High-Throughput Mechanobiology Screening Platform Using Micro- and Nanotopography.

Area of Science:

• Biotechnology
• Cell Biology
• Materials Science

Background:

• Existing high-throughput platforms have limitations in testing multiple factors and assays.
• Micro- and nanotopographies can influence cell behavior, but screening methods are limited.
• Cell activation and proliferation are critical processes in immunology and regenerative medicine.

Purpose of the Study:

• To develop and validate a versatile 96-well plate platform for high-throughput screening of micro- and nanotopographies.
• To investigate the effects of specific topographies and drug-topography combinations on T cell activation and proliferation.
• To identify optimal surface features for enhancing T cell expansion for clinical applications.

Main Methods:

• Development of a 96-well plate platform compatible with liquid handling, high-resolution imaging, and multiwell plate instrumentation.
• Fabrication of "trench-grid" surfaces with varying dimensions (e.g., 100 nm, 200 nm width; <0.5 μm, >2.3 μm pitch).
• Coating surfaces with anti-CD3 and anti-CD28 antibodies to stimulate T cells and assaying for interleukin-2 (IL-2) secretion.

Main Results:

• The platform successfully screened topography effects on T cell activation and proliferation.
• Optimal IL-2 secretion was observed at 200 nm trench width and >2.3 μm grating pitch, while suppression occurred at 100 nm width and <0.5 μm pitch.
• The 200 nm grid pattern, combined with blebbistatin to reduce cell contractility, tripled T cell numbers in long-term expansion.

Conclusions:

• The developed 96-well plate platform is a powerful tool for high-throughput screening of micro- and nanotopographies.
• Specific nanotopographies can significantly modulate T cell activation, proliferation, and cytokine production.
• The findings demonstrate potential for optimizing surface properties in adoptive immunotherapy to enhance T cell expansion.