The study of cells-substrate interaction became a stringent subject in the past decades, since an increasing
variety of new materials and methods have been involved in tissue engineering or implants techniques. The investigation
of this interaction using optical methods is a challenge, especially since these substrates are not optically polished. Due
to their roughness in the micrometric or submicrometric range, the polymeric substrates offers good conditions for cells
adhesion, but the characterization of cells properties can be hindered. In this study, we use Polypyrrole thin films, acting
as substrates for cultured osteoblast-like MG63 cells having applications in tissue engineering for in vivo-like scaffolds.
As characterization technique, we chose digital holographic microscopy, a single-shot technique, to obtain quantitative
information about the sample features in a plane perpendicular to the substrate. Different parameters were tested in the
experimental setup with the aim of finding the optimal conditions for details visualization. The reconstructed 3D images
were filtered using a combination of analytical and implicit functions from MATLAB to exclude small/large objects,
which correspond to Polypyrrole droplets to clearly identify the cells contour for quantitative measurements regarding
their dimensions. These data were correlated with the effects on osteoblasts viability and differentiation. Also, the
thickness and the refractive index of the substrate were determined using the decoupling procedure.