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He mea koʻikoʻi loa nā photosensitizers no ka hoʻohana nui ʻana o ka phototherapy.Eia nō naʻe, pilikia nā photosensitizer maʻamau i ka absorption lōʻihi hawewe pōkole, lawa ʻole ka photostability, haʻahaʻa haʻahaʻa o nā ʻano oxygen reactive (ROS), a me ka hoʻopau ʻana o ka ROS i hoʻohui ʻia.Maʻaneʻi mākou e hōʻike nei i kahi kiʻi kiʻi kiʻi supramolecular kokoke-infrared (NIR) i hoʻopili ʻia e ka hui pū ʻana o Ru(II) -arene organometallic complexes i loko o ka wai.Hiki i ka RuDA ke hoʻopuka i ka oxygen singlet (1O2) ma ka mokuʻāina i hōʻuluʻulu ʻia, a e hōʻike ana i ka ʻano o ka hanauna 1O2 i hoʻohui ʻia ma muli o ka piʻi nui ʻana o ke kaʻina crossover ma waena o ka ʻōnaehana singlet-triplet.Ma lalo o ka hana o 808 nm kukui laser, hōʻike ʻo RuDA i kahi hua nui 1O2 o 16.4% (ʻōmaʻomaʻo indocyanine i ʻae ʻia e FDA: ΦΔ=0.2%) a me kahi kiʻekiʻe kiʻekiʻe o ka hoʻololi ʻana i ka photothermal o 24.2% (nā nanorods gula kalepa) me ka photostability maikaʻi loa.: 21.0%, nā nanoshells gula: 13.0%).Eia kekahi, hiki i ka RuDA-NPs me ka biocompatibility maikaʻi ke hōʻiliʻili i nā wahi maʻi tumora, e hoʻoulu ai i ka hoʻihoʻi nui ʻana i ka wā photodynamic therapy me ka hōʻemi ʻana o 95.2% i ka nui o ka tumor i vivo.Hāʻawi kēia hōʻuluʻulu photodynamic therapy i kahi hoʻolālā no ka hoʻomohala ʻana i nā photosensitizers me nā waiwai photophysical a me photochemical maikaʻi.
Hoʻohālikelike ʻia i ka lāʻau maʻamau, ʻo ka photodynamic therapy (PDT) kahi lāʻau maikaʻi no ka maʻi kanesa ma muli o kāna mau pono nui e like me ka mana spatiotemporal pololei, non-invasiveness, negligible drug resistance, a me ka hōʻemi ʻana i nā hopena ʻaoʻao 1,2,3.Ma lalo o ka hoʻomālamalama māmā, hiki ke hoʻohana ʻia nā photosensitizers e hana i nā ʻano oxygen reactive kiʻekiʻe (ROS), e alakaʻi ana i ka apoptosis / necrosis a i ʻole immune responses4,5. Eia nō naʻe, ʻo ka hapa nui o nā photosensitizers maʻamau, e like me nā chlorins, porphyrins, a me nā anthraquinones, loaʻa i ka absorption lōʻihi pōkole (frequency <680 nm), no laila ka hopena i ke komo ʻana o ka mālamalama maikaʻi ʻole ma muli o ka ikaika o ka absorption o nā molekala ola (eg, hemoglobin a me melanin) i loko. ka ʻāpana ʻike ʻia6,7. Eia nō naʻe, ʻo ka hapa nui o nā photosensitizers maʻamau, e like me nā chlorins, porphyrins, a me nā anthraquinones, loaʻa i ka absorption lōʻihi pōkole (frequency <680 nm), no laila ka hopena i ke komo ʻana o ka mālamalama maikaʻi ʻole ma muli o ka ikaika o ka absorption o nā molekala ola (eg, hemoglobin a me melanin) i loko. ka ʻāpana ʻike ʻia6,7. 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. Eia nō naʻe, ʻo ka hapa nui o nā photosensitizers e like me nā chlorins, porphyrins a me nā anthraquinones he pōkole hawewe lōʻihi (<680 nm) ka hopena i ke komo ʻana o ka mālamalama maikaʻi ʻole ma muli o ke komo ikaika ʻana o nā molekala ola (e like me ka hemoglobin a me ka melanin) i loko o ka māhele ʻike ʻia6,7.然而, 大多数 传统 的 光敏剂 下载 的 光敏剂, 如 氢 蒽醌, 波长 吸收 波长 生物 吸收 吸收和) 的 强烈 黑色素 吸收) 的 强烈强烈,导致光穿透性差。然而, 大多数 传统 的 光敏剂 的 光敏剂, 光敏剂 较, 波长 吸收 波长 的 о 的 m分子 (血红 吸收 吸收 黑色素 (因此血红 吸收 黑色素 (因此 吸收 吸收 黑色素) 因此) 的 吸收 吸收吸收 吸收 吸收 吸收 吸收 吸收 HI导致光穿透性差。 Однако большинство традиционных фотосенсибилизаторов, таких как хлорины, порфирины и антрахиноны, имеют относительно коротковолновое поглощение (частота < 680 нм) из-за сильного поглощения биомолекул, таких как гемоглобин и меланин, что приводит к плохому проникновению света. Eia nō naʻe, ʻo ka hapa nui o nā photosensitizer maʻamau e like me nā chlorins, porphyrins a me nā anthraquinones he pōkole hawewe lōʻihi (frequency <680 nm) ma muli o ka hoʻopaʻa ikaika ʻana o nā biomolecules e like me ka hemoglobin a me ka melanin e hopena i ke komo ʻana o ka māmā.Wahi ʻike ʻia 6.7.No laila, kokoke-infrared (NIR) absor photosensitizers i ho'ā 'ia i loko o ka 700-900 nm "therapeutic window" ua kūpono no ka phototherapy.No ka mea, ʻoi aku ka liʻiliʻi o ke kukui infrared kokoke i nā ʻiʻo olaola, hiki iā ia ke alakaʻi i ke komo hohonu a emi ʻole ka photodamage8,9.
ʻO ka mea pōʻino, ʻaʻole maikaʻi nā kiʻi paʻi kiʻi NIR-absorbing, haʻahaʻa haʻahaʻa o ka oxygen (1O2) hiki ke hoʻohua, a me ka hoʻopau ʻana i ka 1O2 hoʻohuihui, e kaupalena ana i kā lākou noi lapaʻau10,11.ʻOiai ua hoʻoikaika nui ʻia e hoʻomaikaʻi i nā waiwai photophysical a me nā kiʻi kiʻi kiʻi o nā photosensitizer maʻamau, i kēia manawa ua hōʻike ʻia kekahi mau hōʻike e hiki i nā photosensitizers-absorbing NIR ke hoʻoponopono i kēia mau pilikia āpau.Eia kekahi, ua hōʻike kekahi mau photosensitizers i ka ʻōlelo hoʻohiki no ka hana pono ʻana o 1O212,13,14 i ka wā i hoʻomālamalama ʻia me ka mālamalama ma luna o 800 nm, no ka mea, ua emi koke ka ikehu photon ma kahi kokoke-IR.ʻO Triphenylamine (TFA) ma ke ʻano he mea hāʻawi electron a me [1,2,5] thiadiazole- [3,4-i] dipyrido [a, c] phenazine (TDP) ma ke ʻano he hui ʻae electron ʻO Donor-acceptor (DA) ʻano wai kala i kahi papa. o nā mea hoʻonalohelohe , e komo ana i ka infrared kokoke, i aʻo nui ʻia no ka bioimaging II kokoke-infrared a me ka photothermal therapy (PTT) ma muli o ko lākou ʻāpana haiki.No laila, hiki ke hoʻohana ʻia nā dyes ʻano DA no PDT me ka hauʻoli kokoke-IR, ʻoiai ʻaʻole lākou i aʻo ʻia e like me nā photosensitizers no PDT.
Ua ʻike maikaʻi ʻia ʻo ka hana kiʻekiʻe o ka intersystem crossing (ISC) o nā photosensitizers e hāpai i ka hoʻokumu ʻana o 1O2.ʻO kahi hoʻolālā maʻamau no ka holomua ʻana i ke kaʻina ISC ʻo ia ka hoʻomaikaʻi ʻana i ka hui ʻana o ka spin-orbit (SOC) o nā photosensitizers ma o ka hoʻokomo ʻana i nā atom kaumaha a i ʻole nā ​​​​moieties organik kūikawā.Eia naʻe, loaʻa i kēia ala kekahi mau hemahema a me nā palena19,20.I kēia mau lā, ua hāʻawi ʻo supramolecular self-assembly i kahi ala naʻauao i lalo no ka hana ʻana i nā mea hana ma ka pae mole, 21,22 me nā pono he nui i ka phototherapy: (1) hiki i nā photosensitizers hui pū ʻia ke hiki ke hana i nā hale lipine.E like me nā hale uila me ka puʻunaue ʻoi aku ka nui o ka ikehu ma muli o ka overlapping orbits ma waena o nā poloka hale.No laila, e hoʻomaikaʻi ʻia ka hoʻokūkū ikaika ma waena o ka mokuʻāina hoʻohiwahiwa singlelet haʻahaʻa (S1) a me ka mokuʻāina hoʻohiwahiwa triplet kokoke (Tn), i mea e pono ai no ke kaʻina hana ISC 23, 24.(2) Supramolecular hui e hoemi non-radiative hoʻomaha ma muli o ka intramolecular motion limitation mechanism (RIM), ka mea e paipai i ka ISC kaʻina 25, 26.(3) Hiki i ka hui supramolecular ke pale aku i nā molekala o loko o ka monomer mai ka hoʻoheheʻe ʻana a me ka degradation, a laila e hoʻomaikaʻi nui ai i ka photostability o ka photosensitizer.Hāʻawi ʻia i nā pōmaikaʻi ma luna, manaʻoʻiʻo mākou hiki i nā ʻōnaehana photosensitizer supramolecular ke lilo i mea hoʻohiki e lanakila ai i nā hemahema o PDT.
