Zikomo pochezera Nature.com.Mtundu wa msakatuli womwe mukugwiritsa ntchito uli ndi chithandizo chochepa cha CSS.Kuti mudziwe zambiri, tikupangira kuti mugwiritse ntchito msakatuli wosinthidwa (kapena kuletsa Compatibility Mode mu Internet Explorer).Pakadali pano, kuti tithandizire kupitilizabe, tidzapereka tsambalo popanda masitayilo ndi JavaScript.
Ma photosensitizer ogwira mtima ndi ofunikira kwambiri pakugwiritsa ntchito kwachipatala kwa phototherapy.Komabe, ma photosensitizer wamba nthawi zambiri amakhala ndi mayamwidwe a kutalika kwa mafunde pang'ono, kusakwanira kwazithunzi, zokolola zochepa za mitundu ya okosijeni (ROS), komanso kuzimitsidwa kwa ROS.Apa tikunena za pafupi-infrared (NIR) supramolecular photosensitizer (RuDA) mkhalapakati ndi self-assembly wa Ru(II) -arene organometallic complexes mu amadzimadzi solution.RuDA ikhoza kutulutsa mpweya wa singlet (1O2) m'malo ophatikizika, ndipo ikuwonetseratu khalidwe la mbadwo wa 1O2 chifukwa cha kuwonjezeka kwakukulu kwa njira yodutsa pakati pa singlet-triplet system.Pansi pa 808 nm laser kuwala, RuDA ikuwonetsa zokolola za 1O2 quantum za 16.4% (zobiriwira za indocyanine zovomerezeka ndi FDA: ΦΔ = 0.2%) ndi kutembenuka kwapamwamba kwa photothermal kwa 24.2% (malonda a golide nanorods) ndi photostability kwambiri.: 21.0%, nanoshell zagolide: 13.0%).Kuphatikiza apo, RuDA-NPs okhala ndi biocompatibility yabwino amatha kudziunjikira m'malo otupa, zomwe zimapangitsa kuti chotupacho chiwonjezeke panthawi ya chithandizo cha Photodynamic ndikuchepetsa 95.2% kuchuluka kwa chotupa mu vivo.Chithandizo cha photodynamic chophatikizira ichi chimapereka njira yopangira ma photosensitizer okhala ndi mawonekedwe owoneka bwino komanso opangira zithunzi.
Poyerekeza ndi mankhwala ochiritsira, photodynamic therapy (PDT) ndi mankhwala ochititsa chidwi a khansa chifukwa cha ubwino wake waukulu monga kuwongolera kolondola kwa spatiotemporal, kusasokoneza, kukana mankhwala osokoneza bongo, ndi kuchepetsa zotsatira 1,2,3.Pansi pa kuwala kwa kuwala, ma photosensitizers omwe amagwiritsidwa ntchito amatha kutsegulidwa kuti apange mitundu yowonjezereka ya okosijeni (ROS), yomwe imatsogolera ku apoptosis / necrosis kapena mayankho a chitetezo cha mthupi4,5. Komabe, ma photosensitizer ambiri wamba, monga ma chlorin, porphyrins, ndi anthraquinones, amakhala ndi mayamwidwe amfupi a kutalika kwa mafunde (mafupipafupi ochepera 680 nm), zomwe zimapangitsa kuti asalowemo bwino chifukwa cha kuyamwa kwambiri kwa mamolekyu achilengedwe (monga hemoglobin ndi melanin) mu chigawo chowoneka6,7. Komabe, ma photosensitizer ambiri wamba, monga ma chlorin, porphyrins, ndi anthraquinones, amakhala ndi mayamwidwe amfupi a kutalika kwa mafunde (mafupipafupi ochepera 680 nm), zomwe zimapangitsa kuti asalowemo bwino chifukwa cha kuyamwa kwambiri kwa mamolekyu achilengedwe (monga hemoglobin ndi melanin) mu chigawo chowoneka6,7. 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. Komabe, ma photosensitizer odziwika bwino monga ma klorini, porphyrins ndi anthraquinones amakhala ndi mayamwidwe amfupi a kutalika kwa mafunde (< 680 nm) zomwe zimapangitsa kuti kuwala kulowetse bwino chifukwa cha kuyamwa kwambiri kwa mamolekyu achilengedwe (monga hemoglobin ndi melanin) kumalo owoneka bwino6,7., 大多数 传统 的 光敏剂, 如 二 卟酚 卟酚 和 相对 短 短 短 短 短 吸收 吸收 对 对 对导致光穿透性差., 大多数 传统 的 光敏剂, 二 卟酚 蒽醌, 具有 的 短 短 频率 频率 频率 频率 频率 频率 频率 频率 频率吸收 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. Komabe, ma photosensitizer ambiri achikhalidwe monga ma klorini, porphyrins ndi anthraquinones amakhala ndi mayamwidwe amfupi a kutalika kwa mafunde (mafupipafupi ochepera 680 nm) chifukwa cha kuyamwa mwamphamvu kwa ma biomolecules monga hemoglobin ndi melanin zomwe zimapangitsa kuti asalowemo bwino.Malo owoneka 6.7.Choncho, pafupi-infrared (NIR) kuyamwa photosensitizers kuti adamulowetsa mu 700-900 nm "zenera achire" ali oyenerera phototherapy.Popeza kuwala kwapafupi ndi infrared ndikocheperako pang'ono ndi minyewa yachilengedwe, kungayambitse kulowa mwakuya komanso kuchepa kwa photodamage8,9.
Tsoka ilo, ma photosensitizer omwe alipo kale a NIR nthawi zambiri amakhala ndi kutsika kosawoneka bwino, kutsika kwa singlet oxygen (1O2) kutulutsa mphamvu, komanso kuphatikizika kwa 1O2 quenching, zomwe zimalepheretsa kugwiritsa ntchito kwawo kwachipatala10,11.Ngakhale kuti khama lalikulu lapangidwa kuti liwongolere mawonekedwe a photophysical ndi photochemical a photosensitizers wamba, mpaka pano malipoti angapo anena kuti ma photosensitizer omwe amamwa NIR amatha kuthetsa mavuto onsewa.Kuphatikiza apo, ma photosensitizer angapo awonetsa kulonjeza kwa m'badwo wabwino wa 1O212,13,14 akayatsidwa ndi kuwala kopitilira 800 nm, popeza mphamvu ya photon imachepa mwachangu kudera lapafupi ndi IR.Triphenylamine (TFA) monga electron donor ndi [1,2,5]thiadiazole-[3,4-i] dipyrido[a,c]phenazine (TDP) monga gulu lovomereza ma elekitironi Donor-acceptor (DA) mtundu amapaka kalasi za utoto, zotengera pafupi-infrared, zomwe zaphunziridwa mozama kwa pafupifupi infrared bioimaging II ndi photothermal therapy (PTT) chifukwa cha bandgap yawo yopapatiza.Chifukwa chake, utoto wamtundu wa DA utha kugwiritsidwa ntchito ku PDT ndi chisangalalo chapafupi ndi IR, ngakhale sanaphunzirepo ngati ma photosensitizer a PDT.
Ndizodziwika bwino kuti kugwiritsa ntchito bwino kwambiri kwa intersystem kuwoloka (ISC) kwa photosensitizers kumalimbikitsa mapangidwe a 1O2.Njira yodziwika bwino yopititsira patsogolo njira ya ISC ndikukulitsa kulumikizana kwa spin-orbit (SOC) kwa ma photosensitizer poyambitsa maatomu olemera kapena magulu apadera achilengedwe.Komabe, njira iyi ikadali ndi zovuta ndi zolepheretsa19,20.Posachedwapa, supramolecular self-assembly yapereka njira yanzeru yopangira zida zogwirira ntchito pamlingo wa maselo, 21,22 ndi ubwino wambiri mu phototherapy: (1) photosensitizers odziphatikiza okha akhoza kukhala ndi kuthekera kopanga mapangidwe a riboni.Zofanana ndi zida zamagetsi zomwe zimakhala ndi mphamvu zambiri zogawa mphamvu chifukwa cha mizere yodutsana pakati pa midadada yomanga.Choncho, mphamvu machesi pakati m'munsi singlet osangalala boma (S1) ndi oyandikana trilet osangalala boma (Tn) adzakhala bwino, amene ali opindulitsa kwa ndondomeko ISC 23, 24 .(2) Msonkhano wa Supramolecular udzachepetsa kupumula kopanda ma radiation pogwiritsa ntchito intramolecular motion limitation mechanism (RIM), yomwe imalimbikitsanso ndondomeko ya ISC 25, 26.(3) Msonkhano wa supramolecular ukhoza kuteteza mamolekyu amkati a monoma ku okosijeni ndi kuwonongeka, potero kumapangitsa kuti photostability ya photosensitizer ikhale yabwino.Poganizira zabwino zomwe zili pamwambazi, timakhulupirira kuti ma supramolecular photosensitizer system atha kukhala njira yabwino yothanirana ndi zolakwika za PDT.
