At the intersection of materials chemistry and biology, biomaterials have been successfully employed in an array of medical applications. From diagnostic tools to targeted drug delivery, the modular physical and chemical properties of these materials provide numerous applications. For example, porous nanoparticles have been widely integrated as vehicles to carry chemotherapeutics to localized tumor sites. By encapsulating these cytotoxic compounds within a porous framework, the commonly associated adverse side effects of conventional chemotherapeutics, such as Doxorubicin, have been greatly reduced. One such material, mesoporous silica, has received widespread attention due to its excellent biocompatibility, high surface area to mass ratio, tunable pore diameters and volumes, and robust surface chemistry. However, recent studies have demonstrated that exposing silica nanoparticles, and other synthetic materials, to biological milieu envelops the particles in layers of proteins and biomolecules. The resulting protein coat, known as the "protein corona", has been shown to have profound effects on bioavailability, cellular targeting, and cytotoxicity. Thus, in order to develop safe and effective particle-based therapies, it is of utmost importance to establish a more thorough understanding of this process.
To examine how changes in surface chemistry influence protein adsorption, monodisperse, spherical mesoporous silica nanoparticles, ca. 50 nm, were modified with a variety of surface functionalizations, -NH2, -COOH, and -PEG. Exposing these materials to biological fluid revealed drastically different protein fingerprints, suggesting a strong correlation between the surface chemistry and the identity and composition of the protein corona. Quantification of the protein corona, i.e. mg protein/mg particles, was then achieved by performing thermogravimetric analysis. These values, in concert with spectral counts obtained by shotgun proteomics, illustrates a method for quantifying individual proteins present in the corona.
Spherical, silica particles of varying diameters, 70-900 nm, were then synthesized to investigate how particle diameter may affect the biomolecular identity of the protein corona. Applying the previously described methods, it was found that mesoporous particles exhibit a higher affinity for low-molecular weight proteins compared to dense silica particles of similar diameters.
Finally, stochastic optical reconstruction microscopy (STORM) was used to map protein adsorption/diffusion throughout as-prepared (pore diameter ~ 30 Å) .and large pore (pore diameter > 60 Å) mesoporous silica particles. By collecting three-dimensional data on the protein-adsorbed materials, a sphere-fitting algorithm could be applied to determine the center and radius of the host particle. This calculation demonstrated that the depth by which specific proteins diffused into the porous framework was a function of both the protein's molecular weight as well as the pore diameter.
| Identifer | oai:union.ndltd.org:uvm.edu/oai:scholarworks.uvm.edu:graddis-1638 |
| Date | 01 January 2016 |
| Creators | Clemments, Alden Michael |
| Publisher | ScholarWorks @ UVM |
| Source Sets | University of Vermont |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Graduate College Dissertations and Theses |
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