Mitochondria-targeting multi-metallic ZnCuO nanoparticles and IR780 for efficient photodynamic and photothermal cancer treatments

doi:10.1016/j.jmst.2021.01.035

Abstract

Abstract:

Chemo-resistance has pushed cancer treatment to the boundary of failure. This challenge has encouraged scientists to look for nanotechnological solutions. In this study, we have taken this goal one step further without depending on chemotherapy. Specifically, hybrid metal, polymer, and lipid nanoparticles that formed an IR780-a photosensitizer and Zinc copper oxide incorporated nanoparticle (ZCNP) nanoparticles were utilized in a combined photothermal and photodynamic therapy. Through the mediation of triphenylphosphonium (TPP) as a mitochondria-targeting moiety, TPP-conjugated polymer-lipid hybrid nanoparticles containing ZCNP/IR-780 significantly enhanced cellular uptake by cancer cells and selectively targeted the mitochondria, which improved the induction of apoptosis. In tumor-bearing mice, the nanoparticles were detected predominantly in tumors rather than in the other principle organs, which did not show notable signs of toxicity. Both in vitro and in vivo results demonstrated a great improvement in photothermal and photodynamic efficacy in combination when compared to either one individually, and a significant inhibition of tumor growth was observed with the combined therapies. In summary, this study describes an effective mitochondria-targeting nanocarrier for the treatment of cancer using combined photothermal and photodynamic therapies.

Key words: Zinc-doped copper, Nanoparticles, Mitochondria, Cancer, Photothermal, Photodynamic

Hima Bindu Ruttala, Thiruganesh Ramasamy, Raghu Ram Teja Ruttala, Tuan Hiep Tran, Jee-Heon Jeong, Han-Gon Choi, Sae Kwang Ku, Chul Soon Yong, Jong Oh Kim. Mitochondria-targeting multi-metallic ZnCuO nanoparticles and IR780 for efficient photodynamic and photothermal cancer treatments[J]. J. Mater. Sci. Technol., 2021, 86: 139-150.

Figures/Tables 7

Scheme 1. Schematic illustration of the mitochondria-targeting TPP-conjugated-ZnCuO NP (ZCNP) and IR780-loaded polymer-lipid hybrid NPs (TPP-ZC/IR-PNPs).

Fig. 1. Characterization of TPP-ZC/IR-PNPs. (a) TEM images of (i) ZCNP and (ii) TPP-ZC-IR-PNPs; (b) AFM images of (i) ZCNP and (ii) TPP-ZC-IR-PNPs; (c) Elemental analysis of Zn and Cu in the TPP-ZC-IR-PNPs performed by energy dispersive X-ray spectroscopy; (d) UV/vis absorption spectra of ZCNP, IR780, and TPP-ZC-IR-PNPs; (e) Photostability of TPP-ZC-IR-PNPs under daylight and darkness at 24 h.

Fig. 2. In vitro cellular uptake of TPP-ZC-IR-PNPs. (a) TPP-ZC-IR-PNP time-dependent uptake was analyzed by FACS at different time points (0.25, 0.5, 1, and 3 h); (b, c) Co-localization of TPP-ZC-IR-PNPs into mitochondria was analyzed by CSLM in HT29 and A549 cells after 1 h of incubation and staining with MitoTracker Red.

Fig. 3. The generation of heat and ROS under laser irradiation and the impact on cancer cells. (a) Qualitative and quantitative thermal images of the temperature elevation curves of PBS, ZCNP, IR780, and TPP-ZC-IR-PNPs under exposure to 808 nm and 1 W/cm2 over a period of 5 min; (b) Photothermal ablation of HT-29 and A549 cancer cells stained with Calcein-AM (green) and EthD-1 (red) upon treatment with laser-irradiated TPP-ZC-IR-PNPs, and the white dashed line indicates the location of laser irradiation; (c) Changes of intracellular ROS levels with ZCNP, IR780, and other formulations with and without laser irradiation; (d) Evaluation of the GSH/GSSG ratio after treatment with different formulations in the HT-29 and A549 cells.

Fig. 4. Cancer cell death in the presence of various formulations with or without laser irradiation. (a) Cell viability analysis of HT-29 and A549 cancer cells was performed by an MTT assay following incubation with various formulations and NIR irradiation for 5 min with a 808 nm laser (1 W/cm2) Data are expressed as means ± SD (n = 6); (b) Nuclear staining assay in HT-29 and A549 cells after treatment with PBS, IR780, Zn-doped CuO NP, ZC-IR-PNPs, and TPP-ZC-IR-PNPs with and without laser irradiation using fluorescence microscopy.

Fig. 5. TPP-ZC-IR-PNPs distributed mainly into tumors and generated heat but remained non-toxic to the organs in animal models. (a) In vivo fluorescence imaging of tumor-bearing Balb/c nude mice after intravenous administration of TPP-ZC-IR-PNPs and the ex vivo organ fluorescence of the tumor, liver, spleen, heart, kidney, and lungs. All the imaged tissues were excised at 24 h; (b) In vivo photothermal effect demonstrated temperature elevation upon NIR irradiation for 5 min (808 nm, 1 W/cm2) for PBS and TPP-ZC-IR-PNPs, (i) Thermo-images of the treatments, (ii) Summary graph of the temperature change; (c) No organ toxicity was recorded in the principal organs after treatment with the various formulations (i) Control, (ii) ZCNP, (iii) ZC-IR-PNP, (iv) TPP-ZC-IR-PNP, (v) IR780 (+NIR), (vi) ZCNP (+NIR), (vii) ZC-IR-PNP (+NIR), and (viii) TPP-ZC-IR-PNP (+NIR).

Fig. 6. In vivo antitumor efficacy of TPP-ZC-IR-PNP. TPP-ZC-IR-PNP under laser irradiation induced a significant reduction of tumor volume and reduced the expression of tumor markers on tissues. (a) Changes in tumor volume and (b) the body weight of nude mice bearing HT-29 xenografts after treatment with the different formulations with and without NIR (808 nm, 5 min, and 1 W/cm2). The formulations were administered via tail vein injection at a fixed dose of 5 mg/kg and 2.5 mg/kg (ZCNP and IR 780, respectively) on day 1, 4, 7, and 10. Data are presented as the mean ± SD (n = 6); (c) Histological and immunohistochemical analysis following different treatments: H&E, Caspase-3, PARP, Ki67, and CD31. (i) Control, (ii) ZCNP, (iii) ZC-IR-PNP, (iv) TPP-ZC-IR-PNP, (v) IR780 (+NIR), (vi) ZCNP (+NIR), (vii) ZC-IR-PNP (+NIR), and (viii) TPP-ZC-IR-PNP (+NIR). *p < 0.05 and ***p < 0.01. Scale bars = 120 μm.

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