生物材料定义与研究内容 (1).pdf

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1、NanoscalePAPERCite this:Nanoscale,2020,12,12508Received 23rd February 2020,Accepted 13th May 2020DOI:10.1039/d0nr01547drsc.li/nanoscaleA HMCuSMnO2nanocomplex responsive tomultiple tumor environmental clues for photoa-coustic/fluorescence/magnetic resonance trimo-dal imaging-guided and enhanced photo

2、thermal/photodynamic therapyQian Li,a,bJunjie Ren,aQiubing Chen,a,bWeiwei Liu,cZhigang Xu,aYang Cao,*cYuejun Kangaand Peng Xue*aHollow mesoporous copper sulfide nanoparticles(HMCuS NPs)are advantageous for loading small-molecule therapeutic drugs coupled with photothermal ablation for synergistic tu

3、mor therapy.However,treatment efficacy mediated by HMCuS NPs is not always satisfactory owing to their insensitivity towardthe tumor microenvironment(TME),and unpredictable drug leakage may also result in deleterious sys-temic toxicity.Here,a novel HMCuSMnO2-based coreshell nanoplatform was develope

4、d as a highlyefficient TME modulator,which could alleviate tumor hypoxia,deplete the level of intracellular glutathione(GSH)and trigger the dissolution of Mn2+.Moreover,MnO2,in situ grown on the surface of HMCuS,mayact as a gatekeeper by forming a stimulus-responsive plug within the mesoporous struc

5、ture,which effec-tively prevented the premature release of encapsulated photosensitizer chlorin e6(Ce6)and was respon-sive to the acidic TME for demand-based drug release.Under the condition of 660/808 nm dual-wave-length laser irradiation,hyperthermia-mediated photothermal therapy(PTT)and reactive

6、oxygen species(ROS)-mediated photodynamic therapy(PDT)can be triggered for tumor eradication,which were furtherenhanced upon the modification of the TME.In the meantime,splendid photoacoustic(PA)/fluorescence(FL)/magnetic resonance(MR)imaging properties of HMCuSMnO2/Ce6(CMC)NPs could enable thereali

7、zation of more precise,reliable and on-demand combination therapy.In a word,this study illustrateda promising approach to strengthen the efficacy of HMCuS-based nanotherapeutics,which woulddefinitely promote the further exploitation of smarter nanoplatforms for synergistic disease management.1.Intro

8、ductionThe imbalance between unregulated tumor growth and dis-torted blood vessels results in an oxygen deficient tumormicroenvironment(TME),and intratumoral hypoxia contrib-utes to tumor heterogeneity,which significantly promotesaggressive metastatic spread.13Moreover,owing to the dis-ordered metab

9、olism and unstable genome phenotype acceler-ated by hypoxia,undesirable angiogenesis and immunosup-pression are dramatically promoted and aggravated,whichthereby makes the tumor cells more resistant to many oxygen-associated treatment modalities.46For instance,the thera-peutic outcome of photodynami

10、c therapy(PDT)strongly relieson the local oxygen level,during which singlet oxygen(1O2)converted from oxygen molecules(O2)under the photoactiva-tion of photosensitizers can selectively induce irreversibleapoptosis of tumor cells.79However,the concentration oflocal oxygen in tumor cells varies from 0

11、.020.2%,which is tre-mendously lower than that of 29%in normal cells.10,11Therestricted oxygen supply undoubtedly attenuates the treatmentefficacy,and PDT may in turn dramatically consume localoxygen to aggravate the hypoxic status.12Thus far,many nano-particle(NP)-based approaches have been used to

12、 effectivelyalleviate tumor hypoxia and improve the treatment outcome,including direct oxygen delivery mediated by the nanocarrierswith high oxygen affinity(e.g.,perfluorocarbon PFC andElectronic supplementary information(ESI)available.See DOI:10.1039/d0nr01547dThese authors contributed equally to t

13、his work.aInstitute for Clean Energy and Advanced Materials,School of Materials and Energy,Southwest University,Chongqing 400715,China.E-mail:bMedical Research Institute,Wuhan University,Wuhan,Hubei 430071,ChinacChongqing Key Laboratory of Ultrasound Molecular Imaging,Institute of UltrasoundImaging,

