Institute of Biomedical Engineering, School of Preclinical and Forensic Medicine, Sichuan University, Chengdu, 610041, PR China.
Langmuir. 2011 Nov 1;27(21):13206-11. doi: 10.1021/la2008107. Epub 2011 Oct 11.
Zinc oxide nanoparticles (NPs) with the size of ∼100 nm were prepared via a facile biomineralization process in the template of silk fibroin (SF) peptide at room temperature. These ZnO NPs have shown the remarkable behavior of low toxicity to gram-positive bacteria (Staphylococcus aureus, Staphylococcus agalactiae), gram-negative bacteria (Escherichia coli), and eukaryotic cells (mouse L929 fibroblasts). Bacteriological testing indicated that ZnO NPs presented a 50% inhibitory effect on Streptococcus agalactiae at the concentrations of >100 mM, whereas at the same concentrations, the growth of Staphylococcus aureus and Escherichia coli were hardly inhibited. On the other hand, a remarkable proliferation of Staphylococcus aureus or Escherichia coli was observed at the concentrations of ZnO NPs <50 mM. Moreover, the cytotoxicity test demonstrated that ZnO NPs mineralized with SF peptide possessed a low toxicity to mouse L929 fibroblasts. The SF peptide coated on the surface of ZnO NPs permitted greater adhesion and consequently greater proliferation of mouse L929 fibroblasts. Besides, from TEM micrographs of the cell ultrastructure, endocytosis of NPs into the cytoplasm can be detected and the ultrastructure of the cell underwent few changes. The cell membrane retained integrity, euchromatin dispersed homogenously inside the cytoplasm, the mitochondrial architecture remained intact, and no intracellular vacuoles were observed. High-resolution transmission electron microscopy images and selected area electron diffraction patterns of ultrathin cell sections indicated that the crystal structure of NPs was not damaged by the organelle or cytoplasm. All these observations indicated that ZnO NPs mineralized with the SF peptide possess good cytocompatibility.
氧化锌纳米粒子(NPs)的尺寸约为 100nm,通过在丝素肽(SF)肽的模板中在室温下进行简便的生物矿化过程制备。这些 ZnO NPs 表现出对革兰氏阳性菌(金黄色葡萄球菌、无乳链球菌)、革兰氏阴性菌(大肠杆菌)和真核细胞(小鼠 L929 成纤维细胞)的低毒性的显著行为。细菌学测试表明,ZnO NPs 在浓度>100mM 时对无乳链球菌表现出 50%的抑制作用,而在相同浓度下,金黄色葡萄球菌和大肠杆菌的生长几乎不受抑制。另一方面,在 ZnO NPs 的浓度<50mM 时观察到金黄色葡萄球菌或大肠杆菌的显著增殖。此外,细胞毒性测试表明,用 SF 肽矿化的 ZnO NPs 对小鼠 L929 成纤维细胞具有低毒性。SF 肽涂覆在 ZnO NPs 的表面上允许更大的粘附,从而导致小鼠 L929 成纤维细胞的更大增殖。此外,从细胞超微结构的 TEM 显微照片中,可以检测到 NPs 被内吞到细胞质中,并且细胞的超微结构几乎没有变化。细胞膜保持完整,常染色质均匀分散在细胞质中,线粒体结构保持完整,并且没有观察到细胞内空泡。超薄细胞切片的高分辨率透射电子显微镜图像和选区电子衍射图谱表明,纳米颗粒的晶体结构没有被细胞器或细胞质破坏。所有这些观察结果表明,用 SF 肽矿化的 ZnO NPs 具有良好的细胞相容性。