Parameters for Optoperforation-Induced Killing of Cancer Cells Using Gold Nanoparticles Functionalized With the C-terminal Fragment of Clostridium Perfringens Enterotoxin

authored by

Annegret Becker, Tina Lehrich, Stefan Kalies, Alexander Heisterkamp, Anaclet Ngezahayo

Abstract

Recently, we used a recombinant produced C-terminus (D194-F319) of the Clostridium perfringens enterotoxin (C-CPE) to functionalize gold nanoparticles (AuNPs) for a subsequent specific killing of claudin expressing tumor cells using the gold nanoparticle-mediated laser perforation (GNOME-LP) technique. For a future in vivo application, it will be crucial to know the physical parameters and the biological mechanisms inducing cell death for a rational adaptation of the system to real time situation. Regarding the AuNP functionalization, we observed that a relationship of 2.5 × 10⁻¹¹ AuNP/mL to 20 µg/mL C-CPE maximized the killing efficiency. Regardingphysical parameters, a laser fluence up to 30 mJ/cm² increased the killing efficiency. Independent from the applied laser fluence, the maximal killing efficiency was achieved at a scanning velocity of 5 mm/s. In 3D matrigel culture system, the GNOME-LP/C-CPE-AuNP completely destroyed spheroids composed of Caco-2 cells and reduced OE-33 cell spheroid formation. At the biology level, GNOME-LP/C-CPE-AuNP-treated cells bound annexin V and showed reduced mitochondria activity. However, an increased caspase-3/7 activity in the cells was not found. Similarly, DNA analysis revealed no apoptosis-related DNA ladder. The results suggest that the GNOME-LP/C-CPE-AuNP treatment induced necrotic than apoptotic reaction in tumor cells.

Organisation(s)

Institute of Cell Biology and Biophysics
Institute of Quantum Optics
Department of Cell Physiology and Biophysics

External Organisation(s)

NIFE - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development
Hannover Medical School (MHH)
Center for Systems Neuroscience Hannover (ZSN)

Type

Article

Journal

International Journal of Molecular Sciences

Volume

20

ISSN

1661-6596

Publication date

01.09.2019

Publication status

Published

Peer reviewed

Yes

ASJC Scopus subject areas

Catalysis, Molecular Biology, Spectroscopy, Computer Science Applications, Physical and Theoretical Chemistry, Organic Chemistry, Inorganic Chemistry

Sustainable Development Goals

SDG 3 - Good Health and Well-being

Electronic version(s)

https://doi.org/10.3390/ijms20174248 (Access: Open)
https://doi.org/10.15488/9296 (Access: Open)