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Gukoresha amafoto meza cyane ni ngombwa cyane cyane mugukoresha amavuriro menshi.Nyamara, fotosensiseri zisanzwe zibabazwa no kwinjizwa kwigihe gito, gufotora bidahagije, umusaruro muke wubwoko bwa ogisijeni ikora (ROS), hamwe no kuzimya ROS.Hano twatanze raporo hafi-ya-infragre (NIR) supramolecular photosensitizer (RuDA) yunganirwa no kwiteranya kwa Ru (II) -arene organometallic complexes mugisubizo cyamazi.RuDA irashobora kubyara gusa ogisijeni imwe (1O2) muri leta yegeranye, kandi irerekana imyitwarire igaragara-iterwa na 1O2 yimyitwarire bitewe nubwiyongere bugaragara mubikorwa byambukiranya hagati ya sisitemu imwe.Mubikorwa byumucyo wa laser ya 808 nm, RuDA yerekana umusaruro wa 1O2 wa kwant ya 16.4% (icyatsi cya indocyanine cyemewe na FDA: ΦΔ = 0.2%) hamwe nuburyo bwiza bwo guhinduranya amafoto ya 24.2% (nanorodo yubucuruzi ya zahabu) hamwe no gufotora neza.: 21.0%, nanoshell ya zahabu: 13.0%).Byongeye kandi, RuDA-NPs ifite biocompatibilité nziza irashobora gukusanyiriza hamwe ahantu h’ibibyimba, bigatuma ibibyimba bigaruka cyane mugihe cyo kuvura Photodynamic hamwe no kugabanuka kwa 95.2% muri vivo.Ubu buryo bwo kongera imbaraga bwa Photodynamic therapy butanga ingamba zo guteza imbere fotosensiseri hamwe na fotokopi nziza na fotokome.
Ugereranije nubuvuzi busanzwe, kuvura Photodynamic (PDT) nubuvuzi bushimishije kuri kanseri kubera ibyiza byayo nko kugenzura neza spatiotemporal, kudatera, kurwanya ibiyobyabwenge, ndetse no kugabanya ingaruka mbi 1,2,3.Mugihe cyo gucana urumuri, fotosensiseri ikoreshwa irashobora gukoreshwa kugirango habeho ubwoko bwa ogisijeni ikora cyane (ROS), biganisha kuri apoptose / necrosis cyangwa ibisubizo byubudahangarwa4,5. Nyamara, ibyinshi mubisanzwe bifotora, nka chlorine, porphirine, na anthraquinone, bifite uburyo bwo kwinjirira mu gihe gito (inshuro <680 nm), bityo bigatuma urumuri rwinjira nabi kubera kwinjiza cyane molekile yibinyabuzima (urugero, hemoglobine na melanine) muri akarere kagaragara6,7. Nyamara, ibyinshi mubisanzwe bifotora, nka chlorine, porphirine, na anthraquinone, bifite uburyo bwo kwinjirira mu gihe gito (inshuro <680 nm), bityo bigatuma urumuri rwinjira nabi kubera kwinjiza cyane molekile yibinyabuzima (urugero, hemoglobine na melanine) muri akarere kagaragara6,7. 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. Nyamara, ibyinshi bifotora nka chlorine, porphirine na anthraquinone bifite umuvuduko muke wo kwinjirira (<680 nm) bigatuma urumuri rwinjira nabi bitewe no kwinjiza cyane molekile yibinyabuzima (urugero nka hemoglobine na melanine) mukarere kagaragara6,7.然而 ， 大多数 传统 的 光敏剂 ， 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6导致 光 穿透性 差。然而 ， 大多数 传统 的 ， ， 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6吸收 吸收 吸收 吸收 吸收 I HI 导致 光 穿透性 差。 Однако большинстныныных фотосенсибилизов, таких как х аины, порфирин и ант киноиноны, имеют относительно коротковолоо поглощение (частота <680 нм) из-за слного поглог? Nyamara, ibyinshi mubifotora nka chlorine, porphirine na anthraquinone bifite umuvuduko muke wo kwinjirira (inshuro <680 nm) bitewe no kwinjiza cyane biomolecules nka hemoglobine na melanine bigatuma urumuri rwinjira nabi.Agace kagaragara 6.7.Kubwibyo, hafi-ya-infragre (NIR) ikurura fotosensiseri ikoreshwa muri 700-900 nm “idirishya ryo kuvura” ikwiranye no gufotora.Kubera ko hafi yumucyo wa infragre ariwo udakoreshwa cyane nuduce twibinyabuzima, birashobora gutuma umuntu yinjira cyane kandi fotodamage 8,9.
Kubwamahirwe make, ibyuma bifata amafoto ya NIR isanzwe ifite ubusanzwe ifotora nabi, ogisijeni nkeya (1O2) itanga ingufu, hamwe no kuzimya 1O2 kuzimya, bigabanya imiti yabo10,11.Nubwo hashyizweho ingufu nyinshi kugirango tunonosore imiterere ya fotofiziki na fotokimiki yifotozi isanzwe, kugeza ubu raporo nyinshi zagiye zivuga ko abakora amafoto ya NIR bashobora gukemura ibyo bibazo byose.Byongeye kandi, amafoto menshi yerekana amafoto yerekana amasezerano yo kubyara neza 1O212,13,14 mugihe akayangana numucyo uri hejuru ya 800 nm, kubera ko ingufu za fotone zigabanuka vuba mukarere kegereye IR.Triphenylamine (TFA) nkumuterankunga wa electron na [1,2,5] thiadiazole- [3,4-i] dipyrido y'amabara, akurura hafi-ya-infragre, yakozweho ubushakashatsi bwimbitse kuri bioimaging ya hafi ya infrarafaride ya II hamwe nubuvuzi bwa Photothermal therapy (PTT) kubera kwifata kwabo.Rero, amarangi yo mu bwoko bwa DA arashobora gukoreshwa kuri PDT hamwe no kwishima hafi ya IR, nubwo bidakunze kwigwa nkamafoto ya PDT.
Birazwi neza ko imikorere ihanitse yo kwambuka intersystem (ISC) ya fotosensiseri iteza imbere gushiraho 1O2.Ingamba zihuriweho zo guteza imbere gahunda ya ISC ni ukuzamura uruziga ruzenguruka (SOC) rwamafoto yerekana uburyo bwo gutangiza atome ziremereye cyangwa imiterere yihariye.Nyamara, ubu buryo buracyafite imbogamizi nimbogamizi19,20.Vuba aha, guterana kwa supramolecular byatanze uburyo bwo hejuru-bwubwenge bwo guhimba ibikoresho bikora kurwego rwa molekile, 21,22 hamwe nibyiza byinshi mumafoto yubuvuzi:Bisa nuburyo bwa elegitoronike hamwe no gukwirakwiza urwego rwingufu bitewe no kuzenguruka kuzenguruka hagati yububiko.Kubwibyo, umukino wingufu hagati ya reta imwe yishimye (S1) hamwe na leta itatu yishimye (Tn) izanozwa, ifitiye akamaro gahunda ya ISC 23, 24...Urebye ibyiza byavuzwe haruguru, twizera ko sisitemu ya supramolecular ya fotosensitizer ishobora kuba ubundi buryo butanga ikizere cyo gutsinda ibitagenda neza bya PDT.
Ibigo bishingiye kuri Ru (II) ni urubuga rwubuvuzi rwizewe rushobora gukoreshwa mugupima no kuvura indwara bitewe nibinyabuzima byihariye kandi bishimishije 28,29,30,31,32,33,34.Byongeye kandi, ubwinshi bwibihugu byishimye hamwe na fotofisiki ya chimique ihindagurika ya Ru (II) ishingiye ku nyubako zitanga inyungu nini zo guteza imbere Ru (II) ishingiye ku mafoto 35,36,37,38,39,40.Urugero rugaragara ni rutheniyumu (II) polypyridyl complex TLD-1433, kuri ubu iri mu bigeragezo byo mu cyiciro cya kabiri cy’amavuriro nka fotosensitiferi yo kuvura kanseri y’uruhago idatera imitsi (NMIBC) 41.Byongeye kandi, ruthenium (II) arene organometallic complexes ikoreshwa cyane nka chimiotherapeutic miti yo kuvura kanseri kubera uburozi buke kandi byoroshye guhinduka 42,43,44,45.Imiterere ya ionic ya Ru (II) -arene organometallic complexes ntishobora gusa kunoza imikoreshereze mibi ya chromofore ya DA mumashanyarazi asanzwe, ahubwo inatezimbere inteko ya chromofore.Byongeye kandi, pseudooctahedral igice cya sandwich cyubatswe ningingo ngengabuzima ya Ru (II) -arenes irashobora gukumira byimazeyo H-guteranya chromofore yo mu bwoko bwa DA, bityo bikorohereza ishyirwaho rya J-guteranya hamwe na bande itukura.Nyamara, ibibi byavutse bya Ru (II) -ibigo byose, nkumutekano muke hamwe na / cyangwa bioavailable nke, birashobora kugira ingaruka kumiti no mubikorwa bya vivo byibigo bya arene-Ru (II).Nyamara, ubushakashatsi bwerekanye ko izo ngaruka zishobora kuneshwa no gukwirakwiza rutheniyumu hamwe na polymers ya biocompatible polymers hamwe na encapsulation physique cyangwa covalent conjugation.
