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Contents
(Abstract) 5
Introduction 7
Materials and methods 9
Materials 9
Degradation of HA 9
Preparation of cisplatin-incorporated nanoparticles of HA 10
Measurement of the drug contents and loading efficiency of nanoparticles of HA 10
Measurement by gel permeation chromatography(GPC) 11
Transmission electron microscopy (TEM) observation 11
Particle size and zeta potential measurement 11
X-ray powder diffractometry (XRD) measurement 12
Drug release study 12
Hyaluronidase extraction from tumor cell lines 12
Zymography of hyaluronic acid 13
Results 15
Characterization of degraded HA 15
Preparation of cisplatin-incorporated nanoparticles 15
Cisplatin release from nanoparticles 16
Discussion 18
Conclusion 20
References 21
〈초록〉 36
Table1. Characterization of cisplatin-incorporated HA nanoparticles 28
Fig. 1. Schematic illustrations of nanoparticle formation by ion complex between HA and cisplatin. Cisplatin incorporated HA nanoparticles are formed by ion complex between carboxyl groups... 29
Fig. 2. GPC chromatogram of degraded HA. the GPC system was composed of a waters 515 HPLC pump, waters 410 differential refractometer, waters 486 tunable absorbance detector,... 30
Fig. 3. Particle size distribution against drug contents. A sample solution prepared by the dialysis method was used for the particle size measurement (concentration: 0.1 w/v). 31
Fig. 4. Morphology of cisplatin-incorporated HA nanoparticles (100/10 of table 1 ) observed by TEM. A drop of nanoparticle suspension was placed onto a carbon film coated on a copper grid for TEM... 32
Fig. 5. XRD patterns of cisplatin-incorporated polymeric micelle of HAgPEG. cisplatin (a), HA (b), cisplatin-incorporated nanoparticles(100/5 in table 1) (c), cisplatin-incorporated nanoparticles (100/10) ... 33
Fig. 6. The effect of drug contents (a) and presence of hyaluronidase (b) on the cisplatin release from HA nanoparticles. the hyaluronidase (100 units/ml ) was added to the dialysis tube containing... 34
Fig.7. HA zymography using U343MG and U87MG cell lines (a). the effect of extracted hyaluronidase from U343MG and U87MG cell lines (b). the hyaluronidase activity was determined in the form... 35
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