Au Nanocages Probes for Mitochondrial Imaging and Photothermal Damage Cells★

doi:10.6023/A23050222

Acta Chimica Sinica ›› 2023, Vol. 81 ›› Issue (10): 1301-1310.DOI: 10.6023/A23050222 Previous Articles Next Articles

Special Issue: 庆祝《化学学报》创刊90周年合辑

Original article

金纳米笼探针用于线粒体成像和光热损伤细胞^★

孙丽^a, 王亚静^a, 李涛^b, 郭英姝^b^,^*(), 张书圣^a^,^*()

a 临沂大学化学化工学院山东临沂 276005
b 齐鲁工业大学(山东省科学院)化学与化工学院济南 250353

投稿日期:2023-05-12 发布日期:2023-07-17
作者简介:
^★庆祝《化学学报》创刊90周年.
基金资助:
国家自然科学基金(22276102); 泰山学者工程(tsqn202211212); 山东省自然科学基金(ZR2023JQ004); 山东省自然科学基金(ZR2022MB024); 济南市引进创新团队(202228027); 山东省高等学校青创科技计划创新团队(2020KJC003); 齐鲁工业大学(山东省科学院)科教产融合试点工程项目(2023PYI002)

Au Nanocages Probes for Mitochondrial Imaging and Photothermal Damage Cells^★

Li Sun^a, Yajing Wang^a, Tao Li^b, Yingshu Guo^b(), Shusheng Zhang^a()

a School of Chemistry and Chemical Engineering, Linyi University, Linyi, Shandong 276005
b School of Chemistry and Chemical Engineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353

Received:2023-05-12 Published:2023-07-17
Contact: *E-mail: yingshug@126.com; shushzhang@126.com
About author:
^★Dedicated to the 90th anniversary of Acta Chimica Sinica.
Supported by:
National Natural Science Foundation of China(22276102); Young Taishan Scholar Program of Shandong Province of China(tsqn202211212); Shandong Provincial Natural Science Foundation(ZR2023JQ004); Shandong Provincial Natural Science Foundation(ZR2022MB024); University Institute Innovation Team of Jinan(202228027); Development Plan of Youth Innovation Team in Colleges and Universities of Shandong Province(2020KJC003); Science, Education and Industry Integration Pilot Project Plan of Qilu University of Technology (Shandong Academy of Sciences)(2023PYI002)

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Abstract

Mitochondria are the key regulatory organelles of many cell behaviors, and the reduction of mitochondrial membrane potential is considered to be one of the earliest events of cell apoptosis. Therefore, mitochondrial imaging and the detection and analysis of mitochondrial membrane potential are of great scientific significance for the detection and treatment of diseases. In this work, AuNCs/PLEL/JC/KLA, a mitochondrial targeted fluorescent nanoprobe, was developed using Au nanocages (AuNCs) mediated photothermal damage combined with temperature-sensitive drug release. At the same time, the temperature-sensitive hydrogel poly(d,l-lactide)-poly(ethylene glycol)-poly(d,l-lactide) (PDLLA-PEG-PDLLA, PLEL) is used as the outer structure to control the release. A mitochondrial targeting peptide (KLAKLAKKLAKLAK, KLA) was introduced as the "pointer" of the nanoprobe to specifically target the mitochondria. The colocalization experiment showed that the nanoprobe was highly colocalized with mitochondria, indicating that the nanoprobe was selectively enriched in mitochondria. It is worth noting that the nanoprobe has excellent photothermal properties, and its photothermal conversion efficiency can be as high as 39.11%. Therefore, under the irradiation of near infrared light, the probe can absorb light energy into heat. Subsequently, the results of Cell Counting Kit 8 (CCK-8) confirmed that the nanoprobes could achieve local photothermal damage at mitochondrial sites, triggering high temperature mediated mitochondrial dysfunction and inducing apoptosis of cancer cells. Meanwhile, rheological analysis and fluorescence curve showed that high temperature promotes the gel-sol transformation of PLEL thermosensitive hydrogel and realizes the release of fluorescent dye (JC-10). The confocal images of the cells showed that the released JC-10 fluorescent dye can display red and green fluorescence signals based on mitochondrial activity. In conclusion, the fluorescence nanoprobe can not only achieve mitochondrial targeted fluorescence imaging and damage cells, but also monitor the changes of mitochondrial membrane potential.