Muri iki gikorwa, turatanga raporo ya DA-conjugated complexes ya Ru (II) -arene (RuDA) hamwe na NIR imbarutso binyuze mu guhuza ibikorwa hagati ya DAD chromofore na Ru (II) -arene.Inganda zavuyemo zirashobora kwiteranya muri vitosupramolecular viticles mumazi kubera imikoranire idahwitse.Ikigaragara ni uko inteko ya supramolecular yahaye RuDA imitungo ya polymerisime iterwa na intersystem yambukiranya imipaka, ibyo bikaba byongereye cyane imikorere ya ISC, byari byiza cyane kuri PDT (Ishusho 1A).Kugirango hongerwe ikibyimba no muri vivo biocompatibilité, Pluronic F127 yemewe na FDA (PEO-PPO-PEO) yakoreshejwe mugukingira RuDA 47,48,49 kugirango habeho nanoparticles ya RuDA-NP (Igicapo 1B) ikora nka PDT / Dual- uburyo bwa PTT.Mu kuvura kanseri (Ishusho 1C), RuDA-NP yakoreshejwe mu kuvura imbeba zambaye ubusa hamwe n’ibibyimba bya MDA-MB-231 kugira ngo bige ku mikorere ya PDT na PTT muri vivo.
Igishushanyo mbonera cyerekana uburyo bwo gufotora bwa RuDA muburyo bwa monomeric kandi bukusanyirijwe hamwe bwo kuvura kanseri, synthesis ya B RuDA-NPs na C RuDA-NPs kuri PDT na PTT ikora NIR.
RuDA, igizwe n'imikorere ya TPA na TDP, yateguwe hakurikijwe uburyo bwerekanwe ku gishushanyo cya 1 (Ishusho 2A), kandi RuDA yaranzwe na 1H na 13C NMR yerekanwe, electrospray ionisation mass spectrometrie, hamwe nisesengura ryibanze (Ishusho yinyongera 2-4 ).Ikarita ya RuDA itandukanya ikarita yo hasi yinzibacyuho yo hasi cyane yabazwe nigitekerezo gishingiye ku gihe gishingiye ku gihe cyimikorere (TD-DFT) kugirango yige uburyo bwo kohereza amafaranga.Nkuko bigaragara mu gishushanyo cya 5, ubwinshi bwa elegitoronike buva cyane cyane kuri triphenylamine kugera muri TDP yakira nyuma yo gufotora, ibyo bikaba bishobora guterwa ninzibacyuho yimikorere idasanzwe (CT).
Imiterere yimiti ya Ore B Absorption spectra ya Ore muruvange rwimibare itandukanye ya DMF namazi.C Indangagaciro zisanzwe zo kwinjiza RuDA (800 nm) na ICG (779 nm) ugereranije nigihe kuri 0.5 W cm-2 ya 808 nm urumuri rwa laser.D Photodegradation ya ABDA yerekanwa na RuDA iterwa na 1O2 ivanze na DMF / H2O ivanze nibintu bitandukanye byamazi bitewe nigikorwa cyimirasire ya laser ifite uburebure bwa 808 nm nimbaraga za 0.5 W / cm2.
Abstract-UV-igaragara yo kwinjiza spekitroscopi yakoreshejwe mu kwiga imiterere-yo guteranya ya Ore ivanze na DMF n'amazi mubice bitandukanye.Nkuko bigaragara ku gishushanyo.2B, RuDA yerekana imirongo ikurura kuva kuri 600 kugeza 900 nm muri DMF hamwe nigitigiri kinini cyo kwinjiza kuri 729 nm.Kongera umubare wamazi byatumye buhoro buhoro ihinduka ryumutuku wa Ore ntarengwa igera kuri 800 nm, ibyo bikaba byerekana J-guteranya Ore muri sisitemu yateranijwe.Photoluminescence yerekanwe ya RuDA mumashanyarazi atandukanye irerekanwa mugice cyinyongera 6. RuDA isa niyerekana luminescence isanzwe ya NIR-II hamwe nuburebure ntarengwa bwa ca.1050 nm muri CH2Cl2 na CH3OH.Ihinduka rinini rya Stoke (hafi 300 nm) ya RuDA ryerekana impinduka zikomeye muri geometrie ya leta yishimye no gushiraho leta zifite ingufu nke.Umusaruro wa luminescence wa Ore muri CH2Cl2 na CH3OH wariyemeje kuba 3,3 na 0,6%.Nyamara, mu ruvange rwa methanol n'amazi (5/95, v / v), hagaragaye ihinduka rito ry'ibyuka bihumanya ikirere no kugabanuka k'umusaruro wa kwant (0.22%), ibyo bikaba bishobora guterwa no kwishyira hamwe kwa Ore .
Kugirango twiyumvire kwishyira hamwe kwa ORE, twakoresheje ingufu za atome imbaraga za microscopi (AFM) kugirango tumenye impinduka zijyanye na morfologiya muri ORE mumuti wa methanol nyuma yo kongeramo amazi.Iyo amazi yari munsi ya 80%, nta guteranya kugaragara byagaragaye (Ishusho yinyongera).Ariko, hamwe n’ubwiyongere bw’amazi bugera kuri 90-95%, hagaragaye nanoparticles ntoya, yerekanaga kwishyira hamwe kwa Ore. Byongeye kandi, imirasire ya laser ifite uburebure bwa 808 nm ntabwo byagize ingaruka ku gukomera kwa RuDA mu mazi. igisubizo (Igishusho 2C hamwe ninyongera Igishusho 8).Ibinyuranye, kwinjiza icyatsi cya indocyanine (ICG nkigenzura) byagabanutse vuba kuri 779 nm, byerekana ifoto nziza ya RuDA.Byongeye kandi, ituze rya RuDA-NPs muri PBS (pH = 5.4, 7.4 na 9.0), 10% FBS na DMEM (glucose ndende) ryasuzumwe na UV-igaragara ya sisitemu ya sprosroscopi mu bihe bitandukanye.Nkuko bigaragara mu gishushanyo cya 9, impinduka nke mumashanyarazi ya RuDA-NP yagaragaye muri PBS kuri pH 7.4 / 9.0, FBS na DMEM, byerekana ko RuDA-NP ihagaze neza.Nyamara, muburyo bwa acide (рН = 5.4) hydrolysis ya Ore yabonetse.Twasuzumye kandi ituze rya RuDA na RuDA-NP dukoresheje uburyo bwo hejuru bwa chromatografiya (HPLC).Nkuko bigaragara mu gishushanyo cya 10, RuDA yari ituje mu ruvange rwa methanol n’amazi (50/50, v / v) ku isaha ya mbere, kandi hydrolysis yagaragaye nyuma y’amasaha 4.Nyamara, impinga nini gusa ya convex yagaragaye kuri RuDA NPs.Kubwibyo, gel permeation chromatografiya (GPC) yakoreshejwe mugusuzuma ituze rya RuDA NPs muri PBS (pH = 7.4).Nkuko bigaragara mu gishushanyo cya 11, nyuma yamasaha 8 yubushakashatsi mubihe byageragejwe, uburebure bwimpinga, ubugari bwimisozi nubuso bwa NP RuDA ntabwo bwahindutse cyane, byerekana ko NP RuDA ihagaze neza.Byongeye kandi, amashusho ya TEM yerekanaga ko morphologie ya nanoparticles ya RuDA-NP itigeze ihinduka nyuma yamasaha 24 muri buffer ya PBS (pH = 7.4, Ishusho yinyongera 12).
Kuberako kwishyira hamwe bishobora gutanga imikorere itandukanye nubumara kuri Ore, twabonye irekurwa rya 9,10-anthracenediylbis (methylene) dimalonic aside (ABDA, icyerekezo 1O2) mvange ya methanol-amazi.Ore ifite amazi atandukanye50.Nkuko bigaragara ku gishushanyo cya 2D n’inyongera Igishushanyo cya 13, nta gutesha agaciro ABDA byagaragaye igihe amazi yari munsi ya 20%.Hamwe n'ubwiyongere bw'ubushyuhe bugera kuri 40%, iyangirika rya ABDA ryabaye, nkuko bigaragazwa no kugabanuka k'uburemere bwa fluorescence ya ABDA.Byagaragaye kandi ko amazi menshi atera kwangirika vuba, byerekana ko kwiteranya kwa RuDA ari ngombwa kandi bifite akamaro mu kwangirika kwa ABDA.Iyi phenomenon iratandukanye cyane na chromofores ya ACQ igezweho.Iyo iraswe na lazeri ifite uburebure bwa 808 nm, umusaruro wa kwant wa 1O2 RuDA muvanga 98% H2O / 2% DMF ni 16.4%, ibyo bikaba bikubye inshuro 82 ugereranije na ICG (ΦΔ = 0.2%) 51, kwerekana imikorere idasanzwe yibisekuruza 1O2 RuDA muburyo bwo kwegeranya.
