Committee:

Alexander Alexeev, Ph.D. (ME, Georgia Institute of Technology)

Scott Danielsen, Ph.D. (MSE, Georgia Institute of Technology)

David Myers, Ph.D. (BME, Georgia Institute of Technology)

Guillem Pratx, Ph.D. (Radiology and Medical Physics, Stanford University) 

Optimization of Forces, Loading Rates and Strain for Cytosolic Delivery using Mechanoporation  

Efficient intracellular delivery of cargo remains a central challenge in biomedical engineering to improve gene editing, cell therapy production, and cell diagnostics. The optimal delivery not only introduces cargo into cells but achieves cytosolic access while preserving cell viability and normal function. Existing approaches often fall short of this balance. Viral and carrier-mediated methods can promote uptake of molecularly encoded genes but are limited by endosomal entrapment and intracellular degradation. Electroporation can disrupt cell membranes but also disturbs normal cellular physiology. These limitations have motivated interest in microfluidic mechanoporation, in which cells are transiently permeabilized through controlled mechanical deformation. However, the mechanism of mechanoporation remains poorly defined and, as such, performance remains difficult to predict and standardize. To bridge the gap, this dissertation establishes a framework connecting device geometry, transient loading dynamics, and delivery outcomes. A key learning is that mechanoporation performance is demonstrated to be better explained by cell loading kinetics than by deformation magnitude alone. Systematic channel width and ridge angle variation demonstrates how tuning force rate, directionality, and loading history improves delivery efficiency, cell survival and experimental consistency. Integration of high-speed trajectory analysis, computational fluid dynamics (CFD), and multivariate modeling further reveals that steady-state descriptors such as strain magnitude are insufficient to explain mechanoporation outcomes, which instead, correlate strongly with strain rate and history-dependent hydrodynamic forces. Extending these insights to nanoparticle delivery, imaging analyses demonstrate that optimized device geometries enable deeper intracellular access and improved cargo retention relative to conventional incubation-based approaches. Together, these results provide a force-based framework for mechanoporation that links device design to biologically meaningful intracellular delivery.