14、Second Affiliated Hospital,Chongqing Medical University,Chongqing400010,China.E-mail:12508|Nanoscale,2020,12,1250812521This journal is The Royal Society of Chemistry 2020Published on 13 May 2020.Downloaded by SUSTech on 6/29/2021 2:26:09 PM.View Article OnlineView Journal|View Issuehemoglobin Hb),13

15、16and the oxygen generation techniqueby decomposing endogenous hydrogen peroxide(H2O2)viananocatalysts(e.g.,manganese dioxide MnO2 and platinumPt).1720In particular,MnO2-based NPs are the most preva-lently employed catalase-like nanoenzymes for alleviatingtumor hypoxia,thanks to their admirable enzy

16、matic stabilityand high economic effectiveness.21,22Taking into accountweak acidic conditions,and high H2O2and glutathione(GSH)levels in the TME,MnO2can be degraded into Mn2+via acido-lysis,which further activates the catalytic decomposition ofH2O2into O2to realize local oxygenation.23Therefore,MnO2

17、-based NPs with versatile designs exhibit a promising prospectsfor realizing highly efficacious oxygen-dependent tumor thera-peutics by regulating the TME.Hollow mesoporous copper sulfide(HMCuS)NPs have beenreported as a category of potent nanoagents for tumor treat-ment,not only owing to their high

18、 photothermal conversionefficiency for hyperthermia-triggered tumor ablation,but alsodue to their large surface area and internal cavity for theencapsulation of small-molecule drugs as well.2427Moreover,in virtue of the strong absorption in the near infrared(NIR)region,HMCuS NPs can also realize hig

19、h performance photoa-coustic(PA)imaging with high spatiotemporal resolution.28,29However,unitaryHMCuS-mediatedphotothermaltherapy(PTT)is not always satisfactory owing to the heterogeneousTME,and adjuvant treatments are absolutely requisite toachieve a complete tumor eradication.3032In addition,unpre

20、-dictable and uncontrollable drug leakage from HMCuS NPsmay result in deleterious systemic toxicity and even multipledrug resistance(MDR).33Therefore,premature release of theencapsulated drug should be resolved before reaching thetumor region,so as to realize the highest tumor-specificenrichmentofsm

21、all-moleculetherapeuticagents.34,35Inanother aspect,chlorin e6(Ce6),as a widely used photosensiti-zer,is able to generate cytotoxic1O2(one typical reactiveoxygenspeciesROS)underlightactivationwithhighquantum yield efficiency and low background toxicity.3638Moreover,intrinsic fluorescence of Ce6 enab

22、les the monitoringof its biodistribution in vivo,through real-time imaging.39However,poor aqueous solubility,incapability of tumor target-ing and rapid degradation during long-time blood circulationof Ce6 significantly attenuate its curative effect,which wouldsignificantly restrict its extended appl

23、ications in disease man-agement.40Despite the compelling progress in the enhance-ment of Ce6 stability through covalent conjugation,the fluo-rescence quantum yield would be tremendously attenuatedbecause of the aggregation caused quenching(ACQ)effect.41In this study,a novel HMCuSMnO2/Ce6(CMC)coreshe

24、llnanoplatform was developed as a highly efficient TME modu-lator,which could effectively relieve tumor hypoxia throughdecomposing high-level local H2O2and lower the reduced glu-tathione(GSH)through glutathione peroxidase-like activity(Fig.1).Moreover,a MnO2nanoshell can also act as a gate-keeper by

25、 forming a stimulus-responsive plug within the meso-porous structure of HMCuS,which is advantageous for mini-mizing the premature release of entrapped Ce6.Upon CMCarriving at the cancerous region on the basis of the enhancedpermeability and retention(EPR)effect,a mildly acidic TMEFig.1Schematic illu

26、stration of applying CMC NPs for enhanced photothermal/photodynamic therapy under multimodal imaging guidance.NanoscalePaperThis journal is The Royal Society of Chemistry 2020Nanoscale,2020,12,1250812521|12509Published on 13 May 2020.Downloaded by SUSTech on 6/29/2021 2:26:09 PM.View Article Onlinew

