In vitro microdistraction of preosteoblasts: distraction promotes proliferation and oscillation promotes differentiation.

Osteoblast biology is influenced in vivo by a 3-dimensional (3D) extracellular matrix that mediates their adhesion and interaction and by a constant state of compressive and tensile forces. To study the role of mechanical stress on osteoblasts in vitro, these parameters must be addressed. Therefore, this study describes the use of a novel, in vitro system that subjects cells to distractive and compressive forces in a 3D environment. This system, termed a microdistractor system, was used to apply linear forces to 3D collagen type I gels containing preosteoblasts. Gels were induced for up to 16 days in osteogenic medium and subjected to either constant linear distraction (distraction gels) or to repeating cycles of distraction and compression (oscillation gels). The effect of these stresses was evaluated over time by measuring proliferation rates, protein synthesis (i.e., cellular activity), and osteogenic differentiation levels. While linear forces in general appeared to increase protein synthesis, force-specific effects on proliferation and differentiation were observed. Specifically, distraction forces appeared to enhance MC3T3 proliferation while distraction/compressive forces appeared to accelerate their osteogenic differentiation program. Therefore, these results suggest that the microdistraction system may be an appropriate in vitro system for the study of mechanobiology in osteoblast phenotype.

Favorable morphologic change of preosteoblasts in a three-dimensional matrix with in vitro microdistraction.

BACKGROUND: Distraction osteogenesis has been used to correct hypoplastic and asymmetric bony deformities in the growing patient, yet its underlying cellular mechanisms are poorly understood. Using a new in vitro model, the microdistractor, morphologic properties of preosteoblasts under mechanical strain were studied. METHODS: Mouse calvarial MC3T3 cells were suspended in a polymerized three-dimensional collagen gel and stressed for 14 days as one of three groups (n = 30): (1) distraction (0.5 mm/day); (2) oscillation (1 mm/day for 2 days alternated with 1 mm/day for 2 days); and (3) control (no force). A computer modeling system, KS-300, was used to record cell shape (aspect ratio) and orientation (deviance from axis of stress). RESULTS: In part I of the study, morphologic cellular changes were found to be even throughout different regions of the gel (central versus peripheral, versus different vertical layers), suggesting the force was evenly applied to all cells in the gel. In addition, when linear distraction forces were applied, morphologic change occurred over time, suggesting a morphologic response to the applied stress. In part II of the study, with different forces applied, morphologic changes occurred over time such that linear distraction forces caused cells to elongate and align in a parallel direction to the force, whereas oscillation caused cells to switch from parallel (with distraction) to perpendicular (with compression) orientation relative to the force applied. CONCLUSION: The authors' data suggest that the microdistractor device is an effective in vitro model for studying the cellular response to distraction stresses. It may be used in future studies to optimize clinical methods of distraction.