[Effects of laser solid forming of porous titanium on proliferation of osteoblast and RANKL/OPG expression].

To evaluate the effect of laser solid forming (LSF) of porous titanium on receptor activator of NF-κB ligand (RANKL)/osteoprorotegerin (OPG) expression and osteoblast cells growth. Methods: The DMEM and sterile saline were used for porous titanium extract. The osteoblast cells were cultured in the extract while equal amount of DMEM and sterile saline were added to the control group. The growth of the cells were observed under an inverted phase contrast microscope. MTT was used to detect the growth inhibitory rates. The adhesion capacity of osteoblasts were measured. The growth in the material surface was examined by the electron microscope, and the expressions of RANKL and OPG were determined by Westen blot. Results: At the first day, the osteoblast proliferation rate was significantly different (P<0.05), at the fourth and seventh day, the osteoblast proliferation rate was not significantly affected in the LSF group (P>0.05); at each time point, the osteoblast proliferation rate were significantly different between the two groups (P<0.05). Compared with the control group, RANKL and OPG protein expression were not significantly different (P>0.05). The laser solid forming of porous titanium showed well bone compatibility. Conclusion: The porous titanium did not affect osteoblast proliferation due to its well bone compatibility. It did not affect the OPG/RANKL/RANK-axis system of bone metabolism, exibiting a wide applicable prospect for tissue engineering.目的：评价激光立体成形技术制备的多孔钛材料对成骨细胞增殖率、骨相容性、成骨细胞核因子κB受体活化因子配体(receptor activator of NF-κB ligand，RANKL)及护骨素(osteoprorotegerin，OPG)表达的影响。方法：以DMEM培养液和无菌生理盐水作为浸提介质，制取激光立体成形多孔钛浸提液，并在加有此浸提液的培养基中培养成骨细胞，对照组加入等量DMEM培养液和无菌生理盐水。MTT法检测细胞增殖抑制情况；电子显微镜下观察成骨细胞在材料表面的生长状况；Western印迹检测RANKL和OPG蛋白的表达。结果：激光立体成形多孔钛组培养1，4， 7 d后，成骨细胞能够很好地附着、生长和分化。激光立体成形多孔钛组和对照组成骨细胞增殖率比较，1 d时差异有统计学意义(P<0.05)，4和7 d时的差异无统计学意义(P>0.05)。两组组内各时间点相比较，成骨细胞增殖率差异有统计学意义(P<0.05)。Western印迹表明，激光立体成形多孔钛组第1，4，7天RANKL蛋白和OPG蛋白表达水平，与对照组比较，差异均无统计学意义(P>0.05)。电子显微镜下观察超微结构，可见成骨细胞在激光立体成形多孔钛表面附着、铺展、增殖良好。结论：激光立体成形多孔钛材料无成骨细胞增殖抑制作用，具有良好的骨相容性，不影响调节骨代谢的OPG/RANKL/RANK轴系统，在医学组织工程领域存在着广阔的应用前景。.