Development of an in vitro model on cellular adhesion on granular natural bone mineral under dynamic seeding...

Adhesion of osteogenic cells on biomaterials can be studied with static in vitro models, whereas models representing dynamic seeding conditions are rare. Herein, we present an in vitro model to study cell adhesion on granular biomaterials under dynamic seeding conditions. Radiolabeled osteogenic MC3T3-E1 cells were allowed to adhere to granules of natural bovine bone mineral (NBM) under constant rotation. Adhesion of MC3T3-E1 cells was determined by liquid scintillation counting, and cell morphology was visualized by scanning electron microscopy. Cell viability was determined by MTT assay under static and dynamic conditions, at room and body temperature, and in the presence or absence of serum. We show here that MC3T3-E1 cells rapidly adhere to NBM, reaching a peak 3 h after seeding. Attached cells display characteristic signs of spreading. Five to ten percent of total radioactivity remained on NBM after the removal of nonadherent cells. Viability is maintained at room temperature and under rotation for upto 3 h. This data suggests that the dynamic in vitro model presented here provides a tool to study cell adhesion on granular biomaterials.