MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, PR China.
Echocardiography and Vascular Ultrasound Centre, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, PR China.
Colloids Surf B Biointerfaces. 2018 Jan 1;161:279-287. doi: 10.1016/j.colsurfb.2017.10.036. Epub 2017 Oct 12.
The microbubble is a kind of clinically applied ultrasound contrast agent in disease diagnosis that can also rupture under sonication to increase membrane permeability and promote gene entry into targeted cells. However, the development of ultrasound-mediated gene delivery might be restricted because genes and microbubbles were separated and would not reach the targeted cells simultaneously. Herein, a kind of crosslinked positive microbubbles (CPMBs) were prepared to load DNA as gene vectors to promote gene delivery efficiency. The BSA shell of the CPMBs was crosslinked with disulfide bonds, which obviously enhanced the stability of the CPMBs. Furthermore, the CPMBs revealed sonoporation effects comparable to those of clinically applied SonoVue microbubbles. As DNA and CPMBs were electrostatically linked as an entirety, they would reach cells simultaneously. Thus, with the aid of ultrasound, these DNA-loaded microbubbles promoted DNA entry into cytoplasm more effectively and obtained higher cellular uptake efficiency and better transfection efficiency than DNA-mixed microbubbles. Confocal microscopy results showed that rupturing of the CPMBs/DNA entire microbubbles under sonication could carry DNA directly into the cytoplasm or nucleus. All results indicated that the cytocompatible DNA-loaded microbubbles have promising prospects in ultrasound-mediated gene delivery.
微泡是一种临床应用的超声造影剂，可在超声作用下破裂，增加细胞膜通透性，促进基因进入靶细胞。然而，超声介导的基因传递的发展可能会受到限制，因为基因和微泡是分离的，不会同时到达靶细胞。在此，制备了一种交联正微泡（CPMBs）作为基因载体来负载 DNA 以提高基因传递效率。CPMBs 的 BSA 外壳通过二硫键交联，明显提高了 CPMBs 的稳定性。此外，CPMBs 显示出与临床应用的 SonoVue 微泡相当的声孔效应。由于 DNA 和 CPMBs 作为一个整体通过静电相互作用连接，它们会同时到达细胞。因此，在超声的辅助下，这些负载 DNA 的微泡更有效地将 DNA 导入细胞质，获得更高的细胞摄取效率和更好的转染效率比 DNA 混合微泡。共聚焦显微镜结果表明，超声作用下 CPMBs/DNA 整个微泡的破裂可以将 DNA 直接带入细胞质或细胞核。所有结果表明，细胞相容性的 DNA 负载微泡在超声介导的基因传递中具有广阔的前景。
