电子束熔融制造医用Ti6Al4V在人工模拟体液中耐腐蚀行为

目的 研究电子束熔融技术（EBM）3D打印制备的医用Ti6Al4V不同打印截面在Hank''s模拟人工体液中的电化学腐蚀行为。方法 采用开路电位和动电位极化曲线方法研究了Ti6Al4V不同打印截面,分别记为EBM-XOY面（垂直于打印方向的钛合金截面）和EBM-YOZ面（平行于打印方向的钛合金截面）,在Hank''s模拟人工体液中的电化学腐蚀行为,利用SEM、XRD和金相显微镜分析了其表面形貌和物相组成及其腐蚀机理,并与传统医用锻造Ti6Al4V进行了对比研究。结果 与传统医用锻造Ti6Al4V合金相比,EBM-YOZ面钛合金在Hank’s模拟人工体液中的开路电位、腐蚀电位和腐蚀电流与之相当,显示出与之接近的耐腐蚀性能;EBM-XOY面钛合金的耐腐蚀性能较EBM-XOZ面和传统锻造钛合金稍差,主要原因是EBM-XOY面的α+β相界面积大,α相含量较高,且被优先溶解,因此,耐腐蚀性能较差。结论 电子束熔融制造的Ti6Al4V不同截面的耐腐蚀性能不同,EBM-YOZ面的耐腐蚀性能与传统医用锻造钛合金相当,EBM-XOY面的耐腐蚀性能稍差。     

新型β钛合金Ti_(35)Nb_3Zr_2Ta在人工关节假体应用中的生物相容性

背景:目前被广泛应用于人体关节置换的钛合金为Ti6Al4V,但其弹体模量高于人体骨,导致人工关节假体的稳定性差。而新型β钛合金Ti35Nb3Zr2Ta弹性模量较低,或许能成为新一代生物相容性较好的人体关节假体材料。目的:探讨新型β钛合金Ti35Nb3Zr2Ta在人工关节假体应用的生物相容性。方法:应用计算机检索自万方数据库、中国知网和Pub Med文献数据库,检索时间范围2010至2015年,以"新型β钛合金;人工关节假体;生物相容性"为检索词,检索医用人工关节假体材料的应用现状及新型β钛合金Ti35Nb3Zr2Ta在人工关节假体应用的生物相容性的研究。结果与结论:与Ti6Al4V相比,Ti35Nb3Zr2Ta表面粗糙度较高,表面接触角较小,碱性磷酸酶活性以及成骨细胞的钙沉积量明显高于Ti6Al4V,具有良好的生物相容性,可以考虑在人工关节假体领域中进一步广泛应用。     

骨髓间充质干细胞经不同时间成骨细胞诱导后细胞性质比较

目的 探讨骨髓间充质干细胞经成骨细胞诱导不同时间后的细胞性质。方法 成人骨髓分离单个核细胞（MNC）,体外培养获得间充质干细胞（MSC）,传至第4代后加入地塞米松（DEX）、抗坏血酸和β-甘油磷酸,诱导成骨分化。诱导7d和14d后的细胞与正常成人成骨细胞分别加入不同浓度骨形态发生蛋白-2（BMP-2）和DEX,48h后检测细胞增殖、碱性磷酸酶（ALP）活性和骨钙素（OCN）的含量。结果 BMP-2和DEX对诱导7d后的细胞有明显的刺激增殖和ALP活性的作用;对诱导14d后的细胞,BMP-2和DEX都不能刺激细胞增殖和ALP活性,但OCN含量明显增加。正常成人成骨细胞对BMP-2和DEX作用的反应与诱导14d的成骨样细胞基本一致,但其OCN绝对含量明显高于MSC来源的成骨细胞。结论 骨髓MSC向成骨诱导7d和14d后细胞分化程度不同。诱导时间在MSC的成骨分化中是一个非常关键的因素,但其机制和最佳诱导时间尚需进一步研究。诱导14d后的MSC的成骨活性低于正常成人成骨细胞。     