Macomplexes a Ru(II) ndi nsanja yodalirika yazachipatala yomwe ingagwiritsidwe ntchito pozindikira ndi kuchiza matenda chifukwa chapadera komanso kukongola kwawo kwachilengedwe28,29,30,31,32,33,34.Kuonjezera apo, kuchuluka kwa mayiko okondwa komanso ma tunable photophysicochemical properties a Ru(II)-based complexes amapereka ubwino waukulu pa chitukuko cha Ru(II)-based photosensitizers35,36,37,38,39,40.Chitsanzo chodziwika bwino ndi ruthenium(II) polypyridyl complex TLD-1433, yomwe pakali pano ili m'mayesero achipatala a Phase II ngati photosensitizer yochizira khansa ya chikhodzodzo yopanda minofu (NMIBC)41.Kuphatikiza apo, ruthenium(II)arene organometallic complexes amagwiritsidwa ntchito kwambiri ngati chemotherapeutic agents pochiza khansa chifukwa cha kawopsedwe kakang'ono komanso kosavuta kusinthidwa42,43,44,45.Ma ionic a Ru(II) -arene organometallic complexes sangangowonjezera kusungunuka kosauka kwa ma chromophores a DA muzosungunulira wamba, komanso kumathandizira kuphatikiza kwa ma chromophores a DA.Kuphatikiza apo, mawonekedwe a pseudooctahedral theka la masangweji a organometallic complexes a Ru(II) -arenes amatha kuletsa mwamphamvu H-aggregation ya DA-type chromophores, potero kumathandizira kupangidwa kwa J-aggregation yokhala ndi mayamwidwe ofiira.Komabe, kuipa kwachilengedwe kwa Ru(II) -arene complexes, monga kukhazikika kochepa komanso/kapena kusapezeka bwino kwa bioavailability, kumatha kusokoneza mphamvu yamankhwala komanso mu vivo ntchito ya arene-Ru(II) complexes.Komabe, kafukufuku wasonyeza kuti zovuta izi zitha kugonjetsedwa ndikuyika ma ruthenium complexes okhala ndi ma polima ogwirizana ndi biocompatible encapsulation kapena covalent conjugation.
Mu ntchitoyi, timapereka ma DA-conjugated complexes a Ru (II) -arene (RuDA) ndi choyambitsa cha NIR kudzera mu mgwirizano wogwirizanitsa pakati pa chromophore ya DAD ndi Ru (II) -arene moiety.Zotsatira zake zimatha kudziunjikira zokha kukhala ma metalosupramolecular vesicles m'madzi chifukwa chosagwirizana.Makamaka, msonkhano supramolecular anapatsa RuDA ndi polymerization-anachititsa intersystem kuwoloka katundu, amene kwambiri kuchuluka ISC dzuwa, amene anali yabwino kwa PDT (mkuyu. 1A).Kuonjezera kuchuluka kwa chotupa komanso mu vivo biocompatibility, FDA-yovomerezeka Pluronic F127 (PEO-PPO-PEO) idagwiritsidwa ntchito kuyika RuDA47,48,49 kupanga RuDA-NP nanoparticles (Chithunzi 1B) chomwe chimagwira ntchito ngati PDT / Dual- Woyimira wa PTT.Mu phototherapy ya khansa (Chithunzi 1C), RuDA-NP idagwiritsidwa ntchito pochiza mbewa zamaliseche ndi zotupa za MDA-MB-231 kuti ziphunzire mphamvu ya PDT ndi PTT mu vivo.
Chithunzi chojambula cha photophysical mechanism ya RuDA mu monomeric ndi mitundu yambiri ya phototherapy ya khansa, kaphatikizidwe ka B RuDA-NPs ndi C RuDA-NPs kwa NIR-activated PDT ndi PTT.
RuDA, yomwe ili ndi ntchito za TPA ndi TDP, inakonzedwa motsatira ndondomeko yomwe ikuwonetsedwa mu Supplementary Figure 1 (Chithunzi 2A), ndipo RuDA imadziwika ndi 1H ndi 13C NMR spectra, electrospray ionization mass spectrometry, ndi kusanthula koyambira (Ziwerengero Zowonjezera 2-4 ).Mapu a RuDA electron density difference ya masinthidwe otsika kwambiri a singlet adawerengedwa ndi chiphunzitso chodalira nthawi (TD-DFT) kuti aphunzire njira yosinthira ndalama.Monga momwe zasonyezedwera mu Supplementary Figure 5, kachulukidwe ka ma elekitironi amasunthika makamaka kuchokera ku triphenylamine kupita ku gawo lolandirira TDP pambuyo pa kutulutsa chithunzithunzi, chomwe chingabwere chifukwa cha kusintha kwa intramolecular charge transfer (CT).
Mapangidwe a Chemical a Ore B Mayamwidwe a Ore mu zosakaniza zosiyanasiyana za DMF ndi madzi.C Mayamwidwe abwinobwino a RuDA (800 nm) ndi ICG (779 nm) motsutsana ndi nthawi ya 0.5 W cm-2 ya 808 nm laser kuwala.D The photodegradation wa ABDA zikusonyeza ndi RuDA-anachititsa mapangidwe 1O2 mu DMF / H2O osakaniza ndi zinthu zosiyanasiyana madzi pansi zochita za laser poizoniyu ndi wavelength 808 nm ndi mphamvu 0,5 W / cm2.
Abstract—UV-visible absorption spectroscopy idagwiritsidwa ntchito pofufuza momwe Ore amadzipangira okha muzosakaniza za DMF ndi madzi mosiyanasiyana.Monga momwe tawonetsera mkuyu.2B, RuDA imasonyeza magulu a mayamwidwe kuchokera ku 600 mpaka 900 nm mu DMF ndi gulu lalikulu la mayamwidwe pa 729 nm.Kuchulukitsa kuchuluka kwa madzi kudapangitsa kusintha kofiira pang'onopang'ono kwa mayamwidwe a Ore mpaka 800 nm, zomwe zikuwonetsa J-aggregation ya Ore mu dongosolo losonkhana.Mawonekedwe a photoluminescence a RuDA mu zosungunulira zosiyanasiyana akuwonetsedwa mu Supplementary Figure 6. RuDA ikuwoneka kuti ikuwonetsa luminescence ya NIR-II yokhala ndi utali wokwanira wotulutsa ca.1050 nm mu CH2Cl2 ndi CH3OH, motsatana.Kusintha kwakukulu kwa Stokes (pafupifupi 300 nm) kwa RuDA kumasonyeza kusintha kwakukulu kwa geometry ya dziko lokondwa ndi mapangidwe a mphamvu zochepa zopatsa mphamvu.Zokolola za luminescence quantum za Ore mu CH2Cl2 ndi CH3OH zidatsimikiziridwa kukhala 3.3 ndi 0.6%, motsatana.Komabe, mu chisakanizo cha methanol ndi madzi (5/95, v/v), kusintha pang'ono kwa mpweya ndi kuchepa kwa zokolola za quantum (0.22%) zinawonedwa, zomwe zingakhale chifukwa cha kudziphatika kwa Ore. .
Kuti tiwone m'maganizo mwathu kudziphatika kwa ORE, tinagwiritsa ntchito madzi a atomic force microscopy (AFM) kuti tiwone kusintha kwa morphological mu ORE mu njira ya methanol mutawonjezera madzi.Pamene madzi anali pansi pa 80%, palibe kuphatikizika bwino komwe kunawonedwa (Supplementary Fig. 7).Komabe, ndi kuwonjezeka kwina kwa madzi okhutira mpaka 90-95%, tinthu tating'onoting'ono tating'ono tating'ono tating'ono tinkawoneka, zomwe zimasonyeza kudzipangira kwa Ore. yankho (mkuyu 2C ndi Supplementary mkuyu. 8).Mosiyana ndi zimenezi, kuyamwa kwa indocyanine wobiriwira (ICG monga ulamuliro) kunatsika mofulumira pa 779 nm, kusonyeza photostability wabwino kwambiri wa RuDA.Kuonjezera apo, kukhazikika kwa RuDA-NPs mu PBS (pH = 5.4, 7.4 ndi 9.0), 10% FBS ndi DMEM (high glucose) anayesedwa ndi UV-yowoneka mayamwidwe spectroscopy pa nthawi zosiyanasiyana.Monga momwe tawonetsera mu Supplementary Figure 9, kusintha pang'ono kwa magulu a mayamwidwe a RuDA-NP kunawonedwa mu PBS pa pH 7.4 / 9.0, FBS ndi DMEM, kusonyeza kukhazikika kwabwino kwa RuDA-NP.Komabe, mu acidic medium (рН = 5.4) hydrolysis ya Ore inapezeka.Tinayesanso kukhazikika kwa RuDA ndi RuDA-NP pogwiritsa ntchito njira za high performance liquid chromatography (HPLC).Monga momwe tawonetsera mu Supplementary Figure 10, RuDA inali yokhazikika mu chisakanizo cha methanol ndi madzi (50/50, v / v) kwa ola loyamba, ndipo hydrolysis inawonedwa pambuyo pa maola 4.Komabe, nsonga yayikulu yokha ya concave-convex idawonedwa kwa RuDA NPs.Choncho, gel permeation chromatography (GPC) idagwiritsidwa ntchito poyesa kukhazikika kwa RuDA NPs mu PBS (pH = 7.4).Monga momwe tawonetsera mu Supplementary Figure 11, pambuyo pa maola 8 a incubation pansi pa mikhalidwe yoyesedwa, kutalika kwa nsonga, m'lifupi mwake ndi nsonga ya NP RuDA sikunasinthe kwambiri, kusonyeza kukhazikika kwakukulu kwa NP RuDA.Kuonjezera apo, zithunzi za TEM zinasonyeza kuti morphology ya RuDA-NP nanoparticles inakhalabe yosasinthika pambuyo pa maola a 24 mumadzimadzimadzi a PBS (pH = 7.4, Supplementary Fig. 12).
