This thesis focuses on the study of nanotechnology and its applications in immunology and mechanosensing using micro- and nano-scale topographies, such as gratings, grids, and pillar substrates. In the past five years, we have developed three types of platforms and explored the influence of nano-patterned substrates on cell morphology, proliferation, protein secretion, and mechanosensing. I will introduce the three generations of Integrated Mechanobiology Platform (IMP) for T cell study, including the fabrication process of each generation of IMP, their advantages and disadvantages, and the comparison with existing High Throughput Screening System (HTSS). For the applications of IMP, I will focus on grating and grid topographies with IMP generation 3 format, and study how these nano-patterned substrates affect T cell morphology, expansion, cytokine secretion, drug-topography combination effects on T cells and long-term expansion for adoptive immunotherapy. I will demonstrate how IMP enables such studies in a high throughput manner.

I also will discuss how Multiple Stiffness Pillar Platform (MSPP) facilitates the study of mechanosensing in cells spanning across different rigidities. First, I will talk about how MSPP is different from existing dual stiffness platforms. Differences include flexibility in distribution of different rigidities, consistency in pillar dimensions and ease of controlling the stiffness fold increase. In the sections of MSPP fabrication and characterization, I will focus on measurements of stiffness change and surface chemistry uniformity. I will then discuss the Mouse Embryonic Fibroblast (MEF) mechanosensing study on dual stiffness pillar substrates, including the preferential localization of rigidity sensing associated proteins (myosin IIA, phosph-myosin, paxillin, and p130CAS), MEFs actomyosin network building, and adhesion formation. These studies revealed previously undiscovered results in MEF mechanosensing, and demonstrate the great potential of MSPP in this research discipline. In the last part of this thesis, I will present on the mass production of thermoplastic nanopatterned molds.

The demonstrated technology can produce large batches of nanostructured molds with decreased fabrication time and expense. In this chapter, I will discuss the necessity of developing such a technology and platform, as well as the design, fabrication, and characterization of the thermoplastic nano-patterned molds.