Effects of calcium-phosphate topography on osteoblast mechanobiology determined using a cytodetacher

Abstract

The Human fetal osteoblast (hFOB) cell morphology, adhesion force, and proliferation on a calcium-phosphate (Ca-P) micropattern surface were investigated and the mechanobiology was investigated by a cytodetachment test. Ca-P-coated groove patterns with 3.0-μm-deep grooves (C3), 4.5-μm-deep grooves (C4), and 5.5-μm-deep grooves (C5) were produced on silicon wafers using photolithography and wet etching techniques. The grooved substrates were coated with a 200-nm-thick layer of titanium (bond coat) and a 200-nm-thick layer of calcium phosphate (top coat) using a sputtering system. Smooth Ca-P-coated Si wafers were used as control surfaces. Analysis of the scanning electron microscopy observations shows that cells on the Ca-P micropattern showed spreading and elongation. The MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay indicated that C3 and C4 specimens had a significantly higher number of cells than did the control group after 5- and 15-day cultures. The cyto-compatibility of specimens was quantitatively evaluated using a cytodetacher, which directly measures the detachment shear force of an individual cell to the substrate. After 30-min culture, the cell adhesion forces were 38.4 nN for the smooth specimen, 140.8 nN for C3, 124.2 nN for C4, and 67.1 nN for C5. The results indicate that the cell adhesion force is influenced by cell shape and the Ca-P grooved patterns affect the cell shape and cytoskeletal structure, thus influence cell proliferation and cell adhesion force. The cytodetachment test with nanonewton resolution is a sensitive method for studying cell–biomaterial interaction.