骨肉瘤（OS）是最常见的原发性恶性骨肿瘤类型。转导功能性TP53基因可以有效抑制OS细胞活性。由聚焦超声（US）介导的聚乳酸-乙醇酸（PLGA）纳米气泡（NB）可以将外源基因引入动物模型中的靶细胞中，但该技术依赖于药物在体内的被动自由扩散。微泡中包含超顺磁性氧化铁 (SPIO)，可实现基于磁性的组织定位。我所研制了低强度聚焦超声（LIFU）仪器，不同强度的LIFU既可以破坏NBs（RLI-LIFU），也可以对靶组织产生杀细胞作用（RHI-LIFU）。基于这些数据，我们进行了 US 磁介导的 TP53-NB 破坏，并研究了其在体外和体内与 LIFU 结合时抑制 OS 生长的能力。制备并表征了几种 SPIO/TP53/PLGA (STP) NB 变体。对于体外实验，HOS 和 MG63 细胞被随机分配到五个处理组。分别通过CCK8、qRT-PCR和Western blotting检测细胞增殖和TP53的表达。在体内，荷瘤裸鼠被随机分为七个治疗组。 Perls 普鲁士蓝染色组织切片的铁分布通过光学显微镜测定。进行TUNEL-DAPI来检查细胞凋亡。通过qRT-PCR和免疫组化检测TP53的表达。成功制备了粒径约为200 nm的SPIO/TP53/PLGA NB。对于体外实验，已经证明超声靶向转染 OS 细胞中 TP53 的过表达以及对 OS 增殖的有效抑制。此外，在荷瘤裸鼠模型中，RLI-LIFU磁介导的SPIO/TP53/PLGA NBs增加了TP53质粒的转染效率，导致细胞凋亡。在治疗方案中添加RHI-LIFU可显着增加体内OS细胞的凋亡。将LIFU与US磁介导的SPIO/TP53/PLGA NB破坏相结合可能是一种新型的OS无创靶向治疗方法。版权所有©2024 Ren，Xiang 、刘、王、周、胡。

Osteosarcoma (OS) is the most common type of primary malignant bone tumor. Transducing a functional TP53 gene can effectively inhibit OS cell activity. Poly lactic acid-glycolic acid (PLGA) nanobubbles (NBs) mediated by focused ultrasound (US) can introduce exogenous genes into target cells in animal models, but this technique relies on the passive free diffusion of agents across the body. The inclusion of superparamagnetic iron oxide (SPIO) in microbubbles allows for magnetic-based tissue localization. A low-intensity-focused ultrasound (LIFU) instrument was developed at our institute, and different intensities of LIFU can either disrupt the NBs (RLI-LIFU) or exert cytocidal effects on the target tissues (RHI-LIFU). Based on these data, we performed US-magnetic-mediated TP53-NB destruction and investigated its ability to inhibit OS growth when combined with LIFU both in vitro and in vivo.Several SPIO/TP53/PLGA (STP) NB variants were prepared and characterized. For the in vitro experiments, HOS and MG63 cells were randomly assigned into five treatment groups. Cell proliferation and the expression of TP53 were detected by CCK8, qRT-PCR and Western blotting, respectively. In vivo, tumor-bearing nude mice were randomly assigned into seven treatment groups. The iron distribution of Perls' Prussian blue-stained tissue sections was determined by optical microscopy. TUNEL-DAPI was performed to examine apoptosis. TP53 expression was detected by qRT-PCR and immunohistochemistry.SPIO/TP53/PLGA NBs with a particle size of approximately 200 nm were prepared successfully. For in vitro experiments, ultrasound-targeted transfection of TP53 overexpression in OS cells and efficient inhibition of OS proliferation have been demonstrated. Furthermore, in a tumor-bearing nude mouse model, RLI-LIFU-magnetic-mediated SPIO/TP53/PLGA NBs increased the transfection efficiency of the TP53 plasmid, resulting in apoptosis. Adding RHI-LIFU to the treatment regimen significantly increased the apoptosis of OS cells in vivo.Combining LIFU and US-magnetic-mediated SPIO/TP53/PLGA NB destruction is potentially a novel noninvasive and targeted therapy for OS.Copyright © 2024 Ren, Xiang, Liu, Wang, Zhou and Hu.