Key words: mitochondrion, fluorescent probe, photothermal damage, fluorescence imaging, membrane potential

Cite this article

Li Sun, Yajing Wang, Tao Li, Yingshu Guo, Shusheng Zhang. Au Nanocages Probes for Mitochondrial Imaging and Photothermal Damage Cells^★[J]. Acta Chimica Sinica, 2023, 81(10): 1301-1310.

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Fig. & Tab. 11

Figure 1. Synthesis of AuNCs/PLEL/JC/KLA nanoprobes and schematic diagram of targeting mitochondria photothermal damage

Figure 2. TEM images of (a) AgNPs, (b) AuNCs, (c) PLEL, (d) AuNCs/PLEL/JC/KLA

Figure 3. (a) Particle size graphs of AgNPs, AuNCs, PLEL, AuNCs/ PLEL/JC and AuNCs/PLEL/JC/KLA. (b) Zeta potentials of AuNCs, PLEL, AuNCs/PLEL, JC, AuNCs/PLEL/JC, KLA and AuNCs/PLEL/ JC/KLA

Figure 4. (a) UV-vis curves of AgNPs, AuNCs, JC and AuNCs/ PLEL/JC; (b) FTIR curves of PLEL and AuNCs/PLEL/JC/KLA

Figure 5. (a) Temperature rise curves of different concentrations of AuNCs solution irradiated with 1.5 W/cm2 NIR light for 10 min; (b) temperature rise curves of H2O, AuNCs, AuNCs/PLEL and AuNCs/ PLEL/JC/KLA under NIR irradiation of 1.5 W/cm2 for 10 minutes; (c) Heating and cooling curves of AuNCs/PLEL/JC/KLA during three near-infrared on-off cycles; (d) Infrared thermogram of AuNCs/ PLEL/JC/KLA at 1.5 W/cm2 NIR irradiation for 10 min

Figure 6. Rheological analysis diagrams of (a) PLEL and (b) AuNCs/ PLEL/JC/KLA

Figure 7. Fluorescence spectra of JC-10 in AuNCs/PLEL/JC/KLA supernatant at 37 and 40 ℃

Figure 8. Cell viability diagrams of (a) Ramos cells treated with different concentrations of AuNCs, AuNCs/PLEL/JC and AuNCs/PLEL/ JC/KLA without NIR laser irradiation; (b) Ramos cells was treated with different concentrations of AuNCs, AuNCs/PLEL/JC and AuNCs/PLEL/ JC/KLA under the irradiation of NIR laser

Figure 9. Colocalization images of Ramos cells incubated with JC-10 and AuNCs/PLEL/JC/KLA probes respectively, with a scale of 50 μm

Figure 10. After incubating the probe with Ramos cells, confocal images of the cells were irradiated with NIR light for different times, with a scale of 10 μm

Figure 11. Confocal image of Ramos cells incubated with AuNCs/PLEL/JC/KLA after being treated with different concentrations of carbonyl chlorophenylhydrazone cyanide (CCCP), with a scale of 50 μm

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金纳米笼探针用于线粒体成像和光热损伤细胞^★

Au Nanocages Probes for Mitochondrial Imaging and Photothermal Damage Cells^★

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金纳米笼探针用于线粒体成像和光热损伤细胞★

Au Nanocages Probes for Mitochondrial Imaging and Photothermal Damage Cells★

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金纳米笼探针用于线粒体成像和光热损伤细胞^★

Au Nanocages Probes for Mitochondrial Imaging and Photothermal Damage Cells^★