Chifukwa kudzipanga nokha kungapereke mawonekedwe osiyanasiyana ogwira ntchito ndi mankhwala pa Ore, tinawona kutulutsidwa kwa 9,10-anthracenediylbis(methylene)dimalonic acid (ABDA, chizindikiro 1O2) mu zosakaniza za methanol.Ore okhala ndi madzi osiyanasiyana50.Monga momwe tawonetsera mu Chithunzi 2D ndi Supplementary Figure 13, palibe kuwonongeka kwa ABDA komwe kunawonedwa pamene madzi anali pansi pa 20%.Ndi kuwonjezeka kwa chinyezi mpaka 40%, kuwonongeka kwa ABDA kunachitika, monga zikuwonekera ndi kuchepa kwamphamvu kwa ABDA fluorescence.Zawonedwanso kuti kuchuluka kwa madzi kumapangitsa kuwonongeka kwachangu, kutanthauza kuti RuDA yodzipangira yokha ndiyofunikira komanso yopindulitsa pakuwonongeka kwa ABDA.Chodabwitsa ichi ndi chosiyana kwambiri ndi ma chromophores amakono a ACQ (aggregation-induced quenching).Mukayatsidwa ndi laser yokhala ndi kutalika kwa 808 nm, zokolola za quantum za 1O2 RuDA mu chisakanizo cha 98% H2O / 2% DMF ndi 16.4%, zomwe ndi 82 nthawi zambiri kuposa ICG (ΦΔ = 0.2%)51, kuwonetsa m'badwo wodabwitsa wa 1O2 RuDA pakuphatikizana.
Electron spins pogwiritsa ntchito 2,2,6,6-tetramethyl-4-piperidinone (TEMP) ndi 5,5-dimethyl-1-pyrroline N-oxide (DMPO) ngati misampha ya resonance spectroscopy (ESR) idagwiritsidwa ntchito kuzindikira mitundu yomwe imayambitsa AFK.ndi RuDA.Monga momwe zasonyezedwera mu Supplementary Figure 14, zatsimikiziridwa kuti 1O2 imapangidwa pa nthawi ya kuwala pakati pa 0 ndi 4 mphindi.Kuonjezera apo, pamene RuDA incubated ndi DMPO pansi pa kuwala, chizindikiro cha EPR cha mizere inayi cha 1: 2: 2: 1 DMPO-OH · adduct chinapezeka, kusonyeza mapangidwe a hydroxyl radicals (OH ·).Ponseponse, zotsatira zomwe zili pamwambazi zikuwonetsa kuthekera kwa RuDA kulimbikitsa kupanga ROS kudzera mu njira yapawiri ya I/II photosensitization.
Kuti mumvetse bwino zamagetsi a RuDA mu mawonekedwe a monomeric ndi ophatikizana, malire a molecular orbitals a RuDA mu mawonekedwe a monomeric ndi dimeric anawerengedwa pogwiritsa ntchito njira ya DFT.Monga momwe tawonetsera mkuyu.3A, okwera kwambiri occupied molecular orbital (HOMO) wa monomeric RuDA ndi delocalized pamodzi ligand msana ndi otsika kwambiri unoccupied molekyulu orbital (LUMO) wakhazikika pa TDP wolandira unit.M'malo mwake, kachulukidwe ka ma elekitironi mu dimeric HOMO amayang'ana pa ligand ya molekyulu imodzi ya RuDA, pomwe kuchuluka kwa ma elekitironi mu LUMO kumangoyang'ana pa gawo lolandila la molekyulu ina ya RuDA, zomwe zikuwonetsa kuti RuDA ili mu dimer.Zithunzi za CT.
A HOMO ndi LUMO a Ore amawerengedwa mu monomeric ndi dimeric mitundu.B Magawo a mphamvu imodzi ndi katatu a Ore mu ma monomers ndi dimers.C Miyezo yoyerekeza ya RuDA ndi njira zomwe zingatheke za ISC monga monomeric C ndi dimeric D. Mivi imasonyeza njira za ISC zomwe zingatheke.
Kugawidwa kwa ma electron ndi mabowo mu singlet low-energy singlet maiko okondwa a RuDA mu mawonekedwe a monomeric ndi dimeric adawunikidwa pogwiritsa ntchito pulogalamu ya Multiwfn 3.852.53, yomwe inawerengedwa pogwiritsa ntchito njira ya TD-DFT.Monga momwe zasonyezedwera pazowonjezera.Monga momwe ziwonetsedwera mu Zithunzi 1-2, mabowo a monomeric RDA nthawi zambiri amachotsedwa pamsana wa ligand m'madera okondwa a singlet, pamene ma electron amapezeka kwambiri mu gulu la TDP, kusonyeza makhalidwe a intramolecular a CT.Kuphatikiza apo, pamagawo osangalatsa a singlet awa, pali kuphatikizika kocheperako pakati pa mabowo ndi ma elekitironi, kutanthauza kuti mayiko okondwa a singlet awa amathandizira kuchokera ku chisangalalo cha komweko (LE).Kwa ma dimers, kuwonjezera pa mawonekedwe a intramolecular CT ndi LE, gawo lina la ma intermolecular CT adawonedwa m'maboma omwewo, makamaka S3, S4, S7, ndi S8, kutengera kusanthula kwa intermolecular CT, ndi CT intermolecular transitions monga zazikulu. (Supplementary Table).3).
Kuti timvetse bwino zotsatira zoyesera, tidafufuzanso zinthu za RuDA zokondweretsa maiko kuti tifufuze kusiyana pakati pa monomers ndi dimers (Supplementary Tables 4-5).Monga momwe chithunzi 3B chikusonyezera, mphamvu za singlet ndi triplet zokondweretsa za dimer ndizowonjezereka kwambiri kuposa za monoma, zomwe zimathandiza kuchepetsa kusiyana kwa mphamvu pakati pa S1 ndi Tn. Zanenedwa kuti kusintha kwa ISC kumatha kuchitika mkati mwa mphamvu yaying'ono (ΔES1-Tn <0.3 eV) pakati pa S1 ndi Tn54. Zakhala zikudziwika kuti kusintha kwa ISC kungathe kuchitika mkati mwa mphamvu yochepa (ΔES1-Tn <0.3 eV) pakati pa S1 ndi Tn54. Сообщалось, что переходы ISC ikhoza kukuthandizani kuti musinthe pakusintha kwazomwe mukuchita (ΔES1-Tn <0,3 эВ) между S1 ndi Tn5. Zanenedwa kuti kusintha kwa ISC kumatha kuchitika mkati mwa mphamvu yaying'ono (ΔES1-Tn <0.3 eV) pakati pa S1 ndi 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 ingathe kubweretsanso kutulutsa kwatsopano komwe kumafunikira (ΔES1-Tn < 0,3 эВ) между S1 ndi Tn54. Zakhala zikudziwika kuti kusintha kwa ISC kungathe kuchitika mkati mwa mphamvu yochepa (ΔES1-Tn <0.3 eV) pakati pa S1 ndi Tn54.Kuphatikiza apo, orbital imodzi yokha, yotanganidwa kapena yopanda anthu, iyenera kusiyanasiyana m'malo amodzi ndi atatu kuti ipereke gawo lopanda ziro la SOC.Chifukwa chake, potengera kuwunika kwa mphamvu yosangalatsa komanso kusintha kwa orbital, njira zonse zomwe zingatheke kusintha kwa ISC zikuwonetsedwa muzithunzi.3C,D.Makamaka, njira imodzi yokha ya ISC imapezeka mu monomer, pomwe mawonekedwe a dimeric ali ndi njira zinayi za ISC zomwe zitha kupititsa patsogolo kusintha kwa ISC.Choncho, ndizomveka kuganiza kuti mamolekyu ambiri a RuDA akuphatikizidwa, njira za ISC zopezeka mosavuta.Choncho, RuDA aggregates akhoza kupanga zida ziwiri zamagetsi zamagetsi mu singlet ndi triplet states, kuchepetsa kusiyana kwa mphamvu pakati pa S1 ndi Tn yomwe ilipo, potero kuonjezera mphamvu ya ISC kuti ithandize 1O2 m'badwo.
Kuti tipitirize kufotokozera njira yomwe ili pansi pake, tinapanga gulu lofotokozera la arene-Ru (II) complex (RuET) posintha magulu awiri a ethyl ndi magulu awiri a triphenylamine phenyl mu RuDA (Mkuyu 4A, kuti mudziwe zambiri, onani ESI, Supplementary 15). -21 ) Kuchokera kwa wopereka (diethylamine) kuti alandire (TDF), RuET ili ndi zofanana za intramolecular CT monga RuDA.Monga zikuyembekezeredwa, mayamwidwe sipekitiramu wa RuET mu DMF anasonyeza otsika mphamvu mlandu kutengerapo gulu ndi amphamvu mayamwidwe pafupi infuraredi dera m'dera la 600-1100 nm (mkuyu. 4B).Kuonjezera apo, kuphatikizika kwa RuET kunawonedwanso ndi kuchuluka kwa madzi, zomwe zinawonetsedwa mu redshift ya mayamwidwe pazipita, zomwe zinatsimikiziridwa ndi madzi AFM imaging (Supplementary Fig. 22).Zotsatira zikuwonetsa kuti RuET, monga RuDA, ikhoza kupanga mayiko a intramolecular ndikudzisonkhanitsa okha m'magulu ophatikizana.
Mtengo wa magawo RuET.B Mayamwidwe amtundu wa RuET mu zosakaniza zosiyanasiyana za DMF ndi madzi.Malo C EIS Nyquist a RuDA ndi RuET.Mayankho a Photocurrent D a RuDA ndi RuET pansi pa ma radiation a laser okhala ndi kutalika kwa 808 nm.