27、ould degrade the MnO2component into Mn2+for tumor oxy-genation and magnetic resonance(MR)imaging.Afterwards,the disintegration of the MnO2nanoshell would further exposethe mesopores on HMCuS for the release of encapsulated Ce6,accompanying the fluorescence recovery for high resolutionfluorescence(FL

28、)imaging.Upon 660/808 nm dual-wavelengthlaserirradiation,hyperthermia-mediatedPTTandROS-mediated PDT can be triggered for tumor eradication,whichwas further enhanced in this hypoxia-alleviated and GSH-depleted environment.Under the guidance of PA/FL/MR tri-modal imaging,more precise and reliable PTT

29、/PDT combi-nation therapy would be implemented by using the as-devel-oped CMC NPs.Here,physiochemical properties of CMC NPswere characterized through morphology observation,determi-nation of chemical composition,size distribution measure-ment,etc.Thereafter,the admirable theranostic capacity ofCMC N

30、Ps was demonstrated both in vitro and in vivo.2.Materials and methods2.1.ChemicalsCopper chloride dihydrate(CuCl22H2O)was ordered fromSinopharm Chemical Reagent Co.,Ltd(China).Isopropylalcohol,sodium hydroxide(NaOH),polyvinylpyrrolidone(PVPK30),hydrazine monohydrate(N2H4H2O),ammonium sulfidesolution

31、(NH42S,20 wt%),and potassium permanganate(KMnO4)wereobtainedfromChongqingChuandongChemical(Group)Co.,Ltd(China).Poly(allylamine hydro-chloride)(PAH,Mw=15 000)was ordered from Sigma-Aldrich(USA).N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide hydro-chloride crystalline,3-(4,5-dimethylthiazol-2-yl)-2,5-

32、diphenylte-trazolium bromide(MTT)and dimethyl sulfoxide(DMSO)weresupplied by Shanghai Aladdin Bio-Chem Technology Co.,Ltd(China).Chlorin e6(Ce6)was ordered from Frontier Scientific,Inc.(USA).Dulbeccos modified Eagles medium(DMEM),PenStrep(100),fetal bovine serum(FBS),TrypLE ExpressEnzyme,a LIVE/DEAD

33、 Viability/Cytotoxicity Assay Kit,4,6-di-amidino-2-phenylindole(DAPI)and an Alexa Fluor 488annexin V/Dead Cell Apoptosis Kit were provided by ThermoFisher Scientific,Inc.(USA).A TUNEL Apoptosis Assay Kit,Hoechst33342,dichloro-dihydro-fluoresceindiacetate(DCFH-DA)assayandaJC-1MitochondrialMembranePot

34、entialDetectionKitwerepurchasedfromBeyotimeInstitute of Biotechnology(China).All the reagents were ofanalytical grade and directly used without additional purifi-cation.Deionized(DI)water was taken from a water purifi-cation system(Millipore Milli-Q Synthesis A10,France).2.2.Synthesis of HMCuS NPsHM

35、CuS NPs were prepared at room temperature under mildconditions according to a previous report.42Specifically,CuCl22H2O(17 mg)was dissolved in isopropyl alcohol(60 mL),followed by introduction of 0.2 g PVP(K30)and 2 mLNaOH(0.5 mM)into the previous solution.After vortexing for10 min,0.4 mL N2H4H2O(35%

36、)was dropwise added and thereaction was allowed to proceed for another 10 min.The for-mation of orange floccules indicated the successful synthesisof Cu2O,which were centrifugally isolated(8000 rpm,15 min)and further purified with ethanol/H2O at least three times.Toprepare HMCuS NPs,(NH4)2S(35 L,20

37、wt%)was introducedinto the aqueous dispersion of Cu2O(30 mL),followed by stir-ring for 1 h.When the reaction system became dark green,pre-cipitates were centrifugally isolated(8000 rpm,15 min)andfurther purified with ethanol/DI water at least three times.Finally,the as-synthesized HMCuS NPs were deh

