Air Fixation and AFM: A Comparative Study of Nanoparticle-Induced Topographical Changes in Lung Cells
Yıl 2024,
Cilt: 28 Sayı: 5, 1086 - 1092, 25.10.2024
Gamze Yeşilay
Öz
Gold nanoparticles (AuNPs) have emerged as promising agents in biomedical applications due to their unique physicochemical properties. This study investigates the cellular interactions of AuNPs with A549 (non-small cell lung adenocarcinoma) and BEAS-2B (normal bronchial epithelial) cell lines. AuNPs were synthesized via the citrate reduction method, resulting in 20, 50, and 70 nm particles.
Cells were incubated with AuNPs for increasing durations (30 minutes, 4 hours, and 24 hours). Post-incubation, cells were washed with PBS, air-fixed, and subsequently analyzed using Atomic Force Microscopy (AFM) to obtain detailed topographical maps. AFM imaging revealed distinct interactions between AuNPs and the two cell lines.
A549 cells displayed darker regions on the cell surface, indicative of topographical depressions likely resulting from nanoparticle-induced membrane collapse. In contrast, BEAS-2B cells did not exhibit such depressions, which is consistent with the literature that suggests cancer cells are mechanically softer than normal cells.
The surface roughness analysis results indicated that the preservation of surface integrity post-fixation validates the air-fixation methodology for obtaining reliable mechanical data from AFM analyses.
Etik Beyan
The author of the paper declare that they comply with the scientific, ethical and quotation rules of SAUJS in all processes of the paper and that she does not make any falsification on the data collected. In addition, she declares that Sakarya University Journal of Science and its editorial board have no responsibility for any ethical violations that may be encountered, and that this study has not been evaluated in any academic publication environment other than Sakarya University Journal of Science.
Teşekkür
The author would like to thank University of Health Sciences, Experimental Medicine Application & Research Center, Validebag Research Park for Access to research infrastructure.
Kaynakça
- X. Deng, F. Xiong, X. Li, B. Xiang, Z. Li, X. Wu, C. Guo, X. Li, Y. Li, G. Li, W. Xiong, Z. Zeng, “Application of atomic force microscopy in cancer research,” Journal of Nanobiotechnology, vol. 16, p. 102, 2018.
- F. Xia, K. Youcef-Toumi, “Review: Advanced Atomic Force Microscopy Modes for Biomedical Research,” Biosensors, vol. 12, Art. no. 12, 2022.
- A. Massey, J. Stewart, C. Smith, C. Parvini, M. McCormick, K. Do & A. X. Cartagena-Rivera, "Mechanical properties of human tumour tissues and their implications for cancer development," Nature Reviews Physics, vol. 6, p. 269-282, 2024.
- S. O. Konorov, H. G. Schulze, J. M. Piret, R. F. B. Turner, M. W. Blades, “Evidence of marked glycogen variations in the characteristic Raman signatures of human embryonic stem cells,” Journal of Raman Spectroscopy, vol. 42, pp. 1135–1141, 2011.
- M. M. Mariani, P. Lampen, J. Popp, B. R. Wood, V. Deckert, “Impact of fixation on in vitro cell culture lines monitored with Raman spectroscopy,” Analyst, vol. 134, pp. 1154–1161, 2009.
- B. Q. Huang, E. C. Yeung, “Chemical and Physical Fixation of Cells and Tissues: An Overview,” in Plant Microtechniques and Protocols, E. C. T. Yeung, C. Stasolla, M. J. Sumner, B. Q. Huang, Eds., Cham: Springer International Publishing, 2015, pp. 23–43.
- L. Andolfi, E. Bourkoula, E. Migliorini, A. Palma, A. Pucer, M. Skrap, G. Scoles, A.P. Beltrami, D. Cesselli, M. Lazzarino, “Investigation of Adhesion and Mechanical Properties of Human Glioma Cells by Single Cell Force Spectroscopy and Atomic Force Microscopy,” PLOS ONE, vol. 9, p. e112582, 2014.
- C. Huang, T. Ozdemir, L. C. Xu, P. J. Butler, C. A. Siedlecki, J. L. Brown & S. Zhang, "The role of substrate topography on the cellular uptake of nanoparticles," Journal of Biomedical Materials Research Part B: Applied Biomaterials, vol. 104, pp. 488-495, 2016.
- J. Turkevich, P. C. Stevenson, J. Hillier, “A study of the nucleation and growth processes in the synthesis of colloidal gold,” Discussions of the Faraday Society, vol. 11, pp. 55–75, 1951.
- W. Haiss, N. T. K. Thanh, J. Aveyard, D. G. Fernig, “Determination of Size and Concentration of Gold Nanoparticles from UV−Vis Spectra,” Analytical Chemistry, vol. 79, pp. 4215–4221, 007.
- S. Suresh, “Biomechanics and biophysics of cancer cells,” Acta Biomaterialia, vol. 3, pp. 413–438, 2007.
- C. Lara-Cruz, J. J. Salazar, E. Ramón-Gallegos, P. Damian-Matsumura, N. Batina, “Increasing roughness of the human breast cancer cell membrane through incorporation of gold nanoparticles,” International Journal of Nanoscience, vol. 11, pp. 5149–5161, 2016.
- B. D. Chithrani, A. A. Ghazani, W. C. W. Chan, “Determining the Size and Shape Dependence of Gold Nanoparticle Uptake into Mammalian Cells,” Nano Letters, vol. 6, pp. 662–668, 2006.