Kujambula kwa ABDA pamaso pa RuET kunayesedwa ndi kuwala ndi laser yokhala ndi kutalika kwa 808 nm.Chodabwitsa n'chakuti, palibe kuwonongeka kwa ABDA komwe kunawonedwa m'magulu osiyanasiyana a madzi (Supplementary Fig. 23).Chifukwa chotheka ndi chakuti RuET silingapange bwino mawonekedwe amagetsi omangika chifukwa unyolo wa ethyl sulimbikitsa kusamutsa kwapakati kwapakati.Choncho, electrochemical impedance spectroscopy (EIS) ndi miyeso yochepa ya photocurrent inkachitidwa kuti afanizire zinthu za photoelectrochemical za RuDA ndi RuET.Malingana ndi chiwembu cha Nyquist (Chithunzi 4C), RuDA imasonyeza utali wochepa kwambiri kuposa RuET, zomwe zikutanthauza kuti RuDA56 ili ndi maulendo othamanga a intermolecular electron komanso conductivity yabwino.Kuonjezera apo, kachulukidwe ka chithunzi cha RuDA ndipamwamba kwambiri kuposa RuET (Mkuyu 4D), kutsimikizira kuti RuDA57 imayenda bwino.Chifukwa chake, gulu la phenyl la triphenylamine mu Ore limagwira ntchito yofunika kwambiri popereka kutengerapo kwa intermolecular komanso kupanga mawonekedwe amagetsi omangika.
Kuti tiwonjezere kuchuluka kwa chotupa komanso mu vivo biocompatibility, tidaphatikizanso RuDA ndi F127.Pafupifupi hydrodynamic awiri a RuDA-NPs anatsimikiza kukhala 123.1 nm ndi yopapatiza kugawa (PDI = 0.089) ntchito mphamvu kuwala kubalalika (DLS) njira (Figure 5A), amene kulimbikitsa chotupa kudzikundikira ndi kuwonjezeka permeability ndi kusunga.EPR) zotsatira.Zithunzi za TEM zidawonetsa kuti Ore NPs ali ndi mawonekedwe ozungulira ofanana ndi mainchesi a 86 nm.Mwachidziwitso, kuyamwa kwakukulu kwa RuDA-NPs kunawonekera pa 800 nm (Supplementary Fig. 24), kusonyeza kuti RuDA-NPs ikhoza kusunga ntchito ndi katundu wodzipangira okha RuDAs.Kuwerengera kwa ROS quantum zokolola za NP Ore ndi 15.9%, zomwe zikufanana ndi Ore.Monga momwe tawonetsera mkuyu.5B, C, gulu lolamulira (PBS kokha) linawonjezeka pang'ono kutentha, pamene kutentha kwa njira ya RuDA-NPs kunakula mofulumira ndi kutentha kwakukulu (ΔT) mpaka 15.5, 26.1, ndi 43.0 ° C.Zolemba zapamwamba zinali 25, 50, ndi 100 µM, motero, zomwe zimasonyeza mphamvu ya photothermal ya RuDA NPs.Kuonjezera apo, miyeso ya kutentha / kuzizira inatengedwa kuti iwonetsetse kukhazikika kwa photothermal kwa RuDA-NP ndikuyerekeza ndi ICG.Kutentha kwa Ore NPs sikunachepe pambuyo pa maulendo asanu otentha / ozizira (mkuyu 5D), zomwe zimasonyeza kukhazikika kwa photothermal kwa Ore NPs.Mosiyana ndi zimenezi, ICG imasonyeza kutsika kwa photothermal monga momwe zimawonekera kuchokera kukuwonekera kwa kutentha kwa kutentha kwa photothermal pansi pazikhalidwe zomwezo.Malingana ndi njira yapitayi58, photothermal conversion efficient (PCE) ya RuDA-NP inawerengedwa ngati 24.2%, yomwe ili pamwamba kuposa zipangizo zamakono zomwe zilipo monga golide nanorods (21.0%) ndi nanoshells zagolide (13.0%)59.Chifukwa chake, NP Ore imawonetsa zinthu zabwino kwambiri za Photothermal, zomwe zimawapangitsa kukhala othandizira a PTT.
Kusanthula kwa zithunzi za DLS ndi TEM za RuDA NPs (inset).B Matenthedwe zithunzi zosiyanasiyana woipa wa RuDA NPs poyera ndi laser cheza pa wavelength wa 808 nm (0,5 W cm-2).C Photothermal kutembenuka zokhota pa zosiyanasiyana woipa wa ore NPs, amene kachulukidwe deta.B. D Kuwonjezeka kwa kutentha kwa ORE NP ndi ICG pa 5 kutentha-kuzizira kozungulira.
Photocytotoxicity ya RuDA NPs motsutsana ndi MDA-MB-231 maselo a khansa ya m'mawere ya anthu adayesedwa mu vitro.Monga momwe tawonetsera mkuyu.6A, B, RuDA-NPs ndi RuDA adawonetsa cytotoxicity yosasamala pakalibe kuwala, kutanthauza kuchepa kwa mdima wakuda wa RuDA-NPs ndi RuDA.Komabe, pambuyo pokhudzana ndi ma radiation a laser pamtunda wa 808 nm, RuDA ndi RuDA NPs adawonetsa photocytotoxicity yamphamvu motsutsana ndi maselo a khansa a MDA-MB-231 okhala ndi IC50 values (half-maximum inhibitory concentration) ya 5.4 ndi 9.4 μM, motero, kusonyeza kusonyeza IC50. kuti RuDA-NP ndi RuDA ali ndi kuthekera kwa phototherapy ya khansa.Kuonjezera apo, photocytotoxicity ya RuDA-NP ndi RuDA inafufuzidwanso pamaso pa vitamini C (Vc), ROS scavenger, kuti afotokoze ntchito ya ROS mu cytotoxicity yopangidwa ndi kuwala.Mwachiwonekere, mphamvu zama cell zidawonjezeka pambuyo pa kuwonjezera kwa Vc, ndi IC50 mfundo za RuDA ndi RuDA NPs zinali 25.7 ndi 40.0 μM, motero, zomwe zimatsimikizira ntchito yofunikira ya ROS mu photocytotoxicity ya RuDA ndi RuDA NPs.Light-induced cytotoxicity ya RuDA-NPs ndi RuDA mu MDA-MB-231 maselo a khansa ndi moyo / akufa cell staining ntchito calcein AM (green fluorescence kwa maselo amoyo) ndi propidium iodide (PI, red fluorescence kwa maselo akufa).kutsimikiziridwa ndi ma cell) ngati ma probes a fulorosenti.Monga momwe tawonetsera pa Chithunzi 6C, maselo omwe amathandizidwa ndi RuDA-NP kapena RuDA anakhalabe otheka popanda kuwala, monga umboni wa fulorosisi yobiriwira kwambiri.M'malo mwake, pansi pa kuwala kwa laser, fluorescence yofiira yokha inawonedwa, yomwe imatsimikizira kuti photocytotoxicity yabwino ya RuDA kapena RuDA NPs.Ndizofunikira kudziwa kuti fluorescence yobiriwira idawonekera pakuwonjezera kwa Vc, zomwe zikuwonetsa kuphwanya kwa photocytotoxicity ya RuDA ndi RuDA NPs.Zotsatira izi zimagwirizana ndi kuyesa kwa in vitro photocytotoxicity.
Mlingo wodalira mphamvu ya maselo A RuDA- ndi B RuDA-NP m'maselo a MDA-MB-231 pamaso pa Vc (0.5 mM), motero.Mipiringidzo yolakwika, imatanthawuza ± kupatuka kokhazikika (n = 3). Mayeso a t osaphatikizidwa, a mbali ziwiri *p <0.05, **p <0.01, ndi ***p <0.001. Mayeso a t osaphatikizidwa, a mbali ziwiri *p <0.05, **p <0.01, ndi ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 ndi ***p <0,001. Mayeso opanda michira iwiri *p<0.05, **p<0.01, ndi ***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 ndi ***p <0,001. Mayeso opanda michira iwiri *p<0.05, **p<0.01, ndi ***p<0.001.C Kusanthula kwa madontho a ma cell amoyo/akufa pogwiritsa ntchito calcein AM ndi propidium iodide ngati ma probes a fulorosenti.Scale bar: 30 µm.Zithunzi zoyimira za kubwereza katatu kwachilengedwe kuchokera ku gulu lirilonse zikuwonetsedwa.D Zithunzi za Confocal fluorescence za ROS kupanga m'maselo a MDA-MB-231 pansi pazikhalidwe zosiyanasiyana.Green DCF fluorescence imasonyeza kukhalapo kwa ROS.Irradiate ndi laser yokhala ndi kutalika kwa 808 nm ndi mphamvu ya 0.5 W / cm2 kwa mphindi 10 (300 J / cm2).Scale bar: 30 µm.Zithunzi zoyimira za kubwereza katatu kwachilengedwe kuchokera ku gulu lirilonse zikuwonetsedwa.E Flow cytometry RuDA-NPs (50 µM) kapena RuDA (50 µM) kusanthula mankhwala ndi kapena popanda 808 nm laser (0.5 W cm-2) pamaso ndi kusowa kwa Vc (0.5 mM) kwa 10 min.Zithunzi zoyimira za kubwereza katatu kwachilengedwe kuchokera ku gulu lirilonse zikuwonetsedwa.F Nrf-2, HSP70 ndi HO-1 ya maselo a MDA-MB-231 omwe amathandizidwa ndi RuDA-NPs (50 µM) kapena opanda 808 nm laser irradiation (0.5 W cm-2, 10 min, 300 J cm-2) , ma cell amawonetsa 2).Zithunzi zoyimira za kubwereza kwachilengedwe kuwiri kuchokera ku gulu lirilonse zikuwonetsedwa.
