Three-dimensional cell morphometry for the quantification of cell-substrate interactions

The initial attachment of cells to biomaterials is an important indicator of longer term cell-substrate biocompatibility. To study and quantify this interaction, we have developed a protocol for measuring temporal changes in the three-dimensional (3D) morphology of mammalian cells seeded onto different substrates using fluorescence confocal laser scanning microscopy and image processing techniques. This method has been used to investigate how morphology parameters, such as cell thickness, volume, and the footprint area, change over time for osteosarcoma cells on uncoated glass control, fibronectin-coated glass, and titanium substrates. Consistent with other studies, our results show that the presence of a fibronectin coating significantly increases the rate of cell spreading, judged by an increase in the cell footprint area and a decrease in cell thickness, indicating enhanced biocompatibility. Using similar criteria, the same cell line was observed to spread faster on titanium than on uncoated glass. We propose that 3D cell morphometry is a valuable multiparametric tool for quantifying initial cell-substrate interactions providing data which has important applications in quality control for ensuring product/batch consistency and for developing tailored surface finishes.