| 摘要: | Albumin and liposomal nanoparticles (NPs) have gained tremendous interest as therapeutic drug carriers benefitting from their unique physicochemical properties. However, introducing NPs into biological environments would induce layers of protein adsorbed on the surface of the NPs known as protein corona. This in turn might impair the pristine function of NPs and reduce the drug’s efficacy itself. In this regard, we exploited the protein corona behavior on NPs surface through in vitro and in silico study designs. Firstly, the protein corona was studied by determining the size distribution and zeta potential of NPs. The identification of the highly recruited protein profile (hard-corona) was then performed by SDS-PAGE electrophoresis and LC-MS/MS. Furthermore, the interaction between the NPs and the highly recruited proteins, namely albumin and apolipoprotein-C3, was constructed through unbiased molecular dynamics (MD) simulation. As a comparison, the most commonly studied protein corona, apolipoprotein E, was also investigated through MD. Herein, the in silico and experimental observations were carried out to understand the principle and behavior of protein-NPs interaction down to the molecular level. The results obtained from physical characterization showed that the sizes of both types of NPs increased upon being introduced to fetal bovine serum ranging from 130 to 180 nm, with higher negative zeta potential indicated from -0.8 to -13 (for albumin NPs) and +40 to -16 (for liposomal NPs). A diverse protein corona formation was also found on liposomal NPs as opposed to that on albumin NPs. Further, to evaluate the effect of corona formation toward interaction with cells, the hard corona protein and soft corona protein were introduced to albumin NPs. As a result, the uptake efficiency in the two different treatments differs strongly, in association with the different serum concentrations. The effect of serum concentration in regulating the cellular availability was scrutinized by loading the anti-cancer compound into albumin NPs by differing the serum containing media composition (with or without the fetal bovine serum). It shows that higher serum concentration affects highly toward the lower cytotoxicity of Mia-Paca-2 cell lines. Meanwhile, the corona protein interactions with albumin NPs were investigated through MD simulations with Martini-forcefield to gain better understanding beyond the corona protein and albumin NPs phenomena. The result suggests that the most favorable interaction of corona proteins observed in albumin NPs is with corona albumin. More interaction of the albumin-apolipoprotein complex was also observed toward the LDL-receptor, which was further introduced on the membrane surface. This result is associated coherently with the experimental finding that introducing apolipoprotein onto NPs surface is advantageous to enhance the NPs uptake efficiency by looking at the appropriate receptor present on the cancerous membrane surface. In conclusion, the extensive molecular modeling studies potentially provide an excellent agreement with experimental observations and further offering better molecular insights into protein corona formation and interactions. |