ʻO nā complexes e pili ana i ka Ru(II) kahi kahua lapaʻau hoʻohiki no nā noi kūpono i ka maʻi a me ka mālama ʻana i nā maʻi ma muli o kā lākou mau waiwai kūlohelohe a hoihoi28,29,30,31,32,33,34.Eia kekahi, ʻo ka nui o nā mokuʻāina hauʻoli a me nā waiwai photophysicochemical tunable o Ru (II) -based complexes e hāʻawi i nā pono nui no ka hoʻomohala ʻana o nā photosensitizers e pili ana iā Ru (II)35,36,37,38,39,40.ʻO kahi laʻana koʻikoʻi ʻo ka ruthenium(II) polypyridyl complex TLD-1433, aia i kēia manawa i nā hoʻokolohua lapaʻau Phase II ma ke ʻano he photosensitizer no ka mālama ʻana i ka maʻi maʻi maʻi maʻi non-muscle invasive bladder (NMIBC)41.Eia kekahi, hoʻohana nui ʻia nā ruthenium(II)arene organometallic complexes ma ke ʻano he chemotherapeutic agents no ka mālama ʻana i ka maʻi maʻi ma muli o ko lākou haʻahaʻa haʻahaʻa a me ka maʻalahi o ka hoʻololi ʻana42,43,44,45.ʻAʻole hiki i nā waiwai ionic o Ru (II) -arene organometallic complexes ke hoʻomaikaʻi i ka solubility maikaʻi ʻole o nā chromophores DA i nā solvents maʻamau, akā hoʻomaikaʻi pū i ka hui ʻana o nā chromophores DA.Eia kekahi, hiki i ka pseudooctahedral half-sandwich structure o nā organometallic complexes o Ru (II) -arenes ke pale aku i ka H-aggregation o DA-type chromophores, no laila e hoʻomaʻamaʻa i ka hoʻokumu ʻana o ka J-aggregation me nā kaula absorption redshifted.Eia nō naʻe, hiki ke hoʻopilikia i ka maikaʻi o ka lāʻau lapaʻau a me ka hana vivo o arene-Ru (II) paʻakikī, e like me ka haʻahaʻa haʻahaʻa a / a i ʻole ka bioavailability maikaʻi ʻole.Eia nō naʻe, ua hōʻike ʻia nā haʻawina e hiki ke lanakila i kēia mau hemahema ma o ka hoʻopili ʻana i nā complexes ruthenium me nā polymers biocompatible e ka encapsulation kino a i ʻole covalent conjugation.
Ma kēia hana, hōʻike mākou i nā complexes DA-conjugated o Ru (II) -arene (RuDA) me kahi mea hoʻoulu NIR ma o ka pilina paʻa ma waena o ka DAD chromophore a me ka moiety Ru (II) -arene.Hiki i nā mea paʻakikī ke hoʻohui iā lākou iho i loko o nā vesicles metalosupramolecular i loko o ka wai ma muli o nā pilina non-covalent.ʻO ka mea nui, ua hāʻawi ka hui supramolecular iā RuDA me ka polymerization-induced intersystem crossing-over properties, kahi i hoʻonui nui ai i ka pono ISC, he mea maikaʻi loa ia no PDT (Fig. 1A).No ka hoʻonui ʻana i ka hōʻiliʻili ʻana a me ka biocompatibility in vivo, ua hoʻohana ʻia ʻo Pluronic F127 (PEO-PPO-PEO) i ʻae ʻia e FDA e hoʻopili iā RuDA47,48,49 e hana i nā nanoparticles RuDA-NP (Figure 1B) i hana ma ke ʻano he PDT / Dual- mode PTT proxy .I ka phototherapy maʻi 'aʻai (Figure 1C), ua hoʻohana ʻia ʻo RuDA-NP e mālama i nā ʻiole olohelohe me nā ʻōpū MDA-MB-231 e aʻo i ka pono o PDT a me PTT i loko o vivo.
Hōʻike kiʻi kiʻi o ka mīkini photophysical o RuDA i nā ʻano monomeric a i hui pū ʻia no ka phototherapy maʻi maʻi, synthesis o B RuDA-NPs a me C RuDA-NPs no NIR-activated PDT a me PTT.
Ua hoʻomākaukau ʻia ʻo RuDA, me ka TPA a me ka TDP functionality, e like me ke kaʻina hana i hōʻike ʻia ma ke Kiʻi Hoʻohui 1 (Figure 2A), a ua hōʻike ʻia ʻo RuDA e 1H a me 13C NMR spectra, electrospray ionization mass spectrometry, a me ka nānā ʻana i nā elemental (Supplementary Figures 2-4). ).Ua helu ʻia ka palapala ʻokoʻa ʻokoʻa o ka electron electron RuDA o ka hoʻololi hoʻokahi haʻahaʻa loa e ka time-dependent density functional theory (TD-DFT) e aʻo ai i ke kaʻina hana hoʻoili.E like me ka mea i hōʻike ʻia ma ke Kiʻi Pākuʻi 5, ʻoi aku ka nui o ka electron density drifts mai triphenylamine i ka TDP acceptor unit ma hope o ka photoexcitation, hiki ke hoʻopili ʻia i kahi hoʻololi maʻamau intramolecular charge transfer (CT).
ʻO ke kinona o ka ʻOre B B Absorption spectra of Ore i loko o nā hui ʻana o nā lakio like ʻole o DMF a me ka wai.C Nā waiwai absorption maʻamau o RuDA (800 nm) a me ICG (779 nm) me ka manawa ma 0.5 W cm-2 o 808 nm kukui laser.D Hōʻike ʻia ke kiʻi paʻi kiʻi o ABDA e ka RuDA-i hoʻokumu ʻia o 1O2 i nā hui DMF/H2O me nā ʻano wai like ʻole ma lalo o ka hana o ka radiation laser me kahi hawewe o 808 nm a me ka mana o 0.5 W/cm2.
Abstract—UV-visible absorption spectroscopy i hoʻohana ʻia no ke aʻo ʻana i nā waiwai hoʻohuihui ponoʻī o Ore i loko o nā hui ʻana o DMF a me ka wai ma nā ʻano like ʻole.E like me ka hoike ana ma ka fig.2B, hōʻike ʻo RuDA i nā kaula absorption mai 600 a 900 nm ma DMF me kahi kaula absorption kiʻekiʻe loa ma 729 nm.ʻO ka hoʻonui ʻana i ka nui o ka wai i alakaʻi ʻia i kahi hoʻololi ʻulaʻula lohi o ka ore absorption maximum i 800 nm, e hōʻike ana i ka J-aggregation o Ore i ka ʻōnaehana hui.Hōʻike ʻia ke kiʻi kiʻi photoluminescence o RuDA i nā mea hoʻoheheʻe like ʻole ma ke Kiʻi 6. Hōʻike ʻia ʻo RuDA e hōʻike i ka luminescence NIR-II maʻamau me ka lōʻihi o ka hawewe hoʻokuʻu ʻana o ca.1050 nm i CH2Cl2 a me CH3OH.ʻO ka hoʻololi nui o Stokes (e pili ana i 300 nm) o RuDA e hōʻike ana i kahi loli nui i ka geometry o ka moku'āina hauʻoli a me ka hoʻokumu ʻana i nā ʻāina hauʻoli haʻahaʻa.Ua hoʻoholo ʻia ka nui o ka luminescence quantum yield o Ore ma CH2Cl2 a me CH3OH he 3.3 a me 0.6%.Eia naʻe, i loko o ka hui ʻana o ka methanol a me ka wai (5/95, v/v), ua ʻike ʻia kahi ʻulaʻula iki o ka hoʻokuʻu ʻana a me ka emi ʻana o ka hua nui (0.22%), ma muli paha o ka hui pū ʻana o Ore. .
No ka ʻike ʻana i ka hui pū ʻana o ORE, ua hoʻohana mākou i ka microscopy atomic ikaika wai (AFM) e ʻike i nā loli morphological i ORE i ka hopena methanol ma hope o ka hoʻohui ʻana i ka wai.Ke emi ka wai ma lalo o 80%, ʻaʻole i ʻike ʻia kahi hōʻuluʻulu maʻemaʻe (Supplementary Fig. 7).Eia naʻe, me ka hoʻonui hou ʻana o ka wai i ka 90-95%, ua ʻike ʻia nā nanoparticles liʻiliʻi, e hōʻike ana i ka hui pū ʻana o Ore. hoʻonā (Fig. 2C a me ka Fig. 8 Pākuʻi).I ka hoʻohālikelike ʻana, ua hāʻule koke ka indocyanine green (ICG as control) ma 779 nm, e hōʻike ana i ka photostability maikaʻi loa o RuDA.Eia kekahi, ua nānā ʻia ke kūpaʻa o RuDA-NPs ma PBS (pH = 5.4, 7.4 a me 9.0), 10% FBS a me DMEM (high glucose) e ka UV-visible absorption spectroscopy i nā manawa like ʻole.E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 9, ua ʻike ʻia nā loli liʻiliʻi o nā kaula absorption RuDA-NP ma PBS ma pH 7.4 / 9.0, FBS a me DMEM, e hōʻike ana i ke kūpaʻa maikaʻi o RuDA-NP.Eia naʻe, i loko o kahi mea ʻawaʻawa (рН = 5.4) ua loaʻa ka hydrolysis o Ore.Ua loiloi hou mākou i ka paʻa o RuDA a me RuDA-NP me ka hoʻohana ʻana i nā hana chromatography wai kiʻekiʻe (HPLC).E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 10, ua kūpaʻa ʻo RuDA i ka hui ʻana o ka methanol a me ka wai (50/50, v / v) no ka hola mua, a ʻike ʻia ka hydrolysis ma hope o 4 mau hola.Eia naʻe, ʻike wale ʻia kahi piko concave-convex ākea no nā RuDA NPs.No laila, ua hoʻohana ʻia ka gel permeation chromatography (GPC) e loiloi i ke kūpaʻa o RuDA NPs i PBS (pH = 7.4).E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 11, ma hope o 8 mau hola o ka incubation ma lalo o nā kūlana i hoʻāʻo ʻia, ʻaʻole i loli nui ke kiʻekiʻe kiʻekiʻe, ka laulā kiʻekiʻe a me ka wahi kiʻekiʻe o NP RuDA, e hōʻike ana i ke kūpaʻa maikaʻi o NP RuDA.Eia kekahi, hōʻike nā kiʻi TEM i ka morphology o nā nanoparticles RuDA-NP i hoʻololi ʻole ʻia ma hope o nā hola 24 i ka diluted PBS buffer (pH = 7.4, Supplementary Fig. 12).