图像分析方法研究DLC膜/Ti6Al4V梯度材料的血液相容性稳定性

对 DL C膜 / Ti6Al4V梯度材料和 Ti6Al4V钛合金进行了血液相容性稳定性研究。用数字图像分析方法分别测定了材料生物摩擦磨损实验前后的血小板消耗率。研究表明 ,在 Hank' s溶液润滑条件下 ,经过 2 0 0 0 m摩擦磨损实验后 ,Ti6Al4V钛合金血小板消耗率显著增加约 5 0 % ,而 DL C膜 / Ti6Al4V梯度材料血小板消耗率没有明显增加。说明 DL C膜 / Ti6Al4V梯度材料在使用条件下具有很好的血液相容性稳定性     

人骨髓间充质干细胞复合人脐静脉内皮细胞构建预血管化成骨细胞膜片

背景:如何使工程化骨组织移植后能够成活并发挥功能,前提条件是必须含有充足有效的血液供应,而这也正是工程化组织应用于临床的主要障碍之一。目的:探索新的体外构建血管网络的方法以解决工程化骨组织预血管化的问题。方法:人骨髓间充质干细胞以9×10^4/cm^2高密度接种,经成骨诱导连续培养14 d形成细胞膜片,镜下观察细胞形貌,并进行碱性磷酸酶染色和茜素红染色。然后将培养的人脐静脉内皮细胞以5×10^4/cm^2密度接种到上述细胞膜片上,继续培养至7 d。通过镜下观察、CD31免疫荧光染色、Ⅰ型胶原免疫荧光染色评估血管网形成情况及膜片特性。结果与结论:①经连续培养可获得完整的成骨诱导细胞膜片,茜素红染色和碱性磷酸酶染色呈阳性;②倒置相差显微镜观察发现人脐静脉内皮细胞接种在成骨诱导细胞膜片上逐渐重排,CD31染色显示进行性管腔形成;③接种人脐静脉内皮细胞后构建的预血管化成骨诱导细胞膜片Ⅰ型胶原染色阳性;④结果表明,经成骨诱导后可形成具有成骨特性的细胞膜片,将人脐静脉内皮细胞培养在该膜片上可体外形成预血管化成骨细胞膜片,为优化构建预血管化的骨组织提供了新的理论指导。     

缺氧状态下体外培养成骨细胞血管内皮生长因子和骨形态发生蛋白2的表达及意义

目的研究缺氧对体外培养成骨细胞血管内皮生长因子（VEGF）和骨形态发生蛋白2（BMP-2）表达的影响及意义。方法采用酶消化法获取人成骨细胞并进行原代培养，倒置显微镜下观察成骨细胞的生长形态；采用Gomori改良钙钴法和免疫组织化学SABC法分别检测原代培养成骨细胞碱性磷酸酶（ALP）和骨钙素（OCN）的表达，进行原代培养成骨细胞鉴定。细胞传至第2代时，按继续培养条件的不同将细胞分为缺氧组（氧分压〈4．8kPa、氧容积比〈5％）和常氧组，在传代培养24、48、72h时分别收集2组细胞，采用免疫组织化学SABC法检测VEGF和BMP-2的表达，应用图像分析系统进行半定量检测，结果以平均灰度值表示，平均灰度值低示表达水平高。结果原代培养成骨细胞可表达ALP和OCN，初步确定经体外培养获得了具有生物活性的成骨细胞。在培养24、48、72h时，缺氧组成骨细胞VEGF表达水平均明显高于常氧组（平均灰度值分别为123．53±7．38 vs 141．21±6．03、116．45±6．34 vs 138．37±5．04、108．11±5．37 vs 136．65±6．54，均P〈0．01），且缺氧组成骨细胞VEGF表达水平随缺氧时间延长而逐渐升高。缺氧组成骨细胞BMP-2表达水平在培养24h时明显高于常氧组（平均灰度值为143．28±2．82 vs 146．91±2．06，P〈0．01），而在培养48、72h时与常氧组比较，差异均无统计学意义。结论缺氧可诱导成骨细胞VEGF的表达上调；而缺氧延长则不利于成骨细胞BMP-2的表达。缺氧对VEGF和BMP的早期表达调控可能与骨修复的启动相关。     

