Multilayered magnetic nanoparticles fabricated by nanoimprint lithography for magnetomechanical treatment of cancer

Thesis (Ph.D.)--University of Washington, 2017-06Fe3O4-magetite nanoparticles have received wide interest as prominent agents for various biomedical applications, ranging from target-specific cancer treatment, gene therapy, and Magnetic Particle Imaging (MPI). However, Fe3O4-magnetite nanoparticles, synthesized by chemical methods beyond a certain size, present challenges in controlling size distribution and shape. Similarly, Fe3O4-magnetite nanoparticles fabricated by conventional top-down lithographic methods present difficulty of controlling defects and lead to agglomeration due to large size. In order to overcome the difficulties associated with the conventional chemical and top-down lithographic methods, it is critical to develop a fabrication method which produces homogeneous nanoparticles in large quantities with the control of size, defects, and structure. Furthermore, the concept of cell death induced by mechanical perturbation has received wide attention as a way to maximize the cancer cell death with minimal side effects. Previous study has proposed the use of permalloy disk-shaped vortex state microparticles, in order to create cancer cell death by mechanical force. However, insufficient biocompatibility, inadequate mechanical force created by vortex switching, and inability to control the particle size have been critical issues to be further researched and proceeded for in vivo application. Hence, we studied physical and magnetic properties of Fe3O4 as a material in thin film form and proceeded to develop Fe3O4 based synthetic antiferromagnetic (SAF) thin films. Then, we combined these favorable physical/magnetic properties with nanoimprint lithography to fabricate homogeneously patterned synthetic antiferromagnetic (SAF) nanoparticles (wafer area >1 x 1 cm2) with the control of size, shape and structure. Then we demonstrated the release of these particles in an aqueous environment. The fabrication process combines a tetrafluoroethylene (ETFE) "working stamp", a bi-layer resist lift-off, defect-free nanoimprint and sputtering in order to fabricate synthetic antiferromagnetic (SAF) nanoparticles. SAF nanoparticles are composed of alternating magnetic/non-magnetic multilayers to prevent any agglomeration in spite of the ferromagnetic nature of the particles. This heterostructure gives rise to nearly zero magnetic remanence and coercivity values and also prevents possible oxidation of Fe3O4. The superparamagnet-like behavior (nearly zero remanence and coercivity) of SAF nanoparticles suggests that the SAF nanoparticles with favorable geometry fabricated by top-down methods have potential for biomedical application. In order to prove the suitability of SAF nanoparticles for biomedical application, we initially controlled the movement of these SAF nanoparticles with A.C magnetic field, and mechanically rotated them in solution. After we have studied field frequency dependence on mechanical rotation, these SAF nanoparticles were implemented in in vitro environment to test the biocompatibility of these SAF nanoparticlesn, and also to confirm the effectiveness of mechanical force created by A.C magnetic field in order to kill cancer cells. This proof of concept successfully eradicated cancer cells with these SAF nanoparticles. We have demonstrated the effective cancer death after 16 minutes of exposure to mechanically rotating SAF nanoparticles under frequency of 1 Hz (>92% cell death). Furthermore, under the same frequency and exposure time, we have shown that up to 1:4 (nanoparticles:cell) concentration, the mechanical perturbation is effective to kill cancer cells (>80% cell death). However, we suggest to further study the biological mechanism of cancer cell death by mechanical perturbation to truly understand this phenomenon