| Summary: | 碩士 === 國立臺灣大學 === 醫學工程學研究所 === 94 === Abstract
The mechanism of the cells grow in the three dimensional environment (3D) is more complicate than that two dimensional surface (2D), The behavior of the cells is different, in terms of cell attachment, cell aggregation, cell migration, cell shape, cell proliferation and differentiation, gene expression, the interaction between cell and extracellular matrix etc.
In living tissue, the cells mostly exist and connect with ECM components, which integrated into 3D framework. To simulate the growth of the cells in vivo, we cultured the MG63 osteoblastic cells in 3D scaffold, which is polycaprolactone (PCL) and fabricated via fused deposition method. The diameter of PCL fiber of 3D scaffolds is 130um. The size of interconnected pore of 3D scaffold is expressed as the distance between two adjacent PCL fibers and three sizes of #300um, #250um, and #200um were designed.
The morphology of MG63 cells seeded into PCL scaffold were examined by SEM. The MG63 cells cultured on 2D surface, including tissue culture plate and PCL membrane is flat and spindle in shape. However, the cells cluster intermingled with extracellular matrix was noted in 3D scaffold.
The cell proliferation and connection of the cells attached on adjacent PCL fibers increased with culture period, followed by filling-in of the superficial pores of 3D scaffolds.
The cell viability was aided by MTT assay, which indicated the growth of MG63 cells in our study. The results of MTT assay demonstrated the cellular growth in #200um scaffold was the best among the three scaffolds with different pore-sizes in our study.
To provide the substrate for cell adhesion in the 2-dimensional culture surface, the cells grown on the PCL membrane were covered with another PCL membrane, which made MG63 cells interposed between upper and lower membranes. The cell viability and cell attachment to the upper membrane was detected. Moreover, the total viability on upper and lower membranes was higher than that on single membrane.
The gene expression of osteogenic marker, alkaline phosphatase, of MG63 cells was assayed by polymerase chain reaction (PCR) technique, after the MG63 cells were cultured in osteogenic medium for 2 weeks and 4 weeks. Our preliminary results showed that the expression of ALP increased with culture time and was significantly higher in 3D PCL scaffold than on PCL membrane.
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