School of Pure and Applied Physics, Mahatma Gandhi University, P.D Hills (P.O), Kottayam, Kerala, 686 560, India.
Bharata Mata College, Thrikkakara, Ernakulam, Kerala, 682032, India.
Sci Rep. 2023 Jun 3;13(1):9045. doi: 10.1038/s41598-023-34997-w.
The influence of nanoparticles inside the human body and their interactions with biological macromolecules need to be explored/studied prior to specific applications. The objective of this study is to find the potential of camptothecin functionalised silver nanoparticles (CMT-AgNPs) in biomedical applications. This article primarily investigates the binding stratagem of CMT-AgNPs with calf thymus DNA (ctDNA) through a series of spectroscopic and calorimetric methods and then analyses the anticancer activity and cytotoxicity of CMT-AgNPs. The nanoparticles were synthesized using a simple one pot method and characterized using UV-Visible, fourier transform infrared (FTIR) spectroscopy, X-ray diffraction and high-resolution transmission electron microscopy (HRTEM). The average size of CMT-AgNPs is 10 ± 2 nm. A group of experimental techniques such as UV-Visible spectrophotometry, fluorescence dye displacement assay, circular dichroism (CD) and viscosity analysis unravelled the typical groove binding mode of CMT-AgNPs with ctDNA. The CD measurement evidenced the minor conformational alterations of double helical structure of ctDNA in the presence of CMT-AgNPs. The information deduced from the isothermal titration calorimetry (ITC) experiment is that the binding was exothermic and spontaneous in nature. Moreover, all the thermodynamic binding parameters were extracted from the ITC data. The binding constants obtained from UV absorption experiments, fluorescence dye displacement studies and ITC were consistently in the order of 10 Mol. All these results validated the formation of CMT-AgNPs-ctDNA complex and the results unambiguously confirm the typical groove binding mode of CMT-AgNPs. An exhaustive in vitro MTT assay by CMT-AgNPs and CMT against A549, HT29, HeLa and L929 cell lines revealed the capability of CMT-AgNPs as a potential anticancer agent.
在将纳米粒子应用于特定领域之前,需要研究其在人体内的影响及其与生物大分子的相互作用。本研究的目的是探索喜树碱功能化银纳米粒子(CMT-AgNPs)在生物医学中的应用潜力。本文主要通过一系列光谱和量热法研究 CMT-AgNPs 与小牛胸腺 DNA(ctDNA)的结合策略,然后分析 CMT-AgNPs 的抗癌活性和细胞毒性。纳米粒子采用简单的一锅法合成,并通过紫外-可见分光光度法、傅里叶变换红外(FTIR)光谱、X 射线衍射和高分辨率透射电子显微镜(HRTEM)进行表征。CMT-AgNPs 的平均粒径为 10±2nm。一组实验技术,如紫外-可见分光光度法、荧光染料置换法、圆二色性(CD)和粘度分析,揭示了 CMT-AgNPs 与 ctDNA 的典型沟结合模式。CD 测量表明,在 CMT-AgNPs 的存在下,ctDNA 的双螺旋结构发生了轻微的构象变化。等温滴定量热法(ITC)实验得出的信息表明,结合是放热和自发的。此外,所有热力学结合参数都从 ITC 数据中提取出来。从紫外吸收实验、荧光染料置换研究和 ITC 获得的结合常数依次为 10 Mol。所有这些结果都验证了 CMT-AgNPs-ctDNA 复合物的形成,并且结果明确证实了 CMT-AgNPs 的典型沟结合模式。通过 CMT-AgNPs 和 CMT 对 A549、HT29、HeLa 和 L929 细胞系进行的详尽体外 MTT 测定表明,CMT-AgNPs 具有作为潜在抗癌剂的能力。