益骨胶囊含药血清对大鼠成骨细胞IL-6 mRNA及其蛋白表达的影响

目的：从mRNA及蛋白水平探讨益骨胶囊含药血清对成骨细胞表达IL-6的影响。方法：分离、培养新生SD大鼠成骨细胞，传代后分为3组即含药血清组；空白血清组；DMEM组。采用RT-PCR法检测IL-6 mRNA相对表达量，用放射免疫法检测成骨细胞培养上清中的IL-6含量。结果：益骨胶囊含药血清组IL-6 mRNA相对表达量显著低于对照组（P<0.05）；益骨胶囊含药血清组IL-6蛋白表达量也低于对照组（P<0.05）。结论：益骨胶囊含药血清能下调成骨细胞IL-6 mRNA 表达；亦能在蛋白水平降低成骨细胞分泌骨吸收因子IL-6，这可能是益骨胶囊防治骨质疏松症的机制之一。     

过表达BMP-4基因的BMSCs复合生物陶瓷骨在同种异体骨缺损模型中的研究

目的 研究骨形态发生蛋白BMP-4转染骨髓间充质干细胞后与生物陶瓷骨形成复合材料对兔桡骨缺损的修复作用和机制。方法 离体分离培养鉴定兔BMSCs,构建过表达BMP-4基因的慢病毒载体并转染BMSCs细胞,并通过流式细胞术和Western blot鉴定;制备复合生物陶瓷骨,ALP试剂盒检测桡骨组织ALP活性;HE染色观察桡骨组织形态变化;依据Lane-Sandhu法评价受体兔挠骨改善情况;采用免疫荧光检测桡骨组织Col I、OC和OPN表达;Western blot检测桡骨骨膜VEGF和EGF水平。结果 成功分离兔BMSCs细胞,并获得BMP-4基因过表达的BMSCs细胞。BMP-4过表达的BMSCs复合生物陶瓷骨能够显著修复缺损的兔桡骨组织形态,提高组织的Lane-Sandhu评分,并上调挠骨的I型胶原、骨钙素和骨桥素的表达水平和骨膜中血管内皮生长因子（VEGF）和表皮生长因子（EGF）的表达。结论 BMP-4转染BMSCs与生物陶瓷骨复合材料能显著修复兔骨缺损,并增加骨膜组织中VEGF和EGF的表达。     

Ti－6A1—4V医用钛合金表面载银涂层的制备和抗菌性能研究

目的通过两步微弧氧化法在医用Ti－6Al－4V表面制备载银多孔涂层．研究其抗菌性能。方法选用医用Ti－6A1—4V,在其表面先采用250V低电压微弧氧化法制备TiO：预涂层,然后将样品浸入到浓度1mol／L的AgNO,溶液浸泡2h．再经紫外光照射3．5h,以实现银颗粒在预涂层上的固定．最后在含银（浓度O．2mol/L）电解液中经过350V正向电压的二次微弧氧化处理制备出载银多孔涂层。利用扫描电子显微镜和能量分散谱仪对涂层表面形貌、载银量进行了表征,并通过金黄色葡萄球菌检验了涂层的抗菌性能。结果Ti-6Al-4V表面预涂层呈多孔状,孔洞呈火山口形,平均孔径0．71μm,而载银涂层表面平均孔径约为1．57μm。银颗粒分布在微孔表面和孔洞内壁,涂层平均银含量约为8％。抗菌试验结果显示．当载银涂层与细菌的接触1d后,对金黄色葡萄球菌的杀菌率均达到96．94％．7d后抑菌率仍达到71-27％。结论通过两步微弧氧化法可在医用Ti-6A1—4V钛合金表面制备出具有良好抗菌活性的载银涂层。     

钛合金Ti6Al4V表面钛酸锶生物薄膜的水热合成

采用氢氧化钠和硝酸锶混合溶液对Ti6Al4V合金进行水热处理,温度180℃,时间1、3和6 h。X射线衍射、扫描电镜、电子能谱和X射线光电子能谱分析表明,水热处理后钛合金表面形成了钛酸锶的纳米颗粒薄膜,颗粒尺寸80～230 nm,薄膜中含有极少量铝和钒元素。在无钙Hank’s平衡盐液中的动电位极化和电化学阻抗实验表明,与抛光试样相比,水热处理3h试样的耐蚀性大幅提高。显微硬度压入实验表明,钛酸锶薄膜具有良好的附着性和内聚力。水热处理制备钛酸锶薄膜的方法可用于钛合金Ti6Al4V骨科植入体的表面改性。     

