Aquasomes are nanocarrier systems consist of three distinctive layers; an inner core, a polyhydroxy carbohydrate layer and an outer layer of an API (Kossovsky et al., 1991). Aquasomes have a unique structure and ability to carry active molecules through a non-covalent bounding and provide superior stability, especially for proteins and peptides (Masatoshi and Yongning, 1998; Kim and Kim, 2002; Khopade et al., 2002). Different core and coating materials were used to prepare aquasomes under different conditions to investigate the relationship between preparation conditions and loading efficiency. In terms of loading efficiency, hydroxyapatite aquasomes, with either lactose or trehalose as a coating material, had the highest BSA loading (40%-60%) when compared to DSPA aquasomes. While DCPA aquasomes, with either lactose or trehalose as a coating material, had the lowest BSA loading (8%-16%). To investigate the interaction of the three layers of aquasomes, Surface analysis, docking and MD simulations were performed. Surface analysis performed by Discovery Studio showed that HA and trehalose interact by hydrogen bonding with the later acting as a hydrogen acceptor, while BSA displayed almost complete SAS and that there are numerous targets for trehalose attachments (no specific active site). MD simulations of BSA performed by AMBER 12 showed a stable MD simulation of BSA for 5 ns. Total energy analysis of BSA on the two conditions performed (300K and 280K) support the experimental data of lower BSA loadings of aquasomes prepared at 400C compared to those manufactured at 250C (p5.5 and steadily release for 6 hr. Cell culture studies were conducted to demonstrate the controlled release effect of aquasomes using Caco-2 cell lines. The release of metronidazole (model drug) from aquasomes post 2 hr started to slow gradually until it reached its highest difference at 6 hr (p<0.05) when compared to the control
