Biophysical Manipulation of Olfactory Ensheathing Cells by Static Magnetic Field

Background and Aim: Olfactory ensheathing cells (OEC) have beensuccessfully used in transplantation experiments, however, theirbiophysical properties have remained unknown. Their motility andmigratory properties root into their cytoskeleton activities that lead tofilopodia formation, elongation, and orientation and so on, playingimportant role in regenerative medicine. In this study, OEC were isolatedfrom mouse olfactory bulbs and exposed to 10mT SMF. The motilitycharacteristics (velocity and directness) in the presence and absence ofSMF were analyzed to address possible application of the approach inthe treatment of spinal cord injury (SCI).Methods: The olfactory bulbs were isolated and digested using collagenasetype I and trypsin 0.1%. The sediment was cultured in DMEM: F12 15%FBS and 2 mM L-Glutamine for 18 hours, followed by 48-hour culturein the replaced supernatant, and eventual culture in new poly L-lysinetreated plates for 96 hours. Characterization of cells and induction meansof sphere formation was carried out in the presence of BFGF and EGFbut the absence of FBS, and analyzed by immunocytochemistry (ICC)technique using P75 and GFAP antibodies. Then, the positive cells werecultured and their migration was studied by time-lapse experimentsrecording their activities and morphology at 5-minute intervals in thepresence and absence of 10mT SMF for 48 hours. The characteristicsof the membrane and cytosolic organelles recorded in high-resolutionphotos were assessed by ImageJ and Gradientech software and thebiophysical effects of SMF were investigated at cellular and molecularlevels.Results: About 95% of cells attached to the poly L-lysine coated plateswere found to be OEC which used for further characterizations. TheICC positive cells were used for scratch assay test and their migratoryproperties were studied. The velocity of the cells in the presence andabsence of the 10 mT SMF was about 10.48 μm/hour and 6.97 μm/h,respectively. The directness of the cell in the presence and absence of the10 mT SMF with respect to the N pole was 0.76 and 0.65, respectively.The exposure of the OEC to SMF increased the rate of velocity andorientated the directness of the cells towards the N pole of the magnet incomparison to control group which showed less velocity and a randomdirectness. Furthermore, the microtubule arrangement in the presence ofSMF showed a long stretch in comparison to control group.Conclusion: Along with all the efforts of science, the lack of a biophysicalperspective for stem cell therapy is palpable. The promising resultsobtained here prove the potential effect of the magnetic field andrepresent it as a biophysical modality to manipulate stem cells withpossible application towards the treatment of spinal cord injury andapplication in the regenerative medicine in the near future. This is whyfurther studies should be conducted to investigate the matter at variousintensities of SMF in order to obtain comprehensive information tofabricate an optimum non-invasive treatment regime.

کلیدواژه ها:

Static Magnetic Field ، Olfactory ensheathing cells ، Biophysical treatment