[1]邹卫娟,郝俊年,吴建荣,等.氧化铱纳米载药复合物的制备及其用于肿瘤声动力-化疗协同治疗的研究[J].陆军军医大学学报(原第三军医大学学报),2021,43(22):2395-2406.
　ZOU Weijuan,HAO Junnian,WU Jianrong,et al.Preparation of drug-loaded iridium oxidenanocomposites and its application in synergetic sonodynamic/chemotherapeutic therapy against cancer[J].J Amry Med Univ (J Third Mil Med Univ),2021,43(22):2395-2406.
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氧化铱纳米载药复合物的制备及其用于肿瘤声动力-化疗协同治疗的研究(/HTML )
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陆军军医大学学报（原第三军医大学学报）[ISSN:1000-5404/CN:51-1095/R]

卷:
43卷
期数:
2021年第22期
页码:
2395-2406
栏目:
基础医学
出版日期:
2021-11-30

文章信息/Info

Title:
Preparation of drug-loaded iridium oxidenanocomposites and its application in synergetic sonodynamic/chemotherapeutic therapy against cancer
作者:
邹卫娟; 郝俊年; 吴建荣; 郑元义
重庆医科大学超声分子影像重庆市重点实验室；上海交通大学附属第六人民医院，上海超声医学研究所
Author(s):
ZOU Weijuan;  HAO Junnian;  WU Jianrong;  ZHENG Yuanyi
Chongqing Key Laboratory of Ultrasound Molecular Imaging, Chongqing Medical University, Chongqing, 400010; 2Shanghai Institute of Ultrasound Medicine, Sixth People’s Hospital affiliated to Shanghai Jiaotong University, Shanghai, 200233, China
 
关键词:
氧化铱; 声动力治疗; 化疗; 协同治疗; 肿瘤
Keywords:
iridium oxide;  sonodynamic therapy;  chemotherapy;  synergistic therapy;  tumor
分类号:
R454.3; R730.58; R944.9
文献标志码:
A
摘要:

目的制备氧化铱（iridium oxide，IrOX）作为纳米声敏剂，用以负载化疗药物阿霉素（doxorubicin，DOX）后得到具有pH和超声辐照双响应的纳米载药复合物（IrOX@DOX），验证其用于肿瘤声动力治疗（sonodynamic therapy，SDT）-化疗协同治疗肿瘤的效果。方法采用热水解法制备IrOX纳米颗粒，并负载DOX。采用透射电子显微镜（transmission electron microscope, TEM）、X射线光电子能谱（X-ray photoelectron spectroscopy, XPS）、紫外-可见分光光度计（ultraviolet-visible spectrophotometer, UV-vis）、Zeta 电位分析仪等手段对纳米颗粒的形貌及理化性质进行相关表征；采用UV-Vis和酶标仪考察载药量及药物释放行为；用电子顺磁共振波谱仪（lectron spin-resonance spectroscopy, ESR）定性评价其活性氧产生性能；激光共聚焦显微镜评价IrOX@DOX在超声辐照下的细胞摄取行为；荧光显微镜和流式细胞仪探究细胞内活性氧产生；CCK-8法及流式凋亡检测评价IrOX@DOX的声动力治疗-化疗协同治疗效果。建立 Hepa1-6肝癌裸鼠皮下移植瘤模型，并分为生理盐水组、IrOX组、IrOX+US组、IrOX@DOX组、IrOX@DOX+US组（n=3），观察各组荷瘤鼠肿瘤的体积、超声造影成像结果与病理学变化，计算肿瘤体积抑制率评价其体内协同治疗效果。结果 成功制备IrOX纳米颗粒，粒径大约为5 nm。当IrOX与DOX的比值为4.0时，载药量可达18.7%；IrOX@DOX在pH =7.4时释放的DOX量不足10%，证实了其良好的稳定性；pH=5.5 时， DOX的累积释放量可达39.9%。在超声辐照下释放量进一步增加达69.9 %；ESR 和 UV-Vis 实验结果表明活性氧产量与辐照功率和时间呈正相关；激光共聚焦结果显示超声能促进细胞对IrOX@DOX的摄取；细胞毒性实验表明，与单纯化疗组相比，协同治疗组存活率降至19.4%，证明IrOX@DOX在超声辐照下可以实现良好的SDT-化疗协同治疗效果。动物实验表明与生理盐水组相比，IrOX+US组、IrOX@DOX组和IrOX @DOX+US组的肿瘤抑制率分别为27%、57%及76%，差异具有统计学意义（P<0.05）。这与超声造影和HE染色结果一致。结论 成功制备了具有pH和超声辐照双重刺激响应的IrOX@DOX纳米复合物，在超声辐照下可以实现SDT-化疗协同治疗。