Kupanga kwa ROS kwa intracellular m'maselo a MDA-MB-231 kunayesedwa pogwiritsa ntchito njira yowonongeka ya 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA).Monga momwe tawonetsera mkuyu.6D, ma cell omwe amathandizidwa ndi RuDA-NPs kapena RuDA amawonetsa fluorescence yobiriwira yobiriwira akayatsidwa ndi 808 nm laser, zomwe zikuwonetsa kuti RuDA-NPs ndi RuDA ali ndi luso lopanga ROS.M'malo mwake, popanda kuwala kapena pamaso pa Vc, chizindikiro chochepa cha fulorosenti cha maselo chinawonedwa, chomwe chinasonyeza kupanga pang'ono kwa ROS.Miyezo ya ROS ya intracellular m'maselo a RuDA-NP ndi maselo a MDA-MB-231 opangidwa ndi RuDA adatsimikiziridwanso ndi kutuluka kwa cytometry.Monga momwe tawonetsera mu Supplementary Figure 25, mphamvu ya fluorescence (MFI) yopangidwa ndi RuDA-NPs ndi RuDA pansi pa 808 nm laser irradiation inawonjezeka kwambiri pafupifupi 5.1 ndi 4.8 nthawi, motero, poyerekeza ndi gulu lolamulira, kutsimikizira mapangidwe awo abwino a AFK.mphamvu.Komabe, ma intracellular ROS m'maselo a RuDA-NP kapena MDA-MB-231 omwe amachitidwa ndi RuDA ankangofanana ndi maulamuliro opanda kuwala kwa laser kapena pamaso pa Vc, mofanana ndi zotsatira za kufufuza kwa confocal fluorescence.
Zasonyezedwa kuti mitochondria ndiyo chandamale chachikulu cha Ru(II) -arene complexes60.Choncho, malo a subcellular a RuDA ndi RuDA-NPs adafufuzidwa.Monga momwe zasonyezedwera mu Supplementary Figure 26, RuDA ndi RuDA-NP akuwonetsa mbiri yofananira yogawa ma cell ndi kuchuluka kwa mitochondria (62.5 ± 4.3 ndi 60.4 ± 3.6 ng / mg mapuloteni, motsatana).Komabe, zochepa chabe za Ru zinapezeka mu zigawo za nyukiliya za Ore ndi NP Ore (3.5 ndi 2.1%, motsatira).Gawo lotsalira la selo linali ndi ruthenium yotsalira: 31.7% (30.6 ± 3.4 ng / mg mapuloteni) kwa RuDA ndi 42.9% (47.2 ± 4.5 ng / mg mapuloteni) kwa RuDA-NPs.Mwambiri, Ore ndi NP Ore makamaka amawunjikana mu mitochondria.Kuti tiwone kusayenda bwino kwa mitochondrial, tidagwiritsa ntchito madontho a JC-1 ndi MitoSOX Red kuti tiwone kuthekera kwa nembanemba ya mitochondrial ndi mphamvu yopanga superoxide, motsatana.Monga momwe tawonetsera mu Supplementary Fig. 27, chobiriwira kwambiri (JC-1) ndi chofiira (MitoSOX Red) fluorescence chinawonedwa m'maselo omwe amachitidwa ndi RuDA ndi RuDA-NPs pansi pa 808 nm laser irradiation, kusonyeza kuti RuDA ndi RuDA-NPs kwambiri fulorosenti. Imatha kupangitsa kuti mitochondrial membrane depolarization ndi kupanga superoxide.Kuphatikiza apo, njira ya kufa kwa maselo idatsimikiziridwa pogwiritsa ntchito kusanthula kwa annexin V-FITC/propidium iodide (PI).Monga momwe tawonetsera pa Chithunzi 6E, pamene adawunikiridwa ndi 808 nm laser, RuDA ndi RuDA-NP zinapangitsa kuti apoptosis ayambe kuwonjezeka kwambiri (m'munsi mwa quadrant) m'maselo a MDA-MB-231 poyerekeza ndi PBS kapena PBS kuphatikizapo laser.kukonzedwa maselo.Komabe, pamene Vc inawonjezeredwa, mlingo wa apoptosis wa RuDA ndi RuDA-NP unatsika kwambiri kuchokera ku 50.9% ndi 52.0% mpaka 15.8% ndi 17.8%, motero, zomwe zimatsimikizira udindo wofunikira wa ROS mu photocytotoxicity ya RuDA ndi RuDA-NP..Kuphatikiza apo, maselo ang'onoang'ono a necrotic adawonedwa m'magulu onse oyesedwa (kumtunda kumanzere kwa quadrant), kutanthauza kuti apoptosis ikhoza kukhala mtundu waukulu wa kufa kwa maselo oyambitsidwa ndi RuDA ndi RuDA-NPs.
Popeza kuwonongeka kwa oxidative kumapangitsa kuti pakhale vuto lalikulu la apoptosis, nyukiliya yokhudzana ndi erythroid 2, factor 2 (Nrf2) 62, yolamulira kwambiri ya antioxidant system, inafufuzidwa mu MDA-MB-231 ya RuDA-NPs.Njira zamachitidwe a RuDA NPs opangidwa ndi kuwala.Panthawi imodzimodziyo, kufotokoza kwa mapuloteni otsika a heme oxygenase 1 (HO-1) adapezekanso.Monga momwe tawonetsera mu Chithunzi 6F ndi Supplementary Figure 29, RuDA-NP-mediated phototherapy inachulukitsa Nrf2 ndi HO-1 mawu ofotokozera poyerekeza ndi gulu la PBS, kusonyeza kuti RuDA-NPs ikhoza kulimbikitsa njira zowonetsera kupsinjika kwa okosijeni.Kuonjezera apo, kuti muphunzire zotsatira za photothermal za RuDA-NPs63, mawu a mapuloteni a kutentha kwa Hsp70 adayesedwanso.Zikuwonekeratu kuti ma cell omwe amathandizidwa ndi RuDA-NPs + 808 nm laser radiation adawonetsa kuchuluka kwa Hsp70 poyerekeza ndi magulu ena awiri, kuwonetsa kuyankha kwa ma cell ku hyperthermia.
Zotsatira zochititsa chidwi za mu vitro zidatipangitsa kuti tifufuze momwe RuDA-NP imagwirira ntchito mu mbewa zamaliseche ndi zotupa za MDA-MB-231.Kugawidwa kwa minofu ya RuDA NPs kunaphunziridwa pozindikira zomwe zili mu ruthenium mu chiwindi, mtima, ndulu, impso, mapapo, ndi zotupa.Monga momwe tawonetsera mkuyu.7A, pazipita zili Ore NPs mu ziwalo yachibadwa anaonekera koyamba kuonerera nthawi (4 h), pamene pazipita zili anatsimikiza mu minofu chotupa 8 hours pambuyo jekeseni, mwina chifukwa Ore NPs.Zotsatira za EPR za LF.Malinga ndi zotsatira zogawa, nthawi yoyenera ya chithandizo ndi NP ore idatengedwa maola 8 mutatha makonzedwe.Kuti afotokoze ndondomeko ya kudzikundikira kwa RuDA-NPs m'malo otupa, photoacoustic (PA) katundu wa RuDA-NPs ankayang'aniridwa polemba zizindikiro za PA za RuDA-NPs nthawi zosiyanasiyana pambuyo pa jekeseni.Choyamba, chizindikiro cha PA cha RuDA-NP mu vivo chinayesedwa polemba zithunzi za PA za malo otupa pambuyo pa jekeseni wa intratumoral wa RuDA-NP.Monga momwe tawonetsera mu Supplementary Figure 30, RuDA-NPs inasonyeza chizindikiro cha PA cholimba, ndipo panali mgwirizano wabwino pakati pa ndende ya RuDA-NP ndi mphamvu ya chizindikiro cha PA (Supplementary Figure 30A).Kenako, mu vivo PA zithunzi za chotupa malo zinalembedwa pambuyo mtsempha jekeseni wa RuDA ndi RuDA-NP pa osiyana nthawi mfundo pambuyo jekeseni.Monga momwe tawonetsera pa Chithunzi 7B, chizindikiro cha PA cha RuDA-NPs kuchokera kumalo otupa chotupa chinawonjezeka pang'onopang'ono ndi nthawi ndipo chinafika pamtunda pa maola 8 pambuyo pa jekeseni, mogwirizana ndi zotsatira za kugawa kwa minofu zomwe zimatsimikiziridwa ndi kusanthula kwa ICP-MS.Ponena za RuDA (Supplementary Fig. 30B), kuchuluka kwa chizindikiro cha PA kunawonekera maola 4 pambuyo pa jekeseni, kusonyeza kuti RuDA imalowa mofulumira mu chotupacho.Kuonjezera apo, khalidwe la excretory la RuDA ndi RuDA-NPs linafufuzidwa pozindikira kuchuluka kwa ruthenium mu mkodzo ndi ndowe pogwiritsa ntchito ICP-MS.Njira yayikulu yochotsera RuDA (Supplementary Fig. 31) ndi RuDA-NPs (mkuyu 7C) imadutsa ndowe, ndipo chilolezo chogwira ntchito cha RuDA ndi RuDA-NPs chinawonedwa pa nthawi ya maphunziro a 8, zomwe zikutanthauza kuti RuDA ndi RuDA-NPs akhoza efficiently inathetsedwa m'thupi popanda kawopsedwe yaitali.
