Quan Luya, Wang Mengzhu, Wang Zhigang, Du Zhiyu
Department of Ophthalmology, The Guizhou Provincial People's Hospital, Guiyang, Guizhou, 550002, People's Republic of China.
Department of Ultrasound, Chongqing Key Laboratory of Ultrasound Molecular Imaging, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, People's Republic of China.
Int J Nanomedicine. 2025 Apr 3;20:4085-4103. doi: 10.2147/IJN.S506179. eCollection 2025.
To overcome the limitations of traditional therapies in treating retinoblastoma, like low efficiency, systematic toxicity and poor biocompatibility.
PPFG (PLGA-PFH-FeO-GOx) nanoparticles were synthesized by ultrasound double emulsification method and characterized by dynamic laser scattering, ultraviolet spectrometry, confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Phase transition by low-intensity focused ultrasound (LIFU) was observed by microscope and ultrasound imaging. Cellular uptake was compared between Y79 and HUVEC cells. ROS production was detected by 2',7'-dichlorofluorescin diacetate (DCFH-DA). Cell apoptosis was detected by flow cytometry. In vivo therapeutic effects were verified by tumor volume, HE staining, TUNEL and PCNA staining. The in vivo bio-safety was detected by serum biochemistry.
PPFG NPs possesses good stability, biocompatibility and tumor-preferred uptake, with a core-shell spherical structure and an average size of 255.6nm which increases to over 100μm under LIFU irradiation. LIFU was utilized as a stimuli, by which PPFG NPs undergoes a sequential reaction starting with phase transition of PFH causing the release of the oxygen carried by PFH and GOx/SPIO carried by PPFG NPs, followed by the supplemented oxygen facilitating the enzymatic activity of glucose consumption by GOx in tumor cells (tumor starvation), the HO produced during the enzymatic activity can further participate in SPIO NPs-mediated Fenton reaction (CDT), generating massive ROS. The continuously generated ROS together with the cut down of tumor nutrients by GOx effectively inhibited the progression of tumors, and synergistically enhanced ROS production together with tumor starvation promoted cell apoptosis and ultimately kills the tumour cells. No off-site injuries was detected in other major organs.
In this study, PPFG nanoparticles were synthesized to conduct LIFU-triggered combinational therapy on the basis of the cascade reaction among PFH, GOx and SPIO to treat retinoblastoma in vitro/vivo. It showed great potentials in combating retinoblastoma.
克服传统疗法在治疗视网膜母细胞瘤方面的局限性，如效率低、全身毒性和生物相容性差等问题。
采用超声双乳化法合成PPFG（聚乳酸-聚氟代己烷-氧化亚铁-葡萄糖氧化酶）纳米颗粒，并通过动态激光散射、紫外光谱、共聚焦激光扫描显微镜（CLSM）、透射电子显微镜（TEM）和扫描电子显微镜（SEM）对其进行表征。通过显微镜和超声成像观察低强度聚焦超声（LIFU）诱导的相变。比较Y79细胞和人脐静脉内皮细胞（HUVEC）对纳米颗粒的摄取情况。用2',7'-二氯荧光素二乙酸酯（DCFH-DA）检测活性氧（ROS）的产生。通过流式细胞术检测细胞凋亡。通过肿瘤体积、苏木精-伊红（HE）染色、末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）和增殖细胞核抗原（PCNA）染色验证体内治疗效果。通过血清生化检测体内生物安全性。
PPFG纳米颗粒具有良好的稳定性、生物相容性和肿瘤优先摄取特性，呈核壳球形结构，平均粒径为255.6nm，在LIFU照射下粒径增大至100μm以上。利用LIFU作为刺激因素，使PPFG纳米颗粒发生一系列反应，首先是聚氟代己烷的相变，导致聚氟代己烷携带的氧气以及PPFG纳米颗粒携带的葡萄糖氧化酶/超顺磁性氧化铁纳米颗粒（GOx/SPIO）释放，随后补充的氧气促进肿瘤细胞中葡萄糖氧化酶消耗葡萄糖的酶活性（肿瘤饥饿），酶活性过程中产生的羟基自由基（HO）可进一步参与超顺磁性氧化铁纳米颗粒介导的芬顿反应（化学动力学疗法，CDT），生成大量ROS。持续产生的ROS以及葡萄糖氧化酶导致的肿瘤营养物质减少有效抑制了肿瘤进展，ROS产生与肿瘤饥饿协同增强，促进细胞凋亡并最终杀死肿瘤细胞。在其他主要器官未检测到异位损伤。
本研究合成了PPFG纳米颗粒，基于聚氟代己烷、葡萄糖氧化酶和超顺磁性氧化铁纳米颗粒之间的级联反应进行LIFU触发的联合治疗，以在体外/体内治疗视网膜母细胞瘤。其在对抗视网膜母细胞瘤方面显示出巨大潜力。