Abstract:

ObjectiveTo prepare iridium oxide (IrOX) as a nano-sonosensitizer to load chemotherapy drug doxorubicin (DOX) in order to obtain nanocomposites IrOX@DOX with dual response of pH/ultrasound (US) irradiation, and to verify its efficacy in sonodynamic therapy (SDT)-chemotherapy for tumor treatment. MethodsIrOX nanoparticles were prepared by thermal hydrolysis and then loaded with DOX. The morphological, chemical and physical properties of the nanoparticles were characterized by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), ultraviolet-visible spectrophotometer (UV-Vis) and Zeta potential analyzer. The drug loading efficiency and drug release behavior were determined with UV-vis spectrometry and ELISA. The generation of reactive oxygen species (ROS) by IrOX@DOX nanoparticles was evaluated usingelectron spin-resonance spectroscopy (ESR). The uptake of IrOX@DOX by 4T1 cells under ultrasonic irradiation was evaluated with laser confocal scanning microscopy (LCSM), and the level of intracellular ROS was investigated using fluorescence microscopy and flow cytometry. The synergistic efficacy of IrOX@DOX in SDT-chemotherapy against 4T1 cells was evaluated by CCK-8 assay and flow cytometry. In addition, the Hepa1-6 xenograft tumor model in nude mice was built and divided into 5 groups (n=3 in each group) as follows: saline, IrOX, IrOX+US, IrOX@DOX, and IrOX @DOX+US. The tumor volume, contrast-enhanced US imaging and pathological changes of each group were observed, and the inhibitory rate of tumor volume was calculated to evaluate the effect of synergistic therapy in vivo. Results IrOX nanoparticles were successfully prepared, at a diameter of about 5.0 nm, and when the ratio of IrOX to DOX was 4.0, the drug loading reached 18.7%. Prepared IrOX@DOX released less than 10% of DOX at pH 7.4, which confirmed its good stability, while the cumulative release of DOX reached 39.9% at pH 5.5, and further increased to 69.9% under US irradiation. Both ESR and UV-vis experiments demonstrated that ROS production was positively correlated with irradiation power and time. Meanwhile, LCSM showed that US promoted the uptake of IrOX@DOX by tumor cells. In vitro cytotoxicity experiments indicated the survival rate of tumor cells in the synergistic treatment group (IrOX @DOX+US) was reduced to 19.4%, significantly lower than that of the chemotherapy group alone (P<0.05), which proved that IrOX@DOX nanocomposites achieved an excellent synergistic effect of SDT-chemotherapy under ultrasound irradiation. Furthermore, in vivo experiments showed that as compared with saline group, the tumor-inhibiting rate of IrOX+US, IrOX@DOX and IrOX@DOX+US groups were 27%, 57% and 76%, respectively, indicating significant difference (P<0.05), and this was consistent with contrast US imaging and H&E staining results. Conclusion IrOX@DOX nanocomposites with dual stimulation response of pH and US irradiation are successfully developed, and the synergistic treatment of SDT-chemotherapy can be achieved under US irradiation.

参考文献/References:

[1]SUNG H, FERLAY J, SIEGEL R L, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries[J]. CA Cancer J Clin, 2021, 71(3): 209-249. DOI: 10.3322/caac.21660.
 
[2]MILLER K D, SIEGEL R L, LIN C C, et al. Cancer treatment and survivorship statistics, 2016[J]. CA Cancer J Clin, 2016, 66(4): 271-289. DOI: 10.3322/caac.21349.
 
[3]KONG F H, YE Q F, MIAO X Y, et al. Current status of sorafenib nanoparticle delivery systems in the treatment of hepatocellular carcinoma[J]. Theranostics, 2021, 11(11): 5464-5490. DOI: 10.7150/thno.54822.
 
