Title Page
Contents
ABSTRACT 11
1. Introduction 13
2. Materials and Methods 18
2.1. Materials 18
2.2. Synthesis of MA-Azo-Azo-Dox monomer 18
2.3. Synthesis of ferrocene-doxorubicin-containing polymers (FDPs) 23
2.4. Preparation of the FDNPs 24
2.5. Stability analysis of the FDNPs 24
2.6. Characterization of the tumor microenvironments-responsive properties of the FDNPs 24
2.7. Tumor microenvironments responsive release profiles of drugs from the FDNPs 25
2.8. Cell cultures and viability assays 27
2.9. Microscopic analysis of cells 27
3. Results and Discussion 28
3.1. Synthesis and characterization of the MA-Azo-Azo-Dox monomer 28
3.2. Characterization of the ferrocene-doxorubicin-containing polymers (FDPs) 30
3.3. Characterization of FDNPs 32
3.4. Stability analysis of the FDNPs 34
3.5. Tumor microenvironments-responsiveness of the FDNPs 36
3.6. In vitro drug release analysis of tumor microenvironments sensitive FDNPs 40
3.7. Cell viability assays 44
4. Conclusion 51
5. References 52
Table. 1. RP-HPLC conditions for MA-Azo-Azo-Dox monomer analysis. 21
Table. 2. Semi-preparative RP-HPLC conditions for MA-Azo-Azo-Dox monomer analysis. 22
Graphical abstract. Scheme of ferrocene-doxorubicin-containing nanoparticles (FDNPs) for tumor microenvironmentally reactive release of drugs. 12
Figure 1. HPLC peak of purified MA-Azo-Azo-Dox monomer. Analytical reverse-phase high-performance liquid chromatography (RP-HPLC) was... 29
Figure 2. ¹H NMR spectra of the ferrocene-containing polymer and ferrocene-doxorubicin-containing polymer. 31
Figure 3. Characterization of FDNPs synthesized using a ferrocene-doxorubicin-containing polymer. a) Diameter of FDNPs. b) PDI value of FDNPs. c) zeta... 33
Figure 4. Measurement of stability of FDNPs. a) TEM image of FDNPs at week 0. b) TEM image of FDNPs after 4 weeks. c) The diameter of two FDNP groups. d)... 35
Figure 5. Characterization of the tumor microenvironments-responsive properties of FDNPs. A) Change in diameter of FDNPs with time after H₂O₂ treatment. B)... 38
Figure 6. a) Fluorescence intensity standard curve according to the concentration of doxorubicin dispersed in PBS (pH 7.4). b) Graphs of drug... 42
Figure 7. In vitro cytotoxicity of W/O drug loading ferrocene-doxorubicin-containing nanoparticles and DOX loading ferrocene-doxorubicin-containing... 46
Figure 8. In vitro cytotoxicity evaluation of ferrocene-doxorubicin-containing nanoparticles in cancer cells in normoxia and hypoxia environments. (a) HeLa... 48
Figure 9. Micrographs of control and ferrocene-doxorubicin-containing nanoparticles treated cancer cell lines. a) HeLa cells, b) A549 cells. 49
Figure 10. Fluorescence intensity quantification graph of H33342 dye after treatment with ferrocene-doxorubicin-containing nanoparticles. 50
Scheme 1. The detailed synthesis method of ferrocene-doxorubicin-containing nanoparticles (FDNPs) for tumor microenvironments-responsive release of target drugs. 17