38、ydrated undervacuum for 12 h prior to use.2.3.Synthesis of CMC NPs10 mL Ce6 solution(1 mg mL1)was mixed with 10 mLHMCuS NP aqueous dispersion(2 mg mL1).Subsequent to12 h mixing,10 mL KMnO4(2 mg mL1)was dropwise addedinto the above dispersion under stirring.Thereafter,PAH(20 mg),as a reducing agent,w

39、as added into the reactingsystem.The reaction proceeded for 15 min until the solutionchanged color from purple to brown.The product was centri-fugally collected(8000 rpm,15 min)and further purified withDI water thrice.Finally,the as-synthesized CMC NPs were de-hydrated under vacuum overnight ahead o

40、f use.The encapsu-lation efficiency(EE)and loading capacity(LC)of Ce6 in CMCNPs were determined on the basis of the standard curvederived from the fluorescence spectrum of Ce6(ex:640 nm,em:720 nm).And,these characteristic parameters were calcu-lated according to formulas(1)and(2).EE%Weightof encapsu

41、latedCe6=Weightof devotedCe6?100%1LC%Weightof encapsulatedCe6=Weightof CMCNPs?100%22.4.CharacterizationMorphological features of the as-synthesized products werecharacterized by using a LIBRA120 PLUS transmission elec-tron microscope(TEM)and a JSM-7800F field emission scan-ning electron microscope(F

42、ESEM).The elemental compo-sition of samples was determined by using energy dispersivespectroscopy(EDS).The hydrodynamic size and surface poten-tial were determined by using a Zetasizer Nano ZS90(MalvernPanalytical,USA).UV-vis-NIR absorption spectra were recordedwith a Shimadzu UV-1800 UV/Visible sca

43、nning spectrophoto-meterandfluorescencespectrawereacquiredusingaRF-5301PCspectrofluorophotometer.The X-ray diffraction(XRD)pattern was measured using a Shimadzu XRD-7000X-ray diffractometer.Valence states and chemical compo-sitions of products were determined by X-ray photoelectronspectroscopy(XPS)u

44、sing a Thermo ESCALAB250Xi X-rayphotoelectron spectrometer.Nitrogen sorption isotherms weremeasured using a surface area and pore size analyzer(NOVAPaperNanoscale12510|Nanoscale,2020,12,1250812521This journal is The Royal Society of Chemistry 2020Published on 13 May 2020.Downloaded by SUSTech on 6/2

45、9/2021 2:26:09 PM.View Article Online1200e,QuantachromeInstruments)basedonBrunauerEmmettTeller(BET)theory.2.5.Assessment of photothermal performanceAn aqueous dispersion of CMC NPs(3 mL,concentrationrange:0100 g mL1)in cuvettes was illuminated using acontinuous-wave diode NIR laser(808 nm,1.5 W cm2)

46、for10 min.Real-time temperature of different samples wasdynamically measured every 20 s using a digital thermo-meter.NIR laser power dependent temperature elevation wassimilarly measured in accordance with the same procedure.Thermographic images were acquired in 2 min intervals byusingathermalinfrar

47、edimager(FlukeTiS55,USA).Photothermal conversion stability of CMC NPs was deter-mined via on/offlaser irradiation for five cycles(10 minexposure and cooling to room temperature prior to the fol-lowing irradiation period).2.6.Drug release in vitroThe kinetics of Ce6 release from CMC NPs was determine

48、d inaccordance with the standard dialysis method.43Briefly,1 mLaqueous dispersion of CMC NPs(1 mg mL1)in a dialysis bag(Mw=3500)was submerged in 30 mL saline buffer as therelease medium.Distinct pH conditions(pH=7.4 or 6.8)wereapplied to simulate the physiological environment and TME,respectively.Mo

49、reover,100 M H2O2was used to reflect theunique condition in the TME for the“H2O2(+)”group.Thewhole setup was located in a water bath with a fixed tempera-ture of 37 C in the dark.At predefined time points,1 mLrelease medium was sampled,and the releasing system wasreplenished with saline buffer at th

50、e same volume.Thecontent of released Ce6 was determined using fluorescencespectrophotometry as previously mentioned,and cumulativerelease of Mn2+was quantified through inductively coupledplasmamassspectrometry(ICP-MS;XSeriesII,ThermoScientific).2.7.O2generation activated by H2O2Briefly,different sam