Scanning electron microscopy, Immunofluorescence staining and statistical analysis of cells after Cytochalasin-D treatment on nanodots with different dot diameter.

<p>(a): Scanning electron microscopy of cells with/without Cytochalasin-D treatment. Cells displayed a round morphology due to actin de-polymerization by Cytochalasin-D. Very few microfilaments were seen on normal and Cytochalasin-D treated cells showing that even after the morphology transition, cells could not establish adhesions with the nanodots due to a greater nanodot height and inter-dot spacing. Normal represents untreated cells on nanosurfaces. Scale bar = 100μm. (b): Immunofluorescence staining of MG63 cells. From left to right: (i) Vinculin was stained with FITC conjugated goat anti rabbit antibodies (Green). (ii) Cytoskeleton was stained with Phalloidin (Red), and far right displays (iii) Merged images. Scale bars: 20X = 75μm, 40X = 50μm, 63X = 25μm. (c): Statistical analysis of cell area and cell viability of cells seeded on nanodots with 50 and 100nm dot diameter after Cytochalasin-D treatment. Cells displayed a smaller cell area on nanosurfaces with 100nm dot diameter as compared to the nanosurfaces with 50nm dot diameter. Cells also displayed a lower viability on the nanodots with 100nm dot diameter. Cells failed to form focal adhesions with the 100nm nanodots due to a greater height and suffered cell death through Anoikis.