Electron izunguruka ikoresheje 2,2,6,6-tetramethyl-4-piperidinone (TEMP) na 5,5-dimethyl-1-pyrroline N-oxide (DMPO) nk'imitego izunguruka Resonance spectroscopy (ESR) yakoreshejwe kugirango hamenyekane amoko yavuyemo AFK.na RuDA.Nkuko bigaragara mu gishushanyo cya 14, byemejwe ko 1O2 ikorwa mugihe cyo kurasa hagati yiminota 0 na 4.Byongeye kandi, igihe RuDA yashizwemo na DMPO mu gihe cyo kurasa, hagaragaye ibimenyetso bine by'imirongo ine ya EPR ya 1: 2: 2: 1 DMPO-OH · inyongeramusaruro, byerekana ko habayeho hydroxyl radicals (OH ·).Muri rusange, ibisubizo byavuzwe haruguru byerekana ubushobozi bwa RuDA yo kuzamura umusaruro wa ROS binyuze muburyo bubiri bwa I / II bwo gufotora.
Kugirango usobanukirwe neza imiterere ya elegitoronike ya RuDA muburyo bwa monomeric kandi yegeranijwe, imipaka ya molekulire ya orbitale ya RuDA muburyo bwa monomeric na dimeric yabazwe hakoreshejwe uburyo bwa DFT.Nkuko bigaragara ku gishushanyo.3A, molekulire ya orbital nini cyane (HOMO) ya monomeric RuDA irekuwe kuruhande rwumugongo wa ligand kandi orbital yo hasi cyane idakoreshwa cyane (LUMO) ishingiye kumurongo wakira TDP.Ibinyuranye, ubucucike bwa electron muri dimeric HOMO bwibanze kuri ligand ya molekile imwe ya RuDA, mugihe ubucucike bwa electron muri LUMO bwibanda cyane kubice byakira indi molekile ya RuDA, byerekana ko RuDA iri mubihe.Ibiranga CT.
A HOMO na LUMO ya Ore ibarwa muburyo bwa monomeric na dimeric.B Urwego rumwe ningufu eshatu za Ore muri monomers na dimers.C Ikigereranyo cyurwego rwa RuDA nibishoboka ISC imiyoboro nka monomeric C na dimeric D. Imyambi yerekana imiyoboro ishoboka ya ISC.
Ikwirakwizwa rya electron nu mwobo muburyo buke bwimbaraga za leta ya RuDA muburyo bwa monomeric na dimeric bwasesenguwe hakoreshejwe software ya Multiwfn 3.852.53, yabazwe hakoreshejwe uburyo bwa TD-DFT.Nkuko bigaragara kuri label yinyongera.Nkuko bigaragara ku gishushanyo cya 1-2, umwobo wa RDA monomeric usanga ahanini ucibwa mu mugongo wa ligand muri ibi bihugu byishimye, mu gihe electron ziba ziri mu itsinda rya TDP, zikerekana imiterere ya CT.Mubyongeyeho, kuri ibi bihugu byishimye, hariho byinshi cyangwa bike byuzuzanya hagati y’imyobo na electron, byerekana ko ibyo bihugu byishimye bitanga umusanzu uturutse ku byishimo byaho (LE).Ku bipimo, usibye ibiranga CT na LE biranga, igice runaka cyimiterere ya CT cyaragaragaye muri leta zibishinzwe, cyane cyane S3, S4, S7, na S8, hashingiwe ku isesengura rya CT hagati, hamwe ninzibacyuho ya CT nkibyingenzi. (Imbonerahamwe y'inyongera).3).
Kugirango dusobanukirwe neza ibisubizo byubushakashatsi, twakomeje gushakisha imiterere ya RuDA yishimye kugirango tumenye itandukaniro riri hagati ya monomers na dimers (Imbonerahamwe yinyongera 4-5).Nkuko bigaragara ku gishushanyo cya 3B, urwego rwingufu za single na triplet zishimishije za dimer ziba nyinshi cyane ugereranije na monomer, ifasha kugabanya icyuho cyingufu hagati ya S1 na Tn. Byavuzwe ko inzibacyuho ya ISC zishobora kugerwaho mu cyuho gito (ΔES1-Tn <0.3 eV) hagati ya S1 na Tn54. Byavuzwe ko inzibacyuho ya ISC zishobora kugerwaho mu cyuho gito (ΔES1-Tn <0.3 eV) hagati ya S1 na Tn54. Сообщалось, что переходы ISC могут быть реданованя в пределах небольшой ишгетической щели (ΔES1-Tn <0,3 эВ) между S1 na Tn54. Byatangajwe ko inzibacyuho ya ISC zishobora kugerwaho mu cyuho gito (ΔES1-Tn <0.3 eV) hagati ya S1 na Tn54.据 报道 ， ISC 跃迁 可以 在 S1 和 Tn54 之间 的 小 能 隙 Δ 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 Δ ΔES1-Tn <0.3 eV） 内 实现。 Сообщалось, что переход ISC может быть реданован в пределах небольшой ишгетической щели (ΔES1-Tn <0,3 эВ) между S1 и Tn54. Byatangajwe ko inzibacyuho ya ISC ishobora kugerwaho mugihe gito cyingufu (ΔES1-Tn <0.3 eV) hagati ya S1 na Tn54.Mubyongeyeho, orbital imwe gusa, itwarwa cyangwa idatuwe, igomba gutandukana muburyo bumwe hamwe na leta eshatu kugirango itange ibice bitari zeru SOC.Rero, hashingiwe ku isesengura ryingufu zishimishije ninzibacyuho ya orbital, inzira zose zishoboka zinzibacyuho ya ISC zerekanwa mubishushanyo.3C, D.Ikigaragara ni uko umuyoboro umwe gusa wa ISC uraboneka muri monomer, mugihe ifishi ya dimeric ifite imiyoboro ine ISC ishobora kuzamura inzibacyuho ya ISC.Kubwibyo, birakwiye gutekereza ko uko molekile ya RuDA yegeranijwe, niko imiyoboro ya ISC izagerwaho.Kubwibyo, igiteranyo cya RuDA kirashobora gukora ibice bibiri bya elegitoroniki muburyo bumwe kandi butatu, kugabanya itandukaniro ryingufu hagati ya S1 na Tn iboneka, bityo bikongera imikorere ya ISC kugirango byorohereze 1O2.
Kugirango turusheho gusobanura uburyo bwibanze, twashushanyijeho ibice byerekeranye na arene-Ru (II) complexe (RuET) dusimbuza amatsinda abiri ya Ethyl hamwe nitsinda rya triphenylamine fenil muri RuDA (Igishusho 4A, kugirango ubiranga byuzuye, reba ESI, Umugereka wa 15 -21) Kuva ku baterankunga (diethylamine) kugeza kubakira (TDF), RuET ifite imiterere ya CT imeze nka RuDA.Nkuko byari byitezwe, uburyo bwo kwinjiza RuET muri DMF bwerekanye umurongo muke wohereza ingufu hamwe no kwinjiza cyane mukarere kegereye infragre mu karere ka nm 600-100 (Ishusho 4B).Byongeye kandi, igiteranyo cya RuET nacyo cyagaragaye hamwe n’amazi yiyongera, ibyo bikaba byaragaragaye mu itukura ry’imisemburo ntarengwa, ibyo bikaba byarashimangiwe n’amashusho y’amazi ya AFM (Ishusho yinyongera 22).Ibisubizo byerekana ko RuET, kimwe na RuDA, ishobora gukora imiterere idasanzwe no kwishyira hamwe muburyo rusange.
Imiterere yimiti ya RuET.B Absorption spectra ya RuET muruvange rwimibare itandukanye ya DMF namazi.Ibibanza C EIS Nyquist kuri RuDA na RuET.Ibisubizo bifotora D ya RuDA na RuET munsi yumuriro wa laser hamwe nuburebure bwa 808 nm.
Photodegradation ya ABDA imbere ya RuET yasuzumwe na irrasiyo hamwe na laser ifite uburebure bwa 808 nm.Igitangaje ni uko nta gutesha agaciro ABDA byagaragaye mu bice bitandukanye by'amazi (Ishusho y'inyongera 23).Impamvu ishoboka nuko RuET idashobora gukora neza uburyo bwa elegitoronike ihujwe kuko urunigi rwa Ethyl rudateza imbere ihererekanyabubasha ryimikorere ya intermolecular.Kubwibyo rero, hakozwe ibipimo bya electrochemical impedance spectroscopy (EIS) hamwe nipima ryigihe gito ryamafoto yakozwe kugirango ugereranye imiterere ya foto ya elegitoroniki ya RuDA na RuET.Dukurikije umugambi wa Nyquist (Igicapo 4C), RuDA yerekana radiyo ntoya kuruta RuET, bivuze ko RuDA56 ifite umuvuduko wa elegitoronike ya elegitoronike kandi ikagenda neza.Byongeye kandi, ubwinshi bwamafoto ya RuDA buri hejuru cyane ugereranije na RuET (Igishusho 4D), byemeza uburyo bwiza bwo kohereza amafaranga ya RuDA57.Rero, fenyl groupe ya triphenylamine muri Ore igira uruhare runini mugutanga imiyoboro yimikorere no gushiraho ibikoresho bya elegitoroniki.