No ka mea hiki i ka hui ʻana iho ke hāʻawi i nā ʻano hana like ʻole a me nā ʻano kemika ma Ore, ua ʻike mākou i ka hoʻokuʻu ʻia ʻana o 9,10-anthracenediylbis(methylene)dimalonic acid (ABDA, kuhikuhi 1O2) i loko o nā hui methanol-wai.ʻOka me ka wai like ʻole50.E like me ka mea i hōʻike ʻia ma ke Kiʻi 2D a me ke Kiʻi Hoʻohui 13, ʻaʻohe degradation o ABDA i ʻike ʻia i ka wā ma lalo o 20%.Me ka piʻi ʻana o ka haʻahaʻa i 40%, ua hōʻemi ʻia ka ABDA, e like me ka hōʻike ʻana i ka emi ʻana o ka ikaika o ka fluorescence ABDA.Ua ʻike pū ʻia ʻo ka hopena o ka wai kiʻekiʻe i ka hoʻohaʻahaʻa wikiwiki ʻana, e manaʻo ana he pono ka hui pū ʻana o RuDA a pono no ka hōʻino ʻana o ABDA.He ʻokoʻa loa kēia ʻano mai nā chromophores ACQ hou (aggregation-induced quenching).Ke hoʻoulu ʻia me ka laser me ka lōʻihi o ka nalu o 808 nm, ʻo ka hua nui o 1O2 RuDA i ka hui ʻana o 98% H2O/2% DMF he 16.4%, ʻo ia ka 82 mau manawa kiʻekiʻe ma mua o ka ICG (ΦΔ = 0.2%)51, e hōʻike ana i ka maikaʻi o ka hanauna kupaianaha 1O2 RuDA i ke kūlana o ka hōʻuluʻulu.
Hoʻohana ʻia nā electron me ka hoʻohana ʻana i ka 2,2,6,6-tetramethyl-4-piperidinone (TEMP) a me 5,5-dimethyl-1-pyrroline N-oxide (DMPO) ma ke ʻano he spin traps Ua hoʻohana ʻia ʻo Resonance spectroscopy (ESR) e ʻike i nā ʻano hopena. AFK.na RuDA.E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 14, ua hōʻoia ʻia ua hana ʻia ka 1O2 i nā manawa irradiation ma waena o 0 a me 4 mau minuke.Eia kekahi, i ka wā i hoʻokomo ʻia ai ʻo RuDA me DMPO ma lalo o ka irradiation, ua ʻike ʻia kahi hōʻailona EPR maʻamau ʻehā o 1: 2: 2: 1 DMPO-OH · hoʻohui, e hōʻike ana i ka hoʻokumu ʻana o nā radical hydroxyl (OH·).Ma keʻano holoʻokoʻa, hōʻike nā hopena i luna i ka hiki o RuDA e hoʻoulu i ka hana ROS ma o ke kaʻina hana photosensitization ʻano ʻelua ʻano I/II.
No ka hoʻomaopopo maikaʻi ʻana i nā waiwai uila o RuDA ma nā ʻano monomeric a i hui pū ʻia, ua helu ʻia nā orbital molekala mua o RuDA ma nā ʻano monomeric a me nā dimeric me ke ʻano DFT.E like me ka hoike ana ma ka fig.3A, ua wehe ʻia ka orbital molecular kiʻekiʻe loa (HOMO) o ka RuDA monomeric ma ka iwi kuamoʻo ligand a ʻo ka orbital molecular haʻahaʻa loa i noho ʻole ʻia (LUMO) e kau ʻia ma ka ʻāpana ʻae TDP.ʻO ka mea ʻē aʻe, ʻo ka nui o ka electron i loko o ka dimeric HOMO ua kau ʻia ma luna o ka ligand o kahi mole RuDA, aʻo ka nui o ka electron density i ka LUMO ka mea nui i ka ʻae ʻana o kahi mole RuDA ʻē aʻe, e hōʻike ana aia ʻo RuDA i ka dimer.Nā hiʻohiʻona o CT.
A Ua helu ʻia ka HOMO a me ka LUMO o Ore ma nā ʻano monomeric a me dimeric.B Singlet a me triplet ikehu pae o ka Ore ma monomers a me dimer.C Nā kiʻekiʻe i manaʻo ʻia o RuDA a me nā kaha ISC hiki ke like me ka monomeric C a me ka dimeric D. Hōʻike nā pua i nā kaha ISC hiki ke hiki.
ʻO ka māhele ʻana o nā electrons a me nā lua i loko o nā mokuʻāina hauʻoli haʻahaʻa haʻahaʻa o RuDA ma nā ʻano monomeric a me dimeric i kālai ʻia me ka polokalamu Multiwfn 3.852.53, i helu ʻia me ke ʻano TD-DFT.E like me ka mea i hōʻike ʻia ma ka lepili hou.E like me ka mea i hōʻike ʻia ma nā Kiʻi 1-2, hoʻokaʻawale ʻia nā puka RDA monomeric ma ka iwi kuamoʻo i loko o kēia mau mokuʻāina hoʻohiwahiwa, ʻoiai ʻo nā electrons ka hapa nui o ka hui TDP, e hōʻike ana i nā ʻano intramolecular o CT.Eia kekahi, no kēia mau mokuʻāina ʻoliʻoli hoʻokahi, ʻoi aʻe a liʻiliʻi paha ma waena o nā lua a me nā electrons, e manaʻo ana e hāʻawi kēia mau mokuʻāina ʻoliʻoli i kekahi haʻawina mai ka hauʻoli kūloko (LE).No nā dimer, ma waho aʻe o nā hiʻohiʻona intramolecular CT a me LE, ua ʻike ʻia kekahi ʻāpana o nā hiʻohiʻona intermolecular CT ma nā mokuʻāina ʻē aʻe, ʻo ia hoʻi ʻo S3, S4, S7, a me S8, e pili ana i ka nānā ʻana i ka intermolecular CT, me ka CT intermolecular transitions ʻo ia nā mea nui. (Pakaukau Hoʻohui).3).
No ka hoʻomaopopo maikaʻi ʻana i nā hopena hoʻokolohua, ua ʻimi hou mākou i nā waiwai o nā mokuʻāina hauʻoli ʻo RuDA e ʻimi i nā ʻokoʻa ma waena o nā monomer a me nā dimer (Nā Papa Hoʻohui 4-5).E like me ka mea i hōʻike ʻia ma ke Kiʻi 3B, ʻoi aku ka nui o nā pae ikehu o ka mokuʻāina hoʻohiwahiwa hoʻokahi a me triplet o ka dimer ma mua o ko ka monomer, e kōkua ana e hōʻemi i ka ʻokoʻa ikehu ma waena o S1 a me Tn. Ua hōʻike ʻia e hiki ke hoʻokō ʻia nā hoʻololi ISC i loko o ka ʻāpana ikehu liʻiliʻi (ΔES1-Tn <0.3 eV) ma waena o S1 a me Tn54. Ua hōʻike ʻia e hiki ke hoʻokō ʻia nā hoʻololi ISC i loko o kahi wahi ikehu liʻiliʻi (ΔES1-Tn <0.3 eV) ma waena o S1 a me Tn54. Сообщалось, что переходы ISC могут быть реализованы в пределах небольшой энергетической щели (ΔES1-Tn <0,3 эВду) s. Ua hōʻike ʻia e hiki ke hoʻokō ʻia nā hoʻololi ISC i loko o kahi ʻāpana ikehu liʻiliʻi (ΔES1-Tn <0.3 eV) ma waena o S1 a me 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 эВду) s4 м Ua hōʻike ʻia e hiki ke hoʻokō ʻia ka hoʻololi ISC i loko o kahi ʻāpana ikehu liʻiliʻi (ΔES1-Tn <0.3 eV) ma waena o S1 a me Tn54.Eia kekahi, hoʻokahi wale nō orbital, noho ʻia a i ʻole i noho ʻole ʻia, pono e ʻokoʻa i nā mokuʻāina hoʻopaʻa ʻia a me nā mokuʻāina ʻekolu e hāʻawi i kahi integral non-zero SOC.No laila, ma muli o ka nānā ʻana i ka ikehu excitation a me ka hoʻololi orbital, ua hōʻike ʻia nā ala āpau āpau o ka hoʻololi ISC ma Fig.3C,D.ʻO ka mea nui, hoʻokahi wale nō kaila ISC i loaʻa i ka monomer, aʻo ka dimeric form he ʻehā ISC channel e hiki ke hoʻonui i ka hoʻololi ISC.No laila, he mea kūpono ke manaʻo e ʻoi aku ka nui o nā molekala RuDA i hoʻohui ʻia, ʻoi aku ka maʻalahi o nā kahawai ISC.No laila, hiki i ka RuDA aggregates ke hana i nā hale uila ʻelua-band i ka singlet a me triplet states, e hōʻemi ana i ka ʻokoʻa ikehu ma waena o S1 a me Tn i loaʻa, ma laila e hoʻonui ai i ka pono o ka ISC e hoʻomaʻamaʻa i ka hanauna 1O2.
No ka hoʻomaʻamaʻa hou ʻana i ke ʻano kumu, ua hoʻohui mākou i kahi pūhui kuhikuhi o ka arene-Ru (II) complex (RuET) ma ka hoʻololi ʻana i ʻelua hui ethyl me ʻelua mau hui phenyl triphenylamine i RuDA (Fig. -21 ) Mai ka mea hāʻawi (diethylamine) i ka ʻae (TDF), loaʻa iā RuET nā ʻano intramolecular CT like me RuDA.E like me ka mea i manaʻo ʻia, ua hōʻike ʻia ka spectrum absorption o RuET ma DMF i kahi hui hoʻoili uila haʻahaʻa me ka absorption ikaika ma ka ʻāpana infrared kokoke i ka ʻāpana o 600-1100 nm (Fig. 4B).Eia kekahi, uaʻike pūʻia ka hōʻuluʻulu RuET me ka hoʻonuiʻana i ka wai, iʻikeʻia i ka redshift o ka absorption maximum, i hōʻoia houʻia e ka wai AFM imaging (Supplementary Fig. 22).Hōʻike nā hopena i hiki iā RuET, e like me RuDA, ke hana i nā mokuʻāina intramolecular a hoʻohui iā ia iho i loko o nā hale hoʻohui.
ʻO ke kinona o ka RuET.B Absorption spectra o RuET i loko o nā hui ʻana o nā lakio like ʻole o DMF a me ka wai.Nā ʻāpana C EIS Nyquist no RuDA a me RuET.ʻO nā pane kiʻi kiʻi D o RuDA a me RuET ma lalo o ka hana o ka radiation laser me ka lōʻihi o 808 nm.
