Nanoparticle
delivery of polynucleic acids traditionally relies
on the modulation of surface interactions to achieve loading and release.
This work investigates the additional role of confinement in mobility
of dsRNA (84 and 282 base pair (bp) sequences of Spodoptera
frugiperda) as a function of silica nanopore size
(nonporous, 3.9, 8.0, and 11.3 nm). Amine-functionalized nanoporous
silica microspheres (NPSMs, ∼10 μm) are used to directly
visualize the loading and exchange of fluorescently labeled dsRNA.
Porous particles are fully accessible to both lengths of dsRNA by
passive diffusion, except for 282 bp dsRNA in 3.9 nm pores. The stiffness
of dsRNA suggests that encapsulation occurs by threading into nanopores,
which is inhibited when the ratio of dsRNA length to pore size is
large. The mobility of dsRNA at the surface and in the core of NPSMs,
as measured by fluorescence recovery after photobleaching, is similar.
The mobility increases with pore size (from 0.0002 to 0.001 μm2/s for 84 bp dsRNA in 3.9–11.3 nm pores) and decreases
with the length of dsRNA. However, when the dsRNA is unable to load
into the pores (on nonporous particles and for 282 bp dsRNA in 3.9
nm pores), surface mobility is not detectable. The pore structure
appears to serve as a “source” to provide a mobile network
of dsRNA at the particle surface. The importance of mobility is demonstrated
by exchange experiments, where NPSMs saturated with mobile dsRNA can
exchange dsRNA with the surrounding solution, while immobile dsRNA
is not exchanged. These results indicate that nanoparticle synthesis
techniques that provide pores large enough to take up polynucleic
acids internally (and not simply on the external surface of the particle)
can be harnessed to design polynucleic acid/nanoporous silica combinations
for controlled mobility as a path forward toward effective nanocarriers.