[4]HU H, FENG W, QIAN X Q, et al. Emerging nanomedicine-enabled/enhanced nanodynamic therapies beyond traditional photodynamics[J]. Adv Mater, 2021, 33(12): e2005062. DOI: 10.1002/adma.202005062.
 
[5]REN Z G, SUN S C, SUN R R, et al. A metal-polyphenol-coordinated nanomedicine for synergistic cascade cancer chemotherapy and chemodynamic therapy[J]. Adv Mater, 2020, 32(6): e1906024. DOI: 10.1002/adma.201906024.
 
[6]ZHAO N X, ZENG Z H, ZU Y L. Self-assembled aptamer-nanomedicine for targeted chemotherapy and gene therapy[J]. Small, 2018, 14(4): 1702103. DOI: 10.1002/smll.201702103.
 
[7]QIAN X Q, ZHENG Y Y, CHEN Y. Micro/nanoparticle-augmented sonodynamic therapy (SDT): breaking the depth shallow of photoactivation[J]. Adv Mater, 2016, 28(37): 8097-8129. DOI: 10.1002/adma.201602012.
 
[8]PAN X T, WANG H Y, WANG S H, et al. Sonodynamic therapy (SDT): a novel strategy for cancer nanotheranostics[J]. Sci China Life Sci, 2018, 61(4): 415-426. DOI: 10.1007/s11427-017-9262-x.
 
[9]MA A Q, CHEN H Q, CUI Y H, et al. Metalloporphyrin complex-based nanosonosensitizers for deep-tissue tumor theranostics by noninvasive sonodynamic therapy[J]. Small, 2019, 15(5): e1804028. DOI: 10.1002/smll.201804028.
 
[10]SON S, KIM J H, WANG X W, et al. Multifunctional sonosensitizers in sonodynamic cancer therapy[J]. Chem Soc Rev, 2020, 49(11): 3244-3261. DOI: 10.1039/c9cs00648f.
 
[11]LEI B, CHEN M, WANG Y, et al. Double security drug delivery system DDS constructed by multi-responsive (pH/redox/US) microgel[J]. Colloids Surf B Biointerfaces, 2020, 193: 111022. DOI: 10.1016/j.colsurfb.2020.111022.
 
[12]KARIMI M, GHASEMI A, SAHANDI ZANGABAD P, et al. Smart micro/nanoparticles in stimulus-responsive drug/gene delivery systems[J]. Chem Soc Rev, 2016, 45(5): 1457-1501. DOI: 10.1039/c5cs00798d.
 
[13]MENG Z Q, ZHOU X F, SHE J L, et al. Ultrasound-responsive conversion of microbubbles to nanoparticles to enable background-free in vivo photoacoustic imaging[J]. Nano Lett, 2019, 19(11): 8109-8117. DOI: 10.1021/acs.nanolett.9b03331.
 
[14]QIAN X Q, HAN X X, CHEN Y. Insights into the unique functionality of inorganic micro/nanoparticles for versatile ultrasound theranostics[J]. Biomaterials, 2017, 142: 13-30. DOI: 10.1016/j.biomaterials.2017.07.016.
 
[15]YANG S W, WANG X L, HE P, et al. Graphene quantum dots with pyrrole N and pyridine N: superior reactive oxygen species generation efficiency for metal-free sonodynamic tumor therapy[J]. Small, 2021, 17(10): e2004867. DOI: 10.1002/smll.202004867.
 
[16]OSMINKINA L A, NIKOLAEV A L, SVIRIDOV A P, et al. Porous silicon nanoparticles as efficient sensitizers for sonodynamic therapy of cancer[J]. Micropor Mesopor Mater, 2015, 210: 169-175. DOI: 10.1016/j.micromeso.2015.02.037.
 
[17]ZHAO Y, ZHU Y C. Synergistic cytotoxicity of low-energy ultrasound and innovative mesoporous silica-based sensitive nanoagents[J]. J Mater Sci, 2014, 49(10): 3665-3673. DOI: 10.1007/s10853-014-8073-y.
 
[18]BAI S, YANG N, WANG X, et al. Ultrasmall iron-doped titanium oxide nanodots for enhanced sonodynamic and chemodynamic cancer therapy[J]. ACS Nano, 2020, 14(11): 15119-15130. DOI: 10.1021/acsnano.0c05235.
 