ʻO ka photodegradation o ABDA ma ke alo o RuET ua loiloi ʻia e ka irradiation me ka laser me ka nalu o 808 nm.ʻO ka mea kupanaha, ʻaʻole i ʻike ʻia ka hoʻohaʻahaʻa ʻana o ABDA i nā hakina wai like ʻole (Supplementary Fig. 23).ʻO ke kumu paha, ʻaʻole hiki i ka RuET ke hoʻokumu i kahi hoʻolālā uila i hoʻopaʻa ʻia no ka mea ʻaʻole hoʻoikaika ke kaulahao ethyl i ka hoʻoili ʻana o ka uku intermolecular.No laila, ua hana ʻia ka electrochemical impedance spectroscopy (EIS) a me nā ana photocurrent transient e hoʻohālikelike i nā waiwai photoelectrochemical o RuDA a me RuET.Wahi a ka ʻāpana Nyquist (Figure 4C), hōʻike ʻo RuDA i kahi radius ʻoi aku ka liʻiliʻi ma mua o RuET, ʻo ia hoʻi, ʻoi aku ka wikiwiki o ka lawe ʻana i nā electron intermolecular RuDA56 a ʻoi aku ka maikaʻi o ka conductivity.Eia kekahi, ʻoi aku ka kiʻekiʻe o ke kiʻi kiʻi kiʻi o RuDA ma mua o ka RuET (Fig. 4D), e hōʻoia ana i ka maikaʻi o ka hoʻoili kālā ʻana o RuDA57.No laila, he kuleana koʻikoʻi ka hui phenyl o triphenylamine ma Ore i ka hāʻawi ʻana i ka hoʻoili kālā intermolecular a me ka hoʻokumu ʻana i kahi ʻano uila uila.
No ka hoʻonui ʻana i ka hōʻiliʻili ʻana a me ka biocompatibility in vivo, ua hoʻopili hou mākou iā RuDA me F127.ʻO ka awelika hydrodynamic anawaena o RuDA-NPs ua hoʻoholo ʻia he 123.1 nm me ka hāʻawi liʻiliʻi (PDI = 0.089) me ka hoʻohana ʻana i ke ʻano dynamic light scattering (DLS) (Figure 5A), ka mea i hāpai i ka hōʻiliʻili ʻana o ka maʻi ma o ka hoʻonui ʻana i ka permeability a me ka paʻa.EPR) hopena.Ua hōʻike ʻia nā kiʻi TEM he ʻano spherical like ko Ore NPs me ka awelika anawaena o 86 nm.ʻIke ʻia, ʻike ʻia ka nui absorption o RuDA-NPs ma 800 nm (Supplementary Fig. 24), e hōʻike ana e hiki i nā RuDA-NP ke mālama i nā hana a me nā waiwai o nā RuDA hoʻohui ponoʻī.ʻO ka ROS quantum yield i helu ʻia no NP Ore he 15.9%, kahi i hoʻohālikelike ʻia me Ore. Ua aʻo ʻia nā waiwai photothermal o RuDA NPs ma lalo o ka hana o ka radiation laser me ka lōʻihi o ka hawewe o 808 nm me ka hoʻohana ʻana i kahi kamera infrared.E like me ka hoike ana ma ka fig.5B,C, ʻo ka hui hoʻomalu (PBS wale nō) i ʻike i kahi piʻi iki o ka mahana, ʻoiai ua piʻi wikiwiki ka mahana o ka solution RuDA-NPs me ka piʻi ʻana o ka mahana (ΔT) i 15.5, 26.1, a me 43.0°C.ʻO 25, 50, a me 100 µM nā manaʻo kiʻekiʻe, e hōʻike ana i kahi hopena photothermal ikaika o RuDA NPs.Eia kekahi, ua lawe ʻia nā ana pōʻaiapuni hoʻomehana / hoʻomaha no ka loiloi i ka paʻa photothermal o RuDA-NP a hoʻohālikelike me ICG.ʻAʻole i emi ka mahana o ka Ore NPs ma hope o ʻelima mau hoʻomehana wela / hoʻomaha (Fig. 5D), e hōʻike ana i ka paʻa photothermal maikaʻi loa o Ore NPs.ʻO ka ʻokoʻa, hōʻike ka ICG i ke kūpaʻa photothermal haʻahaʻa e like me ka ʻike ʻia mai ka nalo ʻana o ka pā wela photothermal ma lalo o nā kūlana like.E like me ka mea ma mua method58, ka photothermal conversion efficiency (PCE) o RuDA-NP ua helu ʻia he 24.2%, ʻoi aku ka kiʻekiʻe ma mua o nā mea photothermal i loaʻa e like me nā nanorods gula (21.0%) a me nā nanoshells gula (13.0%)59 .No laila, hōʻike ʻo NP Ore i nā waiwai photothermal maikaʻi loa, kahi e hoʻohiki ai lākou i nā ʻoihana PTT.
Ka nānā 'ana i nā ki'i DLS a me TEM o RuDA NPs (inset).B Nā kiʻi wela o nā manaʻo like ʻole o nā RuDA NP i hōʻike ʻia i ka radiation laser ma kahi hawewe o 808 nm (0.5 W cm-2).C Nā kiʻi hoʻololi Photothermal o nā manaʻo like ʻole o nā NP oka, ʻo ia ka ʻikepili quantitative.B. D Ke pi'i 'ana o ka wela o ORE NP a me ICG ma luna o 5 mau pō'ai ho'omehana wela.
Photocytotoxicity o RuDA NPs kū'ē MDA-MB-231 kanaka maʻi 'aʻai maʻi 'aʻai i loiloi 'ia in vitro.E like me ka hoike ana ma ka fig.Ua hōʻike ʻo 6A, B, RuDA-NPs a me RuDA i ka cytotoxicity haʻahaʻa i ka nele o ka irradiation, e hōʻike ana i ka pōʻeleʻele haʻahaʻa o RuDA-NPs a me RuDA.Eia nō naʻe, ma hope o ka ʻike ʻana i ka radiation laser ma kahi hawewe nalu o 808 nm, ua hōʻike ʻo RuDA a me RuDA NPs i ka photocytotoxicity ikaika e kūʻē i nā maʻi maʻi maʻi MDA-MB-231 me nā waiwai IC50 ​​(ka hapalua o ka nui o ka inhibitory concentration) o 5.4 a me 9.4 μM, e hōʻike ana. i loaʻa iā RuDA-NP a me RuDA no ka phototherapy maʻi kanesa.Eia kekahi, ua noiʻi hou ʻia ka photocytotoxicity o RuDA-NP a me RuDA ma ke alo o ka vitamina C (Vc), kahi scavenger ROS, e wehewehe i ke kuleana o ROS i ka cytotoxicity i hoʻomālamalama ʻia.ʻIke loa, ua hoʻonui ʻia ka cell viability ma hope o ka hoʻohui ʻana o Vc, a ʻo nā waiwai IC50 o RuDA a me RuDA NPs he 25.7 a me 40.0 μM, kēlā me kēia, e hōʻike ana i ka hana nui o ROS i ka photocytotoxicity o RuDA a me RuDA NPs.ʻO ka cytotoxicity māmā o RuDA-NPs a me RuDA i loko o MDA-MB-231 mau mea maʻi maʻi ma o ke ola / make cell staining me ka calcein AM (ka'ōmaʻomaʻo'ōmaʻomaʻo no nā pūnaewele ola) a me ka propidium iodide (PI, kaʻulaʻulaʻulaʻula no nā pūnaewele make).hoʻopaʻa ʻia e nā cell) ma ke ʻano he fluorescent probes.E like me ka mea i hōʻike ʻia ma ka Figure 6C, ua mau nā cell i mālama ʻia me RuDA-NP a i ʻole RuDA me ka ʻole o ka irradiation, e like me ka ʻike ʻia e ka fluorescence ʻōmaʻomaʻo ikaika.Ma ka ʻokoʻa, ma lalo o ka laser irradiation, ʻike ʻia ka fluorescence ʻulaʻula wale nō, e hōʻoia ana i ka photocytotoxicity kūpono o RuDA a i ʻole RuDA NPs.He mea koʻikoʻi ua ʻike ʻia ka fluorescence ʻōmaʻomaʻo ma ka hoʻohui ʻana o Vc, e hōʻike ana i ka hōʻino ʻana i ka photocytotoxicity o RuDA a me RuDA NPs.Kūlike kēia mau hopena me nā hōʻike in vitro photocytotoxicity.
ʻO ka hiki ke hilinaʻi i ka hopena o nā pūnaewele A RuDA- a me B RuDA-NP i nā pūnaewele MDA-MB-231 i ke alo a i ʻole ka nele o Vc (0.5 mM), kēlā me kēia.ʻO nā kī kuhi hewa, mean ± ka hoʻokaʻawale maʻamau (n = 3). ʻAʻole i hoʻohui ʻia, ʻelua ʻaoʻao t hōʻike *p <0.05, **p <0.01, a me ***p <0.001. ʻAʻole i hoʻohui ʻia, ʻelua ʻaoʻao t hōʻike *p <0.05, **p <0.01, a me ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 и ***p <0,001. ʻO nā ho'āʻo t-huelo ʻelua i hui ʻole ʻia *p<0.05, **p<0.01, a me ***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 и ***p <0,001. ʻO nā ho'āʻo t-huelo ʻelua i hui ʻole ʻia *p<0.05, **p<0.01, a me ***p<0.001.C Ka hoʻopaʻa ʻana i ke kala ʻana o ke kelepona make me ka hoʻohana ʻana i ka calcein AM a me ka propidium iodide ma ke ʻano he ʻimi fluorescent.ʻO ka pā unahi: 30 µm.Hōʻike ʻia nā kiʻi i hōʻike ʻia o ʻekolu mau mea olaola mai kēlā me kēia hui.D Confocal fluorescence kiʻi o ka hana ROS ma MDA-MB-231 pūnaewele ma lalo o nā kūlana lapaʻau like ʻole.Hōʻike ka ʻōmaʻomaʻo DCF fluorescence i ka hele ʻana o ROS.Iradiate me ka laser me ka nalu o 808 nm me ka mana o 0.5 W/cm2 no 10 minuke (300 J/cm2).ʻO ka pā unahi: 30 µm.Hōʻike ʻia nā kiʻi i hōʻike ʻia o ʻekolu mau mea olaola mai kēlā me kēia hui.E Flow cytometry RuDA-NPs (50 µM) a i ʻole RuDA (50 µM) ka nānā ʻana i ka mālama ʻana me ka laser 808 nm a i ʻole (0.5 W cm-2) i mua a me ka ʻole o Vc (0.5 mM) no 10 min.Hōʻike ʻia nā kiʻi i hōʻike ʻia o ʻekolu mau mea olaola mai kēlā me kēia hui.F Nrf-2, HSP70 a me HO-1 o nā pūnaewele MDA-MB-231 i mālama ʻia me RuDA-NPs (50 µM) me ka 808 nm laser irradiation a i ʻole (0.5 W cm-2, 10 min, 300 J cm-2), hōʻike nā pūnaewele 2).Hōʻike ʻia nā kiʻi i hōʻike ʻia o ʻelua mau mea olaola mai kēlā me kēia hui.
