Al-Ani Lina A, Kadir Farkaad A, Hashim Najihah M, Julkapli Nurhidayatullaili M, Seyfoddin Ali, Lu Jun, AlSaadi Mohammed A, Yehye Wageeh A
Institute of Advanced Studies, Nanotechnology & Catalysis Research Centre (NANOCAT), University of Malaya, Kuala Lumpur, Malaysia.
Department of Anatomy and Medical Imaging, Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand.
Heliyon. 2020 Nov 2;6(11):e05360. doi: 10.1016/j.heliyon.2020.e05360. eCollection 2020 Nov.
Natural plants derivatives have gained enormous merits in cancer therapy applications upon formulation with nanomaterials. Curcumin, as a popular research focus has acquired such improvements surpassing its disadvantageous low bioavailability. To this point, the available research data had confirmed the importance of nanomaterial type in orienting cellular response and provoking different toxicological and death mechanisms that may range from physical membrane damage to intracellular changes. This in turn underlines the poorly studied field of nanoformulation interaction with cells as the key determinant in toxicology outcomes. In this work, curcumin-AuNPs-reduced graphene oxide nanocomposite (CAG) was implemented as a model, to study the impact on cellular membrane integrity and the possible redox changes using colon cancer cell lines (HT-29 and SW-948), representing drug-responsive and resistant subtypes. Morphological and biochemical methods of transmission electron microscopy (TEM), apoptosis assay, reactive oxygen species (ROS) and antioxidants glutathione and superoxide dismutase (GSH and SOD) levels were examined with consideration to suitable protocols and vital optimizations. TEM micrographs proved endocytic uptake with succeeding cytoplasm deposition, which unlike other nanomaterials studied previously, conserved membrane integrity allowing intracellular cytotoxic mechanism. Apoptosis was confirmed with gold-standard morphological features observed in micrographs, while redox parameters revealed a time-dependent increase in ROS accompanied with regressive GSH and SOD levels. Collectively, this work demonstrates the success of graphene as a platform for curcumin intracellular delivery and cytotoxicity, and further highlights the importance of suitable methods to be used for nanomaterial validation.
天然植物衍生物与纳米材料制成制剂后，在癌症治疗应用中展现出诸多优势。姜黄素作为一个热门研究焦点，已取得诸多改进，克服了其生物利用度低这一不利因素。至此，现有研究数据证实了纳米材料类型在引导细胞反应以及引发不同毒理学和死亡机制方面的重要性，这些机制可能从物理膜损伤到细胞内变化不等。这反过来凸显了纳米制剂与细胞相互作用这一研究较少的领域，它是毒理学结果的关键决定因素。在这项工作中，采用姜黄素 - 金纳米粒子 - 还原氧化石墨烯纳米复合材料（CAG）作为模型，利用代表药物敏感和耐药亚型的结肠癌细胞系（HT - 29和SW - 948），研究其对细胞膜完整性的影响以及可能的氧化还原变化。考虑到合适的方案和重要的优化措施，对透射电子显微镜（TEM）、凋亡检测、活性氧（ROS）以及抗氧化剂谷胱甘肽和超氧化物歧化酶（GSH和SOD）水平等形态学和生化方法进行了检测。TEM显微照片证明了细胞通过内吞作用摄取并随后在细胞质中沉积，与之前研究的其他纳米材料不同，它保持了膜的完整性，从而允许细胞内细胞毒性机制的发生。通过显微照片中观察到的金标准形态特征证实了细胞凋亡，而氧化还原参数显示ROS随时间增加，同时GSH和SOD水平呈下降趋势。总的来说，这项工作证明了石墨烯作为姜黄素细胞内递送和细胞毒性平台的成功，并进一步强调了用于纳米材料验证的合适方法的重要性。