[19]WU J R, WILLIAMS G R, NIU S W, et al. Biomineralized bimetallic oxide nanotheranostics for multimodal imaging-guided combination therapy[J]. Theranostics, 2020, 10(2): 841-855. DOI: 10.7150/thno.40715.
 
[20]ZHEN W Y, LIU Y, LIN L, et al. BSA-IrO2: catalase-like nanoparticles with high photothermal conversion efficiency and a high X-ray absorption coefficient for anti-inflammation and antitumor theranostics[J]. Angew Chem Int Ed, 2018, 57(32): 10309-10313. DOI: 10.1002/anie.201804466.
 
[21]WU J R, NIU S W, BREMNER D H, et al. A tumor microenvironment-responsive biodegradable mesoporous nanosystem for anti-inflammation and cancer theranostics[J]. Adv Healthc Mater, 2020, 9(2): e1901307. DOI: 10.1002/adhm.201901307.
 
[22]LI Q L, LIU D L, XU L, et al. Wire-in-tube IrOx architectures: alternative label-free immunosensor for amperometric immunoassay toward α-fetoprotein[J]. ACS Appl Mater Interfaces, 2015, 7(40): 22719-22726. DOI: 10.1021/acsami.5b07895.
 
[23]ZHEN W Y, LIU Y, ZHANG M C, et al. Multi-caged IrO x for facile preparation of “six-in-one” nanoagent for subcutaneous and lymphatic tumors inhibition against recurrence and metastasis[J]. Adv Funct Mater, 2020, 30(24): 2002274. DOI: 10.1002/adfm.202002274. 
 
[24]ZHEN W Y, LIU Y, WANG W, et al. Specific “unlocking” of a nanozyme-based butterfly effect to break the evolutionary fitness of chaotic tumors[J]. Angew Chem Int Ed, 2020, 59(24): 9491-9497. DOI: 10.1002/anie.201916142.
 
[25]郭丹, 汪星月, 谭米肖, 等. 声敏型脂质纳米粒联合低功率聚焦超声对HepG2细胞增殖的影响及其体外CT成像的实验研究[J]. 第三军医大学学报, 2018, 40(23): 2180-2189. DOI: 10.16016/j.1000-5404.201806153. 
 
GUO D, WANG X Y, TAN M X, et al. Effect of sound sensitive nano-liposomes combined with low intensity focused ultrasound on proliferation of HepG2 cells and its CT imaging in vitro[J]. J Third Mil Med Univ, 2018, 40(23): 2180-2189. DOI: 10.16016/j.1000-5404.201806153. 
 
[26]李倩茹, 郝兰, 王艺睿, 等. 基于DVDMS/IR780复合纳米粒抗肿瘤声动力实验研究[J]. 第三军医大学学报, 2021, 43(7): 606-613. DOI: 10.16016/j.1000-5404.202011241.
 
LI Q R, HAO L, WANG Y R, et al. Preparation of sinoporphyrin sodium and IR780-based composite nanoparticles and its anti-tumor efficacy in sonodynamic therapy[J]. J Third Mil Med Univ, 2021, 43(7): 606-613. DOI: 10.16016/j.1000-5404.202011241. 
 
[27]WANG X W, ZHONG X Y, BAI L X, et al. Ultrafine titanium monoxide (TiO1+x) nanorods for enhanced sonodynamic therapy[J]. J Am Chem Soc, 2020, 142(14): 6527-6537. DOI: 10.1021/jacs.9b10228.
 
[28]CHEN H J, ZHOU X B, GAO Y, et al. Recent progress in development of new sonosensitizers for sonodynamic cancer therapy[J]. Drug Discov Today, 2014, 19(4): 502-509. DOI: 10.1016/j.drudis.2014.01.010.
 
[29]COSTLEY D, MC EWAN C, FOWLEY C, et al. Treating cancer with sonodynamic therapy: a review[J]. Int J Hyperther, 2015, 31(2): 107-117. DOI: 10.3109/02656736.2014.992484.
 
[30]GEERS B, LENTACKER I, SANDERS N N, et al. Self-assembled liposome-loaded microbubbles: The missing link for safe and efficient ultrasound triggered drug-delivery[J]. J Control Release, 2011, 152(2): 249-256. DOI: 10.1016/j.jconrel.2011.02.024.

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更新日期/Last Update: 2021-11-23