Ua nānā ʻia ka hana ROS intracellular ma MDA-MB-231 me ka hoʻohana ʻana i ke ʻano staining 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA).E like me ka hoike ana ma ka fig.6D, nā pūnaewele i mālama ʻia me RuDA-NPs a i ʻole RuDA i hōʻike ʻia i ka fluorescence ʻōmaʻomaʻo ʻokoʻa i ka wā i hoʻomālamalama ʻia me ka laser 808 nm, e hōʻike ana i ka RuDA-NPs a me RuDA i hiki ke hana i ka ROS.ʻO ka mea ʻē aʻe, i ka ʻole o ka mālamalama a i ʻole ke alo o Vc, ʻike wale ʻia kahi hōʻailona fluorescent nāwaliwali o nā cell, e hōʻike ana i kahi hoʻokumu liʻiliʻi o ROS.Ua hoʻoholo hou ʻia nā pae ROS Intracellular i nā pūnaewele RuDA-NP a me nā ʻāpana MDA-MB-231 i mālama ʻia e RuDA e ka cytometry kahe.E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 25, ʻo ka mean fluorescence intensity (MFI) i hana ʻia e RuDA-NPs a me RuDA ma lalo o 808 nm laser irradiation ua hoʻonui nui ʻia ma kahi o 5.1 a me 4.8 mau manawa, i hoʻohālikelike ʻia i ka pūʻulu mana, e hōʻoia ana i kā lākou hoʻokumu maikaʻi ʻana AFK.hiki.Eia nō naʻe, hoʻohālikelike nā pae intracellular ROS ma RuDA-NP a i ʻole MDA-MB-231 mau mea i mālama ʻia me RuDA me nā mana me ka ʻole o ka laser irradiation a i ʻole ke alo o Vc, e like me nā hopena o ka nānā ʻana i ka fluorescence confocal.
Ua hōʻike ʻia ʻo ka mitochondria ka pahuhopu nui o Ru (II) -arene complexes60.No laila, ua noiʻi ʻia ka subcellular localization o RuDA a me RuDA-NPs.E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 26, hōʻike ʻo RuDA a me RuDA-NP i nā ʻano like like me ka hoʻonui ʻana i ka mitochondria (62.5 ± 4.3 a me 60.4 ± 3.6 ng / mg protein, kēlā me kēia).Eia naʻe, he wahi liʻiliʻi wale nō o Ru i loaʻa i nā hakina nuklea o Ore a me NP Ore (3.5 a me 2.1%, kēlā me kēia).ʻO ke koena o ka hakina cell i loko o ke koena ruthenium: 31.7% (30.6 ± 3.4 ng/mg protein) no RuDA a me 42.9% (47.2 ± 4.5 ng/mg protein) no RuDA-NPs.Ma keʻano laulā, ʻo Ore a me NP Ore ka mea i hōʻiliʻili nui ʻia i ka mitochondria.No ka loiloi i ka mitochondrial dysfunction, ua hoʻohana mākou i ka JC-1 a me ka MitoSOX Red staining e loiloi i ka mana mitochondrial membrane a me ka mana hana superoxide, kēlā me kēia.E like me ka mea i hōʻike ʻia ma ka Fig 27, ʻike ʻia ka fluorescence ʻōmaʻomaʻo ikaika (JC-1) a me ka ʻulaʻula (MitoSOX Red) i loko o nā cell i mālama ʻia me nā RuDA a me RuDA-NPs ma lalo o 808 nm laser irradiation, e hōʻike ana i ka nui o ka fluorescent RuDA a me RuDA-NPs. Hiki iā ia ke hoʻoulu i ka depolarization membrane mitochondrial a me ka hana superoxide.Eia kekahi, ua hoʻoholo ʻia ke ʻano o ka make cell me ka hoʻohana ʻana i ka flow cytometry based analysis of annexin V-FITC/propidium iodide (PI).E like me ka mea i hōʻike ʻia ma ka Figure 6E, i ka wā i hoʻomālamalama ʻia me ka laser 808 nm, ua hoʻonui ʻo RuDA a me RuDA-NP i ka nui o ka apoptosis mua (haʻahaʻa ʻākau haʻahaʻa) i nā cell MDA-MB-231 i hoʻohālikelike ʻia me PBS a i ʻole PBS me ka laser.nā pūnaewele i hoʻoponopono ʻia.Eia naʻe, i ka wā i hoʻohui ʻia ai ʻo Vc, ua emi nui ka apoptosis o RuDA a me RuDA-NP mai 50.9% a me 52.0% a i 15.8% a me 17.8%, e hōʻoia ana i ka hana nui o ROS i ka photocytotoxicity o RuDA a me RuDA-NP..Eia kekahi, ua ʻike ʻia nā cell necrotic liʻiliʻi i nā hui āpau i hoʻāʻo ʻia (ka ʻaoʻao hema hema), e hōʻike ana ʻo ka apoptosis paha ke ʻano nui o ka make cell i hoʻokomo ʻia e RuDA a me RuDA-NPs.
No ka mea, ʻo ka pōʻino koʻikoʻi oxidative ka mea hoʻoholo nui o ka apoptosis, ua noiʻi ʻia ka mea nuklea e pili ana me ka erythroid 2, factor 2 (Nrf2) 62, kahi mea hoʻoponopono koʻikoʻi o ka ʻōnaehana antioxidant, ma RuDA-NPs-treated MDA-MB-231.ʻO ka hana o ka hana a nā RuDA NP i hoʻoulu ʻia e ka irradiation.I ka manawa like, ua ʻike pū ʻia ka ʻōlelo o ka protein downstream heme oxygenase 1 (HO-1).E like me ka mea i hōʻike ʻia ma ka Figure 6F a me ka Figure 29, RuDA-NP-mediated phototherapy i hoʻonui i nā pae hōʻike Nrf2 a me HO-1 i hoʻohālikelike ʻia i ka pūʻulu PBS, e hōʻike ana e hiki i ka RuDA-NP ke hoʻoulu i nā ala hōʻailona koʻikoʻi oxidative.Eia kekahi, no ke aʻo ʻana i ka hopena photothermal o RuDA-NPs63, ua loiloi pū ʻia ka ʻōlelo o ka protein haʻalulu wela Hsp70.He mea maopopo loa ua hōʻike ʻia nā cell i mālama ʻia me RuDA-NPs + 808 nm laser irradiation i hoʻonui ʻia i ka hōʻike o Hsp70 i hoʻohālikelike ʻia i nā pūʻulu ʻelua ʻē aʻe, e hōʻike ana i kahi pane cellular i hyperthermia.
ʻO nā hopena in vitro kupaianaha i hoʻoikaika iā mākou e noiʻi i ka hana in vivo o RuDA-NP i nā ʻiole olohelohe me nā ʻōpū MDA-MB-231.Ua aʻo ʻia ka hāʻawi ʻana i nā kiko o nā RuDA NP ma ka hoʻoholo ʻana i ka ʻike o ka ruthenium i loko o ke ake, puʻuwai, spleen, puʻupaʻa, māmā, a me nā ʻōpū.E like me ka hoike ana ma ka fig.7A, ʻike ʻia ka ʻike nui loa o ka Ore NPs i nā kino maʻamau i ka manawa nānā mua (4 h), ʻoiai ʻo ka ʻike nui loa i hoʻoholo ʻia i loko o nā ʻiʻo tumora 8 mau hola ma hope o ka hoʻopaʻa ʻana, ma muli paha o Ore NPs.Ka hopena EPR o LF.Wahi a nā hopena hoʻoili, ua lawe ʻia ka lōʻihi o ka mālama ʻana me ka NP ore i 8 mau hola ma hope o ka hoʻokele.No ka hōʻike ʻana i ke kaʻina hana o ka hōʻiliʻili ʻana o nā RuDA-NPs i nā pūnaewele tumora, ua nānā ʻia nā waiwai photoacoustic (PA) o RuDA-NPs ma ka hoʻopaʻa ʻana i nā hōʻailona PA o RuDA-NPs i nā manawa like ʻole ma hope o ka injection.ʻO ka mea mua, ua loiloi ʻia ka hōʻailona PA o RuDA-NP i vivo e ka hoʻopaʻa ʻana i nā kiʻi PA o kahi kahua maʻi ma hope o ka hoʻokomo intratumoral o RuDA-NP.E like me ka mea i hōʻike ʻia ma ke Kiʻi Hoʻohui 30, ua hōʻike ʻo RuDA-NP i kahi hōʻailona PA ikaika, a aia kahi pilina maikaʻi ma waena o ka manaʻo RuDA-NP a me ka ikaika o ka hōʻailona PA (Supplementary Figure 30A).A laila, i loko o vivo PA nā kiʻi o nā wahi maʻi maʻi i hoʻopaʻa ʻia ma hope o ka intravenous injection o RuDA a me RuDA-NP i nā manawa like ʻole ma hope o ka injection.E like me ka mea i hōʻike ʻia ma ka Figure 7B, ua hoʻonui nui ʻia ka hōʻailona PA o RuDA-NPs mai ka pūnaewele tumora me ka manawa a hiki i kahi pāpū ma 8 mau hola post-injection, e like me nā hopena hoʻoili kiko i hoʻoholo ʻia e ka ICP-MS analysis.E pili ana i ka RuDA (Supplementary Fig. 30B), ua ʻike ʻia ka ikaika o ka hōʻailona PA kiʻekiʻe ma hope o 4 mau hola ma hope o ka injection, e hōʻike ana i ka wikiwiki o ke komo ʻana o RuDA i loko o ka ʻōpū.Eia kekahi, ua noiʻi ʻia ka hana excretory o RuDA a me RuDA-NPs ma ka hoʻoholo ʻana i ka nui o ka ruthenium i loko o ka mimi a me nā faeces e hoʻohana ana i ka ICP-MS.ʻO ke ala nui o ka hoʻopau ʻana no RuDA (Supplementary Fig. 31) a me RuDA-NPs (Fig. 7C) ma o nā faeces, a ua ʻike ʻia ka hoʻomaʻemaʻe maikaʻi ʻana o RuDA a me RuDA-NPs i ka wā aʻo 8-lā, ʻo ia hoʻi ʻo RuDA a me ka RuDA-NPs hiki ke hoʻopau maikaʻi ʻia mai ke kino me ka ʻole o ka ʻawaʻawa lōʻihi.
