An acoustic strategy for gold nanoparticle loading in platelets as biomimetic multifunctional carriers

In recent years, a wide variety of bioinspired colloidal particles with novel cell mimetic functions have been the subject of extensive research in materials science, chemistry, biology, physics, and engineering. However, most of the approaches are derived from natural cell membrane coatings, which are still too primitive compared with living cells. In this study, we have chosen gold nanoparticles (GNPs) to explore the bioactivity response of living platelets and nanoparticle loading efficiency under different ultrasonic intensity and frequency treatment conditions. The results show that GNPs with no surface modification could be easily loaded into intra-platelets by both incubation (30 min) and ultrasonic exposure (1 min) methods. The amount of GNP loading was (4.4 ± 0.9) × 10^-3 and (5.8 ± 2.4) × 10^-3 pg per platelet upon incubation and acoustic triggering (1 MHz, 0.25 W cm^-2), respectively. Although the other US treatment intensities (0.75, 1.50 and 2.25 W cm^-2) also promoted higher amounts of GNPs in the platelets, the higher US intensity might bring about partial damage of the platelet membrane. Compared with 1 MHz ultrasonic exposure, the change of the GNP loading amount was not significantly higher upon ultrasonic frequency treatment of 45, 80 or 100 kHz. Therefore, it has been found that an US intensity of 0.25 W cm^-2 could facilitate the intra-platelet delivery efficacy of the GNPs without damaging the biological activity. Furthermore, two possible pathways of GNPs entering into platelets upon US treatment are presented: one is the endocytosis/open canalicular system (OCS), and the other is cell membrane permeability enhancement, which is proved by the SEM and TEM results. Finally, the GNP-loaded platelets have been demonstrated as useful probes for photoacoustic imaging (PAI) and dark-field microscopy (DFM)-based imaging, which might allow a wide range of potential applications in diagnostics and therapy of platelet-related diseases.