Application of a static magnetic field as a complementary aid to healing in an in vitro wound model

Abstract

Static magnetic field (SMF) has long been used as a therapeutic means, though its effects on the activity of cells and the mechanism(s) involved remain unknown. The purpose of this study is to determine the effect of a moderate-intensity SMF on the activity, growth and migration of mouse embryonic fibroblast (NIH 3T3), aiming to mimic wound healing and to study it in real time. A cell-free area (a scratch with a 200-500µm width) was formed in NIH 3T3 cultured cells and used as a wound model. The effects of a SMF (10, 50, 80 and 100mT) on the survival rate (MTT assay), integrity of cell membranes (lactate dehydrogenase (LDH) assay), the morphology of the cell (circularity, number and length of filopodia), cell orientation, and migration (speed, direction, rate) were studied as a function of the incubation time in a time-lapse manner. The exposure of cells to SMF at all intensities had no cytotoxic effect, as revealed by the MTT assay. The integrity of the membranes of the SMF-treated cells studied by the LDH assay test showed no effects. The structure of the membrane at the leading edge of the cells changed and showed several filopodia extended parallel to the field direction. The exposure to the SMF elongated the cells and decreased their circularity at SMF 10mT. The migration of the cells from one edge of the gap towards the other was affected by the applied SMF. The maximum and minimum effects were monitored at 80mT and 10mT, respectively. Analysis of cell migration revealed an average directness of 0.73, 0.66, 0.78, 0.78 and 0.69 under SMF 10, 50, 80, 100mT and control, respectively. The morphological and functional changes of the cells in the presence of SMF revealed particular effects on the membrane and cytoskeleton. Cells were affected by physicochemical changes caused by the applied SMF, though the extent of the incurred effects was not a linear function of the field intensity. This low cost, non-invasive approach can be used as a magneto-manipulative means to tailor a practical, independent or complementary means of manipulating the activities of cells and tissues for clinical purposes.