In vitro biological effects of titanium rough surface obtained by calcium phosphate grid blasting

Surface roughness modulates the osseointegration of orthopaedic and dental titanium implants. High surface roughness are currently obtained by blasting of titanium implants with silica or aluminium oxide abrasive particles. This process may cause the release of cytotoxic silicium or aluminium ions in the peri-implant tissue. To generate a biocompatible roughened titanium surface, we currently develop an innovative grid-blasting process using biphasic calcium phosphate (BCP) particles. Titanium alloy (Ti6Al4V) discs were either polished, BCP grid-blasted or left as-machined. BCP grid-blasting created an average surface roughness of 1.57 +/- 0.07 microm compared to the original machined surface of 0.58 +/- 0.05 microm. X-ray photoelectron spectroscopy indicated traces of calcium and phosphorus and relatively less aluminium on the BCP grid-blasted surface than on the initial titanium specimen. Scanning electronic microscopy observations and measurement of mitochondrial activity (MTS assay) showed that osteoblastic MC3T3-E1 cells were viable in contact with the BCP grid-blasted titanium surface. In addition, our results indicate that MC3T3-E1 cells expressed ALP activity and conserved their responsiveness to bone morphogenetic protein BMP-2. The overall results clearly indicate that this calcium phosphate grid-blasting technique increases the roughness of titanium implants and provides a non-cytotoxic surface with regard to mouse osteoblasts.     

Osteoblast response to plasma-spray porous Ti6Al4V coating on substrates of identical alloy

We have evaluated the in-vitro biocompatibility of Ti6Al4V alloy coated by plasma spraying with an identical alloy. These surfaces are widely used in cementless prosthetic components, although osteoblasts behavior on this treated alloy has not been evaluated to date. Cross sectional examination revealed a thick and rough coating of identical composition without sign of delamination. Within the coating, small discontinuities and nonconnected pores were observed. Osteoblast response was evaluated by assessing cell adhesion, proliferation, and differentiation of primary cultures of human osteoblastic cells. Compared to the polished alloy, osteoblast adhesion measured as cell attachment and actin network reorganization was delayed on the plasma-sprayed surface. Cell proliferation and viability were also impaired on the rough surface. Several informative markers of osteoblastic differentiation such as procollagen I peptide, alkaline phosphatase, osteocalcin, osteoprotegerin, and mineralized nodule formation were evaluated and indicated that the plasma-sprayed alloy favored a more differentiated phenotype than polished alloy. Taken together, our in vitro results indicate that successful osseointegration of plasma spraying of Ti6Al4V with an identical alloy is mediated by modulation of osteoblastic differentiation and mineralization.     

Thermal oxidation enhances early interactions between human osteoblasts and alumina blasted Ti6Al4V alloy

Oxidation of Ti6Al4V at 500 degrees C for 1 h in air results in the formation of an outer ceramic layer that improves osteoblast behavior and decreases Ti and Al ion release. In this work, alumina blasted Ti6Al4V alloy has been thermally treated and its in vitro biocompatibility has been assessed. Roughness of the blasted alloy was not found significantly altered after heat treatment while chemical surface analysis indicated an increase in stable TiO(2) and Al(2)O(3) oxides. Cell attachment, spreading, cytoskeleton organization as well as cell proliferation, viability, and procollagen I peptide secretion of human primary osteoblasts, impaired on alumina blasted Ti6Al4V, were found to be greatly enhanced on the thermally oxidized blasted alloy. Other informative markers of the osteoblastic phenotype such as alkaline phosphatase, osteocalcin, osteoprotegerin, and mineralized nodule formation were evaluated and indicated that osteoblasts responded at the same extent on untreated and thermally treated blasted alloys. Taken together, our in vitro results indicate that thermal oxidation of alumina blasted Ti6Al4V may favor successful osseointegration by promoting early interactions of osteoblastic cells and the modified surface alloy.     

