The effects of short-term stimulation on fibroblast spreading in an in vitro 3D system

Although the short-term effects of mechanical and biochemical stimulation on cell protein and DNA production have recently begun to be evaluated using 3D models, the effects that such stimulation have on cell morphology and adaptation remains unclear. Using a custom-built bioreactor, we were able to create a systematic model to examine the short-term effects of stimulation on cell morphology in a 3D model, specifically by evaluating cell spreading as the short-term indicator of cell adaptation. Fibroblasts were seeded on a porous poly(L-lactic acid) scaffold and cultured in a computerized bioreactor for 24 h under various uniaxial strains (0, 0.6, 3, 6%) and frequencies (0.0125, 0.125 Hz). Also, the effects of transforming growth factor (TGF-beta1) (1, 10, 100 ng/mL) were examined on static, nonstimulated cells-scaffold constructs after 24 h. Fibroblasts that had been subjected to mechanical stretching were found to exhibit significantly more spreading than the static control group. Conversely, TGF-beta1 between 1 and 100 ng/mL did not produce any significant difference in fibroblast spreading from the control groups after 24 h. Collectively, the findings suggest that cell morphology and adaptation may be affected by short-term mechanical stimulation, as seen by increased cell spreading by the fibroblasts under these experimental conditions.