| Summary: | 碩士 === 國立東華大學 === 物理學系 === 107 === In recent years, cancer has become one of the leading deaths worldwide. To develop a new anti-cancer agent, the cytotoxicity test of drugs is traditionally determined by two dimensional (2D) cell culture before animal and clinical studies. However, it cannot adequately mimic the real tumor environment. It provides the different results for the animal and clinical studies. To mimic the drug response with real tumor environment and reduce the gap between the traditional cell experiment and animal study, various combination of three dimensional (3D) tumor spheroid culture and co-culture systems are developed. In this study, we use 3D co-culture model to provide a suitable platform for the development and assessment of the new anti-cancer agent.
Nanodiamond (ND) is considered a feasible platform in bio-imaging and drug delivery application owing to its physical/chemical properties and biocompatibility. ND aggregated easily in buffer solution, so we used human serum albumin (HSA) adsorbed on the ND surface to get a well dispersed ND. Then the ND-HSA was used to conjugate doxorubicin (DOX) to receive ND-HSA-DOX complex. Utilize UV-Visible spectrometer to estimate the adsorption of DOX on ND. The particle size and functionalized surface of ND were characterized using dynamic light scattering analyzer (DLS) and Fourier Transform Infrared spectrometer (FTIR). Moreover, the DOX release from ND-HSA-DOX at different pH conditions were discussed. At lower pH environment, the DOX has higher release from ND-HSA-DOX. Furthermore, the cytotoxicity effects of DOX and ND-HSA-DOX were assessed in 2D- and 3D- co-culture models via MTT assay and growth inhibition assay. Under the observation of confocal fluorescence microscope, the intracellular location of DOX and ND-HSA-DOX were observed in 2D co-culture model, the fluorescence of DOX and ND-HSA-DOX were both revealed in the cell nucleus. The results show ND-HSA-DOX has a similar effect to pure DOX in 2D co-culture model. We further study the growth inhibition of DOX and ND-HSA-DOX in 3D co-culture MCTS. Growth inhibition assay showed the ND-HSA-DOX have greater efficient to pure DOX in co-culture MCTS. We characterized ND-HSA-DOX and showed it has better cancer inhibiting efficacy compare to DOX in 3D co-culture MCTS.
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