  In recent years, extracellular vesicles such have become promising carriers
as the next-generation drug delivery platforms. Effective loading of exogenous
cargos without compromising the extracellular vesicle membrane is a major
challenge. Rapid squeezing through nanofluidic channels is a widely used
approach to load exogenous cargoes into the EV through the nanopores generated
temporarily on the membrane. However, the exact mechanism and dynamics of
nanopores opening, as well as cargo loading through nanopores during the
squeezing process remains unknown and is impossible to be visualized or
quantified experimentally due to the small size of the EV and the fast
transient process. This paper developed a systemic algorithm to simulate
nanopore formation and predict drug loading during extracellular vesicle (EV)
squeezing by leveraging the power of coarse-grain (CG) molecular dynamics
simulations with fluid dynamics. The EV CG beads are coupled with implicit
Fluctuating Lattice Boltzmann solvent. Effects of EV property and various
squeezing test parameters, such as EV size, flow velocity, channel width, and
length, on pore formation and drug loading efficiency are analyzed. Based on
the simulation results, a phase diagram is provided as a design guidance for
nanochannel geometry and squeezing velocity to generate pores on membrane
without damaging the EV. This method can be utilized to optimize the
nanofluidic device configuration and flow setup to obtain desired drug loading
into EVs
