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Ingantattun na'urorin daukar hoto suna da mahimmanci musamman don yaduwar amfani da aikin hoto na asibiti.Koyaya, na'urorin daukar hoto na al'ada gabaɗaya suna fama da ɗan gajeren raƙuman raƙuman ruwa, rashin isasshen hoto, ƙarancin adadin adadin nau'in iskar oxygen (ROS), da kashe ROS mai haifar da tarawa.Anan mun bayar da rahoton wani kusa-infrared (NIR) supramolecular photosensitizer (RuDA) matsakanci ta hanyar taron kai na Ru(II) -arene organometallic complexes a cikin ruwa mai ruwa bayani.RuDA na iya samar da iskar oxygen guda ɗaya (1O2) kawai a cikin yanayin da aka tara, kuma yana nuna a fili tari-induced halin tsararrun 1O2 saboda haɓakar haɓakar tsari tsakanin tsarin guda-triplet.A ƙarƙashin aikin hasken laser na 808 nm, RuDA yana nuna yawan amfanin ƙasa na 1O2 na 16.4% (FDA-approved indocyanine kore: ΦΔ = 0.2%) da kuma babban ingancin juyawa na photothermal na 24.2% (nanorods na zinariya na kasuwanci) tare da kyakkyawan hoto.: 21.0%, zinariya nanoshells: 13.0%).Bugu da ƙari, RuDA-NPs tare da kyakkyawan yanayin halitta na iya fi dacewa tarawa a wuraren ciwon daji, yana haifar da raguwa mai mahimmanci a lokacin maganin photodynamic tare da raguwar 95.2% a cikin ƙwayar ƙwayar cuta a cikin vivo.Wannan haɓakar haɓakar haɓakar haɓakar hoto yana ba da dabara don haɓaka na'urorin daukar hoto tare da kyawawan kaddarorin hoto na jiki da na hoto.
Idan aka kwatanta da na al'ada far, photodynamic far (PDT) ne mai ban sha'awa jiyya ga ciwon daji saboda da gagarumin abũbuwan amfãni kamar m spatiotemporal iko, rashin invasiveness, m magani juriya, da kuma rage girman illa 1,2,3.A ƙarƙashin hasken haske, ana iya kunna masu daukar hoto da aka yi amfani da su don samar da nau'in oxygen mai saurin amsawa (ROS), wanda ke haifar da apoptosis / necrosis ko martani na rigakafi4,5. Duk da haka, yawancin na'urorin daukar hoto na al'ada, irin su chlorins, porphyrins, da anthraquinones, suna da ɗan gajeren lokaci mai tsayi (mita <680 nm), don haka yana haifar da mummunan shigar da haske saboda tsananin ɗaukar kwayoyin halitta (misali, haemoglobin da melanin) a ciki. yankin da ake gani6,7. Duk da haka, yawancin na'urorin daukar hoto na al'ada, irin su chlorins, porphyrins, da anthraquinones, suna da ɗan gajeren lokaci mai tsayi (mita <680 nm), don haka yana haifar da mummunan shigar da haske saboda tsananin ɗaukar kwayoyin halitta (misali, haemoglobin da melanin) a ciki. yankin da ake gani6,7. 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. Koyaya, mafi yawan na'urorin daukar hoto na yau da kullun kamar chlorins, porphyrins da anthraquinones suna da ɗan gajeren raƙuman raƙuman ruwa (<680 nm) wanda ke haifar da ƙarancin shigar hasken haske saboda tsananin ɗaukar ƙwayoyin halitta (misali haemoglobin da melanin) zuwa yankin da ake iya gani6,7.然而, 大多数 的 的 的光敏剂, 如如 的 卟酚, 卟啉卟啉 相对蒽醌, 具具 <680 nm), 因此 波长 <680 nm), 因此具 <680 nm), 因此具 <680 nm), 因此具 <680 nm), 因此具 <680 nm), 因此具 <680 nm)导致光穿透性差。然而, 大多数大多数 的 的光敏剂, 二二, 具具 相对相对 相对相对 相对相对 的吸收 的 的波长 (频率 频率吸收) 因此,, 的 680 nm) 因此,, 的 680 nm) 因此 ,吸收 的 的因此 (频率 蛋白蛋白) 因此 ,吸收 的吸收 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. Duk da haka, yawancin na'urorin daukar hoto na gargajiya irin su chlorins, porphyrins da anthraquinones suna da ɗan gajeren raƙuman raƙuman ruwa (yawanci <680 nm) saboda tsananin sha na kwayoyin halitta irin su haemoglobin da melanin wanda ke haifar da ƙarancin shigar haske.Yanki mai gani 6.7.Saboda haka, kusa-infrared (NIR) masu ɗaukar hotuna masu ɗaukar hoto waɗanda aka kunna a cikin 700-900 nm "taga na warkewa" sun dace da phototherapy.Tunda kusa da hasken infrared shine mafi ƙarancin shanyewar kyallen jikin halitta, yana iya haifar da zurfafa shiga ciki da ƙarancin lalacewar hoto8,9.
Abin baƙin ciki, data kasance NIR-absorbing photosensitiizers gaba daya suna da rashin ingancin photostability, low singlet oxygen (1O2) samar da iya aiki, da aggregation-induced 1O2 quenching, wanda iyakance su asibiti aikace-aikace10,11.Ko da yake an yi ƙoƙari sosai don inganta halayen hoto na jiki da na hoto na na'urorin daukar hoto na al'ada, ya zuwa yanzu rahotanni da dama sun ba da rahoton cewa na'urorin daukar hoto na NIR na iya magance duk waɗannan matsalolin.Bugu da kari, da yawa photosensitizers sun nuna alƙawarin ga ingantaccen tsara na 1O212,13,14 lokacin da hasashe da haske sama da 800 nm, tun da photon makamashi rage sauri a kusa-IR yankin.Triphenylamine (TFA) a matsayin mai ba da gudummawar lantarki da [1,2,5] thiadiazole-[3,4-i] dipyrido [a,c] phenazine (TDP) a matsayin ƙungiyar masu karɓar electron Donor-acceptor (DA) nau'in dyes a aji. na dyes, shafe kusa-infrared, wanda aka yi nazari da yawa don kusa-infrared bioimaging II da kuma photothermal far (PTT) saboda kunkuntar bandgap.Don haka, za'a iya amfani da rini na nau'in DA don PDT tare da jin daɗin kusa-IR, kodayake ba a cika yin nazarin su azaman masu ɗaukar hoto don PDT ba.
An san cewa babban inganci na tsaka-tsakin tsaka-tsaki (ISC) na masu daukar hoto yana inganta samuwar 1O2.Dabarar gama gari don ci gaba da tsarin ISC shine haɓaka haɗin keɓaɓɓu (SOC) na masu ɗaukar hoto ta hanyar gabatar da atom masu nauyi ko ƙwayoyin halitta na musamman.Duk da haka, wannan hanyar har yanzu tana da wasu lahani da iyakoki19,20.Kwanan nan, haɗin kai na supramolecular ya ba da wata hanya ta hankali ta ƙasa don ƙirƙira kayan aiki a matakin ƙwayoyin cuta, 21,22 tare da fa'idodi masu yawa a cikin phototherapy: (1) masu ɗaukar hoto masu haɗa kai na iya samun yuwuwar samar da sifofin ribbon.Kama da sifofin lantarki tare da ɗimbin rarraba matakan makamashi saboda cunkushewar kewayawa tsakanin tubalan gini.Sabili da haka, za a inganta wasan makamashi tsakanin ƙananan ƙananan jin dadi (S1) da kuma maƙwabcin maƙwabta na uku (Tn), wanda ke da amfani ga tsarin ISC 23, 24.(2) Ƙungiyar Supramolecular za ta rage jinkirin da ba a yi amfani da shi ba bisa ga tsarin ƙuntataccen motsi na intramolecular (RIM), wanda kuma yana inganta tsarin ISC 25, 26.(3) Ƙungiyar supramolecular na iya kare ƙwayoyin ciki na monomer daga oxidation da lalata, don haka inganta yanayin hotuna na photosensitizer.Idan aka ba da fa'idodin da ke sama, mun yi imanin cewa tsarin supramolecular photosensitizer na iya zama kyakkyawan zaɓi don shawo kan gazawar PDT.
