Bio-X Center, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China;; Institute of Biomaterials and Marine Biological Resources, College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China.
Regen Biomater. 2016 Mar;3(1):57-63. doi: 10.1093/rb/rbv029. Epub 2016 Jan 13.
In this study, calcium carbonate (CaCO3) nanoparticles with spherical structure were regulated by arginine and successfully synthesized via a facile co-precipitation method. The average particle size of as-prepared CaCO3 was about 900 nm. The properties of nanostructured CaCO3 particles were characterized by scanning electron microscope, Fourier transform infrared spectroscopy, X-ray diffraction and size distribution. After modified with polyethyleneimine (PEI), the ability of PEI-CaCO3 nanoparticles to carry GFP-marked p53 gene (pEGFP-C1-p53) into cancer cells to express P53 protein were studied. Meanwhile, the cytotoxicity, transfection efficiency, cells growth inhibition and the ability to induce apoptosis by expressed P53 protein were conducted to evaluate the performances of PEI-CaCO3 nanoparticles. The results show that prepared PEI-CaCO3 nanoparticles had good biocompatibility and low cytotoxicity in a certain concentration range. PEI-CaCO3 effectively transfected pEGFP-C1 gene into epithelial-like cancer cells. And with the expression of GFP-P53 fusion protein, pEGFP-C1-p53-gene-loaded PEI-CaCO3 particles significantly reduced the proliferation of cancer cells. These findings indicate that our PEI-modified CaCO3 nanoparticles are potential to be successfully used as carriers for gene therapy.
在这项研究中,通过简便的共沉淀法,用精氨酸调控具有球形结构的碳酸钙(CaCO3)纳米粒子,并成功合成。所制备的 CaCO3 的平均粒径约为 900nm。采用扫描电子显微镜、傅里叶变换红外光谱、X 射线衍射和粒径分布对纳米结构 CaCO3 粒子的性能进行了表征。用聚乙烯亚胺(PEI)对其进行修饰后,研究了 PEI-CaCO3 纳米粒子将 GFP 标记的 p53 基因(pEGFP-C1-p53)带入癌细胞表达 P53 蛋白的能力。同时,通过细胞毒性、转染效率、细胞生长抑制和表达的 P53 蛋白诱导细胞凋亡的能力来评估 PEI-CaCO3 纳米粒子的性能。结果表明,所制备的 PEI-CaCO3 纳米粒子在一定浓度范围内具有良好的生物相容性和低细胞毒性。PEI-CaCO3 能有效将 pEGFP-C1 基因转染入上皮样癌细胞。并且,随着 GFP-P53 融合蛋白的表达,载有 pEGFP-C1-p53 基因的 PEI-CaCO3 颗粒显著降低了癌细胞的增殖。这些发现表明,我们的 PEI 修饰的 CaCO3 纳米粒子有潜力成功用作基因治疗的载体。