Kugirango twongere ibibyimba no muri vivo biocompatibilité, twakomeje RuDA hamwe na F127.Impuzandengo ya diametre ya hydrodynamic ya RuDA-NPs yariyemeje kuba 123.1 nm hamwe no kugabura kugufi (PDI = 0.089) hakoreshejwe uburyo bwo gukwirakwiza urumuri (DLS) (Igicapo 5A), cyateje imbere ikibyimba cyiyongera kandi kigakomeza.EPR) ingaruka.Amashusho ya TEM yerekanaga ko Ore NPs ifite imiterere imwe ya sherfike ifite impuzandengo ya diameter ya 86 nm.Ikigaragara ni uko umubare ntarengwa wa RuDA-NPs wagaragaye kuri 800 nm (Ishusho yinyongera 24), byerekana ko RuDA-NPs ishobora kugumana imikorere numutungo wo guteranya RuDAs.Umusaruro wa ROS ubarwa kuri NP Ore ni 15.9%, ugereranije na Ore.Imiterere yumuriro wa RuDA NPs yakozwe hifashishijwe imishwarara ya laser ifite uburebure bwa 808 nm ikoresheje kamera ya infragre.Nkuko bigaragara ku gishushanyo.5B, C, itsinda rishinzwe kugenzura (PBS gusa) ryiyongereyeho ubushyuhe buke, mugihe ubushyuhe bwumuti wa RuDA-NPs bwiyongereye vuba hamwe nubushyuhe bwiyongera (ΔT) bugera kuri 15.5, 26.1, na 43.0 ° C.Ubushuhe bwinshi bwari 25, 50, na 100 µM, byerekana ingaruka zikomeye zifotora ya RuDA NPs.Hiyongereyeho, ibipimo byo gushyushya / gukonjesha byafashwe kugirango hasuzumwe ingufu za RuDA-NP no kugereranya na ICG.Ubushyuhe bwa Ore NPs ntabwo bwagabanutse nyuma yinshuro eshanu zo gushyushya / gukonjesha (Igicapo 5D), cyerekana uburyo bwiza bwo gufotora bwa Ore NPs.Ibinyuranye na byo, ICG igaragaza ubushyuhe buke bwo gufotora nkuko bigaragara kuva ibura ryibura ryubushyuhe bwa fototerique mubihe bimwe.Ukurikije uburyo bwabanjirije 58, imikorere yo guhindura amafoto (PCE) ya RuDA-NP yabazwe nka 24.2%, ikaba isumba ibikoresho bisanzwe bifotora nka nanorodo ya zahabu (21.0%) na nanoshell ya zahabu (13.0%) 59.Rero, NP Ore yerekana ibintu byiza cyane bifotora, bigatuma basezerana abakozi ba PTT.
Isesengura rya DLS na TEM amashusho ya RuDA NPs (inset).B Amashusho yubushyuhe yibintu bitandukanye bya RuDA NPs ihura nimirasire ya laser kumuraba wa 808 nm (0.5 W cm-2).C Photothermal ihinduranya imirongo yibice bitandukanye byamabuye ya NP, namakuru yumubare.B. D Ubushyuhe bwiyongera bwa ORE NP na ICG hejuru yubushyuhe 5 bwo gukonjesha.
Photocytotoxicity ya RuDA NPs irwanya MDA-MB-231 ingirangingo za kanseri y'ibere ya muntu yasuzumwe muri vitro.Nkuko bigaragara ku gishushanyo.6A, B, RuDA-NPs na RuDA bagaragaje cytotoxicitike idahwitse mugihe hatabayeho imirasire, bivuze uburozi bwijimye bwa RuDA-NPs na RuDA.Nyamara, nyuma yo guhura nimirasire ya lazeri ku burebure bwa 808 nm, RuDA na RuDA NPs berekanye imbaraga za Photocytotoxicitike zirwanya kanseri ya MDA-MB-231 ifite agaciro ka IC50 (igice cya kabiri ntarengwa cyo kubuza) ingana na 5.4 na 9.4 μM. ko RuDA-NP na RuDA bifite ubushobozi bwo gufotora kanseri.Byongeye kandi, Photocytotoxicity ya RuDA-NP na RuDA yarushijeho gukorwaho iperereza imbere ya vitamine C (Vc), icyuma cya ROS, kugira ngo isobanure uruhare rwa ROS muri cytotoxicite iterwa n'umucyo.Ikigaragara ni uko imbaraga za selile ziyongereye nyuma yo kongerwaho Vc, kandi indangagaciro za IC50 za RuDA na RuDA NP zari 25.7 na 40.0 μM, ibyo bikaba byerekana uruhare rukomeye rwa ROS mu mafoto ya RuDA na RuDA NPs.Cytotoxicity iterwa n'umucyo wa RuDA-NPs na RuDA muri selile ya MDA-MB-231 ukoresheje selile nzima / yapfuye ikoresheje calcein AM (icyatsi kibisi cya fluorescence ku ngirabuzimafatizo nzima) na iyode ya propidium (PI, fluorescence itukura ku ngirabuzimafatizo zapfuye).byemejwe na selile) nka fluorescent probe.Nkuko bigaragara ku gishushanyo cya 6C, selile zavuwe na RuDA-NP cyangwa RuDA zagumye kubaho neza nta kurasa, nkuko bigaragazwa na fluorescence ikabije.Ibinyuranye na byo, hifashishijwe imirasire ya laser, hagaragaye gusa fluorescence itukura, ibyo bikaba byemeza ko Photocytotoxicity ya RuDA cyangwa RuDA NPs.Birashimishije kubona fluorescence yicyatsi yagaragaye nyuma yo kongerwaho Vc, byerekana kurenga ku mafoto ya Photocytotoxicity ya RuDA na RuDA NPs.Ibisubizo birahuye na vitro Photocytotoxicity assays.
Dose-iterwa nubuzima bwa selile ya RuDA- na B RuDA-NP muri selile MDA-MB-231 imbere ya Vc (0.5 mM) cyangwa idahari.Utubari twibeshya, bivuze iation gutandukana bisanzwe (n = 3). Kudakorwa, ibizamini byimpande zombi * p <0.05, ** p <0.01, na *** p <0.001. Kudakorwa, ibizamini byimpande zombi * p <0.05, ** p <0.01, na *** p <0.001. Непарные дву Кононние t-критерии * p <0,05, ** p <0,01 и *** p <0,001. T-ibipapuro bibiri-umurizo t-ibizamini * p <0.05, ** p <0.01, na *** 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 дву Кононние t-Тыы * p <0,05, ** p <0,01 и *** p <0,001. T-ibipapuro bibiri-umurizo t-ibizamini * p <0.05, ** p <0.01, na *** p <0.001.C Kubaho / gupfira ingirabuzimafatizo ukoresheje calcein AM na iyode ya propidium nka fluorescent.Umwanya munini: 30 µm.Amashusho yerekana amashusho atatu asubiramo buri tsinda arerekanwa.D Amashusho ya fluorescence yerekana umusaruro wa ROS muri selile MDA-MB-231 mubihe bitandukanye byo kuvura.Icyatsi DCF fluorescence yerekana ko ROS ihari.Kurasa na lazeri ifite uburebure bwa 808 nm n'imbaraga za 0.5 W / cm2 muminota 10 (300 J / cm2).Umwanya munini: 30 µm.Amashusho yerekana amashusho atatu asubiramo buri tsinda arerekanwa.E Flow cytometrie RuDA-NPs (50 µM) cyangwa RuDA (50 µM) isesengura ryokuvura hamwe na 808 nm lazeri (0.5 W cm-2) imbere ya Vc (0.5 mM) idahari.Amashusho yerekana amashusho atatu asubiramo buri tsinda arerekanwa.F Nrf-2, HSP70 na HO-1 ya selile MDA-MB-231 ivurwa na RuDA-NPs (50 µM) hamwe na 808 nm ya lazeri ya lazeri (0.5 W cm-2, min 10, 300 J cm-2), selile zigaragaza 2).Amashusho yerekana amashusho abiri asubiramo muri buri tsinda arerekanwa.
