The Combined Effects Of Extracorporeal Shockwave And RhIGF-Ⅰon Promoting Proliferation And Expression Of BMP-2 Of MG63 Cells

Qbjective To demonstrate the combined effects of extracorporeal shockwave (ESW)and recombinant human insulin-like growth factor-1(rhIGF-Ⅰ) on proliferation and expression of bone morphogenetic protein-2(BMP-2) of MG63 cells.Mthods Human osteoblastlike cell lines (MG63) were cultivated in vitro and resuspended in medium at a concentration of 1×106 cells/mL in 1.5mL screw-cap cryotubes . The prepared cell suspension model is divided into A: ESW group, B: IGF-Ⅰgroup, C: control group, D: ESW combined rhIGF-Ⅰgroup, B, D Group added cytokines rhIGF-Ⅰ,with the concentration of 100ng/ml. A, D group were accepteded 5KV, 1000 frequency of shock wave interference, the control group was not handled. Cell survival rate was assessed by trypan blue after the shock wave interference. Morphology of the cell membrane changes after the shock waves was observed by Scanning electron microscopy(SEM).Cell proliferation was measured by methylthiazol tetrazolium (MTT) method till 4 hours,24hours,48hours after the intervention. Collected 24 hours, 48 hours cell culture supernatant after the ESW treatment, the expression of BMP-2 was measured by using Human BMP-2 ELISA kit.Results MG63 cells treated with ESW (5KV,1000times)showed market effects of promoting proliferation (p<0.01) and increasing the expression of BMP-2 (p<0.05) when compared with contral group after ESW application 24hours. After the shock wave interference, porous membrane structure changes occurred on MG63 cells which leads to membrane permeability, but did not change the cytoskeleton structure.MTT results showed that: by the rhIGF-Ⅰconcentration of 100ng/ml, the proliferation of MG63 cells in the maximum activity.100ng/ml rhIGF-Ⅰwas also able to promote proliferation (p<0.01),increasing the expression of BMP-2(p<0.05).The results of MTT and the expression of BMP-2 were similar ,when mearly used ESW or rhIGF-1(100ng/ml),no significant difference(p>0.05).ESW plus rhIGF-Ⅰshowed stronger effects on increasing the expression of BMP-2 and last 48hours later ,however no significant changes in promoting proliferation between using a certain energy of ESW or a certain dosage of rhIGF-1and ESW plus rhIGF-Ⅰ(p=0.16).Conclusion The application of ESW increased membrane permeability, cytoskeletal structure does not change.Both ESW (5kv , 2000times) and rhIGF-Ⅰ(100ng/ml) can promote MG63 cell proliferation and expression of BMP-2,ESWand rhIGF-1 have synergistical effects on promoting cell expression of BMP-2 but no significant effects on promoting cell proliferation.