A. Kugawa kwa Ex vivo kwa RuDA-NP mu minofu ya mbewa kunatsimikiziridwa ndi Ru okhutira (peresenti ya mlingo woyendetsedwa wa Ru (ID) pa gramu ya minofu) pa nthawi zosiyana pambuyo pa jekeseni.Deta imatanthawuza ± kupatuka kokhazikika (n = 3). Mayeso a t osaphatikizidwa, a mbali ziwiri *p <0.05, **p <0.01, ndi ***p <0.001. Mayeso a t osaphatikizidwa, a mbali ziwiri *p <0.05, **p <0.01, ndi ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 ndi ***p <0,001. Mayeso opanda michira iwiri *p<0.05, **p<0.01, ndi ***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 ndi ***p <0,001. Mayeso opanda michira iwiri *p<0.05, **p<0.01, ndi ***p<0.001.B PA zithunzi za mu vivo chotupa malo pa 808 nm chisangalalo pambuyo mtsempha makonzedwe a RuDA-NPs (10 µmol kg-1) pa nthawi zosiyanasiyana.Pambuyo poyambitsa mtsempha wa RuDA NPs (10 µmol kg-1), C Ru adatulutsidwa kuchokera ku mbewa ndi mkodzo ndi ndowe nthawi zosiyanasiyana.Deta imatanthawuza ± kupatuka kokhazikika (n = 3).
Kutentha kwamphamvu kwa RuDA-NP mu vivo kunaphunziridwa mu mbewa zamaliseche ndi MDA-MB-231 ndi zotupa za RuDA poyerekeza.Monga momwe tawonetsera mkuyu.8A ndi mkuyu wowonjezera 32, gulu lolamulira (saline) linasonyeza kusintha kochepa kwa kutentha (ΔT ≈ 3 ° C) pambuyo pa maminiti a 10 akuwonekera mosalekeza.Komabe, kutentha kwa RuDA-NPs ndi RuDA kunakula mofulumira ndi kutentha kwakukulu kwa 55.2 ndi 49.9 ° C, motero, kupereka hyperthermia yokwanira mu vivo chithandizo cha khansa.Kuwonjezeka kwakukulu kwa kutentha kwa RuDA NPs (ΔT ≈ 24 ° C) poyerekeza ndi RuDA (ΔT ≈ 19 ° C) kungakhale chifukwa cha kutsekemera kwake komanso kudzikundikira m'matumbo a chotupa chifukwa cha zotsatira za EPR.
Zithunzi zotentha za infrared za mbewa zokhala ndi zotupa za MDA-MB-231 zoyatsidwa ndi laser 808 nm nthawi zosiyanasiyana maola 8 mutatha jekeseni.Zithunzi zoimira zamoyo zinayi zobwerezabwereza kuchokera ku gulu lirilonse zikuwonetsedwa.B Wachibale chotupa kuchuluka ndi C Avereji chotupa kuchuluka kwa magulu osiyanasiyana a mbewa pa chithandizo.D Zokhotakhota zolemera thupi zamagulu osiyanasiyana a mbewa.Irradiate ndi laser yokhala ndi kutalika kwa 808 nm ndi mphamvu ya 0.5 W / cm2 kwa mphindi 10 (300 J / cm2).Mipiringidzo yolakwika, imatanthawuza ± kupatuka kokhazikika (n = 3). Mayeso a t osaphatikizidwa, a mbali ziwiri *p <0.05, **p <0.01, ndi ***p <0.001. Mayeso a t osaphatikizidwa, a mbali ziwiri *p <0.05, **p <0.01, ndi ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 ndi ***p <0,001. Mayeso opanda michira iwiri *p<0.05, **p<0.01, ndi ***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 ndi ***p <0,001. Mayeso opanda michira iwiri *p<0.05, **p<0.01, ndi ***p<0.001. E H & E yodetsa zithunzi za ziwalo zazikulu ndi zotupa zochokera m'magulu osiyanasiyana a mankhwala, kuphatikizapo Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, ndi RuDA-NPs + Laser magulu. E H & E yodetsa zithunzi za ziwalo zazikulu ndi zotupa zochokera m'magulu osiyanasiyana a mankhwala, kuphatikizapo Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, ndi RuDA-NPs + Laser magulu. Изображения окрашивания E H&E основных органов и опухолей из разных групп лечения, включая группы физиологического раствора, физиологического раствора + лазера, RuDA, RuDA + Laser, RuDA-NPs и RuDA-NPs + Laser. E H & E yodetsa zithunzi za ziwalo zazikulu ndi zotupa zochokera m'magulu osiyanasiyana a mankhwala, kuphatikizapo saline, saline + laser, RuDA, RuDA + Laser, RuDA-NPs, ndi RuDA-NPs + Laser magulu.来自不同治疗组的主要器官和肿瘤的E H&E 染色图像，包括盐水、盐水+ 激光、RuDA、RuDA +和激光光、RuDA +和激光 NPs、RuDA + 激光、RuDA来自不同治疗组的主要器官和肿瘤的E H&E Окрашивание E H&E основных органов и опухолей из различных групп лечения, включая физиологический раствор, + Rula, +DA, Rule, +DA, Rule, +DA, +DAKDA, +DA E H & E kudetsa kwa ziwalo zazikulu ndi zotupa zochokera m'magulu osiyanasiyana ochizira monga saline, saline + laser, RuDA, RuDA + laser, RuDA-NPs, ndi RuDA-NPs + laser.Scale bar: 60 µm.
Zotsatira za phototherapy mu vivo ndi RuDA ndi RuDA NPs zinayesedwa momwe mbewa zamaliseche ndi zotupa za MDA-MB-231 zinalowetsedwa ndi RuDA kapena RuDA NPs pa mlingo umodzi wa 10.0 µmol kg-1 kudzera mtsempha wa mchira, ndiyeno 8 maola pambuyo jekeseni.kuwala kwa laser ndi kutalika kwa 808 nm.Monga momwe tawonetsera pa Chithunzi 8B, kuchuluka kwa chotupa kunawonjezeka kwambiri m'magulu a saline ndi laser, zomwe zimasonyeza kuti saline kapena laser 808 ili ndi mphamvu zochepa pa kukula kwa chotupa.Monga gulu la saline, kukula kofulumira kwa chotupa kunawonedwanso mu mbewa zothandizidwa ndi RuDA-NPs kapena RuDA popanda kuwala kwa laser, kusonyeza kuopsa kwawo kwakuda.Mosiyana ndi zimenezi, pambuyo pa kuwala kwa laser, chithandizo cha RuDA-NP ndi RuDA chinachititsa kuti chotupa chiwonongeke kwambiri ndi kuchepetsa kuchuluka kwa chotupa cha 95.2% ndi 84.3%, motero, poyerekeza ndi gulu la mankhwala a saline, zomwe zimasonyeza PDT yabwino kwambiri., mkhalapakati ndi mphamvu ya RuDA/CHTV.Poyerekeza ndi RuDA, RuDA NPs inasonyeza bwino phototherapeutic effect, yomwe inali makamaka chifukwa cha zotsatira za EPR za RuDA NPs.Zotsatira zolepheretsa kukula kwa chotupa zinayesedwanso ndi kulemera kwa chotupa kuchotsedwa pa tsiku la 15 la chithandizo (mkuyu 8C ndi Supplementary Fig. 33).The zikutanthauza chotupa misa mu RuDA-NP ankachitira mbewa ndi RuDA ankachitira mbewa anali 0,08 ndi 0,27 g motero, amene anali opepuka kwambiri kuposa gulu ulamuliro (1.43 g).
Kuonjezera apo, kulemera kwa thupi la mbewa kunalembedwa masiku atatu aliwonse kuti aphunzire poizoni wakuda wa RuDA-NPs kapena RuDA mu vivo.Monga momwe tawonetsera pa Chithunzi 8D, palibe kusiyana kwakukulu kwa kulemera kwa thupi komwe kunawonedwa kwa magulu onse a chithandizo. Kuphatikiza apo, kudetsa kwa hematoxylin ndi eosin (H&E) kwa ziwalo zazikulu (mtima, chiwindi, ndulu, mapapo, ndi impso) zochokera m'magulu osiyanasiyana ochizira zidapangidwa. Kuonjezera apo, kutayira kwa hematoxylin ndi eosin (H & E) kwa ziwalo zazikulu (mtima, chiwindi, ndulu, mapapo, ndi impso) kuchokera m'magulu osiyanasiyana ochiritsira anachitidwa. Кроме того, было проведено окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селезенки, легких и порганов). Kuonjezera apo, kutayira kwa hematoxylin ndi eosin (H & E) kwa ziwalo zazikulu (mtima, chiwindi, ndulu, mapapo, ndi impso) kuchokera kumagulu osiyanasiyana ochizira.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 разлихупень. Kuonjezera apo, kutayira kwa hematoxylin ndi eosin (H & E) kwa ziwalo zazikulu (mtima, chiwindi, ndulu, mapapo, ndi impso) zinkachitidwa m'magulu osiyanasiyana a mankhwala.Monga momwe tawonetsera mkuyu.8E, zithunzi zodetsa za H & E za ziwalo zazikulu zisanu zochokera kumagulu a RuDA-NPs ndi RuDA siziwonetsa zowoneka bwino kapena kuwonongeka kwa ziwalo. 8E, zithunzi zodetsa za H & E za ziwalo zazikulu zisanu zochokera kumagulu a RuDA-NPs ndi RuDA siziwonetsa zowoneka bwino kapena kuwonongeka kwa ziwalo.Monga momwe tawonetsera mkuyu.8E, andzobraжения окрашивания H&E пяти основных органов органов из групп RuDA-NPs ndi RuDA sanayambe kusokoneza ntchito zapakhomo. Zithunzi za 8E, H & E zodetsa za ziwalo zazikulu zisanu kuchokera kumagulu a RuDA-NPs ndi RuDA siziwonetsa zolakwika za chiwalo kapena zotupa.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 pa рисунке 8E, ndikuyambitsa ukadaulo wa H&E pakupanga zinthu zatsopano komanso RuDA-NPs ndi RuDA sizinaphule kanthu. Monga momwe tawonetsera pa Chithunzi 8E, zithunzi zodetsa za H & E za ziwalo zazikulu zisanu kuchokera ku magulu a RuDA-NPs ndi RuDA sizinawonetse zowoneka bwino kapena kuwonongeka kwa ziwalo.Zotsatirazi zinasonyeza kuti RuDA-NP kapena RuDA sanasonyeze zizindikiro za poizoni mu vivo. Komanso, H & E kudetsa zithunzi zotupa anasonyeza kuti onse RuDA + Laser ndi RuDA-NPs + Laser magulu akhoza kuwononga kwambiri khansa maselo, kusonyeza bwino mu vivo phototherapeutic efficacy wa RuDA ndi RuDA-NPs. Komanso, H & E kudetsa zithunzi zotupa anasonyeza kuti onse RuDA + Laser ndi RuDA-NPs + Laser magulu akhoza kuwononga kwambiri khansa maselo, kusonyeza bwino mu vivo phototherapeutic efficacy wa RuDA ndi RuDA-NPs.Kuonjezera apo, zithunzi za chotupa za hematoxylin-eosin zimasonyeza kuti magulu onse a RuDA + Laser ndi RuDA-NPs + Laser angapangitse chiwonongeko choopsa cha maselo a khansa, kusonyeza mphamvu yapamwamba ya phototherapeutic ya RuDA ndi RuDA-NPs mu 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，说多坏，话市明经坏，话市明明的智慧。此外 , 肿瘤 的 & e 染色 显示 , ruda + laser ndi ruda-nps + laser 组均 导致 的 癌细胞 破坏 , 证明 光的。 ...Kuonjezera apo, zithunzi za chotupa za hematoxylin ndi eosin zimasonyeza kuti magulu onse a RuDA + Laser ndi RuDA-NPs + Laser adayambitsa kuwonongeka kwakukulu kwa maselo a khansa, kusonyeza mphamvu yapamwamba ya phototherapeutic ya RuDA ndi RuDA-NPs mu vivo.