Ru(II) - tushen hadaddun ƙayyadaddun ƙayyadaddun kayan aikin likita ne don yuwuwar aikace-aikace a cikin ganewar asali da jiyya na cututtuka saboda na musamman da kyawawan kaddarorin halittu28,29,30,31,32,33,34.Bugu da kari, yawan jahohi masu sha'awa da abubuwan da za'a iya amfani da su na photophysicochemical na Ru(II) -based complexes suna ba da fa'ida mai girma ga ci gaban Ru(II) na tushen hotuna35,36,37,38,39,40.Babban misali shine ruthenium (II) polypyridyl hadaddun TLD-1433, wanda a halin yanzu yana cikin gwajin gwaji na asibiti na Phase II a matsayin mai daukar hoto don maganin ciwon daji na mafitsara mara tsoka (NMIBC)41.Bugu da ƙari, ruthenium (II) arene organometallic complexes ana amfani da su sosai a matsayin magungunan chemotherapeutic don maganin ciwon daji saboda ƙananan guba da sauƙi na gyare-gyare42,43,44,45.Abubuwan ionic na Ru (II) -arene organometallic complexes ba za su iya inganta ƙarancin solubility na DA chromophores a cikin kaushi na kowa ba, amma kuma inganta taron DA chromophores.Bugu da ƙari, tsarin pseudooctahedral rabin-sandwich tsarin na organometallic complexes na Ru (II) -arenes na iya sterically hana H-aggregation na DA-nau'in chromophores, game da shi sauƙaƙa samuwar J-aggregation tare da redshifted sha makada.Koyaya, rashin lahani na ru(II) -arene hadaddun, kamar ƙarancin kwanciyar hankali da/ko rashin lafiyar rayuwa, na iya shafar tasirin warkewa da ayyukan vivo na rukunin arene-Ru(II).Koyaya, binciken ya nuna cewa ana iya shawo kan waɗannan lahani ta hanyar haɗa rukunin ruthenium tare da polymers masu dacewa ta hanyar ɗaukar hoto ta jiki ko haɗin haɗin gwiwa.
A cikin wannan aikin, mun bayar da rahoton DA-conjugated complexes na Ru(II) -arene (RuDA) tare da jawo NIR ta hanyar haɗin kai tsakanin DAD chromophore da Ru (II) -arene moiety.Abubuwan da aka samo asali na iya haɗa kansu zuwa cikin vesicles na metalosupramolecular a cikin ruwa saboda hulɗar da ba ta da alaƙa.Musamman ma, taron supramolecular ya ba RuDA tare da abubuwan da ke tattare da tsarin haɗin gwiwar polymerization, wanda ya ƙara yawan ƙimar ISC, wanda ya fi dacewa ga PDT (Fig. 1A).Don haɓaka tarin ƙwayar cuta da kuma in vivo biocompatibility, FDA-approved Pluronic F127 (PEO-PPO-PEO) da aka yi amfani da su encapsulate RuDA47,48,49 don ƙirƙirar RuDA-NP nanoparticles (Figure 1B) wanda ya yi aiki a matsayin mai inganci PDT / Dual- yanayin PTT wakili.A cikin maganin cutar kansa (Hoto 1C), RuDA-NP an yi amfani da shi don bi da mice tsirara tare da ciwace-ciwacen MDA-MB-231 don nazarin ingancin PDT da PTT a cikin vivo.
Misali na tsari na tsarin photophysical na RuDA a cikin nau'i-nau'i na monomeric da tarawa don maganin ciwon daji, haɗin B RuDA-NPs da C RuDA-NPs don NIR- kunna PDT da PTT.
RuDA, wanda ya ƙunshi ayyukan TPA da TDP, an shirya su bisa ga tsarin da aka nuna a Ƙarin Hoto 1 (Hoto 2A), kuma RuDA yana da siffa ta 1H da 13C NMR, electrospray ionization mass spectrometry, da bincike na farko (Ƙarin Figures 2-4). ).An ƙididdige taswirar bambancin ƙarfin lantarki na RuDA na mafi ƙanƙanta canji guda ɗaya ta ka'idar aiki mai dogaro da lokaci (TD-DFT) don nazarin tsarin canja wurin caji.Kamar yadda aka nuna a Ƙarin Hoto na 5, yawan ƙwayar lantarki yana motsawa musamman daga triphenylamine zuwa sashin karɓa na TDP bayan hoto, wanda za'a iya dangana shi ga canjin cajin intramolecular (CT).
Siffar sinadarai na Ore. B Bakan Bakin Ore a cikin gaurayawan ma'auni daban-daban na DMF da ruwa.C daidaitattun ƙimar sha na RuDA (800 nm) da ICG (779 nm) tare da lokaci a 0.5 W cm-2 na hasken laser 808 nm.D Ana nuna lalatawar hoto na ABDA ta hanyar RuDA-induced samuwar 1O2 a cikin gaurayawan DMF/H2O tare da abun ciki na ruwa daban-daban a ƙarƙashin aikin radiation laser tare da tsayin tsayin 808 nm da ƙarfin 0.5 W/cm2.
Abstract-An yi amfani da duban gani na shaye-shaye na UV don nazarin abubuwan haɗin kai na Ore a cikin gaurayawan DMF da ruwa a cikin ma'auni daban-daban.Kamar yadda aka nuna a cikin fig.2B, RuDA yana nuna makaɗaɗɗen sha daga 600 zuwa 900 nm a cikin DMF tare da matsakaicin adadin sha a 729 nm.Ƙara yawan adadin ruwa ya haifar da motsin jan hankali na Ore mafi girma zuwa 800 nm, wanda ke nuna J-aggregation na Ore a cikin tsarin da aka haɗa.Bakan hoton hoto na RuDA a cikin abubuwan kaushi daban-daban ana nuna su a Ƙarin Hoto 6. RuDA yana bayyana yana nuna haske na NIR-II na yau da kullun tare da matsakaicin tsayin hayaki na ca.1050 nm a cikin CH2Cl2 da CH3OH, bi da bi.Babban jujjuyawar Stokes (kimanin 300 nm) na RuDA yana nuna babban canji a cikin juzu'i na yanayin jin daɗi da samuwar jahohi masu ƙarancin kuzari.Abubuwan da ake samu na luminescence na Ore a cikin CH2Cl2 da CH3OH an ƙaddara su zama 3.3 da 0.6%, bi da bi.Duk da haka, a cikin cakuda methanol da ruwa (5/95, v / v), an sami ɗan ja-jajayen fitarwa da raguwar yawan amfanin ƙasa (0.22%), wanda zai iya kasancewa saboda haɗin kai na Ore. .
Don ganin taron kai na ORE, mun yi amfani da microscopy mai ƙarfi na atomatik (AFM) don ganin canje-canjen morphological a cikin ORE a cikin maganin methanol bayan ƙara ruwa.Lokacin da abun cikin ruwa ya kasance ƙasa da 80%, ba a lura da ƙayyadaddun ƙira ba (Ƙarin Hoton 7).Duk da haka, tare da ƙarin karuwa a cikin ruwa zuwa 90-95%, ƙananan ƙananan nanoparticles sun bayyana, wanda ya nuna haɗin kai na Ore. Bugu da ƙari, hasken wuta na laser tare da tsawon 808 nm ba ya shafar ƙarfin sha na RuDA a cikin ruwa mai ruwa. bayani (Fig. 2C da Ƙarin Hoto 8).Sabanin haka, ɗaukar indocyanine kore (ICG a matsayin sarrafawa) ya ragu da sauri a 779 nm, yana nuna kyakkyawan yanayin hoto na RuDA.Bugu da ƙari, kwanciyar hankali na RuDA-NPs a cikin PBS (pH = 5.4, 7.4 da 9.0), 10% FBS da DMEM (glucose mai girma) an bincikar su ta hanyar UV-bayyana shayar spectroscopy a lokuta daban-daban.Kamar yadda aka nuna a Ƙarin Hoto na 9, ƴan canje-canje a cikin ruda-NP na shaye-shaye an lura da su a cikin PBS a pH 7.4/9.0, FBS da DMEM, yana nuna kyakkyawan kwanciyar hankali na RuDA-NP.Koyaya, a cikin matsakaiciyar acidic (рН = 5.4) an sami hydrolysis na Ore.Mun kuma ƙara kimanta kwanciyar hankali na RuDA da RuDA-NP ta amfani da manyan ayyuka na chromatography (HPLC).Kamar yadda aka nuna a Ƙarin Hoto na 10, RuDA ta kasance tsayayye a cikin cakuda methanol da ruwa (50/50, v/v) na sa'a ta farko, kuma an lura da hydrolysis bayan 4 hours.Duk da haka, kawai an lura da kololuwar kololuwa-convex don RuDA NPs.Sabili da haka, an yi amfani da gel permeation chromatography (GPC) don tantance kwanciyar hankali na RuDA NPs a cikin PBS (pH = 7.4).Kamar yadda aka nuna a Ƙarin Hoto 11, bayan sa'o'i 8 na shiryawa a ƙarƙashin yanayin da aka gwada, tsayin kololuwa, faɗin kololuwa da yanki mafi girma na NP RuDA bai canza sosai ba, yana nuna kyakkyawan kwanciyar hankali na NP RuDA.Bugu da ƙari, Hotunan TEM sun nuna cewa ilimin halittar jini na RuDA-NP nanoparticles ya kasance kusan canzawa bayan sa'o'i 24 a cikin buffer PBS diluted (pH = 7.4, Ƙarin Fig. 12).