A. Ua hoʻoholo ʻia ka hāʻawi ʻana o Ex vivo o RuDA-NP i loko o nā ʻiole e ka ʻike Ru (ka pākēneka o ka hoʻohana ʻia ʻana o Ru (ID) i kēlā me kēia gram o ka ʻiʻo) i nā manawa like ʻole ma hope o ka injection.ʻO keʻano o kaʻikepili ± ka haʻalele maʻamau (n = 3). ʻAʻole i hoʻohui ʻia, ʻelua ʻaoʻao t hōʻike *p <0.05, **p <0.01, a me ***p <0.001. ʻAʻole i hoʻohui ʻia, ʻelua ʻaoʻao t hōʻike *p <0.05, **p <0.01, a me ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 и ***p <0,001. ʻO nā ho'āʻo t-huelo ʻelua i hui ʻole ʻia *p<0.05, **p<0.01, a me ***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 и ***p <0,001. ʻO nā ho'āʻo t-huelo ʻelua i hui ʻole ʻia *p<0.05, **p<0.01, a me ***p<0.001.Nā kiʻi B PA o nā wahi maʻi maʻi vivo ma 808 nm excitation ma hope o ka hoʻokele intravenous o RuDA-NPs (10 µmol kg-1) i nā manawa manawa like ʻole.Ma hope o ka lawelawe intravenous o RuDA NPs (10 µmol kg-1), ua hoʻokuʻu ʻia ʻo C Ru mai nā ʻiole me ka mimi a me nā faeces i nā manawa like ʻole.ʻO keʻano o kaʻikepili ± ka haʻalele maʻamau (n = 3).
Ua aʻo ʻia ka mana wela o RuDA-NP in vivo i nā ʻiole olohelohe me MDA-MB-231 a me nā maʻi maʻi RuDA no ka hoʻohālikelike.E like me ka hoike ana ma ka fig.8A a me ka Fig 32, ua hōʻike ka pūʻulu mana (saline) i ka liʻiliʻi o ka hoʻololi ʻana o ka mahana (ΔT ≈ 3 °C) ma hope o 10 mau minuke o ka hoʻomau mau.Eia naʻe, ua piʻi wikiwiki ka mahana o RuDA-NPs a me RuDA me nā mahana kiʻekiʻe loa o 55.2 a me 49.9 °C, e hāʻawi ana i ka hyperthermia lawa no ka maʻi maʻi maʻi maʻi vivo.ʻO ka piʻi ʻana o ka mahana kiʻekiʻe no nā RuDA NPs (ΔT ≈ 24 ° C) i hoʻohālikelike ʻia me RuDA (ΔT ≈ 19 ° C) ma muli paha o kona permeability maikaʻi a me ka hōʻiliʻili ʻana i nā ʻiʻo tumora ma muli o ka hopena EPR.
ʻO nā kiʻi wela infrared o nā ʻiole me nā ʻōpū MDA-MB-231 i hoʻomālamalama ʻia me ka laser 808 nm i nā manawa like ʻole 8 mau hola ma hope o ka injection.Hōʻike ʻia nā kiʻi i hōʻike ʻia o ʻehā mau hana hou mai kēlā me kēia hui.B Ka nui o ka ma'i ma'i a me C Ka nui o ka ma'i ma'amau o nā pū'ulu iole like 'ole i ka wā o ka mālama 'ana.D Nā ʻeke o nā paona kino o nā hui ʻiole like ʻole.Iradiate me ka laser me ka nalu o 808 nm me ka mana o 0.5 W/cm2 no 10 minuke (300 J/cm2).ʻO nā kī kuhi hewa, mean ± ka hoʻokaʻawale maʻamau (n = 3). ʻAʻole i hoʻohui ʻia, ʻelua ʻaoʻao t hōʻike *p <0.05, **p <0.01, a me ***p <0.001. ʻAʻole i hoʻohui ʻia, ʻelua ʻaoʻao t hōʻike *p <0.05, **p <0.01, a me ***p <0.001. Непарные двусторонние t-критерии *p <0,05, **p <0,01 и ***p <0,001. ʻO nā ho'āʻo t-huelo ʻelua i hui ʻole ʻia *p<0.05, **p<0.01, a me ***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 и ***p <0,001. ʻO nā ho'āʻo t-huelo ʻelua i hui ʻole ʻia *p<0.05, **p<0.01, a me ***p<0.001. E H&E hoʻopaʻa i nā kiʻi o nā kino nui a me nā ʻōpū mai nā hui lapaʻau like ʻole, e like me Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, a me nā hui RuDA-NPs + Laser. E H&E hoʻopaʻa i nā kiʻi o nā kino nui a me nā ʻōpū mai nā hui lapaʻau like ʻole, e like me Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, a me nā hui RuDA-NPs + Laser. 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 hoʻopaʻa i nā kiʻi o nā kino nui a me nā ʻōpū mai nā hui lapaʻau like ʻole, me ka paakai, saline + laser, RuDA, RuDA + Laser, RuDA-NPs, a me nā hui RuDA-NPs + Laser.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、守光、RuDA-Rus、RuDA-Rus、RuDA-RU来自不同治疗组的主要器官和肿瘤的E H&E ОКии ИМгиновнннки и азка E H&E hoʻopaʻa ʻia o nā ʻōpū nui a me nā ʻōpū mai nā hui lapaʻau like ʻole me ka saline, saline + laser, RuDA, RuDA + laser, RuDA-NPs, a me RuDA-NPs + laser.ʻO ka pā unahi: 60 µm.
Ua loiloi ʻia ka hopena o ka phototherapy in vivo me RuDA a me RuDA NPs kahi i hoʻopaʻa ʻia ai nā ʻiole me ka MDA-MB-231 maʻi maʻi me ka RuDA a i ʻole RuDA NPs ma kahi pākahi o 10.0 µmol kg-1 ma o ka huelo vein, a laila 8 mau hola ma hope o ka hoʻokomo ʻana.irradiation laser me ka nalu o 808 nm.E like me ka mea i hōʻike ʻia ma ka Figure 8B, ua hoʻonui nui ʻia ka nui o nā tumora i ka saline a me nā pūʻulu laser, e hōʻike ana he liʻiliʻi ka hopena o ka saline a i ʻole ka laser 808 irradiation i ka ulu ʻana o ka tumor.E like me ka hui paʻakai, ʻike ʻia ka ulu wikiwiki ʻana i nā ʻiole i mālama ʻia me RuDA-NPs a i ʻole RuDA me ka ʻole o ka laser irradiation, e hōʻike ana i ko lākou haʻahaʻa haʻahaʻa ʻeleʻele.Ma ka hoʻohālikelike, ma hope o ka laser irradiation, ʻo RuDA-NP a me RuDA lapaʻau i hoʻoulu i ka regression koʻikoʻi me ka hōʻemi ʻana o ka nui o ka tumo o 95.2% a me 84.3%, i hoʻohālikelike ʻia i ka hui mālama saline, e hōʻike ana i ka synergistic PDT maikaʻi loa., i hoʻopili ʻia e ka hopena RuDA/CHTV.- NP a i ʻole Ore. Hoʻohālikelike ʻia me RuDA, ua hōʻike ʻo RuDA NP i kahi hopena phototherapeutic ʻoi aku ka maikaʻi, ʻo ia ka mea ma muli o ka hopena EPR o RuDA NPs.Ua loiloi hou ʻia nā hopena o ka hoʻoulu ʻana o ka tumo e ke kaumaha o ka tumo i hoʻopau ʻia i ka lā 15 o ka mālama ʻana (Fig. 8C and Supplementary Fig. 33).ʻO ka mean tumor mass ma RuDA-NP i mālama ʻia nā ʻiole a me RuDA mālama ʻia nā ʻiole ʻo 0.08 a me 0.27 g, ʻoi aku ka māmā ma mua o ka pūʻulu mana (1.43 g).
Eia kekahi, ua hoʻopaʻa ʻia ke kaumaha o ke kino o nā ʻiole i kēlā me kēia lā ʻekolu e aʻo ai i ka ʻona ʻeleʻele o RuDA-NPs a i ʻole RuDA in vivo.E like me ka mea i hōʻike ʻia ma ka Figure 8D, ʻaʻohe ʻokoʻa koʻikoʻi o ke kaumaha o ke kino i ʻike ʻia no nā hui lapaʻau āpau. Eia kekahi, ua hoʻopaʻa ʻia ka hematoxylin a me ka eosin (H&E) o nā kino nui (naʻau, ate, spleen, ʻāmāmā, a me nā puʻupaʻa) mai nā hui lapaʻau like ʻole. Eia kekahi, ua hoʻopaʻa ʻia ka hematoxylin a me ka eosin (H&E) o nā mea nui (naʻau, ate, spleen, ʻāmāmā, a me nā puʻupaʻa) mai nā hui lapaʻau like ʻole. Кроме того, было проведено окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, гиплени, злочник) Eia hou, ua hoʻopaʻa ʻia ka hematoxylin a me ka eosin (H&E) o nā mea nui (naʻau, ate, spleen, māmā, a me nā puʻupaʻa) mai nā hui lapaʻau like ʻole.此外，对不同治疗组的主要器官（心脏、肝脏、脾脏、肺和肾脏）进行苏和石行苏月石行苍 (H&E) Кроме того, проводили окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селехленкрач, селехликрач) Eia hou, ua hoʻopaʻa ʻia ka hematoxylin a me ka eosin (H&E) o nā kino nui (naʻau, ate, spleen, ʻāmāmā, a me nā puʻupaʻa) i nā hui lapaʻau like ʻole.E like me ka hōʻike ʻana ma ka Fig.8E, ʻaʻole hōʻike nā kiʻi ʻoniʻoni H&E o ʻelima mau kino nui mai nā hui RuDA-NP a me RuDA ʻaʻole i hōʻike ʻia nā mea ʻino a i ʻole nā ​​pōʻino kino. 8E, ʻaʻole hōʻike nā kiʻi ʻoniʻoni H&E o ʻelima mau kino nui mai nā hui RuDA-NP a me RuDA ʻaʻole i hōʻike ʻia nā mea ʻino a i ʻole nā ​​pōʻino kino.E like me ka hoike ana ma ka fig.8E, изображения окрашивания H&E пяти основных органов из групп RuDA-NPs a me RuDA ʻaʻohe mea hoʻohana i ka hoʻokele waiwai. 8E, H&E nā kiʻi ʻoniʻoni o ʻelima mau kino nui mai nā hui RuDA-NP a me RuDA ʻaʻole hōʻike i nā mea ʻino ʻole a i ʻole nā ​​​​liona.RuDA-NPs如图8E 所示，来自RuDA-NPs 和RuDA 组的五个主要器官的H&E E kiʻi i ka рисунке 8E, e hāʻawi iā H&E i ka hoʻopaʻa inoa ʻana a me ka RuDA-NPs a me ka RuDA ʻaʻole e hoʻohana i ka manaʻo. E like me ka mea i hōʻike ʻia ma ka Figure 8E, ʻaʻole i hōʻike ʻia nā kiʻi ʻoniʻoni H&E o nā kino nui ʻelima mai nā hui RuDA-NPs a me RuDA ʻaʻole i hōʻike ʻia i nā mea ʻino a i ʻole ka pōʻino kino.Ua hōʻike ʻia kēia mau hopena ʻaʻole i hōʻike ʻo RuDA-NP a i ʻole RuDA i nā hōʻailona o ka make i loko o vivo. Eia kekahi, ua hōʻike ʻia nā kiʻi ʻoniʻoni H&E o nā maʻi maʻi i hiki i nā hui ʻelua RuDA + Laser a me RuDA-NPs + Laser ke hōʻeha nui i ka maʻi kanesa, e hōʻike ana i ka maikaʻi o ka vivo phototherapeutic efficacy o RuDA a me RuDA-NPs. Eia kekahi, ua hōʻike ʻia nā kiʻi ʻoniʻoni H&E o nā maʻi maʻi i hiki i nā hui ʻelua RuDA + Laser a me RuDA-NPs + Laser ke hōʻeha nui i ka maʻi kanesa, e hōʻike ana i ka maikaʻi o ka vivo phototherapeutic efficacy o RuDA a me RuDA-NPs.Eia kekahi, hematoxylin-eosin stained tumor kiʻi i hōʻike i ka RuDA + Laser a me RuDA-NPs + Laser pūʻulu hiki ke hoʻoulu i ka luku nui ʻana o nā maʻi kanesa, e hōʻike ana i ka maikaʻi phototherapeutic o RuDA a me RuDA-NPs in vivo.H&E此外 ， 肿瘤 的 & e 染色 显示 ， ruda + laser 和 ruda-nps + laser 组均 导致 的 癌细胞 破坏 ， 证明 和 。 。 ..Eia kekahi, hematoxylin a me eosin stained tumor kiʻi i hōʻike i nā hui ʻelua RuDA + Laser a me RuDA-NPs + Laser i hopena i ka luku nui ʻana o nā maʻi maʻi maʻi, e hōʻike ana i ka pono phototherapeutic kiʻekiʻe o RuDA a me RuDA-NPs in vivo.
