Lee Sun Young, Choi Jae Won, Lee Tae Geol, Heo Min Beom, Son Jin Gyeong
Nanobio Measurement Group, Division of Biomedical Metrology, Korea Research Institute of Standards and Science 267 Gajeong-ro, Yuseong-gu Daejeon 34113 Republic of Korea
Department of Biomedical Science and Technology, Graduate School, Kyung Hee University Seoul 02447 Republic of Korea.
Nanoscale Adv. 2024 Oct 10;6(22):5585-97. doi: 10.1039/d4na00743c.
Reliable characterization of protein coronas (PCs) that form when nanomaterials are introduced into biological fluids is a critical step in the development of safe and efficient nanomedicine. We observed that bovine serum albumin (BSA)-coated multi-walled carbon nanotubes (MWCNTs) do not induce cytotoxicity, but have different cellular uptake rates depending on the BSA pretreatment concentration. To determine how these slight differences affect A549 cell responses and intracellular changes, we conducted spectroscopic (circular dichroism and Fourier-transform infrared) and spectrometric (nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry) analyses. The various characterization techniques conducted in this study reveal the following. (i) The composition ratio of PCs on MWCNTs differs depending on the BSA concentration. (ii) Analysis of the secondary structure of the proteins revealed that the α-helix structure increased with increasing BSA concentration. (iii) Proteomic analysis showed that different biological pathways were activated at levels higher and lower than 5 mg mL. Such combined spectroscopic and spectrometric approaches provide an integrated understanding of PC composition as well as how nano/bio-interface states are linked to cellular-level responses. Our results can support reliable and practical applications of nanomedicine development.
当纳米材料引入生物流体中时，对形成的蛋白质冠层（PCs）进行可靠表征是安全高效纳米医学发展的关键一步。我们观察到，牛血清白蛋白（BSA）包覆的多壁碳纳米管（MWCNTs）不会诱导细胞毒性，但根据BSA预处理浓度的不同，其细胞摄取率也不同。为了确定这些细微差异如何影响A549细胞反应和细胞内变化，我们进行了光谱分析（圆二色性和傅里叶变换红外光谱）和光谱测定（纳流液相色谱 - 电喷雾电离 - 串联质谱）分析。本研究中采用的各种表征技术揭示了以下几点：（i）MWCNTs上PCs的组成比例因BSA浓度而异；（ii）蛋白质二级结构分析表明，α - 螺旋结构随着BSA浓度的增加而增加；（iii）蛋白质组学分析表明，在高于和低于5 mg/mL的水平上激活了不同的生物途径。这种光谱和光谱测定相结合的方法能够全面了解PC的组成，以及纳米/生物界面状态如何与细胞水平的反应相关联。我们的结果可为纳米医学发展的可靠和实际应用提供支持。