Saboda haɗin kai na iya ba da halaye daban-daban na aiki da sinadarai akan Ore, mun lura da sakin 9,10-anthracenediylbis (methylene) dimalonic acid (ABDA, mai nuna alama 1O2) a cikin gaurayawan methanol-ruwa.Ore mai ruwa daban-daban50.Kamar yadda aka nuna a Hoto na 2D da Ƙarin Hoto na 13, ba a sami raguwar ABDA ba lokacin da abun cikin ruwa ya kasance ƙasa da 20%.Tare da karuwa a cikin zafi zuwa 40%, lalata ABDA ta faru, kamar yadda shaida ta raguwa a cikin ƙarfin ABDA fluorescence.Har ila yau, an lura da cewa mafi girma abun ciki na ruwa yana haifar da raguwa da sauri, yana nuna cewa haɗin kai na RuDA yana da mahimmanci kuma yana da amfani ga lalata ABDA.Wannan al'amari ya sha bamban da na zamani ACQ (aggregation-induced quenching) chromophores.Lokacin da aka lalata tare da laser tare da tsawon 808 nm, yawan adadin 1O2 RuDA a cikin cakuda 98% H2O / 2% DMF shine 16.4%, wanda shine sau 82 mafi girma fiye da na ICG (ΦΔ = 0.2%)51, yana nuna ingantaccen ƙarni na 1O2 RuDA a cikin yanayin tarawa.
Electron yana jujjuyawa ta amfani da 2,2,6,6-tetramethyl-4-piperidinone (TEMP) da 5,5-dimethyl-1-pyrroline N-oxide (DMPO) kamar yadda aka yi amfani da tarko Resonance spectroscopy (ESR) don gano nau'ikan da suka haifar. AFK.da RuDA.Kamar yadda aka nuna a Ƙarin Hoto na 14, an tabbatar da cewa ana samar da 1O2 a lokutan haskakawa tsakanin mintuna 0 zuwa 4.Bugu da ƙari, lokacin da aka shigar da RuDA tare da DMPO a ƙarƙashin hasken wuta, an gano siginar EPR guda hudu na yau da kullum na 1: 2: 2: 1 DMPO-OH · adduct, yana nuna samuwar hydroxyl radicals (OH ·).Gabaɗaya, sakamakon da ke sama yana nuna ikon RuDA don haɓaka samar da ROS ta hanyar sarrafa hotuna na nau'in I/II dual.
Don samun mafi kyawun fahimtar kaddarorin lantarki na Ruda a Monomeric da kuma siffofin orbulling na Ruda a monomeric da aka lissafta su ta amfani da hanyar Dft.Kamar yadda aka nuna a cikin fig.3A, mafi girman ma'aunin kwayoyin halitta (HOMO) na monomeric RuDA an ƙaddamar da shi tare da kashin baya na ligand kuma mafi ƙanƙanci maras nauyi na kwayoyin halitta (LUMO) yana dogara ne akan sashin mai karɓar TDP.Sabanin haka, yawan electron da ke cikin dimeric HOMO ya ta’allaka ne a kan ligand na kwayoyin RuDA guda daya, yayin da karfin lantarkin da ke cikin LUMO ya fi ta’allaka ne a kan bangaren karban wani kwayar halittar RuDA, wanda ke nuni da cewa RuDA tana cikin dimer.Siffofin CT.
A HOMO da LUMO na Ore ana ƙididdige su a cikin nau'ikan monomeric da dimeric.B Singlet da matakan makamashi uku na Ore a cikin monomers da dimers.C Ƙimar matakan RuDA da yiwuwar tashoshin ISC kamar yadda monomeric C da dimeric D. Kibiyoyi suna nuna yiwuwar tashoshin ISC.
Rarraba electrons da ramuka a cikin jahohin RuDA masu ƙarancin kuzari a cikin nau'ikan monomeric da dimeric an bincika ta amfani da software na Multiwfn 3.852.53, waɗanda aka ƙididdige su ta amfani da hanyar TD-DFT.Kamar yadda aka nuna akan ƙarin lakabin.Kamar yadda aka nuna a cikin Figures 1-2, ramukan RDA na monomeric yawanci suna raguwa tare da kashin baya na ligand a cikin waɗannan jihohi masu jin dadi, yayin da electrons galibi suna cikin rukunin TDP, suna nuna halayen intramolecular na CT.Bugu da ƙari, ga waɗannan jihohi masu jin daɗi, akwai ƙarin ko žasa da juna tsakanin ramuka da electrons, yana nuna cewa waɗannan jihohi masu sha'awar jima'i suna ba da gudummawa ta gida (LE).Don dimers, ban da siffofin CT na intramolecular CT da LE, an lura da wani nau'i na nau'in CT na intermolecular a cikin jihohi daban-daban, musamman S3, S4, S7, da S8, dangane da nazarin CT na intermolecular, tare da CT intermolecular miƙa mulki a matsayin manyan. (Karin Teburin).3).
Don ƙarin fahimtar sakamakon gwaji, mun ƙara bincika kaddarorin jihohi masu sha'awar RuDA don gano bambance-bambance tsakanin monomers da dimers (Ƙarin Tables 4-5).Kamar yadda aka nuna a cikin Hoto na 3B, matakan makamashi na singlet da uku masu jin dadi na dimer sun fi na monomer, wanda ke taimakawa wajen rage tazarar makamashi tsakanin S1 da Tn. An ba da rahoton cewa za a iya samun canji na ISC a cikin ƙaramin ƙarfin makamashi (ΔES1-Tn <0.3 eV) tsakanin S1 da Tn54. An ba da rahoton cewa za a iya samun canji na ISC a cikin ƙaramin ƙarfin makamashi (ΔES1-Tn <0.3 eV) tsakanin S1 da Tn54. Собщалось, что переходы ISC za su iya yin amfani da realyzovanы в прелах немольший Богой ческий щели (ΔES1-4). An ba da rahoton cewa ana iya samun canjin ISC a cikin ƙaramin rata na makamashi (ΔES1-Tn <0.3 eV) tsakanin S1 da 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 перехоd ISC может быtь relylizovan в прелах немольшой эnerghetycheskoy эnerghetycheskoy ISC можете да (ΔES1-3). An ba da rahoton cewa ana iya samun canjin ISC a cikin ƙaramin rata na makamashi (ΔES1-Tn <0.3 eV) tsakanin S1 da Tn54.Bugu da kari, orbital daya kacal, wanda aka shagaltar da shi ko wanda ba a ciki, dole ne ya bambanta a cikin bound singlet da jihohi uku don samar da haɗin SOC mara sifili.Don haka, dangane da nazarin makamashi mai ban sha'awa da sauye-sauye na orbital, duk tashoshi masu yiwuwa na ISC ana nuna su a cikin Fig.3C,D.Musamman ma, tashar ISC ɗaya kawai tana samuwa a cikin monomer, yayin da nau'in dimeric yana da tashoshin ISC guda huɗu waɗanda zasu iya haɓaka canjin ISC.Sabili da haka, yana da kyau a ɗauka cewa yawancin ƙwayoyin RuDA suna tarawa, yawancin tashoshin ISC za su kasance.Saboda haka, RuDA aggregates iya samar da biyu-band lantarki Tsarin a cikin singlet da triplet jihohi, rage makamashi rata tsakanin S1 da samuwa Tn, game da shi ƙara da inganci na ISC don sauƙaƙe 1O2 tsara.
Don ƙarin fayyace tsarin da ke ƙasa, mun haɗa wani ma'anar tunani na hadaddun arene-Ru(II) (RuET) ta maye gurbin ƙungiyoyin ethyl guda biyu tare da ƙungiyoyin phenyl triphenylamine guda biyu a cikin RuDA (Fig. 4A, don cikakkiyar sifa, duba ESI, Ƙarin 15). -21) Daga mai ba da gudummawa (diethylamine) zuwa mai karɓa (TDF), RuET yana da halaye iri ɗaya na CT na intramolecular kamar RuDA.Kamar yadda ake tsammani, nau'in shayarwa na RuET a cikin DMF ya nuna ƙananan cajin cajin wutar lantarki tare da karfi mai karfi a cikin yankin da ke kusa da infrared a cikin yanki na 600-1100 nm (Fig. 4B).Bugu da ƙari, an kuma lura da haɓakar RuET tare da ƙara yawan ruwa, wanda aka nuna a cikin jajayen mafi yawan sha, wanda aka kara tabbatar da shi ta hanyar hoton AFM na ruwa (Ƙarin Hoton 22).Sakamakon ya nuna cewa RuET, kamar RuDA, na iya samar da jahohin intramolecular da kuma haɗa kai cikin tsarin da aka tara.
Tsarin sinadarai na RuET.B bakan shayarwa na RuET a cikin gaurayawan ma'auni daban-daban na DMF da ruwa.Plots C EIS Nyquist na RuDA da RuET.Amsoshin Photocurrent D na RuDA da RuET a ƙarƙashin aikin radiation na laser tare da tsawon 808 nm.
