The Preliminary Study Of Nanosecond Pulsed Electric Fields Stimulation On Osteoblastic Cells In Vitro

Various Oral diseases need bone remodeling in clinical treatment. Based on common clinical conditions, bone remodeling is a long process.So,we need new technology to improve the present situation.Using electrical stimulation to treat bone disease have a history more than 50 years. Nanosecond pulsed electric fields is a high field and short pulse width pulses,which have special chief bioeffect.Hence ,researching the transduction of biological effect of nsPEFs stimulate on osteoblasts is very necessary.At present, there have some reports about nsPEFs exist"window effect"。However,the variation of nsPEFs parameter t is unclear.Methods: osteoblast-like cells were cultured in F-12 with 10% FBS,then nsPEFs stimulate on osteoblast-like cells to research the variation of"window effect"with Orthogonal Design. Subsequently selected fitting combination to research the biological effect of nsPEFs stimulate on osteoblasts osteoblast-like cells.We want to use MTT assay to r easerch transduction of cell proliferation detection , use a flow cytometer to r easerch transduction ofFAS antigens. using AO/EB immunofluorescence assay to observe morphologica changes.Based on these findings,our results are as follows:1"window effect"is exist which the variation of nsPEFs parameter is 10-15ns ,5-15kv/cm. The chief biological effect is apoptosis2 Fitting combination of nsPEFs parameter is 50ns , 5kv ,50Hz ,through Orthogonal experiment . Analysis the results,we find the relation of nsPEFs parameters : pulse width> field strength > frequency.3 nsPEFs stimulation can inhibiting osteoblast-like cells proliferation,which are time dependent.After 4 hours ,we find osteoblast-like cells recovery. Compare control group and experiment group,this difference can be proved by statistics(p<0.05).4 nsPEFs stimulation can strengthenthe number of FAS antigens which have time dependent. After 24 hours, the number of FAS antigensstart to drop off . Compare control group and experiment group ,this difference can be proved by statistics(p<0.05).From 4h to24h, the number of FAS is stability, which can't be proved by statistics.5 Compared with the cells are normal in appearance, osteoblast-like cells membrane by nsPEFs stimulation is integrity .but the nucleus is unlear. It prove that osteoblast-like cells is apoptosis.