Yefimova Svetlana, Onishchenko Anatolii, Klochkov Vladimir, Myasoedov Valeriy, Kot Yurii, Tryfonyuk Liliya, Knigavko Oleksandr, Maksimchuk Pavel, Kökbaş Umut, Kalashnyk-Vakulenko Yuliia, Arkatov Andrii, Khanzhyn Vladyslav, Prokopyuk Volodymyr, Vyshnytska Iryna, Tkachenko Anton
Institute for Scintillation Materials, National Academy of Sciences of Ukraine, 60 Nauky ave, 61072 Kharkiv, Ukraine.
Research Institute of Experimental and Clinical Medicine, Kharkiv National Medical University, 4 Nauky ave, 61022 Kharkiv, Ukraine.
Nanotechnology. 2023 Mar 1;34(20). doi: 10.1088/1361-6528/acbb7f.
. Rare-earth orthovanadate nanoparticles (ReVO:Eu, Re = Gd, Y or La) are promising agents for photodynamic therapy of cancer due to their modifiable redox properties. However, their toxicity limits their application.. The aim of this research was to elucidate pro-eryptotic effects of GdVO:Euand LaVO:Eunanoparticles with identification of underlying mechanisms of eryptosis induction and to determine their pharmacological potential in eryptosis-related diseases.. Blood samples (= 9) were incubated for 24 h with 0-10-20-40-80 mg lGdVO:Euor LaVO:Eunanoparticles, washed and used to prepare erythrocyte suspensions to analyze the cell membrane scrambling (annexin-V-FITC staining), cell shrinkage (forward scatter signaling), reactive oxygen species (ROS) generation through 2',7'-dichlorodihydrofluorescein diacetate (H2DCFDA) staining and intracellular Calevels via FLUO4 AM staining by flow cytometry. Internalization of europium-enabled luminescent GdVO:Euand LaVO:Eunanoparticles was assessed by confocal laser scanning microscopy.Both nanoparticles triggered eryptosis at concentrations of 80 mg l. ROS-mediated mechanisms were not involved in rare-earth orthovanadate nanoparticles-induced eryptosis. Elevated cytosolic Caconcentrations were revealed even at subtoxic concentrations of nanoparticles. LaVO:Eunanoparticles increased intracellular calcium levels in a more pronounced way compared with GdVO:Eunanoparticles. Our data disclose that the small-sized (15 nm) GdVO:Eunanoparticles were internalized after a 24 h incubation, while the large-sized (∼30 nm) LaVO:Eunanoparticles were localized preferentially around erythrocytes.Both internalized GdVO:Euand non-internalized LaVO:Eunanoparticles (80 mg l) promote eryptosis of erythrocytes after a 24 h exposurevia Casignaling without involvement of oxidative stress. Eryptosis is a promising model for assessing nanotoxicity.
稀土正钒酸盐纳米颗粒（ReVO₄:Eu，Re = Gd、Y 或 La）因其可调节的氧化还原特性，是癌症光动力治疗的有前景的药物。然而，它们的毒性限制了其应用。本研究的目的是阐明 GdVO₄:Eu 和 LaVO₄:Eu 纳米颗粒的促红细胞凋亡作用，确定红细胞凋亡诱导的潜在机制，并确定它们在红细胞凋亡相关疾病中的药理潜力。采集 9 份血样，分别与 0 - 10 - 20 - 40 - 80 mg/L 的 GdVO₄:Eu 或 LaVO₄:Eu 纳米颗粒孵育 24 小时，洗涤后用于制备红细胞悬液，通过流式细胞术分析细胞膜磷脂酰丝氨酸外翻（ annexin-V-FITC 染色）、细胞皱缩（前向散射信号）、通过 2',7'-二氯二氢荧光素二乙酸酯（H₂DCFDA）染色检测活性氧（ROS）生成以及通过 FLUO4 AM 染色检测细胞内钙水平。通过共聚焦激光扫描显微镜评估含铕的发光 GdVO₄:Eu 和 LaVO₄:Eu 纳米颗粒的内化情况。两种纳米颗粒在浓度为 80 mg/L 时均引发红细胞凋亡。稀土正钒酸盐纳米颗粒诱导的红细胞凋亡不涉及 ROS 介导的机制。即使在纳米颗粒的亚毒性浓度下，也观察到细胞溶质钙浓度升高。与 GdVO₄:Eu 纳米颗粒相比，LaVO₄:Eu 纳米颗粒更显著地提高细胞内钙水平。我们的数据表明，小尺寸（15 nm）的 GdVO₄:Eu 纳米颗粒在孵育 24 小时后被内化，而大尺寸（约 30 nm）的 LaVO₄:Eu 纳米颗粒优先定位在红细胞周围。内化的 GdVO₄:Eu 和未内化的 LaVO₄:Eu 纳米颗粒（80 mg/L）在暴露 24 小时后均通过钙信号传导促进红细胞凋亡，且不涉及氧化应激。红细胞凋亡是评估纳米毒性的一个有前景的模型。