An ƙididdige ɓarkewar hoto na ABDA a gaban RuET ta hanyar iska mai haske tare da laser tare da tsayin 808 nm.Abin mamaki, ba a sami raguwar ABDA ba a cikin ɓangarorin ruwa daban-daban (Ƙarin Hoto 23).Dalili mai yuwuwa shine RuET ba zai iya samar da tsarin lantarki da aka haɗa da kyau ba saboda sarkar ethyl ba ta inganta ingantaccen cajin intermolecular.Saboda haka, electrochemical impedance spectroscopy (EIS) da ma'aunin hoto na wucin gadi an yi don kwatanta kaddarorin photoelectrochemical na RuDA da RuET.Dangane da makircin Nyquist (Hoto 4C), RuDA yana nuna ƙarami radius fiye da RuET, wanda ke nufin cewa RuDA56 yana da saurin jigilar wutar lantarki ta intermolecular da ingantaccen aiki.Bugu da ƙari, yawan nauyin hoto na RuDA ya fi girma fiye da na RuET (Fig. 4D), yana tabbatar da ingantaccen canja wurin cajin RuDA57.Don haka, rukunin phenyl na triphenylamine a cikin Ore yana taka muhimmiyar rawa wajen samar da canja wurin cajin intermolecular da samuwar tsarin lantarki mai banded.
Don haɓaka tarin ƙari da in vivo biocompatibility, mun ƙara sanya RuDA tare da F127.Matsakaicin diamita na hydrodynamic na RuDA-NPs an ƙaddara ya zama 123.1 nm tare da kunkuntar rarraba (PDI = 0.089) ta amfani da hanyar rarraba haske mai ƙarfi (DLS) (Figure 5A), wanda ya inganta haɓakar ƙwayar cuta ta hanyar haɓaka haɓakawa da riƙewa.EPR) sakamako.Hotunan TEM sun nuna cewa Ore NPs suna da siffa mai kamanni iri ɗaya tare da matsakaicin diamita na 86 nm.Musamman ma, matsakaicin ɗaukar RuDA-NPs ya bayyana a 800 nm (Ƙarin Hoton 24), yana nuna cewa RuDA-NPs na iya riƙe ayyuka da kaddarorin RuDAs masu haɗa kai.Ƙididdigar adadin adadin ROS na NP Ore shine 15.9%, wanda yayi daidai da Ore. An yi nazarin kaddarorin photothermal na RuDA NPs a ƙarƙashin aikin radiation na laser tare da tsawon 808 nm ta amfani da kyamarar infrared.Kamar yadda aka nuna a cikin fig.5B, C, ƙungiyar kulawa (PBS kawai) ta sami ɗan ƙara yawan zafin jiki, yayin da zafin jiki na RuDA-NPs bayani ya karu da sauri tare da yawan zafin jiki (ΔT) zuwa 15.5, 26.1, da 43.0 ° C.Maɗaukaki masu girma sun kasance 25, 50, da 100 μM, bi da bi, wanda ke nuna tasirin photothermal mai ƙarfi na RuDA NPs.Bugu da ƙari, an ɗauki ma'auni na sake zagayowar dumama / sanyaya don kimanta kwanciyar hankali na photothermal na RuDA-NP kuma kwatanta da ICG.Yanayin zafin jiki na Ore NPs bai ragu ba bayan zagayowar dumama / sanyaya guda biyar (Fig. 5D), wanda ke nuna kyakkyawan kwanciyar hankali na photothermal Ore NPs.Sabanin haka, ICG yana nuna ƙananan kwanciyar hankali na photothermal kamar yadda aka gani daga bacewar yanayin zafin jiki na photothermal a ƙarƙashin yanayi iri ɗaya.Dangane da hanyar da ta gabata58, an ƙididdige ingancin canjin photothermal (PCE) na RuDA-NP a matsayin 24.2%, wanda ya fi girma fiye da kayan aikin photothermal na yanzu kamar nanorods na zinariya (21.0%) da nanoshells na zinariya (13.0%) 59.Don haka, NP Ore yana nuna kyawawan kaddarorin photothermal, wanda ke sa su ba da tabbacin wakilan PTT.
Binciken hotunan DLS da TEM na RuDA NPs (saka).B Hotunan thermal na ƙididdiga daban-daban na RuDA NPs da aka fallasa zuwa radiation laser a tsawon 808 nm (0.5 W cm-2).C Maɓallin juyawa na Photothermal na ƙididdiga daban-daban na NPs tama, waɗanda bayanai ne masu ƙididdigewa.B. D Haɓaka yanayin zafi na ORE NP da ICG akan zagayowar dumama-sanyi 5.
Photocytotoxicity na RuDA NPs akan MDA-MB-231 ɗan adam ciwon nono an kimanta a cikin vitro.Kamar yadda aka nuna a cikin fig.6A, B, RuDA-NPs da RuDA sun baje kolin cytotoxicity mara kyau idan babu iska mai haske, yana nuna ƙananan guba mai duhu na RuDA-NPs da RuDA.Duk da haka, bayan da aka fallasa zuwa radiation laser a tsawon 808 nm, RuDA da RuDA NPs sun nuna karfi photocytotoxicity akan MDA-MB-231 ciwon daji Kwayoyin tare da IC50 dabi'u (rabin-mafi girman inhibitory taro) na 5.4 da 9.4 μM, bi da bi, nuna. cewa RuDA-NP da RuDA suna da yuwuwar maganin cutar kansa.Bugu da ƙari, an ƙara binciken photocytotoxicity na RuDA-NP da RuDA a gaban bitamin C (Vc), mai lalata ROS, don bayyana rawar ROS a cikin cytotoxicity mai haske.Babu shakka, haɓakar tantanin halitta ya karu bayan ƙari na Vc, kuma ƙimar IC50 na RuDA da RuDA NPs sun kasance 25.7 da 40.0 μM, bi da bi, wanda ke tabbatar da muhimmiyar rawar ROS a cikin photocytotoxicity na RuDA da RuDA NPs.Cytotoxicity mai haske na RuDA-NPs da RuDA a cikin MDA-MB-231 Kwayoyin ciwon daji ta hanyar raye-rayen rayuwa / matattu ta hanyar amfani da calcein AM (koren haske don sel masu rai) da propidium iodide (PI, jan haske ga matattu).an tabbatar da su ta hanyar sel) azaman bincike mai kyalli.Kamar yadda aka nuna a cikin Hoto 6C, ƙwayoyin da aka yi wa magani tare da RuDA-NP ko RuDA sun kasance masu ƙarfi ba tare da sakawa a iska ba, kamar yadda aka tabbatar da tsananin haske.Akasin haka, a ƙarƙashin hasken wuta na Laser, kawai an lura da haske mai haske, wanda ke tabbatar da ingantaccen photocytotoxicity na RuDA ko RuDA NPs.Abin lura ne cewa koren kyalli ya bayyana akan ƙari na Vc, wanda ke nuna cin zarafin photocytotoxicity na RuDA da RuDA NPs.Waɗannan sakamakon sun yi daidai da in vitro photocytotoxicity assays.
Dogaro da adadin ƙwayoyin A RuDA- da B RuDA-NP a cikin ƙwayoyin MDA-MB-231 a gaban ko rashi na Vc (0.5 mM), bi da bi.Kuskuren sanduna, ma'anar ± daidaitaccen karkata (n = 3). Ba a haɗa su ba, gwaje-gwaje masu gefe biyu *p <0.05, **p <0.01, da *** p <0.001. Ba a haɗa su ba, gwaje-gwaje masu gefe biyu *p <0.05, **p <0.01, da *** p <0.001. Непарные двусторонние t-kритерии *p <0,05, **p <0,01 da *** p <0,001. Gwaje-gwaje masu wutsiya biyu waɗanda ba a haɗa su ba *p <0.05, ** p <0.01, da *** 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 da *** p <0,001. Gwaje-gwaje masu wutsiya biyu waɗanda ba a haɗa su ba *p <0.05, ** p <0.01, da *** p<0.001.C Live/matattu tantanin tantanin halitta ta amfani da calcein AM da propidium iodide a matsayin bincike na fluorescent.Matsakaicin girman: 30µm.Ana nuna Hotunan wakilci na maimaitawar halitta guda uku daga kowace ƙungiya.Hotunan kyalli na Confocal na samar da ROS a cikin ƙwayoyin MDA-MB-231 a ƙarƙashin yanayin jiyya daban-daban.Green DCF fluorescence yana nuna kasancewar ROS.Rarraba tare da Laser tare da tsawon 808 nm tare da ikon 0.5 W / cm2 na minti 10 (300 J / cm2).Matsakaicin girman: 30µm.Ana nuna Hotunan wakilci na maimaitawar halitta guda uku daga kowace ƙungiya.E Flow cytometry RuDA-NPs (50 µM) ko RuDA (50 µM) nazarin jiyya tare da ko ba tare da 808 nm laser (0.5 W cm-2) a gaban da rashin Vc (0.5 mM) na 10 min.Ana nuna Hotunan wakilci na maimaitawar halitta guda uku daga kowace ƙungiya.F Nrf-2, HSP70 da HO-1 na MDA-MB-231 Kwayoyin da aka bi da su tare da RuDA-NPs (50 µM) tare da ko ba tare da 808 nm radiation radiation laser (0.5 W cm-2, 10 min, 300 J cm-2), sel express 2).Hotunan wakilci na maimaitawar halittu biyu daga kowace ƙungiya ana nuna su.
