Basic Research Of Shock Wave Assisted With Methotrexate In The Treatment Of Osteosarcoma

Low doses extracorporeal shock wave can affect and increase the permeability ofthe cell membrane.Besides it can induce substances which cannot easily pass the cellmembrane into cells more easily.In vitro experiments showed that extracorporeal shockwave synergistically can force the chemotherapy drugs into the cells and induceapoptosis.This suggests that extracorporeal shock wave can be used to assistchemotherapy drugs in the treatment of cancer.So we assume that extracorporeal shock wave can increase the permeability of cellmembrane and promote chemotherapy drugs to enter into the targeted cells,so we canreduce the amount of chemotherapy drugs, while enhancing the treatment of theosteosarcoma.Thus this study, by showing how to increase the sensitivity ofosteosarcoma cells to chemotherapy drugs, reducing dosage of chemotherapy drug,enhancing tumor targeting therapy, providing a new means and theoretical basis.Objective This paper studies the mechanism of low doses extracorporeal shockwave cooperate with methotrexate in the treatment of osteosarcoma, and whetherlow-dose extracorporeal shock wave can increase the permeability of cellmembranes,so that it can induce the chemotherapy drug-methotrexate which is noteasily pass through the cell membrane to enter into the cells and improve the treatmentof osteosarcoma.Experimental methods This study uses different frequencies，the0.3F capacitor,working voltage of7KV and14KV LDESW to deal with the U2OS cells;uses the trypanblue staining to detect the effect of LDESW on the cell activity;detects thecritical frequency of7KV and14KV LDESW which have no significant effect on U2OScell mortality (P<5%) and increase the permeability of cell membrane. In order to studywhether the shock wave can increase the permeability of osteosarcoma cell membrane,we choose two kind of fluorescent substance (calcein) and (Lucifer Yellow, LY) which can not penetrate the cell membrane and dissolved in the extracellular fluid, andcompare the amount of fluorescent substances in cells between experimental group andthe control group after the cells are processed by low dose shock wave. Finally,in orderto detect whether the extracorporeal shock wave can enhance MTX to enter into thecells, we add the same concentration of MTX into the experimental group and controlgroup, after shock wave is used to deal with the cells,we use the methotrexate kit todetect and compare the concentration of MTX in the cells between experimental groupand the control group.Results This study showed that: capacitance0.3μF, operating voltage7KV ofLDESW less than400times, capacitance0.3μF, operating voltage14KV of LDESWless than150times has no effect on cell viability after the cells are processed by thistwo kinds of LDESW.After the qualitative analysis with the fluorescence microscopyand quantitative analysis with the Fluoroskan Ascent FL, the study finds that theexperimental group has much more quantity of fluorescent substance than the controlgroup after the osteosarcoma cells are processed by LDESW,and the MTX detected bymethotrexate kit in experimental group cells is significantly much more than the controlgroup after the experimental group is processed by LDESW.Conclusion This study demonstrated that LDESW which has no effect on cellviability can increases the permeability of the U2OS osteosarcoma cell membrane,induce much more MTX entering into the cell membrane and assist in the treatment ofosteosarcoma.