltd. (Kunming, China).[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione) kunye ne-4,7-bis[4-(N,N-diphenylamino)phenyl]-5 , I-6-Diamino-2,1,3-benzothiadiazole yenziwe ngokwezifundo zangaphambili64,65.I-NMR spectra irekhodwe kwi-Bruker Avance III-HD 600 MHz spectrometer kwiZiko loVavanyo loVavanyo loHlahlo lweYunivesithi yaseMzantsi-mpuma usebenzisa i-d6-DMSO okanye i-CDCl3 njenge-solvent.Utshintsho lwekhemikhali δ lunikwa nge ppm.Ngokubhekiselele kwi-tetramethylsilane, kunye nentsebenziswano engaguqukiyo J inikwe kumaxabiso apheleleyo kwi-hertz.I-high resolution mass spectrometry (HRMS) yenziwa kwisixhobo se-Agilent 6224 ESI/TOF MS.Uhlalutyo olusisiseko lwe-C, H, kunye no-N lwenziwa kwi-Vario MICROCHNOS i-elemental analyzer (Elementar).Umboniso obonakalayo we-UV walinganiswa kwi-Shimadzu UV3600 spectrophotometer.I-Fluorescence spectra yarekhodwa kwi-Shimadzu RF-6000 spectrofluorimeter.I-spectra ye-EPR yarekhodwa kwisixhobo se-Bruker EMXmicro-6/1.I-morphology kunye nesakhiwo seesampuli ezilungisiweyo zafundwa kwi-FEI Tecnai G20 (TEM) kunye neBruker Icon (AFM) izixhobo ezisebenza kumbane we-200 kV.I-Dynamic light scattering (DLS) yenziwa kwi-Nanobrook Omni analyzer (Brookhaven).Iimpawu ze-Photoelectrochemical zalinganiswa kwi-electrochemical setup (CHI-660, China).Imifanekiso ye-Photoacoustic ifunyenwe kusetyenziswa inkqubo ye-FUJIFILM VisualSonics Vevo® LAZR.Imifanekiso yeConfocal ifunyenwe kusetyenziswa i-Olympus FV3000 confocal microscope.Uhlalutyo lwe-FACS lwenziwa kwi-cytometer ye-BD Calibur.Uvavanyo lwe-chromatography yolwelo oluphezulu (HPLC) lwenziwa kwi-Waters Alliance e2695 inkqubo kusetyenziswa i-2489 UV/Vis detector.Iimvavanyo ze-Gel Permeation Chromatography (GPC) zirekhodwe kwisixhobo se-Thermo ULTIMATE 3000 usebenzisa i-ERC RefratoMax520 i-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, I-N-diphenylamino)phenyl]-5,6-diamino-2,1,3-benzothiadiazole 65 (652.0 mg, 1.0 mmol) kunye ne-glacial acetic acid (30 mL) yaxutywa kwifriji ye-reflux kwiiyure ze-12.Isinyibilikisi saye sasuswa kwi-vacuo kusetyenziswa i-evaporator ejikelezayo.Intsalela ebangelwayo yahlanjululwa nge-flash column chromatography (i-silica gel, CH2Cl2: MeOH = 20: 1) ukufumana i-RuDA njengepowder eluhlaza (isivuno: 877.5 mg, 80%).umva.Kubalwe kwi-C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.Ifunyenwe: 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 (i-150 MHZ, DMSO), δ , 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 yenziwe ngamanyathelo amabini.I-Pd (PPh3) 4 (46 mg, 0.040 mmol) yongezwa kwi-N, N-diethyl-4- (tributylstannyl) aniline (1.05 g, 2.4 mmol) kunye ne-4,7-dibromo-5,6-dinitro isisombululo - 2, I-1,3-benzothiadiazole (0.38 g, 1.0 mmol) kwi-toluene eyomileyo (100 ml).Umxube wawuxutywe kwi-100 ° C kwiiyure ezingama-24.Emva kokususa i-toluene kwi-vacuo, ukuqina okubangelwayo kwahlanjwa nge-petroleum ether.Emva koko umxube wale ngqungquthela (234.0 mg, 0.45 mmol) kunye ne-iron powder (0.30 g, 5.4 mmol) kwi-acetic acid (20 ml) yaxutywa kwi-80 ° C. iiyure ze-4.Umxube reaction wagalelwa emanzini kunye nesiphumo esiqinileyo brown yaqokelelwa ukuhluzwa.Imveliso yahlanjululwa kabini nge-vacuum sublimation ukunika ukuqina okuluhlaza (126.2 mg, 57% isivuno).umva.Kubalwe kwi-C26H32N6S: C 67.79, H 7.00, N 18.24.Ifunyenwe: 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.
Ii-Compounds zalungiswa kwaye zahlanjululwa ngokulandela iinkqubo ezifana ne-RuDA.umva.Kubalwe kwi-C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.Ifunyenwe: 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 (i-151 MHZ, i-CDCL3), δ (i-PPM) 158.20, 135.7.7.71, 135.7.71, 135.7.7.8.8.8.8.0, 84.4.4.4, 84.4.4, 84.4.4.4., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m/z [M-Cl]+ = 905.24.
