Preparation and characterization of multi-functional caspase-3 loaded nanoparticles for folate receptor and CD44 targeting delivery

• Main Authors: Chiu-Yi HUang, 黃久益
• Other Authors: Fwu-Long Mi
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/18994941615758134983

碩士 === 萬能科技大學 === 工程科技研究所 === 100 === Targeted delivery of drugs to cancer cells has attracted considerable attention in developing new chemotherapeutic modalities. In this study, chitosan (CS) and hyaluronan (HA) based nanoparticles with prominent targeting moieties capable of specific interaction with the CD44 and folate receptors (FR) on colon carcinoma HT-29 cells have been developed for Caspase-3 delivery. In the first part of the experiment, crude Caspase-3 protein was purified using hydrophobic HIC column and dialysis apparatus. The purified protein was subsequently indentified by electrophoresis analysis to obtain the 36kDa molecular weight of Caspase-3 protein. In the second part of the experiment, average particle size, light scattering intensity and zetal potential of the self-assembled CS-FA/HA nanoparticles were all determined by dynamic light scattering particle size analysis (DLS). Caspase-3 was encapsulated in the nanoparticles and the protein loading efficiency was measured. Subsequently, the release profiles of Caspase-3 from the nanoparticles were investigated in phosphate buffered saline (PBS). In the third part of the experiment, the FITC-labeled CS-FA/HA nanoparticles was used as a drug delivery carrier for targeting HT-29 cell (a human colorectal cancer cell) over-expressing CD44 and folate receptors. The intensity of fluorescence observed in HT-29 cell incubated with the CS-FA/HA nanoparticles was stronger than that incubated with the CS/HA nanoparticles. These results indicated that the prepared CS-FA/HA nanoparticles had the higher specific interaction with HT-29 cell via the ligand-receptor-mediated recognition therefore may be suitable for the delivery of Caspase-3 to CD44/folate receptors over-expressed tumor cells. Keyword: chitosan, hyaluronan, Caspase-3, CD44, folate receptor