A new austenitic stainless steel with negligible nickel content: an in vitro and in vivo comparative investigation

New nickel (Ni)-reduced stainless-steel metals have recently been developed to avoid sensitivity to Ni. In the present study, an austenitic Ni-reduced SSt named P558 (P558, B?hler, Milan, Italy) was studied in vitro on primary osteoblasts and in vivo after bone implantation in the sheep tibia, and was compared to ISO 5832-9 SSt (SSt) and Ti6Al4V. Cells were cultured directly on P558 and Ti6Al4V. Cells cultured on polystyrene were used as controls. Osteoblast proliferation, viability and synthetic activity were evaluated at 72 h by assaying WST1, alkaline phosphatase activity (ALP), nitric oxide, pro-collagen I (PICP), osteocalcin (OC), transforming growth factor-beta1 (TGFbeta-1) and interleukin-6 (IL-6) after 1.25(OH)2D3 stimulation. Under general anaesthesia, four sheep were submitted for bilateral tibial implantation of P558, SSt and Ti6Al4V rods. In vitro results demonstrated that the effect of P558 on osteoblast viability, PICP, TGF beta-1, tumor necrosis factor-alpha production did not significantly differ from that exerted by Ti6Al4V and controls. Furthermore, P558 enhanced osteoblast differentiation, as confirmed by ALP and OC levels, and reduced IL-6 production. At 26 weeks, the bone-to-implant contact was higher in P558 than in SSt (28%, p<0.005) and Ti6Al4V (4%, p<0.05), and was higher in Ti6Al4V than in SSt (22%, p<0.005). The tested materials did not affect bone microhardness in pre-existing host bone as evidenced by the measurements taken at 1000 microm from the bone-biomaterial interface (F=1.89, ns). At the bone-biomaterial interface the lowest HV value was found for SSt, whereas no differences in HV were observed between materials (F=1.55, ns). The current findings demonstrate P558 biocompatibility both in vitro and in vivo, and osteointegration processes are shown to be significantly improved by P558 as compared to the other materials tested.     

Synergistic effect of titanium alloy and collagen type I on cell adhesion, proliferation and differentiation of osteoblast-like cells

A number of studies have demonstrated the pivotal role of collagen in modulating cell growth and differentiation. In bone, where the extracellular matrix is composed of approximately 85% type I collagen, cellular interaction with matrix components has been shown to be important in the regulation of the osteoblast phenotype. Preservation or enhancement of normal osteoblast function and appositional bone formation after implant placement represents a strategy that can be useful for the purpose of improving osseointegration. In order to further improve biocompatibility, we combined two known favorable compounds, namely the titanium alloy, Ti6A14V, with type I collagen. We assessed the in vitro behavior of primary osteoblasts grown on both fibrillar collagen-coated and tropocollagen-coated Ti6A14V in comparison with uncoated titanium alloy, using an improved adsorption procedure. As parameters of biocompatibility, a variety of processes, including cell attachment, spreading, cytoskeletal organization, focal contact formation, proliferation and expression of a differentiated phenotype, were investigated. Our results demonstrated for the first time that in comparison to uncoated titanium alloy, collagen-coated alloy enhanced spreading and resulted in a more rapid formation of focal adhesions and their associated stress fibers. Growing on collagen-coated Ti6A14V, osteoblasts had a higher proliferative capacity and the intracellular expression of osteopontin was upregulated compared to uncoated titanium alloy. Type I collagen-coated titanium alloy exhibits favorable effects on the initial adhesion and growth activities of osteoblasts, which is encouraging for its potential use as bone graft material. Moreover, collagen type I may serve as an excellent biocompatible carrier for osteotropic factors such as cell adhesion molecules (e.g. fibronectin) or bone-specific growth factors.     