Umusemburo wa ROS mu ngirabuzimafatizo za MDA-MB-231 wasuzumwe hifashishijwe diacetate ya 2,7-dichlorodihydrofluorescein (DCFH-DA).Nkuko bigaragara ku gishushanyo.6D, selile zavuwe na RuDA-NPs cyangwa RuDA zerekanaga florescence zitandukanye zicyatsi iyo zishushe hamwe na lazeri 808 nm, byerekana ko RuDA-NPs na RuDA bifite ubushobozi bunoze bwo kubyara ROS.Ibinyuranye na byo, mu gihe nta mucyo uhari cyangwa imbere ya Vc, hagaragaye gusa ibimenyetso bya fluorescent nkeya ya selile byagaragaye, byerekana ko ROS yabayeho gato.Urwego ROS rudasanzwe muri selile RuDA-NP na RuDA ivurwa na MDA-MB-231 selile zagenwe na cytometrike.Nkuko bigaragara mu gishushanyo cya 25, ubukana bwa fluorescence (MFI) bwakozwe na RuDA-NPs na RuDA munsi ya 808 nm ya lazeri ya lazeri yiyongereye cyane inshuro zigera kuri 5.1 na 4.8, ugereranije nitsinda rishinzwe kugenzura, byemeza ko ari AFK.ubushobozi.Nyamara, urwego ROS rudasanzwe muri RuDA-NP cyangwa MDA-MB-231 selile zavuwe na RuDA zagereranijwe gusa no kugenzura nta lazeri ya lazeri cyangwa imbere ya Vc, bisa n'ibisubizo by'isesengura rya fluorescence.
Byerekanwe ko mitochondria ari yo ntego nyamukuru ya Ru (II) -arene complexes60.Kubwibyo, iperereza ryibanze rya RuDA na RuDA-NPs ryarakozweho iperereza.Nkuko bigaragara mu gishushanyo cya 26, RuDA na RuDA-NP byerekana imyirondoro isa nogukwirakwiza ingirabuzimafatizo hamwe na mitochondriya (62.5 ± 4.3 na 60.4 ± 3,6 ng / mg proteine).Nyamara, umubare muto wa Ru wabonetse mu bice bya kirimbuzi bya Ore na NP Ore (3.5 na 2,1%).Igice cya selile gisigaye cyarimo rutheniyumu isigaye: 31.7% (30.6 ± 3,4 ng / mg proteine) kuri RuDA na 42.9% (47.2 ± 4.5 ng / mg proteine) kuri RuDA-NPs.Muri rusange, Ore na NP Ore birundanya cyane muri mitochondria.Kugirango dusuzume imikorere mibi ya mitochondrial, twakoresheje JC-1 na MitoSOX Umutuku wirabura kugirango dusuzume ubushobozi bwa mitochondrial membrane nubushobozi bwa superoxide.Nkuko bigaragara ku gishushanyo cya 27, icyatsi kibisi (JC-1) na fluorescence itukura (MitoSOX Red) byagaragaye mu tugari twavuwe na RuDA na RuDA-NPs munsi ya 808 nm ya lazeri, byerekana ko RuDA na RuDA-NPs fluorescent nyinshi Irashobora gutera neza mitochondrial membrane depolarisation hamwe numusaruro wa superoxide.Byongeye kandi, uburyo bwo gupfa kwakagari bwagenwe hifashishijwe isesengura rya cytometrie ishingiye kuri annexine V-FITC / iyode ya propidium (PI).Nkuko bigaragara ku gishushanyo cya 6E, iyo ikozwe na lazeri 808 nm, RuDA na RuDA-NP byatumye ubwiyongere bukabije bwa apoptose hakiri kare (hepfo iburyo bwa quadrant) muri selile MDA-MB-231 ugereranije na PBS cyangwa PBS wongeyeho laser.ingirabuzimafatizo.Ariko, igihe Vc yongeyeho, igipimo cya apoptose ya RuDA na RuDA-NP cyaragabanutse cyane kuva kuri 50.9% na 52.0% kugera kuri 15.8% na 17.8%, ibyo bikaba byemeza uruhare rukomeye rwa ROS mugufotora kwa RuDA na RuDA-NP..Byongeye kandi, ingirabuzimafatizo nkeya zagaragaye mu matsinda yose yapimwe (hejuru y’ibumoso hejuru), byerekana ko apoptose ishobora kuba uburyo bwiganje mu rupfu rwatewe na RuDA na RuDA-NPs.
Kubera ko kwangirika kwa okiside ari ikintu gikomeye kigena apoptose, ibintu bya kirimbuzi bifitanye isano na erythroide 2, ingingo ya 2 (Nrf2) 62, igenzura rikomeye rya sisitemu ya antioxydeant, byakorewe ubushakashatsi muri RuDA-NPs ivurwa na MDA-MB-231.Uburyo bwibikorwa bya RuDA NPs iterwa na irrasiyo.Muri icyo gihe, hagaragaye imvugo ya poroteyine yo hepfo ya heme ogisijene 1 (HO-1).Nkuko bigaragara ku gishushanyo cya 6F n’inyongera Igishushanyo cya 29, RuDA-NP yifashishije imiti yo kuvura yongereye urwego Nrf2 na HO-1 imvugo ugereranije nitsinda rya PBS, byerekana ko RuDA-NPs ishobora gutera imbaraga za okiside yerekana inzira.Hiyongereyeho, kugirango hamenyekane ingaruka zifotora ya RuDA-NPs63, hasuzumwe kandi imvugo ya proteine Hsp70 yubushyuhe.Biragaragara ko selile zavuwe na RuDA-NPs + 808 nm ya lazeri ya laser yerekanaga imvugo ya Hsp70 ugereranije nandi matsinda abiri, bikagaragaza igisubizo cya selile kuri hyperthermia.
Ibidasanzwe mubisubizo bya vitro byaduteye gukora iperereza mubikorwa bya vivo ya RuDA-NP mu mbeba zambaye ubusa hamwe na MDA-MB-231.Ikwirakwizwa rya tissue ya RuDA NPs ryakozwe mu kumenya ibirimo rutheniyumu mu mwijima, umutima, impyiko, impyiko, ibihaha, n'ibibyimba.Nkuko bigaragara ku gishushanyo.7A, ibintu byinshi bya Ore NPs mubice bisanzwe byagaragaye mugihe cyambere cyo kwitegereza (4 h), mugihe ibyinshi byagaragaye mumyanya yibibyimba nyuma yamasaha 8 nyuma yo guterwa, bishoboka bitewe na Ore NPs.Ingaruka ya EPR ya LF.Ukurikije ibisubizo byatanzwe, igihe cyiza cyo kuvura hamwe nubutare bwa NP cyafashwe nyuma yamasaha 8 nyuma yubuyobozi.Kugirango tugaragaze inzira yo kwegeranya RuDA-NPs ahantu h’ibibyimba, imiterere ya fotoacoustic (PA) ya RuDA-NPs yakurikiranwe no kwandika ibimenyetso bya PA bya RuDA-NPs mu bihe bitandukanye nyuma yo gutera inshinge.Ubwa mbere, ikimenyetso cya PA cya RuDA-NP muri vivo cyasuzumwe no gufata amashusho ya PA yikibanza kibyimba nyuma yo gutera inshinge RuDA-NP.Nkuko bigaragara mu gishushanyo cya 30, RuDA-NPs yerekanye ikimenyetso gikomeye cya PA, kandi hariho isano ryiza hagati ya RuDA-NP hamwe nuburemere bwibimenyetso bya PA (Ishusho yinyongera 30A).Noneho, muri vivo PA amashusho yikibanza cyibibyimba yanditswe nyuma yo guterwa imitsi ya RuDA na RuDA-NP ahantu hatandukanye nyuma yo guterwa.Nkuko bigaragara ku gishushanyo 7B, ikimenyetso cya PA cya RuDA-NPs kivuye aho kibyimba cyagiye cyiyongera buhoro buhoro hamwe nigihe kandi kigera kuri plateau mumasaha 8 nyuma yo guterwa inshinge, bihuye nibisubizo byo gukwirakwiza tissue byagenwe nisesengura rya ICP-MS.Kubijyanye na RuDA (Ishusho yinyongera. 30B), ubukana bwa signal bwa PA bwagaragaye nyuma yamasaha 4 nyuma yo guterwa, byerekana umuvuduko wihuse wa RuDA mubyimba.Byongeye kandi, imyitwarire yo gusohora RuDA na RuDA-NPs yakozweho iperereza mu kumenya ingano ya rutheniyumu mu nkari n’umwanda ukoresheje ICP-MS.Inzira nyamukuru yo kurandura RuDA (Ishusho yinyongera 31) na RuDA-NPs (Igicapo 7C) inyura mumyanda, kandi gukuraho neza RuDA na RuDA-NPs byagaragaye mugihe cyiminsi 8 yo kwiga, bivuze ko RuDA na RuDA-NPs birashobora kuvaho neza mumubiri nta burozi bwigihe kirekire.
A. Gukwirakwiza vivo kwa RuDA-NP mubice byimbeba byagenwe nibirimo Ru (ijanisha ryimiti yatanzwe ya Ru (ID) kuri garama ya tissue) mugihe gitandukanye nyuma yo gutera inshinge.Amakuru arasobanutse iation gutandukana bisanzwe (n = 3). Kudakorwa, ibizamini byimpande zombi * p <0.05, ** p <0.01, na *** p <0.001. Kudakorwa, ibizamini byimpande zombi * p <0.05, ** p <0.01, na *** p <0.001. Непарные дву Кононние t-критерии * p <0,05, ** p <0,01 и *** p <0,001. T-ibipapuro bibiri-umurizo t-ibizamini * p <0.05, ** p <0.01, na *** 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 дву Кононние t-Тыы * p <0,05, ** p <0,01 и *** p <0,001. T-ibipapuro bibiri-umurizo t-ibizamini * p <0.05, ** p <0.01, na *** p <0.001.B PA amashusho yibibanza bya vivo kuri 808 nm yishimye nyuma yubuyobozi bwimitsi ya RuDA-NPs (10 µmol kg-1) kumwanya utandukanye.Nyuma yubuyobozi bwimitsi ya RuDA NPs (10 µmol kg-1), C Ru yasohotse mu mbeba zifite inkari n’umwanda mu bihe bitandukanye.Amakuru arasobanutse iation gutandukana bisanzwe (n = 3).