I ka hopena, ua hoʻolālā ʻia ka Ru (II) -arene (RuDA) organometallic complex me DA-type ligands e hoʻomaʻamaʻa i ke kaʻina hana ISC me ka hoʻohana ʻana i ke ʻano hoʻohui.Hiki i ka RuDA Synthesized ke hoʻohui iā ia iho ma o nā pilina non-covalent e hana i nā ʻōnaehana supramolecular i loaʻa iā RuDA, a laila e hoʻomaʻamaʻa i ka hoʻokumu ʻana o 1O2 a me ka hoʻololi photothermal kūpono no ka mālama ʻana i ka maʻi kanesa.Hoʻomaopopo ʻia ʻaʻole i hana ʻo RuDA monomeric i ka 1O2 ma lalo o ka irradiation laser ma 808 nm, akā hiki ke hoʻoulu i ka nui o 1O2 i ka mokuʻāina aggregated, e hōʻike ana i ka noʻonoʻo a me ka pono o kā mākou hoʻolālā.Ua hōʻike ʻia nā haʻawina ma hope e hāʻawi ka supramolecular hui iā RuDA me ka hoʻomaikaʻi ʻana i nā waiwai photophysical a me photochemical, e like me ka redshift absorption a me ka pale ʻana i ka photobleaching, kahi i makemake nui ʻia no ka hana PDT a me PTT.Ua hōʻike ʻia nā hoʻokolohua in vitro a me in vivo ʻo RuDA NPs me ka biocompatibility maikaʻi a me ka hōʻiliʻili maikaʻi ʻana i ka ʻōpū e hōʻike ana i ka hana anticancer maikaʻi loa i hoʻoiho ʻia ma luna o ka laser irradiation ma kahi hawewe o 808 nm.No laila, ʻo RuDA NPs e like me ka bimodal supramolecular PDT/PTW reagents e hoʻonui i ka hoʻonohonoho o nā photosensitizers i hoʻāla ʻia ma nā nalu nalu ma luna o 800 nm.ʻO ka hoʻolālā manaʻo o ka ʻōnaehana supramolecular e hāʻawi i kahi ala kūpono no nā photosensitizer i hoʻāla ʻia e NIR me nā hopena photosensitizing maikaʻi loa.
Ua loaʻa mai nā kemika a me nā mea hoʻoheheʻe mai nā mea kūʻai aku a hoʻohana ʻia me ka hoʻomaʻemaʻe ʻole.Ua kūʻai ʻia ʻo RuCl3 mai Boren Precious Metals Co., Ltd. (Kunming, Kina).[(η6-p-cym)Ru(fendio)Cl]Cl (fendio = 1,10-phenanthroline-5,6-dione) a me 4,7-bis[4-(N,N-diphenylamino)phenyl]-5 ,6-Diamino-2,1,3-benzothiadiazole ua synthesized e like me nā haʻawina mua64,65.Ua hoʻopaʻa ʻia ka NMR spectra ma kahi Bruker Avance III-HD 600 MHz spectrometer ma Southeastern University Analytical Test Center me ka hoʻohana ʻana i ka d6-DMSO a i ʻole CDCl3 ma ke ʻano he solvent.Hāʻawi ʻia nā hoʻololi kemika δ ma ka ppm.e pili ana i ka tetramethylsilane, a ua hāʻawi ʻia nā mea hoʻohālikelike J i nā waiwai piha i hertz.Ua hana ʻia ke kiʻekiʻe resolution mass spectrometry (HRMS) ma kahi mea kani Agilent 6224 ESI/TOF MS.Ua hoʻokō ʻia ka loiloi kumu o C, H, a me N ma kahi mea hoʻohālikelike kumu ʻo Vario MICROCHNOS (Elementar).Ua ana ʻia nā kikoʻī ʻike ʻike UV ma kahi spectrophotometer Shimadzu UV3600.Ua hoʻopaʻa ʻia nā kikoʻī Fluorescence ma kahi kikoʻī Shimadzu RF-6000 spectrofluorimeter.Ua hoʻopaʻa ʻia ka spectra EPR ma kahi mea kani Bruker EMXmicro-6/1.Ua aʻo ʻia ka morphology a me ke ʻano o nā mea hoʻohālike i hoʻomākaukau ʻia ma FEI Tecnai G20 (TEM) a me Bruker Icon (AFM) e hana ana ma ka volta o 200 kV.Hana ʻia ʻo Dynamic light scattering (DLS) ma kahi mea hōʻike Nanobrook Omni (Brookhaven).Ua ana ʻia nā waiwai Photoelectrochemical ma kahi hoʻonohonoho electrochemical (CHI-660, Kina).Loaʻa nā kiʻi photoacoustic me ka hoʻohana ʻana i ka ʻōnaehana FUJIFILM VisualSonics Vevo® LAZR.Loaʻa nā kiʻi confocal me ka hoʻohana ʻana i kahi microscope confocal Olympus FV3000.Hana ʻia ka loiloi FACS ma kahi cytometer kahe BD Calibur.Ua hana ʻia nā hoʻokolohua wai kiʻekiʻe (HPLC) ma kahi ʻōnaehana Waters Alliance e2695 me ka hoʻohana ʻana i kahi mea ʻike 2489 UV/Vis.Ua hoʻopaʻa ʻia nā hoʻokolohua Gel Permeation Chromatography (GPC) ma kahi mea hana Thermo ULTIMATE 3000 me ka hoʻohana ʻana i kahi mea ʻike kuhikuhi kuhikuhi refractive ERC RefratoMax520.
[(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) a me ka waikawa acetic glacial (30 mL) i hoʻoulu ʻia i ka reflux reflux no 12 mau hola.Wehe ʻia ka mea hoʻoheheʻe i loko o ka vacuo me ka hoʻohana ʻana i kahi evaporator rotary.Ua hoʻomaʻemaʻe ʻia ke koena i loaʻa e ka chromatography kolamu flash (silica gel, CH2Cl2: MeOH=20:1) e loaʻa iā RuDA ma ke ʻano he pauka ʻōmaʻomaʻo (hua: 877.5 mg, 80%).anus.Ua helu ʻia no C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.Loaʻa: 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), 12,5.28, 13,5.28, 13,5.28, 13,5.28, 13,5.28, 13,5.29, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49, 103.49 , 103. , 86.52, 84.75, 63.29, 30.90, 22.29, 18.83.ESI-MS: m/z [M-Cl]+ = 1097.25.
Synthesis o 4,7-bis[4-(N,N-diethylamino)phenyl-5,6-diamino-2,1,3-benzothiadiazole (L2): Ua hoʻohui ʻia ʻo L2 i ʻelua mau ʻanuʻu.Pd(PPh3)4 (46 mg, 0.040 mmol) i hoʻohui ʻia i ka N,N-diethyl-4-(tributylstannyl)aniline (1.05 g, 2.4 mmol) a me 4,7-dibromo-5,6-dinitro solution - 2, 1,3-benzothiadiazole (0.38 g, 1.0 mmol) i ka toluene maloʻo (100 ml).Hoʻopili ʻia ka hui ʻana ma 100 ° C no 24 mau hola.Ma hope o ka wehe ʻana i ka toluene i loko o ka vacuo, ua holoi ʻia ka mea paʻa me ka petroleum ether.A laila ua hoʻoulu ʻia kahi hui ʻana o kēia pūhui (234.0 mg, 0.45 mmol) a me ka pauka hao (0.30 g, 5.4 mmol) i ka acetic acid (20 ml) ma 80 ° C. no 4 mau hola.Ua ninini ʻia ka hui ʻana i loko o ka wai a ua ʻohi ʻia ka paʻakai ʻeleʻele e ka kānana.Ua hoʻomaʻemaʻe ʻia ka huahana i ʻelua manawa ma o ka hoʻoheheʻe ʻana i ka lepo e hāʻawi i kahi paʻa ʻōmaʻomaʻo (126.2 mg, 57% hua).anus.Ua helu ʻia no C26H32N6S: C 67.79, H 7.00, N 18.24.Loaʻa: 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.