Pomaliza, Ru (II) -arene (RuDA) organometallic complex ndi DA-mtundu wa ligands adapangidwa kuti athandize ndondomeko ya ISC pogwiritsa ntchito njira yophatikizira.Synthesized RuDA ikhoza kudzisonkhanitsa yokha kupyolera muzosagwirizana ndi covalent kupanga machitidwe a RuDA-derived supramolecular systems, potero kumathandizira kupanga 1O2 ndi kutembenuka kwa photothermal kwabwino kwa chithandizo cha khansa choyambitsa kuwala.Ndizochititsa chidwi kuti RuDA ya monomeric sinapange 1O2 pansi pa laser irradiation pa 808 nm, koma ikhoza kupanga kuchuluka kwa 1O2 mu chikhalidwe chophatikizana, kusonyeza kulingalira ndi luso la mapangidwe athu.Kafukufuku wotsatira wasonyeza kuti msonkhano wa supramolecular umapatsa RuDA ndi luso lojambula bwino komanso lojambula zithunzi, monga kuyamwa kwa redshift ndi photobleaching resistance, zomwe ndizofunikira kwambiri pa PDT ndi PTT processing.Mayesero onse mu m'galasi ndi mu vivo awonetsa kuti RuDA NPs yokhala ndi biocompatibility yabwino komanso kudzikundikira bwino mu chotupacho imawonetsa ntchito yabwino kwambiri yolimbana ndi khansa pakuwala kwa laser pamtunda wa 808 nm.Choncho, RuDA NPs monga ogwira bimodal supramolecular PDT/PTW reagents adzalemeretsa ya photosensitizers adamulowetsa pa wavelengths pamwamba 800 nm.Mapangidwe amalingaliro a supramolecular system amapereka njira yabwino yopangira ma photosensitizer opangidwa ndi NIR okhala ndi zotsatira zabwino kwambiri zopanga zithunzi.
Mankhwala onse ndi zosungunulira zinapezedwa kuchokera kwa ogulitsa malonda ndipo amagwiritsidwa ntchito popanda kuyeretsedwa kwina.RuCl3 inagulidwa ku Boren Precious Metals Co., Ltd. (Kunming, China).[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione) ndi 4,7-bis[4-(N,N-diphenylamino)phenyl]-5 , 6-Diamino-2,1,3-benzothiadiazole inapangidwa malinga ndi maphunziro apitalo64,65.Zowonera za NMR zinajambulidwa pa Bruker Avance III-HD 600 MHz spectrometer ku Southeastern University Analytical Test Center pogwiritsa ntchito d6-DMSO kapena CDCl3 monga zosungunulira.Kusintha kwa mankhwala δ kumaperekedwa mu ppm.mokhudzana ndi tetramethylsilane, ndi zolumikizirana J zimaperekedwa mwatsatanetsatane mu hertz.High resolution mass spectrometry (HRMS) idapangidwa pa chida cha Agilent 6224 ESI/TOF MS.Kusanthula kwapadera kwa C, H, ndi N kunachitika pa Vario MICROCHNOS elemental analyzer (Elementar).Zowoneka bwino za UV zidayesedwa pa Shimadzu UV3600 spectrophotometer.Mawonekedwe a Fluorescence adajambulidwa pa Shimadzu RF-6000 spectrofluorimeter.Zowonera za EPR zidajambulidwa pa chida cha Bruker EMXmicro-6/1.Mapangidwe ndi mapangidwe a zitsanzo zokonzedwa adaphunziridwa pa zida za FEI Tecnai G20 (TEM) ndi Bruker Icon (AFM) zomwe zimagwira ntchito pamagetsi a 200 kV.Dynamic light scattering (DLS) idachitidwa pa Nanobrook Omni analyzer (Brookhaven).Mphamvu za Photoelectrochemical zidayesedwa pakupanga kwa electrochemical (CHI-660, China).Zithunzi za Photoacoustic zidapezedwa pogwiritsa ntchito FUJIFILM VisualSonics Vevo® LAZR system.Zithunzi za Confocal zidapezedwa pogwiritsa ntchito maikulosikopu ya Olympus FV3000 confocal.Kusanthula kwa FACS kunachitika pa BD Calibur flow cytometer.High performance liquid chromatography (HPLC) inachitidwa pa Waters Alliance e2695 system pogwiritsa ntchito 2489 UV/Vis detector.Mayeso a Gel Permeation Chromatography (GPC) adalembedwa pa chida cha Thermo ULTIMATE 3000 pogwiritsa ntchito ERC RefratoMax520 refractive index detector.
[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione)64 (481.0 mg, 1.0 mmol), 4,7-bis[4 -(N, N-diphenylamino)phenyl]-5,6-diamino-2,1,3-benzothiadiazole 65 (652.0 mg, 1.0 mmol) ndi glacial acetic acid (30 mL) anasonkhezeredwa mu reflux firiji kwa maola 12.Chosungunuliracho chinachotsedwa mu vacuo pogwiritsa ntchito evaporator yozungulira.Chotsalira chotsatiracho chinayeretsedwa ndi flash column chromatography (silica gel, CH2Cl2: MeOH = 20: 1) kuti apeze RuDA ngati ufa wobiriwira (zokolola: 877.5 mg, 80%).anus.Kuwerengeredwa kwa C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.Anapezeka: C 67.92, H 4.26, N 9.82.1H NMR (600 MHz, d6-DMSO) δ 10.04 (s, 2H), 8.98 (s, 2H), 8.15 (s, 2H), 7.79 (s, 4H), 7.44 (s, 8H), 7.21 (d, J = 31.2 Hz, 16H), 6.47 (s, 2H), 6.24 (s, 2H), 2.69 (s, 1H), 2 .25 (s, 3H), 0.99 (s, 6H).13c nmr (150 mhz, D6-DMSO), Δ (ppm) 158.03, wazaka 95.82, wazaka 95,51, wazaka. , 103. , 86.52, 84.75, 63.29, 30.90, 22.29, 18.83.ESI-MS: m/z [M-Cl]+ = 1097.25.
Kaphatikizidwe ka 4,7-bis [4-(N,N-diethylamino) phenyl-5,6-diamino-2,1,3-benzothiadiazole (L2): L2 inapangidwa muzitsulo ziwiri.Pd(PPh3)4 (46 mg, 0.040 mmol) adawonjezedwa ku N,N-diethyl-4-(tributylstannyl)aniline (1.05 g, 2.4 mmol) ndi 4,7-dibromo-5,6-dinitro solution - 2, 1,3-benzothiadiazole (0.38 g, 1.0 mmol) mu toluene youma (100 ml).Kusakaniza kunalimbikitsidwa pa 100 ° C kwa maola 24.Pambuyo pochotsa toluene mu vacuo, cholimbacho chinatsukidwa ndi petroleum ether.Kenaka chisakanizo cha mankhwalawa (234.0 mg, 0.45 mmol) ndi ufa wachitsulo (0.30 g, 5.4 mmol) mu acetic acid (20 ml) unagwedezeka pa 80 ° C. kwa maola anayi.Zomwe osakaniza anathiridwa m'madzi ndipo chifukwa bulauni olimba anasonkhanitsidwa ndi kusefera.Mankhwalawa adayeretsedwa kawiri ndi vacuum sublimation kuti apereke cholimba chobiriwira (126.2 mg, 57% zokolola).anus.Kuwerengeredwa kwa C26H32N6S: C 67.79, H 7.00, N 18.24.Anapezeka: 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.