I-RuDA yachithwa kwi-MeOH / H2O (5/95, v / v) kwi-concentration ye-10 μM.I-spectrum ye-absorption ye-RuDA yalinganiswa yonke imizuzu ye-5 kwi-spectrophotometer ye-Shimadzu UV-3600 phantsi kwe-irradiation kunye nokukhanya kwe-laser kunye ne-wavelength ye-808 nm (0.5 W / cm2).I-spectra ye-ICG ibhalwe phantsi kweemeko ezifanayo kunye nomgangatho.
I-spectra ye-EPR irekhodwe kwi-spectrometer ye-Bruker EMXmicro-6/1 enegunya le-microwave ye-20 mW, i-scanning range ye-100 G, kunye ne-modulation yentsimi ye-1 G. 2,2,6,6-tetramethyl-4-piperidone. (TEMP) kunye ne-5,5-dimethyl-1-pyrroline N-oxide (DMPO) isetyenziswe njenge-spin traps.I-Electron spin resonance spectra yabhalwa kwiisombululo ezixubileyo ze-RuDA (50 µM) kunye ne-TEMF (20 mM) okanye i-DMPO (20 mM) phantsi kwesenzo se-laser radiation kunye ne-wavelength ye-808 nm (0.5 W / cm2).
Izibalo ze-DFT kunye ne-TD-DFT ze-RuDA zenziwa kwi-PBE1PBE / 6-31 G *//LanL2DZ amanqanaba kwisisombululo samanzi usebenzisa inkqubo ye-Gaussian 1666,67,68.I-HOMO-LUMO, imbobo kunye nokuhanjiswa kwe-electron ye-low-energy singlet echulumancisayo i-RuDA yacwangciswa ngokusebenzisa inkqubo ye-GaussView (uhlobo lwe-5.0).
Siqale sazama ukulinganisa ukusebenza kakuhle kwesizukulwana se-1O2 RuDA usebenzisa i-spectroscopy ye-UV-ebonakalayo ebonakalayo kunye ne-ICG (ΦΔ = 0.002) njengomgangatho, kodwa ukuchithwa kwe-photodegradation ye-ICG kuchaphazele kakhulu iziphumo.Ngaloo ndlela, isivuno se-quantum se-1O2 RuDA silinganiswe ngokufumanisa utshintsho kwi-intensitet ye-ABDA fluorescence malunga ne-428 nm xa ihlanjululwa nge-laser ene-wavelength ye-808 nm (0.5 W / cm2).Iimvavanyo zenziwa kwi-RuDA kunye ne-RuDA NPs (20 μM) emanzini / i-DMF (98/2, v / v) equkethe i-ABDA (50 μM).Imveliso ye-quantum ye-1O2 ibalwa ngokusebenzisa le fomula ilandelayo: ΦΔ (PS) = ΦΔ (ICG) × (rFS/APS)/(rICG/AICG).I-rPS kunye ne-rICG yimilinganiselo yokusabela kwe-ABDA kunye ne-1O2 efunyenwe kwi-photosensitizer kunye ne-ICG, ngokulandelanayo.I-APS kunye ne-AICG yi-absorbection ye-photosensitizer kunye ne-ICG kwi-808 nm, ngokulandelanayo.
Imilinganiselo ye-AFM iqhutywe kwiimeko zolwelo kusetyenziswa imowudi yokuskena kwi-Bruker Dimension Icon ye-AFM inkqubo.Ukusebenzisa isakhiwo esivulekileyo kunye neeseli zamanzi, iiseli zihlanjwe kabini nge-ethanol kwaye zomiswe ngomlambo we-nitrogen.Faka iiseli ezomileyo kwintloko ye-optical ye-microscope.Kwangoko beka ithontsi lesampulu kwidama lolwelo kwaye ulibeke kwi-cantilever usebenzisa isirinji yeplastiki elahlwayo engenazintsholongwane kunye nenaliti engenanto.Enye i-drop ifakwe ngokuthe ngqo kwisampuli, kwaye xa intloko ye-optical iyancipha, amaconsi amabini adibanisa, enze i-meniscus phakathi kwesampuli kunye ne-liquid reservoir.Imilinganiselo ye-AFM yenziwa kusetyenziswa i-SCANASYST-FLUID V-shaped nitride cantilever (Bruker, hardness k = 0.7 N m-1, f0 = 120–180 kHz).
Iikhromatogram ze-HPLC zafunyanwa kwinkqubo ye-Waters e2695 exhotyiswe ngekholamu ye-phoenix C18 (250×4.6 mm, 5 µm) kusetyenziswa i-2489 UV/Vis detector.Ubude be-detector bungama-650 nm.Izigaba ezihambahambayo ze-A kunye ne-B zazingamanzi kunye ne-methanol, ngokulandelanayo, kunye nenqanaba lokuhamba kwesigaba esihambayo sasiyi-1.0 ml · min-1.Ithambeka (i-solvent B) ibe ngolu hlobo lulandelayo: 100% ukusuka kwi-0 ukuya kwimizuzu emi-4, i-100% ukuya kwi-50% ukusuka kwimizuzu emi-5 ukuya kwengama-30, ize iphinde ibekwe kwi-100% ukusuka kwimizuzu engama-31 ukuya kwengama-40.I-Ore yachithwa kwisisombululo esixubileyo se-methanol kunye namanzi (50/50, ngomthamo) kwi-concentration ye-50 μM.Umthamo wenaliti wawungama-20 μl.