Ua hoʻomākaukau a hoʻomaʻemaʻe ʻia nā mea hoʻohui ma muli o nā kaʻina hana e like me RuDA.anus.Ua helu ʻia no C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.Loaʻa: C, 61.32, H, 5.12, N, 11.81,1H NMR (600 MHz, d6-DMSO), δ (ppm) 10.19 (s, 2H), 9.28 (s, 2H), 8.09 (s, 2H), 7.95 (s, 4H), 6.93 (s, 4H), 6.48 (d, 2H), 6.34 (s, 2H), 3.54 (t, 8H), 2.80 (m, 1H), 2.33 (s, 3H), 1.31 (t, 12H), 1.07 (s, 6H).13c nmr (151 mhz, CDCL3), δ (PPM) 158.20, 153.36, 148.82, 148.14, 138.59, 136.79, 135.75, 134.71, 130.44, 128.87, 128.35, 121.70, 111.84, 110.76, 105.07, 104.23, 87.0, 84.4., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m/z [M-Cl]+ = 905.24.
Ua hoʻoheheʻe ʻia ʻo RuDA ma MeOH / H2O (5/95, v / v) ma kahi ʻano o 10 μM.Ua ana ʻia ka spectrum absorption o RuDA i kēlā me kēia 5 mau minuke ma kahi spectrophotometer Shimadzu UV-3600 ma lalo o ka irradiation me ke kukui laser me kahi hawewe o 808 nm (0.5 W/cm2).Ua hoʻopaʻa ʻia ka spectra ICG ma lalo o nā kūlana like me ka maʻamau.
Ua hoʻopaʻa ʻia ka spectra EPR ma kahi Bruker EMXmicro-6/1 spectrometer me ka mana microwave o 20 mW, kahi scanning range o 100 G, a me kahi modulation kahua o 1 G. 2,2,6,6-tetramethyl-4-piperidone (TEMP) a me 5,5-dimethyl-1-pyrroline N-oxide (DMPO) i hoʻohana ʻia e like me nā pahele wili.Ua hoʻopaʻa ʻia ka spectra electron spin resonance no nā hoʻonā hui ʻia o RuDA (50 µM) a me TEMF (20 mM) a i ʻole DMPO (20 mM) ma lalo o ka hana o ka radiation laser me kahi hawewe o 808 nm (0.5 W/cm2).
Ua hoʻokō ʻia nā helu DFT a me TD-DFT no RuDA ma PBE1PBE/6–31 G*//LanL2DZ pae i loko o ka hoʻonā wai e hoʻohana ana i ka papahana Gaussian 1666,67,68.Ua hoʻolālā ʻia ka HOMO-LUMO, ka lua a me nā puʻu electron o ka mokuʻāina ʻoliʻoli haʻahaʻa haʻahaʻa ʻo RuDA me ka hoʻohana ʻana i ka polokalamu GaussView (version 5.0).
Ua ho'āʻo mua mākou e ana i ka pono o ka hanauna o 1O2 RuDA me ka hoʻohanaʻana i ka spectroscopy UV-visible maʻamau me ICG (ΦΔ = 0.002) ma keʻano he maʻamau, akāʻo ka photodegradation o ICG i hoʻopilikia nui i nā hopena.No laila, ua ana ʻia ka hua nui o 1O2 RuDA ma ka ʻike ʻana i ka hoʻololi ʻana i ka ikaika o ka fluorescence ABDA ma kahi o 428 nm i ka wā i hoʻomālamalama ʻia me ka laser me ka nalu o 808 nm (0.5 W/cm2).Ua hana ʻia nā hoʻokolohua ma RuDA a me RuDA NPs (20 μM) i ka wai / DMF (98/2, v / v) i loaʻa iā ABDA (50 μM).Ua helu ʻia ka hua nui o 1O2 me ka hoʻohana ʻana i kēia ʻano: ΦΔ (PS) = ΦΔ (ICG) × (rFS/APS)/(rICG/AICG).ʻO rPS a me rICG nā helu pane o ABDA me 1O2 i loaʻa mai ka photosensitizer a me ICG.ʻO APS a me AICG ka absorbance o ka photosensitizer a me ICG ma 808 nm, kēlā me kēia.
Hana ʻia nā ana AFM i nā kūlana wai me ka hoʻohana ʻana i ke ʻano scan ma kahi ʻōnaehana Bruker Dimension Icon AFM.Me ka hoʻohana ʻana i kahi ʻano hāmama me nā pūnāwai wai, ua holoi ʻia nā keʻena me ka ethanol a maloʻo me kahi kahawai o ka nitrogen.E hoʻokomo i nā keena maloʻo i loko o ke poʻo opi o ka microscope.E kau koke i kahi kulu o ka laʻana i loko o ka wai wai a kau ma luna o ka cantilever me ka hoʻohana ʻana i kahi syringe plastic disposable a me kahi nila sterile.Hoʻokomo pololei ʻia kekahi hāʻule ma luna o ka hāpana, a i ka wā e hoʻohaʻahaʻa ʻia ai ke poʻo optical, hui pū nā kulu ʻelua, a lilo i meniscus ma waena o ka hāpana a me ka waihona wai.Ua hana ʻia nā ana AFM me ka SCANASYST-FLUID V-shaped nitride cantilever (Bruker, paʻakikī k = 0.7 N m-1, f0 = 120-180 kHz).
Loaʻa nā chromatograms HPLC ma kahi ʻōnaehana Waters e2695 i hoʻolako ʻia me kahi kolamu phoenix C18 (250 × 4.6 mm, 5 µm) me ka hoʻohana ʻana i kahi mea ʻike 2489 UV/Vis.ʻO 650 nm ka lōʻihi o ka nalu.He wai a me ka methanol nā māhele mobile A a me B, a ʻo ke kahe o ka pae kelepona he 1.0 ml·min-1.Penei ka gradient (solvent B): 100% mai 0 a 4 minuke, 100% a 50% mai 5 a 30 minuke, a ho'iho'i i 100% mai 31 a 40 minuke.Ua hoʻoheheʻe ʻia ka Ore i loko o kahi hopena hui ʻia o ka methanol a me ka wai (50/50, ma ka leo) ma kahi ʻano o 50 μM.ʻO ka nui o ka injection he 20 μl.
Ua hoʻopaʻa ʻia nā hōʻike GPC ma kahi mea hana Thermo ULTIMATE 3000 i hoʻolako ʻia me ʻelua kolamu PL aquagel-OH MIXED-H (2 × 300 × 7.5 mm, 8 µm) a me kahi mea ʻike kuhikuhi kuhikuhi refractive ERC RefratoMax520.Ua hoʻokahe ʻia ke kolamu GPC me ka wai ma kahi kahe kahe o 1 ml / min ma 30 ° C.Ua hoʻoheheʻe ʻia nā NP Ore i ka hopena PBS (pH = 7.4, 50 μM), ʻo 20 μL ka nui o ka injection.
Ua ana ʻia nā kiʻi kiʻi ma kahi hoʻonohonoho electrochemical (CHI-660B, Kina).ʻO nā pane optoelectronic i ka wā i hoʻohuli ʻia ai ka laser (808 nm, 0.5 W / cm2) ua ana ʻia i ka volta o 0.5 V i loko o kahi pahu ʻeleʻele.Ua hoʻohana ʻia kahi kelepona ʻekolu-electrode maʻamau me ka L-shaped glassy carbon electrode (GCE) ma ke ʻano he electrode hana, kahi calomel electrode maʻamau (SCE) ma ke ʻano he electrode kuhikuhi, a me kahi diski platinum ma ke ʻano he counter electrode.Ua hoʻohana ʻia kahi hopena 0.1 M Na2SO4 ma ke ʻano he electrolyte.
Ua kūʻai ʻia ka laina maʻi maʻi umauma kanaka MDA-MB-231 mai KeyGEN Biotec Co., LTD (Nanjing, Kina, helu helu: KG033).Ua ulu ʻia nā cell i nā monolayers ma Dulbecco's Modified Eagle's Medium (DMEM, high glucose) i hoʻohui ʻia me ka hopena o 10% fetal bovine serum (FBS), penicillin (100 μg/ml) a me streptomycin (100 μg/ml).Hoʻoulu ʻia nā cell a pau ma 37°C i loko o kahi ea me 5% CO2.
Ua hoʻohana ʻia ka MTT assay no ka hoʻoholo ʻana i ka cytotoxicity o RuDA a me RuDA-NPs i ke alo a me ka ʻole o ka irradiation māmā, me ka ʻole o Vc (0.5 mM).Ua ulu ka MDA-MB-231 i nā pā maʻi maʻi i loko o nā papa 96-well ma kahi kiko o ke kelepona ma kahi o 1 x 105 cell/ml/well a incubated no 12 mau hola ma 37.0 ° C i ka lewa o 5% CO2 a me 95% ea.Hoʻohui ʻia nā RuDA a me RuDA NP i hoʻoheheʻe ʻia i ka wai i nā cell.Ma hope o 12 mau hola o ka incubation, ua hōʻike ʻia nā cell i ka 0.5 W cm -2 laser radiation ma kahi hawewe o 808 nm no 10 mau minuke (300 J cm -2) a laila hoʻokomo ʻia i ka pōʻeleʻele no 24 mau hola.Hoʻopiliʻia nā pūnaewele me MTT (5 mg / ml) no nā hola 5'ē aʻe.ʻO ka hope loa, e hoʻololi i ka media i DMSO (200 µl) e hoʻopau i nā kristal formazan poni.Ua ana ʻia nā waiwai OD me ka mea heluhelu microplate me ka nalu o 570/630 nm.Ua helu ʻia ka waiwai IC50 no kēlā me kēia laʻana me ka hoʻohana ʻana i ka polokalamu SPSS mai nā ʻāpana pane-pane i loaʻa mai ʻekolu mau hoʻokolohua kūʻokoʻa.
Ua mālamaʻia nā pūnaewele MDA-MB-231 me RuDA a me RuDA-NP ma kahi o 50 μM.Ma hope o 12 mau hola o ka incubation, ua hoʻonāʻia nā pūnaewele me ka laser me ka nalu o 808 nm a me ka mana o 0.5 W / cm2 no 10 min (300 J / cm2).I loko o ka hui vitamina C (Vc), ua mālama ʻia nā cell me ka 0.5 mM Vc ma mua o ka hoʻomālamalama ʻana i ka laser.Hoʻopiliʻia nā pūnaewele i loko o ka pōʻeleʻele no nā hola 24 hou, a laila hoʻopaʻaʻia me ka calcein AM a me propidium iodide (20 μg / ml, 5 μl) no 30 mau minuke, a laila holoiʻia me ka PBS (10 μl, pH 7.4).nā kiʻi o nā pūnaewele ʻeleʻele.