Zosakaniza zinakonzedwa ndikuyeretsedwa motsatira njira zofanana ndi RuDA.anus.Kuwerengeredwa kwa C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.Zapezeka: C, 61.32, H, 5.12, N, 11.81,1H NMR (600 MHz, d6-DMSO), δ (ppm) 10.19 (s, 2H), 9.28 (s, 2H), 8.09 (s, 2H), 7.95 (s, 4H), 6.93 (s, 4H), 6.48 (d, 2H), 6.34 (s, 2H) , 3.54 (t, 8H), 2.80 (m, 1H), 2.33 (s, 3H), 1.31 (t, 12H), 1.07 (s, 6H).13C NMR (151 Mhz, CDCL3), Δ (mas) 158.20, 158.8.8: 138.82, 138.82, 138.82, 138.82, 138.87.8., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m/z [M-Cl]+ = 905.24.
RuDA inasungunuka mu MeOH / H2O (5/95, v / v) pamagulu a 10 μM.Kuthamanga kwa RuDA kunayesedwa mphindi iliyonse ya 5 pa Shimadzu UV-3600 spectrophotometer pansi pa kuwala ndi laser kuwala kwa 808 nm (0.5 W / cm2).Mawonekedwe a ICG adalembedwa pansi pamikhalidwe yofanana ndi muyezo.
Mawonekedwe a EPR adalembedwa pa Bruker EMXmicro-6/1 spectrometer yokhala ndi mphamvu ya microwave ya 20 mW, makina ojambulira a 100 G, ndi kusintha kwa 1 G. 2,2,6,6-tetramethyl-4-piperidone (TEMP) ndi 5,5-dimethyl-1-pyrroline N-oxide (DMPO) amagwiritsidwa ntchito ngati misampha yozungulira.Electron spin resonance spectra adalembedwa kuti apange njira zosakanikirana za RuDA (50 µM) ndi TEMF (20 mM) kapena DMPO (20 mM) mothandizidwa ndi ma radiation a laser okhala ndi kutalika kwa 808 nm (0.5 W/cm2).
Kuwerengera kwa DFT ndi TD-DFT kwa RuDA kunachitika pamilingo ya PBE1PBE/6–31 G*//LanL2DZ munjira yamadzi pogwiritsa ntchito pulogalamu ya Gaussian 1666,67,68.The HOMO-LUMO, dzenje ndi ma elekitironi kugawa kwa otsika mphamvu singlet okondwa boma RuDA anakonza pogwiritsa ntchito pulogalamu GaussView (mtundu 5.0).
Tidayesa koyamba kuyeza kuchuluka kwa m'badwo wa 1O2 RuDA pogwiritsa ntchito mawonekedwe owoneka bwino a UV ndi ICG (ΦΔ = 0.002) monga muyezo, koma kujambula kwa ICG kunakhudza kwambiri zotsatira zake.Choncho, zokolola za quantum za 1O2 RuDA zinayesedwa pozindikira kusintha kwa mphamvu ya ABDA fluorescence pafupifupi 428 nm pamene imayatsidwa ndi laser yokhala ndi kutalika kwa 808 nm (0.5 W / cm2).Kuyesera kunachitika pa RuDA ndi RuDA NPs (20 μM) m'madzi / DMF (98/2, v / v) yomwe ili ndi ABDA (50 μM).Kuchuluka kwa zokolola za 1O2 kunawerengedwa pogwiritsa ntchito njira zotsatirazi: ΦΔ (PS) = ΦΔ (ICG) × (rFS/APS)/(rICG/AICG).rPS ndi rICG ndizochita za ABDA ndi 1O2 zopezedwa kuchokera ku photosensitizer ndi ICG, motsatana.APS ndi AICG ndi kuyamwa kwa photosensitizer ndi ICG pa 808 nm, motsatana.
Miyezo ya AFM idachitika m'mikhalidwe yamadzimadzi pogwiritsa ntchito jambulani pa Bruker Dimension Icon AFM system.Pogwiritsa ntchito mawonekedwe otseguka ndi maselo amadzimadzi, maselowo amatsukidwa kawiri ndi ethanol ndikuwumitsa ndi mtsinje wa nayitrogeni.Ikani maselo owuma mu mutu wa kuwala wa maikulosikopu.Nthawi yomweyo ikani dontho lachitsanzo mu dziwe lamadzimadzi ndikuchiyika pa cantilever pogwiritsa ntchito syringe yapulasitiki yosabala komanso singano yosabala.Dontho lina limayikidwa mwachindunji pa chitsanzo, ndipo pamene mutu wa kuwala umatsitsidwa, madontho awiriwo amalumikizana, kupanga meniscus pakati pa chitsanzo ndi madzi osungira madzi.Miyezo ya AFM inkagwiritsidwa ntchito pogwiritsa ntchito SCANASYST-FLUID V-shape nitride cantilever (Bruker, hardness k = 0.7 N m-1, f0 = 120-180 kHz).
Ma chromatogram a HPLC adapezedwa pa Waters e2695 system yokhala ndi phoenix C18 column (250 × 4.6 mm, 5 µm) pogwiritsa ntchito chowunikira cha 2489 UV / Vis.Kutalika kwa detector ndi 650 nm.Magawo am'manja A ndi B anali madzi ndi methanol, motsatana, ndipo kuthamanga kwa gawo la mafoni kunali 1.0 ml · min-1.The gradient (zosungunulira B) zinali motere: 100% kuchokera 0 mpaka 4 mphindi, 100% mpaka 50% kuchokera 5 mpaka 30 mphindi, ndi bwererani ku 100% kuchokera 31 mpaka 40 mphindi.Ore adasungunuka mu njira yosakanikirana ya methanol ndi madzi (50/50, ndi voliyumu) pamlingo wa 50 μM.Kuchuluka kwa jakisoni kunali 20 μl.
Mayesero a GPC adalembedwa pa chida cha Thermo ULTIMATE 3000 chokhala ndi zigawo ziwiri za PL aquagel-OH MIXED-H (2 × 300 × 7.5 mm, 8 µm) ndi ERC RefratoMax520 refractive index detector.Mzere wa GPC unatulutsidwa ndi madzi pamtunda wa 1 ml / min pa 30 ° C.Ore NPs adasungunuka mu yankho la PBS (pH = 7.4, 50 μM), voliyumu ya jekeseni inali 20 μL.
Photocurrents anayesedwa pa electrochemical kukhazikitsa (CHI-660B, China).Mayankho a optoelectronic pamene laser adayatsidwa ndikuzimitsa (808 nm, 0.5 W / cm2) adayesedwa pamagetsi a 0.5 V mubokosi lakuda, motsatana.Selo yodziwika bwino ya ma elekitirodi atatu idagwiritsidwa ntchito ndi magalasi owoneka ngati L opangidwa ndi magalasi a carbon electrode (GCE) ngati electrode yogwira ntchito, muyezo wa calomel electrode (SCE) ngati maelekitirodi owerengera, ndi disiki ya platinamu ngati electrode yowerengera.Njira ya 0.1 M Na2SO4 idagwiritsidwa ntchito ngati electrolyte.
Mzere wa khansa ya m'mawere ya munthu MDA-MB-231 idagulidwa kuchokera ku KeyGEN Biotec Co., LTD (Nanjing, China, nambala yamakatale: KG033).Maselo anakulira mu monolayers mu Dulbecco's Modified Eagle's Medium (DMEM, high glucose) yowonjezeredwa ndi yankho la 10% fetal bovine serum (FBS), penicillin (100 μg/ml) ndi streptomycin (100 μg/ml).Maselo onse adakulitsidwa pa 37 ° C mumlengalenga wachinyontho wokhala ndi 5% CO2.
Kuyesa kwa MTT kunagwiritsidwa ntchito kuti adziwe cytotoxicity ya RuDA ndi RuDA-NPs pamaso ndi kusowa kwa kuwala kwa kuwala, kapena popanda Vc (0.5 mM).Maselo a khansa a MDA-MB-231 adakula mu mbale za 96-chitsime pa kachulukidwe ka selo pafupifupi 1 × 105 maselo/ml/chabwino ndipo amakulungidwa kwa maola 12 pa 37.0 ° C mumlengalenga wa 5% CO2 ndi 95% mpweya.RuDA ndi RuDA NPs zosungunuka m'madzi zinawonjezeredwa ku maselo.Pambuyo pa maola 12 a incubation, maselowo adawonetsedwa ndi 0.5 W cm -2 laser radiation pa wavelength ya 808 nm kwa mphindi 10 (300 J cm -2) kenako amaikidwa mumdima kwa maola 24.Maselowo adayikidwa ndi MTT (5 mg / ml) kwa maola ena asanu.Pomaliza, sinthani sing'angayo kukhala DMSO (200 µl) kuti musungunule makhiristo ofiirira a formazan.Makhalidwe a OD adayesedwa pogwiritsa ntchito chowerengera cha microplate chokhala ndi kutalika kwa 570/630 nm.Mtengo wa IC50 pachitsanzo chilichonse udawerengedwa pogwiritsa ntchito pulogalamu ya SPSS kuchokera kumakhota oyankhira mlingo wopezedwa kuchokera ku zoyeserera zosachepera zitatu.
Maselo a MDA-MB-231 anachitidwa ndi RuDA ndi RuDA-NP pamagulu a 50 μM.Pambuyo pa maola a 12 a incubation, ma cell adayatsidwa ndi laser yokhala ndi kutalika kwa 808 nm ndi mphamvu ya 0.5 W / cm2 kwa mphindi 10 (300 J / cm2).Mu gulu la vitamini C (Vc), maselo amathandizidwa ndi 0.5 mM Vc isanayambe kuyatsa laser.Maselo adakulungidwa mumdima kwa maola owonjezera a 24, kenako amathiridwa ndi calcein AM ndi propidium iodide (20 μg/ml, 5 μl) kwa mphindi 30, kenako amatsukidwa ndi PBS (10 μl, pH 7.4).zithunzi za ma cell odetsedwa.
Nthawi yotumiza: Sep-23-2022
