■骨肉瘤(OS)是最常见的原发性恶性骨肿瘤。转导功能性TP53基因可有效抑制OS细胞活性。在动物模型中,通过聚焦超声(US)介导的聚乳酸-乙醇酸(PLGA)纳米泡(NBs)可以将外源基因引入靶细胞,但是这种技术依赖于药物在体内的被动自由扩散。在微泡中包含超顺磁性氧化铁(SPIO)允许基于磁性的组织定位。我们研究所开发了一种低强度聚焦超声(LIFU)仪器,和不同强度的LIFU可以破坏NB(RLI-LIFU)或对靶组织施加杀细胞作用(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联合使用时抑制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,将HOS和MG63细胞随机分为五个治疗组。CCK8、qRT-PCR和Westernblotting检测细胞增殖和TP53的表达,分别。在体内,荷瘤裸鼠被随机分为7个治疗组.通过光学显微镜确定Perls\'普鲁士蓝染色的组织切片的铁分布。进行TUNEL-DAPI以检查细胞凋亡。通过qRT-PCR和免疫组织化学检测TP53的表达。
■成功制备了粒径约为200nm的SPIO/TP53/PLGANB。对于体外实验,已经证明了在OS细胞中TP53过表达的超声靶向转染和对OS增殖的有效抑制。此外,在荷瘤裸鼠模型中,RLI-LIFU磁性介导的SPIO/TP53/PLGANBs提高了TP53质粒的转染效率,导致细胞凋亡。在治疗方案中加入RHI-LIFU显著增加了体内OS细胞的凋亡。
■LIFU和US-磁性介导的SPIO/TP53/PLGANB破坏联合使用可能是一种新型的OS非侵入性靶向治疗方法。
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: 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.
UNASSIGNED: Combining LIFU and US-magnetic-mediated SPIO/TP53/PLGA NB destruction is potentially a novel noninvasive and targeted therapy for OS.