An yi nazarin samar da ROS na intracellular a cikin ƙwayoyin MDA-MB-231 ta amfani da hanyar 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA).Kamar yadda aka nuna a cikin fig.6D, ƙwayoyin da aka bi da su tare da RuDA-NPs ko RuDA sun nuna nau'in haske mai haske lokacin da aka haskaka tare da laser na 808 nm, yana nuna cewa RuDA-NPs da RuDA suna da ingantaccen ikon samar da ROS.Akasin haka, idan babu haske ko a gaban Vc, kawai an sami alamar siginar kyalli mai rauni na sel, wanda ke nuna ƙarancin samuwar ROS.Matakan ROS na intracellular a cikin ƙwayoyin RuDA-NP da ƙwayoyin MDA-MB-231 da aka yi wa RuDA an ƙara ƙaddara ta hanyar cytometry mai gudana.Kamar yadda aka nuna a Ƙarin Hoto na 25, ma'anar ƙarfin haske (MFI) da RuDA-NPs da RuDA suka haifar a ƙarƙashin 808 nm radiation radiation laser ya karu sosai da kusan 5.1 da 4.8 sau, bi da bi, idan aka kwatanta da ƙungiyar kulawa, yana tabbatar da kyakkyawan samuwar AFK.iya aiki.Koyaya, matakan ROS na intracellular a cikin ƙwayoyin RuDA-NP ko MDA-MB-231 waɗanda aka bi da su tare da RuDA sun kasance kawai kwatankwacin sarrafawa ba tare da saka iska mai iska ba ko a gaban Vc, kama da sakamakon bincike na fluorescence.
An nuna cewa mitochondria shine babban manufa na Ru(II) -arene complexes60.Sabili da haka, an bincika ƙayyadaddun ƙayyadaddun ƙayyadaddun tsarin RuDA da RuDA-NPs.Kamar yadda aka nuna a Ƙarin Hoto na 26, RuDA da RuDA-NP suna nuna irin wannan bayanin martaba na rarraba salula tare da mafi girma a cikin mitochondria (62.5 ± 4.3 da 60.4 ± 3.6 ng/mg protein, bi da bi).Koyaya, ƙaramin adadin Ru kawai aka samo a cikin ɓangarori na makaman nukiliya na Ore da NP Ore (3.5 da 2.1%, bi da bi).Ragowar ƙwayar tantanin halitta ya ƙunshi ragowar ruthenium: 31.7% (30.6 ± 3.4 ng / MG protein) don RuDA da 42.9% (47.2 ± 4.5 ng / MG protein) don RuDA-NPs.Gabaɗaya, Ore da NP Ore galibi suna taruwa a cikin mitochondria.Don tantance dysfunction mitochondrial, mun yi amfani da JC-1 da MitoSOX Red tabo don tantance yuwuwar mitochondrial membrane da ƙarfin samar da superoxide, bi da bi.Kamar yadda aka nuna a Ƙarin Hoto 27, an lura da hasken kore mai tsanani (JC-1) da ja (MitoSOX Red) a cikin sel da aka bi da su tare da RuDA da RuDA-NPs a karkashin 808 nm radiation radiation, yana nuna cewa duka RuDA da RuDA-NPs suna da haske sosai. Yana iya haifar da ƙaddamarwar mitochondrial membrane yadda ya kamata da samar da superoxide.Bugu da ƙari, an ƙaddara tsarin mutuwar tantanin halitta ta amfani da bincike na tushen cytometry na annexin V-FITC/propidium iodide (PI).Kamar yadda aka nuna a cikin Hoto 6E, lokacin da aka ƙone tare da laser na 808 nm, RuDA da RuDA-NP sun haifar da haɓakar haɓakar apoptosis na farko (ƙananan ƙananan dama) a cikin ƙwayoyin MDA-MB-231 idan aka kwatanta da PBS ko PBS tare da laser.Kwayoyin sarrafawa.Koyaya, lokacin da aka ƙara Vc, ƙimar apoptosis na RuDA da RuDA-NP sun ragu sosai daga 50.9% da 52.0% zuwa 15.8% da 17.8%, bi da bi, wanda ya tabbatar da muhimmiyar rawar ROS a cikin photocytotoxicity na RuDA da RuDA-NP..Bugu da ƙari, an lura da ƙananan ƙwayoyin necrotic a cikin duk ƙungiyoyin da aka gwada (ƙananan hagu na sama), suna nuna cewa apoptosis na iya zama babban nau'i na mutuwar kwayar halitta wanda RuDA da RuDA-NP suka haifar.
Tun da lalacewar danniya mai mahimmanci shine babban mahimmanci na apoptosis, nau'in nukiliya da ke hade da erythroid 2, factor 2 (Nrf2) 62, mai mahimmanci mai mahimmanci na tsarin antioxidant, an bincika a cikin RuDA-NPs-magance MDA-MB-231.Hanyar aiwatar da aikin RuDA NPs wanda aka jawo ta hanyar sakawa a iska.A lokaci guda, an kuma gano bayanin furotin na ƙasa heme oxygenase 1 (HO-1).Kamar yadda aka nuna a cikin Hoto 6F da Ƙarin Hoto na 29, RuDA-NP-matsakaici phototherapy ya karu Nrf2 da HO-1 matakan magana idan aka kwatanta da ƙungiyar PBS, yana nuna cewa RuDA-NPs na iya ƙarfafa hanyoyin siginar damuwa na oxidative.Bugu da ƙari, don nazarin tasirin photothermal na RuDA-NPs63, an kuma kimanta bayanin furotin girgiza zafi Hsp70.A bayyane yake cewa sel da aka yi amfani da su tare da ruda-NPs + 808 nm laser irradiation sun nuna ƙarar magana na Hsp70 idan aka kwatanta da sauran ƙungiyoyi biyu, suna nuna martanin salon salula ga hyperthermia.
Sakamakon in vitro na ban mamaki ya sa mu bincika in vivo aikin RuDA-NP a cikin mice tsirara tare da ciwace-ciwacen MDA-MB-231.An yi nazarin rarraba nama na RuDA NPs ta hanyar ƙayyade abubuwan da ke cikin ruthenium a cikin hanta, zuciya, safiya, kodan, huhu, da ciwace-ciwace.Kamar yadda aka nuna a cikin fig.7A, matsakaicin matsakaicin abun ciki na Ore NPs a cikin gabobin al'ada ya bayyana a farkon lokacin kallo (4h), yayin da aka ƙaddara mafi girman abun ciki a cikin kyallen ƙwayar cuta 8 hours bayan allura, mai yiwuwa saboda Ore NPs.Rahoton da aka ƙayyade na LF.Dangane da sakamakon rarraba, an dauki mafi kyawun lokacin jiyya tare da ma'adinai na NP 8 hours bayan gudanarwa.Don kwatanta tsarin tarawar RuDA-NPs a cikin wuraren ciwon daji, ana kula da kaddarorin photoacoustic (PA) na RuDA-NP ta hanyar yin rikodin alamun PA na RuDA-NP a lokuta daban-daban bayan allura.Na farko, an tantance siginar PA na RuDA-NP a cikin vivo ta hanyar yin rikodin hotunan PA na wani wuri mai ƙari bayan allurar intratumoral na RuDA-NP.Kamar yadda aka nuna a Ƙarin Hoto na 30, RuDA-NPs sun nuna siginar PA mai ƙarfi, kuma akwai ingantacciyar alaƙa tsakanin ƙaddamarwar RuDA-NP da ƙarfin siginar PA (Ƙarin Hoto 30A).Sa'an nan, a cikin vivo PA hotunan wuraren ciwon daji an rubuta su bayan allurar RuDA da RuDA-NP a cikin lokaci daban-daban bayan allura.Kamar yadda aka nuna a cikin Hoto 7B, siginar PA na RuDA-NPs daga wurin ciwon daji a hankali ya karu tare da lokaci kuma ya kai ga wani fili a 8 hours bayan allura, daidai da sakamakon rarraba nama wanda aka ƙaddara ta hanyar bincike na ICP-MS.Game da RuDA (Ƙarin Hoton 30B), matsakaicin siginar siginar PA ya bayyana 4 hours bayan allura, yana nuna saurin shigar da RuDA a cikin ƙwayar cuta.Bugu da ƙari, an bincika halayen halayen RuDA da RuDA-NPs ta hanyar ƙayyade adadin ruthenium a cikin fitsari da najasa ta amfani da ICP-MS.Babban hanyar kawar da RuDA (Ƙarin Hoton 31) da RuDA-NPs (Fig. 7C) ta hanyar najasa ne, kuma an lura da tasiri mai tasiri na RuDA da RuDA-NP a lokacin nazarin 8-day, wanda ke nufin cewa RuDA da RuDA-NPs na iya kawar da su da kyau daga jiki ba tare da guba na dogon lokaci ba.