Ubushobozi bwo gushyushya RuDA-NP muri vivo bwakorewe mu mbeba zambaye ubusa hamwe n'ibibyimba bya MDA-MB-231 na RuDA kugirango ubigereranye.Nkuko bigaragara ku gishushanyo.8A hamwe ninyongera Igishushanyo cya 32, itsinda ryigenzura (saline) ryerekanye ihinduka ryubushyuhe buke (ΔT ≈ 3 ° C) nyuma yiminota 10 yo gukomeza guhura.Nyamara, ubushyuhe bwa RuDA-NPs na RuDA bwiyongereye vuba hamwe nubushyuhe ntarengwa bwa 55.2 na 49.9 ° C, butanga hyperthermie ihagije yo kuvura kanseri ya vivo.Ubwiyongere bwagaragaye bwubushyuhe bwo hejuru kuri RuDA NPs (ΔT ≈ 24 ° C) ugereranije na RuDA (ΔT ≈ 19 ° C) bushobora guterwa no kuba bworoshye kandi bukwirakwira mu ngingo z’ibibyimba bitewe n'ingaruka za EPR.
Amashusho yubushyuhe bwimbeba yimbeba hamwe nibibyimba bya MDA-MB-231 byatewe na lazeri 808 nm mugihe gitandukanye nyuma yamasaha 8 nyuma yo guterwa.Amashusho yerekana amashusho ane asubiramo buri tsinda arerekanwa.B Ingano yibibyimba bifitanye isano na C Ugereranije ikibyimba kinini cyimbeba zitandukanye mugihe cyo kuvura.D Imirongo yuburemere bwumubiri wamatsinda atandukanye yimbeba.Kurasa na lazeri ifite uburebure bwa 808 nm n'imbaraga za 0.5 W / cm2 muminota 10 (300 J / cm2).Utubari twibeshya, bivuze iation gutandukana bisanzwe (n = 3). Kudakorwa, ibizamini byimpande zombi * p <0.05, ** p <0.01, na *** p <0.001. Kudakorwa, ibizamini byimpande zombi * p <0.05, ** p <0.01, na *** p <0.001. Непарные дву Кононние t-критерии * p <0,05, ** p <0,01 и *** p <0,001. T-ibipapuro bibiri-umurizo t-ibizamini * p <0.05, ** p <0.01, na *** 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 дву Кононние t-Тыы * p <0,05, ** p <0,01 и *** p <0,001. T-ibipapuro bibiri-umurizo t-ibizamini * p <0.05, ** p <0.01, na *** p <0.001. E H&E yerekana amashusho yingingo n’ibibyimba biva mu matsinda atandukanye yo kuvura, harimo Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, na RuDA-NPs + Laser. E H&E yerekana amashusho yingingo n’ibibyimba biva mu matsinda atandukanye yo kuvura, harimo Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, na RuDA-NPs + Laser. Изображения окрашивания E H&E основных хикәяов һәм опухолы из разных группа лечения, включая группы физиологического раствора, физиологического раствора + лазера, RuDA, RuDA + Laser, RuDA-NPs na RuDA-NPs. E H&E yerekana amashusho yingingo n’ibibyimba biva mu matsinda atandukanye yo kuvura, harimo saline, saline + laser, RuDA, RuDA + Laser, RuDA-NPs, na RuDA-NPs + Laser.来自 不同 治疗 组 的 主要 器官 和 肿瘤 H E H&E 染色 图像 ， 包括 盐水 、 盐水 + 激光 、 RuDA 、 RuDA + 激光 、 RuDA-NPs 和 RuDA-NPs + 激光 组。H 不同 治疗 组 的 主要 器官 和 肿瘤 H E H&E Окрашивание E H&E основных повов һәм опухо у из различных группа лечения, включая физиологический раствор, фииологический раствор + лазер, RuDA, RuDA + лазер, RuDA-NPs na RuDA-NPs + лазер. E H&E kwanduza ingingo nini nibibyimba biva mumatsinda atandukanye yo kuvura harimo saline, saline + laser, RuDA, RuDA + laser, RuDA-NPs, na RuDA-NPs + laser.Umwanya munini: 60 µm.
Ingaruka zo gufotora muri vivo hamwe na RuDA na RuDA NPs zasuzumwe aho imbeba zambaye ubusa zifite ibibyimba bya MDA-MB-231 zatewe mu mitsi hamwe na RuDA cyangwa RuDA NP ku gipimo kimwe cya 10.0 µmol kg-1 binyuze mu mitsi umurizo, hanyuma 8 amasaha nyuma yo guterwa.imirasire ya laser hamwe nuburebure bwa 808 nm.Nkuko bigaragara ku gishushanyo cya 8B, ibibyimba byiyongereye cyane mu matsinda ya saline na laser, byerekana ko imirasire ya saline cyangwa laser 808 ntacyo byagize ku mikurire yikibyimba.Kimwe no mu itsinda rya saline, gukura kw'ibibyimba byihuse byagaragaye no ku mbeba zavuwe na RuDA-NPs cyangwa RuDA mu gihe hatabayeho imirasire ya lazeri, bikerekana uburozi buke bwijimye.Ibinyuranye na byo, nyuma yo gukwirakwizwa na lazeri, kuvura RuDA-NP na RuDA byombi byatumye habaho gusubira inyuma kw'ibibyimba hamwe no kugabanuka kw'ibibyimba byagabanutseho 95.2% na 84.3%, ugereranije n'itsinda rivura saline, byerekana PDT nziza cyane., byahujwe n'ingaruka za RuDA / CHTV.- NP cyangwa Ore. Ugereranije na RuDA, RuDA NPs yerekanye ingaruka nziza yo gufotora, byatewe ahanini n'ingaruka za EPR ya RuDA NPs.Ibisubizo byo gukumira ibibyimba byongeye gusuzumwa nuburemere bwibibyimba byaciwe kumunsi wa 15 wokuvurwa (Igicapo 8C hamwe ninyongera ya 33).Ikibyimba kibyibushye muri RuDA-NP cyavuwe nimbeba na RuDA zavuwe ni 0.08 na 0.27 g, bikaba byari byoroshye cyane ugereranije nitsinda rishinzwe kugenzura (1.43 g).
