抄録
In this study, surface modification method using O_2 plasma-treated polydimethylsiloxane (PDMS) was developed to minimize the influences of fibroblast for bone formation. 3-well chambers consist of polycarbonate wall and glass slide coated with PDMS were prepared. Ultra-thin glass plates coated with PDMS were prepared to measure adhesive force between cell and PDMS surface. Contact angle of PDMS surface was controlled by O_2 plasma treatment (0°, 50° and 100°). Fibroblast and osteoblast-like cell were seeded respectively on the ultra-thin glass plate and 3-well chambers. Adhesive force was measured by detaching a cell from PDMS surface with micropipette and focal adhesion of vinculin was counted with fluorescent observation after 6 hours of cultivation The difference of adhesive force between osteoblast-like cell and fibroblast was highest on the PDMS surface with contact angle of 50°, and the difference of the number of focal adhesions was also highest on the surface. Co-culture of osteoblast-like cells and fibroblasts was carried out on the 3-well chambers. The ratio of osteoblast-like cell to fibroblast was 1:1. In the case of PDMS with contact angle of 50°, the number of adherent osteoblast-like cells was highest according to fluorescent observation after 6 hours of cultivation, and ALP activity was also highest after 9 days of cultivation. These results showed that PDMS surface with contact angle of 50° can maintain ALP activity of osteoblast-like cells with the influence of fibroblasts minimized for co-culture because the difference of cell adhesion between osteoblast-like cell and fibroblast was high on the contact angle.