A. Ex vivo rarraba RuDA-NP a cikin ƙwayoyin linzamin kwamfuta an ƙaddara ta hanyar abun ciki na Ru (kashi na kashi na Ru (ID) a kowace gram na nama) a lokuta daban-daban bayan allura.Bayanai suna nufin ± daidaitaccen karkata (n = 3). Ba a haɗa su ba, gwaje-gwaje masu gefe biyu *p <0.05, **p <0.01, da *** p <0.001. Ba a haɗa su ba, gwaje-gwaje masu gefe biyu *p <0.05, **p <0.01, da *** p <0.001. Непарные двусторонние t-kритерии *p <0,05, **p <0,01 da *** p <0,001. Gwaje-gwaje masu wutsiya biyu waɗanda ba a haɗa su ba *p <0.05, ** p <0.01, da *** 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 da *** p <0,001. Gwaje-gwaje masu wutsiya biyu waɗanda ba a haɗa su ba *p <0.05, ** p <0.01, da *** p<0.001.Hotunan B PA na in vivo wuraren kumburi a 808 nm tashin hankali bayan gudanarwar cikin jini na RuDA-NPs (10 μmol kg-1) a wurare daban-daban.Bayan gudanar da aikin jijiya na RuDA NPs (10 μmol kg-1), an fitar da C Ru daga beraye tare da fitsari da najasa a lokuta daban-daban.Bayanai suna nufin ± daidaitaccen karkata (n = 3).
An yi nazarin ƙarfin dumama na RuDA-NP a cikin vivo a cikin mice tsirara tare da ciwace-ciwacen MDA-MB-231 da RuDA don kwatantawa.Kamar yadda aka nuna a cikin fig.8A da ƙari na 32, ƙungiyar kulawa (saline) ta nuna ƙarancin canjin zafin jiki (ΔT ≈ 3 ° C) bayan minti 10 na ci gaba da nunawa.Koyaya, zafin jiki na RuDA-NPs da RuDA ya karu da sauri tare da matsakaicin yanayin zafi na 55.2 da 49.9 °C, bi da bi, yana ba da isasshen hyperthermia don maganin ciwon daji na vivo.Ƙara yawan zafin jiki mai girma ga RuDA NPs (ΔT ≈ 24 ° C) idan aka kwatanta da RuDA (ΔT ≈ 19 ° C) na iya zama saboda mafi kyawun haɓakawa da tarawa a cikin ƙwayoyin tumor saboda sakamakon EPR.
Hotunan thermal infrared na beraye tare da ciwace-ciwacen daji na MDA-MB-231 da aka haska tare da laser 808 nm a lokuta daban-daban sa'o'i 8 bayan allura.Ana nuna Hotunan wakilci na maimaitawar halittu huɗu daga kowace ƙungiya.B Ƙarar ƙwayar ƙwayar cuta da C Matsakaicin yawan ƙwayar ƙwayar cuta na ƙungiyoyi daban-daban na beraye yayin jiyya.D Lanƙwan nauyin nauyin jiki na ƙungiyoyin mice daban-daban.Rarraba tare da Laser tare da tsawon 808 nm tare da ikon 0.5 W / cm2 na minti 10 (300 J / cm2).Kuskuren sanduna, ma'anar ± daidaitaccen karkata (n = 3). Ba a haɗa su ba, gwaje-gwaje masu gefe biyu *p <0.05, **p <0.01, da *** p <0.001. Ba a haɗa su ba, gwaje-gwaje masu gefe biyu *p <0.05, **p <0.01, da *** p <0.001. Непарные двусторонние t-kритерии *p <0,05, **p <0,01 da *** p <0,001. Gwaje-gwaje masu wutsiya biyu waɗanda ba a haɗa su ba *p <0.05, ** p <0.01, da *** 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 da *** p <0,001. Gwaje-gwaje masu wutsiya biyu waɗanda ba a haɗa su ba *p <0.05, ** p <0.01, da *** p<0.001. E H&E Hotunan hotuna na manyan gabobin da ciwace-ciwace daga kungiyoyin jiyya daban-daban, gami da Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, da RuDA-NPs + Laser kungiyoyin. E H&E Hotunan hotuna na manyan gabobin da ciwace-ciwace daga kungiyoyin jiyya daban-daban, gami da Saline, Saline + Laser, RuDA, RuDA + Laser, RuDA-NPs, da RuDA-NPs + Laser kungiyoyin. 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 Hotunan hotuna na manyan gabobin da ciwace-ciwace daga kungiyoyin jiyya daban-daban, ciki har da saline, saline + laser, RuDA, RuDA + Laser, RuDA-NPs, da RuDA-NPs + Laser kungiyoyin.来自不同治疗组的主要器官和肿瘤的E H&E 染色图像。来自不同治疗组的主要器官和肿瘤的E H&E Окрашивание E H&E основных органов и опухолей из различных групп лечения, включая физиологический раствор, физиологический раствор + лазер, RuDA, RuDA + лазер, RuDA-NPs и RuDA-NPs + лазер. E H & E tabo na manyan gabobin da ciwace-ciwace daga kungiyoyin jiyya daban-daban ciki har da saline, saline + laser, RuDA, RuDA + Laser, RuDA-NPs, da RuDA-NPs + Laser.Matsakaicin girman: 60µm.
An kimanta tasirin phototherapy a cikin vivo tare da RuDA da RuDA NPs wanda aka yi amfani da beraye tsirara tare da ciwace-ciwacen MDA-MB-231 tare da RuDA ko RuDA NPs a kashi ɗaya na 10.0 µmol kg-1 ta hanyar jijiyar wutsiya, sannan 8 awanni bayan allura.Laser a iska mai guba tare da tsawon 808 nm.Kamar yadda aka nuna a cikin Hoto 8B, ƙananan ƙwayar ƙwayar cuta sun karu sosai a cikin saline da kungiyoyin laser, yana nuna cewa saline ko laser 808 irradiation yana da ɗan tasiri akan ci gaban ƙwayar cuta.Kamar yadda yake a cikin rukunin saline, an kuma lura da haɓakar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar ƙwayar cuta ta RuDA-NPs ko RuDA ba tare da rashin hasken laser ba.Sabanin haka, bayan hasarar laser, duka RuDA-NP da RuDA magani sun haifar da raguwar ƙwayar ƙwayar cuta mai mahimmanci tare da raguwar ƙarar ƙwayar cuta na 95.2% da 84.3%, bi da bi, idan aka kwatanta da ƙungiyar da aka yi da saline, yana nuna kyakkyawan haɗin gwiwar PDT., mai shiga tsakani ta tasirin RuDA/CHTV.- NP ko Ore. Idan aka kwatanta da RuDA, RuDA NPs sun nuna sakamako mai kyau na phototherapeutic, wanda ya kasance saboda sakamakon EPR na RuDA NPs.Sakamakon hana ci gaban Tumor an ƙara kimantawa ta hanyar ƙwayar ƙwayar cuta da aka cire a ranar 15 na jiyya (Fig. 8C da Ƙarin Hoto 33).Ma'anar ƙwayar ƙwayar cuta a cikin RuDA-NP da aka yi wa mice da RuDA da aka kula da beraye shine 0.08 da 0.27 g, bi da bi, wanda ya fi sauƙi fiye da a cikin ƙungiyar kulawa (1.43 g).
