脉冲电磁场对新型低弹多孔钛合金表面成骨效应的影响研究

目的:观察脉冲电磁场刺激对新型低弹多孔钛合金支架表面的成骨效应。方法:使用3D打印技术制备多孔钛合金(Ti-24Nb-4Zr-8Sn,Ti2448)支架,并使用扫描电镜对支架的表面形貌进行观察;将成骨细胞接种于支架表面,给予脉冲电磁场刺激的支架为实验(PMEF)组,不给予脉冲电磁场刺激的支架为对照组;使用CCK-8(Cell Counting Kit-8)试剂盒检测两组细胞的增殖能力;使用活/死细胞染色观察两组细胞在支架上的活性;使用扫描电镜观察两组细胞在支架上的形态;通过实时定量PCR观察两组细胞相关成骨基因分化;通过兔股骨外侧髁缺损模型进行体内骨长入分析,分别使用荧光标记和Van Gieson染色观察两组支架的骨长入情况。结果:使用3D打印技术成功制备实验所需的多孔钛合金支架;PMEF组成骨细胞的增殖能力、细胞活性明显高于对照组,差异有统计学意义(P<0.05);PMEF组成骨细胞可见明显细胞伪足,细胞状态良好;PMEF组成骨细胞的成骨相关基因表达明显高于对照组(P<0.05);术后4周和12周,PMEF组新生骨沉积速率(两种荧光间距)均高于对照组(P<0.05),Van Gieson染色发现PMEF组骨长入明显多于对照组(P<0.05)。结论:脉冲电磁场刺激和多孔Ti2448植入物的组合可为骨科、口腔和整形等领域的骨修复重建提供一种新方法和新思路。

Pulsed Electromagnetic Fields Enhance Biological Effects of Osteogenesis on Porous Titanium Alloy Surface

Objective Observe the osteogenic effect of PEMF stimulation on the surface of the new low-elasticity porous titanium alloy scaffold.Methods The new low-elasticity porous titanium alloy(Ti-24Nb-4Zr-8Sn,Ti2448)scaffolds were prepared by 3D printing technology.The surface morphology of the scaffolds were characterized by scanning electron microscopy(SEM).The osteoblast-like cell line MC3T3-E1 was cultured in the absence(control)or presence of PEMF stimulation on porous Ti2448 disc surface,and the adhesion and proliferation of the cells were investigated by SEM,live/dead cell imaging kit and CCK-8 assay.Furthermore,the expression of osteogenesis-related genes was also examined by quantitative real-time PCR.The porous Ti2448 scaffolds for animal experiment were implanted into the lateral femoral epicondyle of female New Zealand white rabbits.All of the 24 rabbits were then randomly divided equally into two groups,one group with PEMF stimulation for 2 h everyday and the other with no stimulation as control.After 4 weeks and 12 weeks,fluorescent markers and Van Gieson staining were used to observe the bone ingrowth of the two groups of scaffolds.Results The porous titanium alloy scaffolds required for the experiment were successfully prepared.The proliferation ability and cell activity of MC3T3-E1 in PMEF group were significantly higher than those in control group(P<0.05).The cells in the PMEF group showed obvious cell pseudopodia,the osteoblast-related genes expression of cells in PMEF group were significantly higher than that in control group(P<0.05).At 4 and 12 weeks after surgery,the rate of new bone deposition(two types of fluorescence spacing)in the PMEF group was higher than that in the control group(P<0.05).Van Gieson staining showed that the bone growth in the PMEF group was significantly more than that in the control group(P<0.05).Conclusion The combination of PMEF and porous Ti2448 implant can provide a new idea for bone repair and reconstruction in the fields of orthopedics,oral cavity and plastic surgery.