Exposure of nanoparticles in a porous medium, such as a hydrogel, to low-intensity ultrasound has been observed to dramatically enhance particle penetration rate. Enhancement of nanoparticle penetration is a key issue affecting applications such as biofilm mitigation and targeted drug delivery in human tissue. The current study used fluorescent imaging to obtain detailed experimental measurements of the effect of ultrasound amplitude and frequency on diffusion of nanoparticles of different diameters in an agarose hydrogel, which is often used as a simulant for biofilms and biological tissues. We demonstrate that the acoustic enhancement occurs via the phenomenon of oscillatory diffusion, in which a combination of an oscillatory flow together with random hindering of the particles by interaction with hydrogel proteins induces a stochastic random walk of the particles. The measured variation of acoustic diffusion coefficients with amplitude and frequency were used to validate a previous statistical theory of oscillatory diffusion based on the continuous time random walk approach.