Bugu da ƙari, an rubuta nauyin jikin beraye kowane kwana uku don nazarin duhun guba na RuDA-NPs ko RuDA a cikin vivo.Kamar yadda aka nuna a cikin Hoto 8D, ba a sami bambance-bambance masu mahimmanci a cikin nauyin jiki ba don duk kungiyoyin jiyya. Bugu da ƙari kuma, an gudanar da aikin hematoxylin da eosin (H&E) na manyan gabobin (zuciya, hanta, saifa, huhu, da koda) daga ƙungiyoyin jiyya daban-daban. Bugu da ƙari kuma, an yi amfani da hematoxylin da eosin (H&E) na manyan gabobin (zuciya, hanta, saifa, huhu, da koda) daga ƙungiyoyin jiyya daban-daban. Кроме того, было проведено окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селезенки, легких и почек) из разных групп лечения. Bugu da ƙari, an yi amfani da hematoxylin da eosin (H & E) na manyan gabobin (zuciya, hanta, safiya, huhu, da kodan) daga kungiyoyin jiyya daban-daban.此外，对不同治疗组的主要器官（心脏、肝脏、脾脏、肺和肾脏）要器官（心脏、肝脏、脾脏、肺和肾脏）要器官（心脏、肝脏、脾脏、肺和肾脏）进行苏咢 (H&E) Кроме того, проводили окрашивание гематоксилином и эозином (H&E) основных органов (сердца, печени, селезенки, легких и почек) в различных группах лечения. Bugu da ƙari, an yi amfani da hematoxylin da eosin (H&E) na manyan gabobin (zuciya, hanta, saifa, huhu, da koda) a cikin ƙungiyoyin jiyya daban-daban.Kamar yadda aka nuna a cikin Fig.8E, hotunan H&E masu lalata manyan gabobi guda biyar daga RuDA-NPs da RuDA ba su nuna rashin daidaituwa ko lahani na gabobin. 8E, hotunan H&E masu lalata manyan gabobi guda biyar daga RuDA-NPs da RuDA ba su nuna rashin daidaituwa ko lahani na gabobin.Kamar yadda aka nuna a cikin fig.8E, изображения H&E пяти основных органов из групп RuDA-NPs da RuDA. 8E, H&E Hotunan Hotuna na manyan gabobin biyar daga RuDA-NPs da RuDA ba su nuna alamun rashin daidaituwa ko raunuka.如图8E 所示，来自RuDA-NPs 和RuDA 组的五个主要器官的H&E如图8E 所示，来自RuDA-NPs 和RuDA 组的五个主要器官的H&E Как показано на рисунке 8E, изображения окрашивания H&E пяти Kamar yadda aka nuna a cikin Hoto 8E, hotunan H & E na manyan gabobin jiki guda biyar daga RuDA-NPs da RuDA ba su nuna rashin daidaituwa ko lalata gabobin ba.Wadannan sakamakon sun nuna cewa babu RuDA-NP ko RuDA da suka nuna alamun guba a cikin vivo. Haka kuma, hotunan H&E na ciwace-ciwacen ciwace-ciwacen ciwace-ciwace sun nuna cewa duka RuDA + Laser da RuDA-NPs + ƙungiyoyin Laser na iya haifar da lalata ƙwayoyin cutar kansa mai tsanani, yana nuna mafi kyawun tasirin tasirin phototherapeutic na RuDA da RuDA-NPs. Haka kuma, hotunan H&E na ciwace-ciwacen ciwace-ciwacen ciwace-ciwace sun nuna cewa duka RuDA + Laser da RuDA-NPs + ƙungiyoyin Laser na iya haifar da lalata ƙwayoyin cutar kansa mai tsanani, yana nuna mafi kyawun tasirin tasirin phototherapeutic na RuDA da RuDA-NPs.Bugu da ƙari, hotunan hematoxylin-eosin da aka lalata sun nuna cewa duka RuDA + Laser da RuDA-NPs + Laser kungiyoyin na iya haifar da mummunar lalata kwayoyin cutar kansa, yana nuna ingantaccen ingancin phototherapeutic na RuDA da RuDA-NPs a cikin vivo.此外，肿瘤的H&E 染色图像显示，RuDA + Laser 和RuDA-NPs + LaserRuda + Laser 和 ruda-nps + Laser 和 ruda-nps + Laser ....Bugu da ƙari, hotunan hematoxylin da eosin da aka lalata sun nuna cewa duka RuDA + Laser da RuDA-NPs + Laser kungiyoyin sun haifar da mummunar lalata kwayoyin cutar kansa, yana nuna ingantaccen ingancin phototherapeutic na RuDA da RuDA-NPs a cikin vivo.
A ƙarshe, Ru (II) -arene (RuDA) organometallic hadaddun tare da DA-type ligands an tsara su don sauƙaƙe tsarin ISC ta amfani da hanyar tarawa.RuDA da aka haɗa ta na iya haɗa kai ta hanyar hulɗar da ba ta dace ba don samar da tsarin supramolecular da aka samu RuDA, ta haka yana sauƙaƙe samuwar 1O2 da ingantaccen canjin photothermal don maganin ciwon daji mai haifar da haske.Abin lura shi ne cewa monomeric RuDA bai samar da 1O2 a ƙarƙashin hasken wuta na laser a 808 nm ba, amma zai iya samar da adadi mai yawa na 1O2 a cikin jihar da aka tara, yana nuna ma'auni da ingancin ƙirar mu.Nazarin da suka biyo baya sun nuna cewa taron supramolecular yana ba da RuDA tare da ingantattun kaddarorin hoto da kayan aikin hoto, irin su jan shift da juriya na hoto, waɗanda ke da sha'awar sarrafa PDT da PTT.Dukansu gwaje-gwajen in vitro da in vivo sun nuna cewa RuDA NPs tare da ingantaccen biocompatibility da kyakkyawar tarawa a cikin ƙwayar cuta suna nuna kyakkyawan aikin rigakafin cutar kansa wanda ke haifar da haske akan hasken laser a tsawon 808 nm.Don haka, RuDA NPs azaman ingantaccen bimodal supramolecular PDT/PTW reagents zai wadatar da saitin na'urar daukar hoto da aka kunna a tsayin tsayi sama da 800 nm.Tsarin ra'ayi na tsarin supramolecular yana ba da ingantacciyar hanya don masu amfani da hotuna na NIR tare da ingantaccen tasirin hotuna.
An samo duk sinadarai da kaushi daga masu samar da kasuwanci kuma an yi amfani da su ba tare da ƙarin tsarkakewa ba.An sayi RuCl3 daga Boren Precious Metals Co., Ltd. (Kunming, China).[(η6-p-cym) Ru (fendio) Cl] Cl (fendio = 1,10-phenanthroline-5,6-dione) da 4,7-bis [4- (N-diphenylamino) phenyl] -5 ,6-Diamino-2,1,3-benzothiadiazole an haɗa shi bisa ga binciken da ya gabata64,65.An yi rikodin bakan NMR akan na'urar sikirin Bruker Avance III-HD 600 MHz a Cibiyar Gwajin Nazarin Jami'ar Kudu maso Gabas ta amfani da d6-DMSO ko CDCl3 azaman sauran ƙarfi.Ana ba da canjin sinadarai a cikin ppm.Game da tetramethylsilane, kuma ana ba da ma'auni na ma'amala a cikin cikakkiyar dabi'u a cikin hertz.An yi babban ƙayyadaddun ƙididdigar taro (HRMS) akan kayan aikin Agilent 6224 ESI/TOF MS.An gudanar da bincike na abubuwan C, H, da N akan na'urar nazari na farko na Vario MICROCHNOS (Elementar).An auna bakan da ake iya gani UV akan ma'aunin sitiriyo na Shimadzu UV3600.An yi rikodin bakan mai walƙiya akan na'urar sikirin spectrofluorimeter Shimadzu RF-6000.An yi rikodin bakan EPR akan kayan aikin Bruker EMXmicro-6/1.An yi nazarin ilimin halittar jiki da tsarin samfuran da aka shirya akan FEI Tecnai G20 (TEM) da kayan aikin Bruker Icon (AFM) waɗanda ke aiki a ƙarfin lantarki na 200 kV.An yi watsawar haske mai ƙarfi (DLS) akan Nanobrook Omni analyzer (Brookhaven).An auna Properties Photoelectrochemical akan saitin electrochemical (CHI-660, China).An samo hotunan hoto ta amfani da tsarin FUJIFILM VisualSonics Vevo® LAZR.An samo hotuna masu banƙyama ta amfani da na'urar hangen nesa ta Olympus FV3000.An yi nazarin FACS akan sitometer kwararar BD Calibur.An yi gwaje-gwajen chromatography na ruwa mai girma (HPLC) akan tsarin Waters Alliance e2695 ta amfani da mai gano UV/Vis 2489.Gel Permeation Chromatography (GPC) an yi rikodin gwaje-gwajen akan kayan aikin Thermo ULTIMATE 3000 ta amfani da na'urar ganowa ta 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) da glacial acetic acid (30 ml) an motsa su a firiji na tsawon awanni 12.Daga nan an cire sauran ƙarfi a cikin injin daskarewa ta amfani da injin rotary evaporator.Sakamakon da aka samu an tsarkake shi ta hanyar chromatography ginshiƙi (silica gel, CH2Cl2: MeOH = 20: 1) don samun RuDA a matsayin foda mai kore (samar: 877.5 mg, 80%).dubura.An ƙididdige shi don C64H48Cl2N8RuS: C 67.84, H 4.27, N 9.89.An samo: 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, 135.88, 120.45, 120.45, 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.
Maganar 4,7-bis [4- (N, N-diethylamino) phenyl-5,6-diamino-2,1,3-benzothiadiazole (L2): L2 an haɗa shi cikin matakai biyu.Pd (PPh3) 4 (46 mg, 0.040 mmol) an ƙara zuwa N, N-diethyl-4- (tributylstannyl) aniline (1.05 g, 2.4 mmol) da 4,7-dibromo-5,6-dinitro bayani - 2, 1,3-benzothiadiazole (0.38 g, 1.0 mmol) a bushe toluene (100 ml).An motsa cakuda a 100 ° C na awa 24.Bayan cire toluene a cikin vacuo, sakamakon da aka samu an wanke shi da ether mai.Sa'an nan kuma cakuda wannan fili (234.0 MG, 0.45 mmol) da baƙin ƙarfe foda (0.30 g, 5.4 mmol) a cikin acetic acid (20 ml) an motsa shi a 80 ° C. na 4 hours.An zuba cakudar da aka yi a cikin ruwa kuma an tattara sakamakon da aka samu ta hanyar tacewa.An tsarkake samfurin sau biyu ta hanyar vacuum sublimation don ba da m kore (126.2 MG, 57% yawan amfanin ƙasa).dubura.An ƙididdige shi don C26H32N6S: C 67.79, H 7.00, N 18.24.An samo: 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.
