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通过原位荧光显微镜研究银纳米片在HeLa细胞中的光热疗法。

Photothermal therapy with silver nanoplates in HeLa cells studied by in situ fluorescence microscopy.

作者信息

Rivas Aiello María Belén, Azcárate Julio C, Zelaya Eugenia, David Gara Pedro, Bosio Gabriela N, Gensch Thomas, Mártire Daniel O

机构信息

Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicadas (INIFTA), Facultad de Ciencias Exactas, Universidad Nacional de La Plata, C. C. 16, Suc. 4, (1900) La Plata, Argentina.

出版信息

Biomater Sci. 2021 Apr 7;9(7):2608-2619. doi: 10.1039/d0bm01952f. Epub 2021 Feb 17.

Abstract

Photothermal therapy (PTT) is a noninvasive treatment for cancer relying on the incorporation of NIR-light absorbing nanomaterials into cells, which upon illumination release heat causing thermally induced cell death. We prove that irradiation of aqueous suspensions of poly(vinylpyrrolidone)-coated silver nanoplates (PVPAgNP) or PVPAgNP in HeLa cells with red or NIR lasers causes a sizeable photothermal effect, which in cells can be visualized with the temperature sensing fluorophore Rhodamine B (RhB) using spinning disk confocal fluorescence microscopy or fluorescence lifetime imaging. Upon red-light irradiation of cells that were incubated with both, RhB and PVPAgNP at concentrations with no adverse effects on cell viability, a substantial heat release is detected. Initiation of cell death by photothermal effect is observed by positive signals of fluorescent markers for early and late apoptosis. Surprisingly, a new nanomaterial-assisted cell killing mode is operating when PVPAgNP-loaded HeLa cells are excited with moderate powers of fs-pulsed NIR light. Small roundish areas are generated with bright and fast (<1 ns) decaying emission, which expand fast and destroy the whole cell in seconds. This characteristic emission is assigned to efficient optical breakdown initiation around the strongly absorbing PVPAgNP leading to plasma formation that spreads fast through the cell.

摘要

光热疗法(PTT)是一种用于癌症的非侵入性治疗方法,它依赖于将吸收近红外光的纳米材料引入细胞,这些纳米材料在光照下释放热量,导致热诱导的细胞死亡。我们证明,用红色或近红外激光照射聚(乙烯基吡咯烷酮)包覆的银纳米板(PVPAgNP)的水悬浮液或HeLa细胞中的PVPAgNP会产生相当大的光热效应,在细胞中可以使用旋转盘共聚焦荧光显微镜或荧光寿命成像,通过温度传感荧光团罗丹明B(RhB)来可视化这种效应。在用对细胞活力无不利影响的浓度的RhB和PVPAgNP孵育的细胞进行红光照射后,检测到大量的热量释放。通过早期和晚期凋亡荧光标记物的阳性信号观察到光热效应引发的细胞死亡。令人惊讶的是,当用中等功率的飞秒脉冲近红外光激发负载PVPAgNP的HeLa细胞时,一种新的纳米材料辅助细胞杀伤模式正在起作用。会产生小的圆形区域,具有明亮且快速(<1 ns)衰减的发射,这些区域迅速扩展并在几秒钟内破坏整个细胞。这种特征发射归因于在强吸收的PVPAgNP周围有效引发光学击穿,导致等离子体形成,并迅速在细胞中扩散。

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