Iimvavanyo ze-GPC zirekhodwe kwisixhobo se-Thermo ULTIMATE 3000 esixhotyiswe ngeekholamu ezimbini ze-PL aquagel-OH MIXED-H (2 × 300 × 7.5 mm, 8 µm) kunye ne-ERC RefratoMax520 i-index refractive index detector.Ikholomu ye-GPC yahlanjululwa ngamanzi kwinqanaba lokuhamba kwe-1 ml / min kwi-30 ° C.Ii-NP ze-Ore zachithwa kwisisombululo se-PBS (pH = 7.4, 50 μM), umthamo we-injection wawuyi-20 μL.
Iifotocurrents zilinganiswe kwi-electrochemical setup (CHI-660B, China).Iimpendulo ze-optoelectronic xa i-laser ivuliwe kwaye ivaliwe (808 nm, 0.5 W / cm2) yalinganiswa kwi-voltage ye-0.5 V kwibhokisi elimnyama, ngokulandelanayo.Iseli esemgangathweni ye-electrode emithathu isetyenziswe nge-L-shaped glassy carbon electrode (GCE) njenge-electrode esebenzayo, i-calomel electrode eqhelekileyo (SCE) njenge-electrode reference, kunye ne-platinum disk njenge-electrode counter.Isisombululo se-0.1 M Na2SO4 sisetyenziswe njenge-electrolyte.
Umgca weseli yomhlaza webele lomntu MDA-MB-231 yathengwa kwi-KeyGEN Biotec Co., LTD (Nanjing, China, inombolo yekhathalogu: KG033).Iiseli zakhuliswa kwi-monolayers kwi-Dulbecco's Modified Eagle's Medium (DMEM, i-glucose ephezulu) yongezwa ngesisombululo se-10% ye-fetal bovine serum (FBS), i-penicillin (100 μg / ml) kunye ne-streptomycin (100 μg / ml).Zonke iiseli zakhuliswa kwi-37 ° C kwindawo efumileyo ene-5% CO2.
Uvavanyo lwe-MTT lwalusetyenziselwa ukumisela i-cytotoxicity ye-RuDA kunye ne-RuDA-NPs phambi kokungabikho kokukhanya kwe-irradiation, kunye okanye ngaphandle kwe-Vc (0.5 mM).Iiseli zomhlaza ze-MDA-MB-231 zakhuliswa kwiipleyiti ze-96-well kwi-cell density malunga ne-1 x 105 iiseli / ml / kakuhle kwaye zifakwe kwiiyure ze-12 kwi-37.0 ° C kwi-atmosphere ye-5% CO2 kunye ne-95% yomoya.I-RuDA kunye ne-RuDA NPs ezichithwe emanzini zongezwa kwiiseli.Emva kweeyure eziyi-12 zokufukanyelwa, iiseli ziye zavezwa kwi-0.5 W cm -2 laser radiation kwi-wavelength ye-808 nm kangangemizuzu eyi-10 (300 J cm -2) emva koko zifukanywe ebumnyameni iiyure ezingama-24.Emva koko iiseli zifakwe kwi-MTT (5 mg / ml) kwezinye iiyure ze-5.Okokugqibela, tshintsha okuphakathi ukuya kwi-DMSO (200 µl) ukunyibilikisa isiphumo sekristale emfusa yeformazan.Amaxabiso e-OD alinganiswa kusetyenziswa i-microplate reader ene-wavelength ye-570/630 nm.Ixabiso le-IC50 kwisampulu nganye libalwe kusetyenziswa isofthiwe ye-SPSS ukusuka kwi-dose-response curves efunyenwe ubuncinane kwiimvavanyo ezintathu ezizimeleyo.
Iiseli ze-MDA-MB-231 zaphathwa nge-RuDA kunye ne-RuDA-NP kwi-concentration ye-50 μM.Emva kweeyure ze-12 zokufakelwa, iiseli zatshatyalaliswa nge-laser ene-wavelength ye-808 nm kunye namandla e-0.5 W / cm2 kwi-10 min (300 J / cm2).Kwiqela le-vitamin C (Vc), iiseli zaphathwa nge-0.5 mM Vc ngaphambi kwe-laser irradiation.Emva koko iiseli zifakwe ebumnyameni kwiiyure ezingama-24 ezongezelelweyo, emva koko zifakwe i-calcein AM kunye nepropidium iodide (20 μg / ml, 5 μl) imizuzu engama-30, emva koko ihlanjwe nge-PBS (10 μl, pH 7.4).imifanekiso yeeseli ezinebala.