Byongeye kandi, uburemere bwumubiri wimbeba bwanditswe buri minsi itatu kugirango bige uburozi bwijimye bwa RuDA-NPs cyangwa RuDA muri vivo.Nkuko bigaragara ku gishushanyo cya 8D, nta tandukaniro rikomeye ryibiro byumubiri byagaragaye kumatsinda yose yo kuvura. Byongeye kandi, hematoxyline na eosine (H&E) byanduza ingingo nkuru (umutima, umwijima, impyiko, ibihaha, nimpyiko) bivuye mu matsinda atandukanye yo kuvura. Byongeye kandi, hakozwe hematoxyline na eosine (H&E) yerekana ingingo nkuru (umutima, umwijima, impyiko, ibihaha, nimpyiko) ziva mu matsinda atandukanye yo kuvura. Кроме того, было проведено окрашивание гемат начилином и эозином (H&E) основных чанов (сердца, пченчени, селезенки, легких и по ки) из разных групп лечения. Byongeye kandi, hakozwe hematoxyline na eosine (H&E) yangiza ingingo zikomeye (umutima, umwijima, impyiko, ibihaha, nimpyiko) ziva mu matsinda atandukanye yo kuvura.此外 ， 对 不同 治疗 的 的 主要 器官 心脏 、 肝脏 、 红 H H H H (H&E) 染色。 (H&E) Кроме того, про କାନили орашивание гемат начилином и эозином (H&E) основных баров (сердца, пченчени, селезенки, легких и по п) в различных грах лечения. Byongeye kandi, hematoxyline na eosine (H&E) byanditseho ingingo zikomeye (umutima, umwijima, impyiko, ibihaha, nimpyiko) byakorewe mu matsinda atandukanye yo kuvura.Nkuko bigaragara ku gishushanyo.8E, H&E yerekana amashusho yingingo eshanu zikomeye zo mumatsinda ya RuDA-NPs na RuDA ntagaragaza ibintu bidasanzwe cyangwa kwangirika kwingingo. 8E, H&E yerekana amashusho yingingo eshanu zikomeye zo mumatsinda ya RuDA-NPs na RuDA ntagaragaza ibintu bidasanzwe cyangwa kwangirika kwingingo.Nkuko bigaragara ku gishushanyo.8E. 8E, H&E yerekana amashusho yingingo eshanu zikomeye zo mu matsinda ya RuDA-NPs na RuDA yerekana ko nta ngingo zigaragara zidasanzwe cyangwa ibikomere.E 8E 所示 ， 来自 RuDA-NPs 和 RuDA 组 的 五个 主要 器官 的 H&E 染色 图像 没有 显示 出 明显 的 异常 或 器官 损伤。如图 8E 所示 ， 来自 RuDA-NPs 和 RuDA 组 的 五个 主要 器官 的 H&E Как покажано на рисунке 8E, изображения окрашивания H&E пяти основных панов из групп РуDA-NPs и RuDA не показали явных аномалий или повреждения Ковов. Nkuko bigaragara ku gishushanyo cya 8E, H&E yerekana amashusho yingingo eshanu zikomeye zo mu matsinda ya RuDA-NPs na RuDA yerekanaga ko bidasanzwe cyangwa kwangirika kwingingo.Ibisubizo byerekanaga ko yaba RuDA-NP cyangwa RuDA yerekanye ibimenyetso byuburozi muri vivo. Byongeye kandi, H&E yerekana amashusho yibibyimba yerekanaga ko amatsinda ya RuDA + Laser na RuDA-NPs + Laser ashobora guteza kanseri ikomeye ya kanseri, bikerekana ibyiza bya vivo Phototherapeutic efficacy ya RuDA na RuDA-NPs. Byongeye kandi, H&E yerekana amashusho yibibyimba yerekanaga ko amatsinda ya RuDA + Laser na RuDA-NPs + Laser ashobora guteza kanseri ikomeye ya kanseri, bikerekana ibyiza bya vivo Phototherapeutic efficacy ya RuDA na RuDA-NPs.Byongeye kandi, hematoxyline-eosine yerekana ibibyimba byerekana ibibyimba byerekanaga ko amatsinda yombi ya RuDA + Laser na RuDA-NPs + Laser ashobora kwangiza cyane ingirabuzimafatizo za kanseri, bikerekana uburyo bwiza bwo gufotora bwa RuDA na RuDA-NPs muri 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 ， 了 DA RuDA 和 RuDA-NPs 的 优异 的 体内 光 疗 功效。。此外 ， 肿瘤 & e 染色 显示 ， ruda + laser 和 ruda-nps + laser 组 均 导致 的 癌细胞 破坏 ， 证明 da ruda 和 ruda-nps 的 的 体内 光 疗 。。。。。。。。。。。。。 。。。Byongeye kandi, hematoxyline na eosine yerekana ibibyimba byerekana ibibyimba byerekanaga ko amatsinda yombi ya RuDA + Laser na RuDA-NPs + Laser yatumye habaho kurimbuka gukabije kwingirangingo za kanseri, byerekana imbaraga za Phototherapeutique ya RuDA na RuDA-NPs muri vivo.
Mu gusoza, uruganda rwa Ru (II) -arene (RuDA) rugizwe na ligande yo mu bwoko bwa DA rwakozwe kugirango rworohereze inzira ya ISC hakoreshejwe uburyo bwo guteranya.Synthesized RuDA irashobora kwishyira hamwe ikoresheje imikoranire idahwitse kugirango ikore sisitemu ya supramolecular ikomoka kuri RuDA, bityo byorohereze gukora 1O2 no guhinduranya neza amafoto yo kuvura kanseri iterwa n'umucyo.Birashimishije kubona RuDA monomeric idatanga 1O2 munsi ya lazeri ya lazeri kuri 808 nm, ariko ishobora kubyara umubare munini wa 1O2 muri leta yegeranye, byerekana gushyira mu gaciro no gukora neza mubishushanyo byacu.Ubushakashatsi bwakurikiyeho bwerekanye ko inteko ya supramolecular iha RuDA uburyo bwiza bwo gufotora no gufotora, nko kwinjiza redshift no kurwanya fotobleaching, bifuzwa cyane gutunganya PDT na PTT.Haba muri vitro ndetse no mubushakashatsi bwa vivo bwerekanye ko RuDA NPs ifite biocompatibilité nziza hamwe no kwegeranya neza mu kibyimba igaragaza ibikorwa byiza biterwa na anticancer iterwa n'umucyo kuri lazeri ya lazeri ku burebure bwa 808 nm.Rero, RuDA NPs nkibimodal supramolecular PDT / PTW reagent bizakungahaza urutonde rwamafoto akoreshwa kumurongo muremure uri hejuru ya 800 nm.Igishushanyo mbonera cya sisitemu ya supramolecular itanga inzira nziza kubakoresha amafoto ya NIR ikora hamwe ningaruka nziza zifotora.
Imiti yose hamwe nu mashanyarazi byabonetse kubatanga ubucuruzi kandi bikoreshwa nta yandi masuku.RuCl3 yaguzwe na Boren Precious Metals Co., Ltd. (Kunming, Ubushinwa).[(η6-p-cym) Ru (fendio) Cl] Cl (fendio = 1,10-phenanthroline-5,6-dione) na 4,7-bis , 6-Diamino-2,1,3-benzothiadiazole yashizwemo ukurikije ubushakashatsi bwabanje 64,65.Ikirangantego cya NMR cyanditswe kuri Bruker Avance III-HD 600 MHz yerekanwe mu kigo cy’ibizamini cya kaminuza y’Amajyepfo y’Amajyepfo ukoresheje d6-DMSO cyangwa CDCl3 nkigisubizo.Guhindura imiti δ bitangwa muri ppm.kubijyanye na tetramethylsilane, hamwe n'imikoranire ihamye J itangwa mumico yuzuye muri hertz.Ikirangantego kinini cyane (HRMS) cyakorewe kuri Agilent 6224 ESI / TOF MS igikoresho.Isesengura ryibanze rya C, H, na N ryakorewe kuri Vario MICROCHNOS isesengura ryibanze (Elementar).UV igaragara igaragara yapimwe kuri Shimadzu UV3600 yerekanwe.Fluorescence yerekanwe kuri Shimadzu RF-6000 spectrofluorimeter.EPR yerekanwe kuri Bruker EMXmicro-6/1 igikoresho.Imiterere n'imiterere y'ibyitegererezo byateguwe byizwe ku bikoresho bya FEI Tecnai G20 (TEM) na Bruker Icon (AFM) bikoresha kuri voltage ya 200 kV.Gukwirakwiza urumuri rutangaje (DLS) byakorewe kuri Nanobrook Omni isesengura (Brookhaven).Ibikoresho bifotora byapimwe ku mashanyarazi (CHI-660, Ubushinwa).Amashusho ya Photoacoustic yabonetse ukoresheje sisitemu ya FUJIFILM VisualSonics Vevo® LAZR.Amashusho yibeshya yabonetse hakoreshejwe microscope ya Olympus FV3000.Isesengura rya FACS ryakorewe kuri BD Calibur itemba cytometero.Ubushakashatsi bukomeye bwa chromatografiya (HPLC) bwakorewe kuri sisitemu ya Waters Alliance e2695 hakoreshejwe 2489 UV / Vis detector.Ibizamini bya Gel Permeation (GPC) byanditswe ku gikoresho cya Thermo ULTIMATE 3000 ukoresheje ERC RefratoMax520 yerekana ibimenyetso byerekana.
[(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) na acide glacial acetike (30 mL) yashizwemo muri firigo ya refleks mu gihe cyamasaha 12.Umuti waje gukurwa muri vacuo ukoresheje moteri izunguruka.Ibisigisigi byavuyemo byahanaguwe na flash colonment chromatografiya (silika gel, CH2Cl2: MeOH = 20: 1) kugirango ibone RuDA nkifu yicyatsi (umusaruro: 877.5 mg, 80%).anus.Kubarwa kuri C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.Byabonetse: 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, 152.81, 149.31, 147.98, 147.16, 139.98, 136.21, 135.57, 134.68, 130.34, 130.02, 128.68, 128.01, 125.51, 124.45, 120.81, 103.45. , 103., 86.52, 84,75, 63.29, 30.90, 22.29, 18.83.ESI-MS: m / z [M-Cl] + = 1097.25.
Synthesis ya 4,7-bis [4- (N, N-diethylamino) phenyl-5,6-diamino-2,1,3-benzothiadiazole (L2): L2 yashizwemo mubyiciro bibiri.Pd (PPh3) 4 (46 mg, 0.040 mmol) yongewe kuri N, N-diethyl-4- (tributylstannyl) aniline (1.05 g, 2,4 mmol) na 4,7-dibromo-5,6-dinitro - 2, 1,3-benzothiadiazole (0,38 g, 1.0 mmol) muri toluene yumye (ml 100).Uruvange rwavanze kuri 100 ° C mumasaha 24.Nyuma yo gukuraho toluene muri vacuo, ibivuyemo byogejwe hamwe na peteroli ya ether.Hanyuma uruvange rwuru ruganda (234.0 mg, 0,45 mmol) nifu ya fer (0,30 g, 5,4 mmol) muri acide acike (ml 20) byavanze kuri 80 ° C. mumasaha 4.Uruvangitirane rwa reaction rwasutswe mumazi hanyuma ibivamo ibara ry'umukara byegeranijwe no kuyungurura.Igicuruzwa cyahanaguweho kabiri na vacuum sublimation kugirango gitange icyatsi kibisi (126.2 mg, umusaruro wa 57%).anus.Kubarwa kuri C26H32N6S: C 67.79, H 7.00, N 18.24.Byabonetse: 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.
