Title Page
Contents
Chapter 1. Development of biodegradable polyhydroxyalkanoate-based nanocarrier for drug release 2
Contents 4
국문초록 6
Abstract 8
Abbreviation 10
I. Introduction 14
II. Purpose 21
III. Materials & Methods 22
1. Materials 22
2. Preparation of paclitaxel loaded nanoparticles 23
3. Entrapment efficiency 23
4. Determination of nanoparticles size and zeta potential 24
5. Storage stability of nanoparticles 25
6. Fourier-Transform Infrared Spectroscopic analysis 25
7. Differential Scanning Calorimetric analysis 26
8. X-ray Diffraction analysis 26
9. Transmission electron microscope of nanoparticles 27
10. In vitro drug release of nanoparticles 28
11. Release kinetics of PTX from nanoparticles 29
12. In vitro cell experiment 31
IV. Results & Discussion 33
1. Physicochemical properties of nanoparticles 33
2. Storage stability 44
3. In vitro Release of nanoparticles 47
4. Release kinetics of PTX from nanoparticles 49
5. Intra cellular uptake of nanoparticles 55
V. Conclusions 57
VI. References 59
Chapter 2. Development of Paclitaxel-loaded polyhydroxyalkanoate nanoparticles for drug-eluting stent 71
Contents 72
국문초록 74
Abstract 76
Abbreviation 79
I. Introduction 83
II. Purpose 89
III. Materials & Methods 90
1. Materials 90
2. Preparation of paclitaxel loaded nanoparticle 91
3. Entrapment efficiency 91
4. Determination of nanoparticle size and zeta potential 92
5. Storage stability of nanoparticle 93
6. Fourier-Transform Infrared Spectroscopic analysis 93
7. Differential Scanning Calorimetric analysis 94
8. X-ray Diffraction analysis 94
9. In vitro drug release of nanoparticle 95
10. Release kinetics of PTX from nanoparticle 96
11. Preparation of Cobalt-chromium alloy for drug eluting stent 98
IV. Results & Discussion 101
1. Physicochemical properties of nanoparticle 101
2. Storage stability 108
3. I n vitro Release of nanoparticles 110
4. Release kinetics of PTX from nanoparticles 110
5. Cobalt-chromium alloy for drug eluting stent 114
V. Conclusions 119
VI. References 120
Chapter 1. Development of biodegradable polyhydroxyalkanoate-based nanocarrier for drug release 11
Table 1. Physicochemical properties of NPs. 35
Table 2. Release kinetics of PTX from NPs at pH 7.4. 50
Chapter 2. Development of Paclitaxel-loaded polyhydroxyalkanoate nanoparticles for drug-eluting stent 80
Table 1. Physicochemical properties of NP. 102
Table 2. Release kinetics of PTX from NP at pH 7.4. 113
Table 3. Physicochemical properties of Chitosan-modified nanoparticle. 115
Chapter 1. Development of biodegradable polyhydroxyalkanoate-based nanocarrier for drug release 12
Figure 1. Chemical structure of poly(latic-co-glycolic acid). 16
Figure 2. Chemical structure of polyhydroxyalkanoate. 18
Figure 3. Chemical structure of paclitaxel. 20
Figure 4. HeLa cell from the Korean Cell Line Bank (KCLB No.1002). 32
Figure 5. TEM images of NPs (a) NP1 (b) NP2 (c) NP3 (d) NP4. 36
Figure 6. FT-IR spectra of NPs (a) NP1, (b) NP2, (c) NP3, (d) NP4. The major peaks are indicated for spectra of each of the NPs. 39
Figure 7. DSC analysis of NPs (a) NP1, (b) NP2, (c) NP3, (d) NP4. The major peaks are indicated for the DSC curve of each NP. 41
Figure 8. X-ray diffraction (XRD) characterization of NPs (a) NP1, (b) NP2, (c) NP3, (d) NP4. Diffraction patterns were scanned... 43
Figure 9a. Storage stability of NPs at 4 ℃ for one week. The size of NPs were monitored every day. 45
Figure 9b. Storage stability of NPs at 4 ℃ for one week. The zeta potential (ζ) values of NPs were monitored every day. 46
Figure 10. In vitro release profile of PTX from NPs at 37 ℃. Amount of PTX released was quantitatively analyzed at pH 7.4.... 48
Figure 11a. PTX release data plotted to various kinetic models(zero-order, first-order, Higuchi, and Hixson-Crowell) of NP1 obtained... 51
Figure 11b. PTX release data plotted to various kinetic models(zero-order, first-order, Higuchi, and Hixson-Crowell) of NP2 obtained... 52
Figure 11c. PTX release data plotted to various kinetic models(zero-order, first-order, Higuchi, and Hixson-Crowell) of NP3 obtained... 53
Figure 11d. PTX release data plotted to various kinetic models(zero-order, first-order, Higuchi, and Hixson-Crowell) of NP4 obtained... 54
Figure 12. Intracellular uptake of NPs into HeLa cells. Fluorescence indicates nucleus (blue), NPs (red), and lysosomes (green), as... 56
Figure 13. Scheme of Nanoparticles. 58
Chapter 2. Development of Paclitaxel-loaded polyhydroxyalkanoate nanoparticles for drug-eluting stent 81
Figure 1. Chemical structure of polyhydroxyalkanoate. 86
Figure 2. Chemical structure of paclitaxel. 88
Figure 3. Cobalt-chromium alloy coating. 100
Figure 4. FT-IR spectra of NP. The major peaks are indicated for spectra of the NP. 104
Figure 5. DSC analysis of The major peaks are indicated for the DSC curve of NP. 105
Figure 6. X-ray diffraction (XRD) characterization of NP. Diffraction patterns were scanned between 2 and 80°/2θ. 107
Figure 7a. Storage stability of NP at 4 ℃ for one week. The size values of NP were monitored every day. 109
Figure 7b. Storage stability of NP at 4 ℃ for one week. The zeta potential (ζ) values of NP were monitored everyday. 109
Figure 8. In vitro release profile of PTX from NP at 37 ℃. Amount of PTX released was quantitatively analyzed at pH 7.4.... 111
Figure 9. PTX release data plotted to various kinetic models(zero-order, first-order, Higuchi, and Hixson-Crowell) of NP... 112
Figure 10a. Storage stability of C-NP at 4 ℃ for one week. The size of C-NP were monitored every day. 117
Figure 10b. Storage stability of C-NP at 4 ℃ for one week. The zeta potential (ζ) values of C-NP were... 117
Figure 11. SEM image of Cobalt-chromium alloy coated with chitosan-modified nanoparticle 118