Objectives: to find a morphological and molecular linkage between outside mechanical force and inside biological signals by examining the effect of force on FAC characteristics.
Methods: human PDL fibroblasts isolated from healthy first premolar teeth. In order to apply force in-vitro, cells were seeded in a special transparent elastic flask "OptiCell". Pressure force with different durations and loads applied on the OptiCell in the incubator. To determine the viability of the cells we used Trypan Blue test and "LIVE/DEAD Cell Vitality Assay Kit" FACS test. To define cell shape, size and FAC characteristics we performed immunohistochemistry and light microscopy directly from the Opticell after application of pressure force.
Results: our results show that cell vitality after application of 100 or 200gr/cm2 for two days showed 95% and 80% respectively. We therefore used 100gr/cm2. Immunohistochemical stain of actin and vinculin showed less organized, unshaped cell under pressure. More podia per cell under pressure by fold of 1.5 and 2.15 for one or two days respectively. Larger cell area by fold of 1.5 in both times.
Additionally, cells under pressure have more mature FA vinculin.
Conclusions: cells under pressure tend to be more unshaped, have more podia, tend to be spread on larger area and have larger amount of vinculin protein.