An shirya abubuwan haɗin gwiwa kuma an tsarkake su ta bin hanyoyin kama da RuDA.dubura.An ƙididdige shi don C48H48Cl2N8RuS: C 61.27, H 5.14, N 11.91.An samo: 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, 135.75, 120.75, 120.75, 120.75, 120.78.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 80.7.70, 84.7.0.7.0.7.70, 80.7.70, 84.7.0.7.70, 80.7.70, 80.7.70, 80.7.70, 84.7.0,50., 38.06, 31.22, 29.69, 22.29, 19.19, 14.98, 12.93.ESI-MS: m/z [M-Cl]+ = 905.24.
An narkar da RuDA a cikin MeOH/H2O (5/95, v/v) a matakin 10 μM.An auna bakan na RuDA kowane minti 5 akan Shimadzu UV-3600 spectrophotometer karkashin iska mai haske tare da hasken Laser tare da tsayin 808 nm (0.5 W/cm2).An yi rikodin bakan ICG a ƙarƙashin yanayi iri ɗaya da ma'auni.
An yi rikodin nau'ikan nau'ikan EPR akan na'urar sikirin Bruker EMXmicro-6/1 tare da ikon microwave na 20 mW, kewayon dubawa na 100 G, da yanayin yanayin filin na 1 G. 2,2,6,6-tetramethyl-4-piperidone. (TEMP) da 5,5-dimethyl-1-pyrroline N-oxide (DMPO) an yi amfani da su azaman tarko.An yi rikodin bakan magana na lantarki don gaurayawan mafita na RuDA (50 µM) da TEMF (20 mM) ko DMPO (20 mM) ƙarƙashin aikin radiation na Laser tare da tsawon 808 nm (0.5 W/cm2).
An gudanar da lissafin DFT da TD-DFT don RuDA a matakan PBE1PBE / 6-31 G *// LanL2DZ a cikin maganin ruwa ta amfani da shirin Gaussian 1666,67,68.HOMO-LUMO, ramuka da kuma rarrabawar lantarki na ƙananan makamashi singlet mai farin ciki na jihar RuDA an tsara su ta hanyar amfani da shirin GaussView (sigar 5.0).
Na farko mun yi ƙoƙari don auna ƙarfin tsararrun 1O2 RuDA ta yin amfani da na al'ada UV-visible spectroscopy tare da ICG (ΦΔ = 0.002) a matsayin ma'auni, amma photodegradation na ICG ya shafi sakamako sosai.Don haka, an auna yawan amfanin ƙasa na 1O2 RuDA ta hanyar gano canji a cikin ƙarfin ƙarfin ABDA fluorescence a kusan 428 nm lokacin da aka haskaka shi tare da laser tare da tsawon 808 nm (0.5 W/cm2).An yi gwaje-gwaje akan RuDA da RuDA NPs (20 μM) a cikin ruwa / DMF (98/2, v / v) dauke da ABDA (50 μM).An ƙididdige yawan adadin adadin 1O2 ta amfani da tsari mai zuwa: ΦΔ (PS) = ΦΔ (ICG) × (rFS/APS)/(rICG/AICG).rPS da rICG sune ƙimar amsawar ABDA tare da 1O2 da aka samu daga photosensitizer da ICG, bi da bi.APS da AICG sune ɗaukar hotuna na hotuna da ICG a 808 nm, bi da bi.
An gudanar da ma'aunin AFM a cikin yanayin ruwa ta amfani da yanayin dubawa akan tsarin Bruker Dimension Icon AFM.Yin amfani da tsari mai buɗewa tare da ƙwayoyin ruwa, an wanke sel sau biyu tare da ethanol kuma an bushe su tare da rafi na nitrogen.Saka busassun sel a cikin kan na gani na mahalli.Nan da nan sanya digo na samfurin a cikin tafkin ruwa kuma sanya shi a kan cantilever ta amfani da sirinji na filastik da ba za a iya zubar da shi ba da kuma allura maras kyau.Ana sanya wani digo kai tsaye a kan samfurin, kuma lokacin da aka saukar da kai na gani, saukowa biyu suna haɗuwa, suna samar da meniscus tsakanin samfurin da tafki na ruwa.An gudanar da ma'aunin AFM ta amfani da SCANASYST-FLUID V mai siffar nitride cantilever (Bruker, hardness k = 0.7 N m-1, f0 = 120-180 kHz).
An sami chromatogram na HPLC akan tsarin Waters e2695 sanye take da ginshiƙin Phoenix C18 (250 × 4.6 mm, 5 µm) ta amfani da mai gano 2489 UV/Vis.Tsawon zangon na'urar ganowa shine 650 nm.Matakan tafi-da-gidanka A da B sune ruwa da methanol, bi da bi, kuma yawan kwararar lokaci ta wayar hannu shine 1.0 ml·min-1.Mai narkewa (mai narkewa B) ya kasance kamar haka: 100% daga mintuna 0 zuwa 4, 100% zuwa 50% daga mintuna 5 zuwa 30, kuma sake saita zuwa 100% daga mintuna 31 zuwa 40.An narkar da Ore a cikin wani gauraye bayani na methanol da ruwa (50/50, ta girma) a wani taro na 50 μM.Adadin allurar shine 20 μl.
An yi rikodin ƙididdigar GPC akan kayan aikin Thermo ULTIMATE 3000 sanye take da ginshiƙan PL aquagel-OH MIXED-H guda biyu (2 × 300 × 7.5 mm, 8 µm) da ERC RefratoMax520 mai gano ma'anar refractive.An lalata ginshiƙin GPC da ruwa a ƙimar 1 ml/min a 30°C.An narkar da Ore NPs a cikin maganin PBS (pH = 7.4, 50 μM), girman allura shine 20 μL.
An auna hotuna a kan saitin sinadaran lantarki (CHI-660B, China).Amsoshin optoelectronic lokacin da aka kunna da kashe Laser (808 nm, 0.5 W/cm2) an auna su a ƙarfin lantarki na 0.5 V a cikin akwatin baƙar fata, bi da bi.An yi amfani da madaidaicin tantanin halitta guda uku tare da na'urar lantarki mai nau'in L-dimbin gilashi (GCE) azaman lantarki mai aiki, ma'aunin lantarki na calomel (SCE) azaman na'urar magana, da faifan platinum azaman na'urar lantarki.An yi amfani da maganin 0.1 M Na2SO4 azaman electrolyte.
An siyi layin kwayar cutar kansar nono na mutum MDA-MB-231 daga KeyGEN Biotec Co., LTD (Nanjing, China, lambar kasida: KG033).An girma kwayoyin halitta a cikin monolayers a Dulbecco's Modified Eagle's Medium (DMEM, high glucose) wanda aka haɓaka tare da maganin 10% fetal bovine serum (FBS), penicillin (100 μg/ml) da streptomycin (100 μg/ml).An haɓaka dukkan sel a 37°C a cikin yanayi mai ɗanɗano mai ɗauke da 5% CO2.
An yi amfani da gwajin MTT don ƙayyade cytotoxicity na RuDA da RuDA-NPs a gaban da rashin hasken haske, tare da ko ba tare da Vc (0.5 mM).MDA-MB-231 Kwayoyin ciwon daji sun girma a cikin faranti 96-rijiya a yawan ƙwayar tantanin halitta na kusan 1 x 105 cell / ml / rijiya kuma an sanya shi don 12 hours a 37.0 ° C a cikin yanayi na 5% CO2 da 95% iska.RuDA da RuDA NPs da aka narkar da su cikin ruwa an ƙara su zuwa sel.Bayan sa'o'i 12 na shiryawa, kwayoyin sun fallasa su zuwa 0.5 W cm -2 laser radiation a tsawon 808 nm na minti 10 (300 J cm -2) sannan kuma a cikin duhu don 24 hours.Daga nan aka sanya ƙwayoyin sel tare da MTT (5 mg/ml) na wani 5 hours.A ƙarshe, canza matsakaici zuwa DMSO (200 µl) don narkar da sakamakon lu'ulu'u na formazan purple.An auna ƙimar OD ta amfani da mai karanta microplate tare da tsawon 570/630 nm.An ƙididdige ƙimar IC50 na kowane samfuri ta amfani da software na SPSS daga matakan amsa kashi da aka samu daga aƙalla gwaje-gwaje masu zaman kansu guda uku.
Kwayoyin MDA-MB-231 an bi da su tare da RuDA da RuDA-NP a wani taro na 50 μM.Bayan sa'o'i 12 na shiryawa, an lalata kwayoyin halitta tare da laser tare da tsayin daka na 808 nm da ikon 0.5 W / cm2 na 10 min (300 J / cm2).A cikin rukuni na bitamin C (Vc), an yi amfani da kwayoyin halitta tare da 0.5 mM Vc kafin hasken laser.Daga nan aka sanya ƙwayoyin sel a cikin duhu don ƙarin sa'o'i 24, sa'an nan kuma an lalata su da calcein AM da propidium iodide (20 μg/ml, 5 μl) na minti 30, sannan a wanke da PBS (10 μl, pH 7.4).hotunan sel masu tabo.

Lokacin aikawa: Satumba-23-2022