Synthesis of large pore sized mesoporous silica nanoparticles for bioapplication

다국어 초록 (Multilingual Abstract)

Mesoporous silica nanoparticles (MSNs) have been applied to delivery carriers for various guest molecules based on their large surface area and high pore volume. However, most MSNs have small pore size (~ 3 nm) which is a limitation to load relatively large sized proteins in many bioapplication. Although there have been reports on the preparation of large pore-sized MSNs, it is still challenging to control pore structure of MSNs with large pores and to combine with functional nanoparticles. In this study, we demonstrate the integration of large pore sized MSNs with superparamagnetic nanoparticles. The large pore-sized MSNs encapsulating magnetic nanoparticles were prepared by using water-dispersed magnetic nanoparticles as cores and co-solvent as pore expander in the silica sol-gel reaction in the presence of structure directing agents. The resulting large pore-sized MSNs have a bimodal pore structure composed of 3 nm sized small mesopores and ~ 30 nm sized large mesopores. The pore size, pore morphology, and surface area of the resulting MSNs could be controlled by changing the ratio of co-solvents and/or the amounts of silica precursor. To enhance the adsorption of several types of biomolecules typically used in cancer immunotherapy, the surface of large pore-sized MSNs were modified with amine groups, which resulted in higher loading of model antigen protein and immune adjuvant compared to pristine MSNs with surface unmodified and small pores. The resulting MSNs were tested as delivery vehicles of antigenic protein and immune adjuvant to dendritic cells, showing the potential of the large pore-sized MSNs as delivery vehicles for cancer immunotherapy.

목차 (Table of Contents)

• Abstract .. 10
• 1.Introduction .. 11
• 1.1 Nanocarrier for biomedical applications .. 11
• 1.2 Issue of cancer vaccine .. 12
• 1.3 Mesoporous silica nanoparticles .. 14
• 1.4 Multi-functional mesoporous silica nanoparticles .. 19
• 1.5 Limitation of the conventional mesoporous silica nanoparticles .. 20
• 1.6 Scope of this work .. 21
• 2. Experiments .. 25
• 2.1 Synthesis of magnetic iron oxide nanoparticles .. 25
• 2.1.1 Materials .. 25
• 2.1.2 Method .. 25
• 2.1.2.1 Synthesis of iron-oleate complex .. 25
• 2.1.2.2 Synthesis of iron oxide nanoparticles .. 26
• 2.2 Synthesis of the large-pore sized mesoporous silica nanoparticles embedded with iron oxide nanoparticles .. 26
• 2.2.1 Materials .. 26
• 2.2.2 Method .. 26
• 2.3 Adsorption of biomolecules on mesoporous silica nanoparticles .. 27
• 2.3.1 Materials .. 27
• 2.3.2 BSA adsorption kinetics .. 28
• 2.3.3 OVA adsorption .. 28
• 2.3.4 DNA adsorption .. 28
• 2.4 In vitro experiment .. 30
• 2.4.1 Materials .. 30
• 2.4.2 Cell line culture .. 30
• 2.4.3 Isolation and culture of bone marrow derived dendritic cells.. 30
• 2.4.4 Cellular uptake .. 31
• 2.4.5 Cytotoxicity study .. 32
• 2.4.6 Intracellular delivery.. 32
• 2.4.7 Activation of BMDCs .. 32
• 2.5 Characterization .. 33
• 3. Results and discussion .. 34
• 3.1 Synthesis of MNP@L_MSN .. 34
• 3.1.1 Comparison of small-pore sized mesoporous silica nanoparticles and large-pore sized mesoprous silica nanoaparticles embedded magnetic nanoparticles .. 34
• 3.1.2 Control of physical properties .. 35
• 3.1.3 Surface modification .. 42
• 3.2 Intracellular delivery of immune adjuvant and model antigen .. 43
• 3.2.1 Loading of the biomolecules .. 43
• 3.2.2 Cellular uptake .. 46
• 3.2.3 Cytotoxicity .. 50
• 3.2.4 Intracellular delivery .. 50
• 3.2.5 Activation of BMDCs .. 53
• 4. Conclusion .. 55
• 5. References .. 56
• 6. Summary in Korean .. 60