In vitro study of homologous targeted multimodal imaging nanoprobes in synergistic photothermal-chemotherapy treatment
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摘要:
目的 制备同源靶向性诊疗一体化纳米探针MnO2@DOX@ICG@CM(MDIC)，观察MDIC纳米探针的同源靶向能力，探讨其多模态成像价值以及光热联合化疗的协同治疗作用。 方法 通过化学裂解和超声匀浆法获取乳腺癌4T1细胞膜，以脂质体挤压法制备细胞膜包被的同时载有化疗药阿霉素和光敏剂吲哚菁绿的MnO2纳米探针；对MDIC的外观形态、粒径和表面电位等性质进行基本表征，并研究其光声成像及磁共振成像的能力；观察不同浓度的MDIC纳米探针的光热升温性能，评估其对4T1细胞的光热-化疗协同治疗的抗肿瘤效果。 结果 成功制备MDIC纳米探针，呈花状结构，平均粒径为183.3 nm，表面电位为-20.6±0.77 mV。透射电镜下观察到细胞膜成功包被；MDIC经激光照射后，溶液温度随着纳米探针浓度的升高而增加，具有较好的光热升温效果；光声和磁共振成像结果显示MDIC具有良好的成像对比能力；激光共聚焦显微镜观察到MDIC具有良好的同源靶向性；细胞毒性实验结果显示光热-化疗协同治疗具有更好的癌细胞杀伤作用。 结论 本研究成功制备了一种同源靶向性的光声/磁共振多模态成像对比探针MDIC，该探针具有良好的光热升温效能和体外抗肿瘤效果，并且可以显著增强光声成像效果和磁共振T1增强效果。 Abstract:Objective To prepare homologous targeting therapeutic integrated nanoprobe MnO2@DOX@ICG@CM(MDIC), observe the homologous targeting ability of MDIC nanoprobe, and explore its multimodal imaging value and the synergistic effect of photothermal combined chemotherapy. Methods The cell membrane of breast cancer 4T1 was obtained by chemical lysis and ultrasonic homogenization. The MnO2 nanoprobe coated with doxorubicin DOX and photosensitizer indocyanine green was prepared by liposome extruding method. The appearance morphology, particle size and surface potential of MDIC were characterized, and its photoacoustic imaging and magnetic resonance imaging capabilities were studied. The photothermal heating properties of MDIC nanoprobes with different concentrations were observed. MTT assay was used to evaluate the antitumor effect of photothermal-chemotherapy synergistic therapy on 4T1 cells. Results MDIC nanoprobes were successfully prepared with flower-like structure, average particle size of 183.3 nm, and surface potential of -20.6±0.77 mV. The membrane was successfully coated under transmission electron microscope. After laser irradiation of MDIC, the solution temperature increases with the increase of nanoprobe concentration, which has a good photothermal heating effect. The results of photoacoustic and magnetic resonance imaging showed that MDIC had good imaging contrast ability. Laser confocal microscopy showed that MDIC had good homology targeting. The results of cytotoxicity test showed that photothermalchemotherapy combined therapy had better cancer cell killing effect. Conclusion A homologous targeting photoacoustic/ magnetic resonance multimodal imaging contrast probe MDIC was successfully prepared. The probe has good photothermal heating efficiency and anti-tumor effect in vitro. It can significantly enhance the photoacoustic imaging effect and magnetic resonance T1 enhancement effect. -
图 1 MDIC制备示意图
A~C: 分别为MnO2、MDI、MDIC的TEM图; D: 动态光散射图; E: 表面电位图; F: 紫外吸收光谱图.
Figure 1. Schematic diagram of preparation of MDIC.
图 3 MDIC纳米探针的光声成像性能
A: 不同浓度纳米探针的光声信号强度值(750 nm); B: MDIC与不同浓度H2O2反应后的光声信号强度值(750 nm); C: 不同浓度纳米探针的光声信号强度值(875 nm); D: MDIC与不同浓度H2O2反应后的光声信号强度值(875 nm); E、F: 不同pH值、H2O2浓度下MDIC的不同激发波长光声信号谱图.
Figure 3. Photoacoustic imaging performance of MDIC nanoprobes.
图 4 MDIC分子探针磁共振成像性能
A：不同浓度MDIC的体外磁共振成像图; B: 体外纵向弛豫率(r1)比较.
Figure 4. Magnetic resonance imaging properties of MDIC molecular probes.
图 5 MDIC分子探针的光热升温性能评估
A: 不同浓度MDIC在808 nm激光下(1.5 W/cm2)照射5 min后的升温曲线(MnO2: 6.25 μg/mL、12.5 μg/mL、25 μg/mL、50 μg/mL和100 μg/mL); B: 不同浓度MDIC经808 nm激光照射后的热成像图.
Figure 5. Evaluation of photothermal performance of MDIC molecular probe
图 6 激光共聚焦显微镜观察靶向纳米探针靶向能力
A: 红色荧光染料DOX标记的MDIC-4T1; B: 蓝色荧光染料DAPI标记的4T1细胞; C: 靶向组纳米粒靶向情况; D: 红色荧光染料DOX标记的MDIC-Hela; E: 蓝色荧光染料DAPI标记的4T1细胞; F: 非靶向组纳米粒靶向情况. 比例尺：20 μm.
Figure 6. The targeting ability of the targeted nanoprobe was observed by laser confocal microscopy.
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