Ibicuruzwa byateguwe kandi bisukurwa muburyo bukurikira busa na RuDA.anus.Kubarwa kuri C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.Byabonetse: 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., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m / z [M-Cl] + = 905.24.
RuDA yasheshwe muri MeOH / H2O (5/95, v / v) kuri concentration ya 10 μM.Ikirangantego cyo kwinjiza RuDA cyapimwe buri minota 5 kuri Shimadzu UV-3600 spekitifotometero munsi ya irrasiyo hamwe nurumuri rwa laser hamwe nuburebure bwa 808 nm (0.5 W / cm2).ICG yerekanwe yanditswe mubihe bimwe nkibisanzwe.
EPR yerekanwe kuri Bruker EMXmicro-6/1 yerekanwe hamwe na microwave ifite ingufu za mW 20, gusikana 100 G, hamwe no guhinduranya umurima wa 1 G. 2,2,6,6-tetramethyl-4-piperidone (TEMP) na 5,5-dimethyl-1-pyrroline N-oxyde (DMPO) yakoreshejwe nk'imitego ya spin.Electron spin resonance spectra yanditswe kubisubizo bivanze bya RuDA (50 µM) na TEMF (20 mM) cyangwa DMPO (20 mM) hifashishijwe imirasire ya laser ifite uburebure bwa 808 nm (0.5 W / cm2).
Ibarura rya DFT na TD-DFT kuri RuDA ryakorewe ku rwego rwa PBE1PBE / 6-31 G * // LanL2DZ mu gisubizo cy’amazi ukoresheje gahunda ya Gaussiya 1666,67,68.HOMO-LUMO, umwobo hamwe na elegitoronike yo gukwirakwiza ingufu nkeya imwe yishimye leta RuDA yateguwe hakoreshejwe gahunda ya GaussView (verisiyo 5.0).
Twabanje kugerageza gupima imikorere ya 1O2 RuDA dukoresheje UV isanzwe igaragara ya spekitroscopi hamwe na ICG (ΦΔ = 0.002) nkibisanzwe, ariko gufotora ICG byagize ingaruka cyane kubisubizo.Gutyo, umusaruro wa kwant wa 1O2 RuDA wapimwe no kumenya impinduka zuburemere bwa ABDA fluorescence igera kuri 428 nm iyo iraswe na lazeri ifite uburebure bwa 808 nm (0.5 W / cm2).Ubushakashatsi bwakorewe kuri RuDA na RuDA NPs (20 μM) mumazi / DMF (98/2, v / v) irimo ABDA (50 μM).Umusaruro wa kwant wa 1O2 wabazwe ukoresheje formula ikurikira: ΦΔ (PS) = ΦΔ (ICG) × (rFS / APS) / (rICG / AICG).rPS na rICG nigipimo cyibisubizo bya ABDA hamwe na 1O2 yabonetse kumafoto na ICG.APS na AICG ninjoro ya fotosensitizer na ICG kuri 808 nm.
Ibipimo bya AFM byakozwe mubihe byamazi hakoreshejwe uburyo bwa scan kuri sisitemu ya Bruker Dimension Icon AFM.Ukoresheje imiterere ifunguye hamwe ningirabuzimafatizo, selile zogejwe kabiri na Ethanol hanyuma zumishwa numugezi wa azote.Shyiramo selile zumye mumutwe wa optique ya microscope.Ako kanya shyira igitonyanga cyicyitegererezo muri pisine yamazi hanyuma ubishyire kuri kantileveri ukoresheje siringi ya plastike ikoreshwa neza hamwe ninshinge ya sterile.Ikindi gitonyanga gishyirwa kumurongo, kandi iyo umutwe wa optique umanuwe, ibitonyanga byombi birahuza, bigakora menisk hagati yicyitegererezo nikigega cyamazi.Ibipimo bya AFM byakozwe hifashishijwe kantileveri ya SCANASYST-FLUID V ya nitride (Bruker, ubukana k = 0.7 N m-1, f0 = 120–180 kHz).
Chromatogramu ya HPLC yabonetse kuri sisitemu y'amazi e2695 ifite inkingi ya phoenix C18 (250 × 4,6 mm, 5 µm) ikoresheje icyuma cya 2489 UV / Vis.Uburebure bwumurongo wa detector ni 650 nm.Icyiciro kigendanwa A na B cyari amazi na methanol, kandi icyiciro cyimikorere ya mobile cyari 1.0 ml · min-1.Icyiciro (solvent B) cyari gikurikira: 100% kuva 0 kugeza 4, 100% kugeza 50% kuva muminota 5 kugeza 30, hanyuma usubire 100% kuva muminota 31 kugeza 40.Ore yashongeshejwe mumuti ivanze ya methanol namazi (50/50, mubunini) murwego rwa 50 μM.Ingano yo gutera inshinge yari 20 μl.
GPC isuzuma ryanditswe ku gikoresho cya Thermo ULTIMATE 3000 gifite ibikoresho bibiri bya PL aquagel-OH MIXED-H (2 × 300 × 7.5 mm, 8 µm) hamwe na ERC RefratoMax520 yerekana ibimenyetso byerekana.Inkingi ya GPC yavuzweho amazi ku kigero cya ml 1 / min kuri 30 ° C.Ore NPs yasheshwe mugisubizo cya PBS (pH = 7.4, 50 μM), inshinge zatewe ni 20 μL.
Amafoto yapimwe ku mashanyarazi (CHI-660B, Ubushinwa).Ibisubizo bya optoelectronic mugihe laser yafunguye ikazimya (808 nm, 0.5 W / cm2) byapimwe kuri voltage ya 0.5 V mumasanduku yumukara.Akagari gasanzwe ka electrode eshatu gakoreshwa hamwe na L karubone ya karubone ya karubone (GCE) nka electrode ikora, electrode isanzwe ya calomel (SCE) nka electrode yerekana, na disiki ya platine nka compte electrode.Igisubizo 0.1 M Na2SO4 cyakoreshejwe nka electrolyte.
Umurongo wa kanseri y'ibere ya muntu MDA-MB-231 waguzwe muri KeyGEN Biotec Co, LTD (Nanjing, Ubushinwa, nomero ya catalog: KG033).Ingirabuzimafatizo zahinzwe muri monolayers muri Dulbecco's Modified Eagle's Medium (DMEM, glucose) yongerewe igisubizo cya 10% serumu y'inda y'inda (FBS), penisiline (100 μg / ml) na streptomycine (100 μg / ml).Ingirabuzimafatizo zose zarezwe kuri 37 ° C mu kirere cyuzuye kirimo CO2 5%.
Isuzuma rya MTT ryakoreshejwe kugirango hamenyekane cytotoxicity ya RuDA na RuDA-NPs imbere no kutagira imirasire yumucyo, hamwe na Vc (0.5 mM) cyangwa idafite.Ingirabuzimafatizo za MDA-MB-231 zahinzwe mu isahani 96 y’iriba ku bucucike bw’utugingo ngengabuzima tugera kuri 1 x 105 / ml / iriba kandi byashyizwe mu masaha 12 kuri 37.0 ° C mu kirere cya 5% CO2 n’umwuka wa 95%.RuDA na RuDA NPs zashonga mumazi zongerewe muri selile.Nyuma yamasaha 12 yubushakashatsi, selile zerekanwe nimirasire ya 0.5 W cm -2 ya lazeri kumuraba wa 808 nm muminota 10 (300 J cm -2) hanyuma ikabikwa mumwijima mumasaha 24.Ingirabuzimafatizo zashyizwemo MTT (5 mg / ml) andi masaha 5.Hanyuma, hindura uburyo kuri DMSO (200 µl) kugirango ushongeshe kristu ya formazan yijimye.Indangagaciro za OD zapimwe hifashishijwe umusomyi wa microplate ufite uburebure bwa 570/630 nm.Agaciro IC50 kuri buri sample yabazwe ukoresheje software ya SPSS uhereye kumurongo-wo gusubiza umurongo wabonetse byibuze byibuze bitatu byigenga.
MDA-MB-231 selile zavuwe hamwe na RuDA na RuDA-NP kuri 50 μM.Nyuma yamasaha 12 yubushakashatsi, selile zashyizwemo lazeri ifite uburebure bwa 808 nm nimbaraga za 0.5 W / cm2 kuminota 10 (300 J / cm2).Mu itsinda rya vitamine C (Vc), selile zavuwe na 0.5 mM Vc mbere yo kurasa kwa laser.Ingirabuzimafatizo zashizwe mu mwijima amasaha 24 yiyongera, hanyuma zisigara hamwe na calcein AM hamwe na iyode ya propidium (20 μg / ml, 5 μl) mu minota 30, hanyuma yozwa na PBS (10 μl, pH 7.4).amashusho yingirabuzimafatizo.
Igihe cyo kohereza: Nzeri-23-2022