Attachment and proliferation of neonatal rat calvarial osteoblasts on Ti6Al4V: effect of surface chemistries of the alloy

This study examined the cell attachment and proliferation of neonatal rat calvarial osteoblasts on Ti6Al4V alloy as affected by the surface modifications. The modifications could alter simultaneously the surface chemistries of the alloy (elemental difference of Ti, Al, V, Cu and Ni about 300-600mum thick examined by EDS) as well as the XPS nano-surface characteristics of oxides on the metal surface (chemistries of oxides, amphoteric OH group adsorbed on oxides, and oxide thickness). Three materials including two from modifications and a control were examined. It is argued that a slight change of the nano-surface characteristics of oxides as a result of the modifications neither alters the in vitro capability of Ca and P ion adsorption nor affects the metal ion dissolution behavior of the alloy. This implies that any influence on the cytocompatibility of the materials should only be correlated to the effect of surface chemistries of the alloy and the associated metal ion dissolution behavior of the alloy. The experimental results suggest that the cell response of neonatal rat calvarial osteoblasts on the Ti6Al4V alloy should neither be affected by the variation of surface chemistries of the alloy in a range studied.     

Collagen type I-coating of Ti6Al4V promotes adhesion of osteoblasts

The initial contact of osteoblasts with implant surfaces is an important event for osseointegration of implants. Osseointegration of Ti6Al4V may be improved by precoating of its surface with collagen type I. In this study, the adhesion of rat calvarial osteoblasts to uncoated and collagen type I-coated titanium alloy was investigated over a period of 24 h. Collagen type I-coating accelerates initial adhesion of osteoblasts in the presence of fetal calf serum. One hour after plating, no differences in the percentage of adherent cells between the surfaces investigated were found. Adhesion of osteoblasts to uncoated surfaces was reduced by the GRGDSP peptide by about 70%, whereas adhesion to collagen type I-coated surfaces remained unaffected by treatment of the cells with the peptide. Cell adhesion to coated materials was reduced by about 80% by anti-integrin beta1 antibody. The integrin beta1 antibody did not influence the adhesion to uncoated titanium alloy. The results suggest that osteoblasts adhere to collagen type I-coated materials via integrin beta1 but not by interacting with RGD peptides, whereas adhesion to uncoated titanium alloy is mediated by RGD sequences but not via integrin beta1. Fibronectin does not seem to be involved in the adhesion of osteoblasts to either coated or uncoated titanium alloy.     

In vitro corrosion behaviour and osteoblast response of thermally oxidised Ti6Al4V alloy

In this work, the influence of thermal oxidation treatments of Ti6Al4V at 500 degrees C and 700 degrees C for 1 h on the in vitro corrosion behaviour and osteoblast response is studied. The potential of these treatments, aimed to improve the wear surface performance as biomaterial, relies in the formation of an outer "ceramic" layer of rutile. The corrosion behaviour was evaluated in simulated human fluids by electrochemical impedance spectroscopy and anodic polarisation tests. The effect of these thermal oxidation treatments on osteoblastic behaviour was studied in primary cultures of human osteoblastic cells. Results show that thermal oxidation treatments do not decrease the high in vitro corrosion resistance of the Ti6Al4V alloy. Osteoblast adhesion studies indicate that thermal oxidation treatments do not impair the material biocompatibility. Moreover, the thermal oxidation at 700 degrees C enhances the in vitro osteoblastic cell attachment compared to the thermal oxidation at 500 degrees C.     

Proliferation and differentiation of rat calvarial osteoblasts on type I collagen-coated titanium alloy.

Several attempts have been made to improve osseointegration of titanium alloy as an implant material by modification of its surface. In the present study, proliferation, differentiation, and mineralization of osteoblasts on type I collagen-coated Ti6Al4V were investigated. The activity of alkaline phosphatase and the accumulation of calcium by osteoblasts grown on titanium alloy were significantly higher compared to cells grown on polystyrene. Precoating of the implant surface with type I collagen did not extensively affect proliferation, the activity of alkaline phosphatase, collagen synthesis, calcium accumulation, or the mRNA levels for collagen I alpha1, osteopontin, osteocalcin, MMP-2, and TIMP-2. Maximum collagen synthesis by osteoblasts was observed at day 4 of culture independent of the type of implant material. The specific activity of alkaline phosphatase reached its maximum at day 18 of culture. Accumulation of calcium and elevated mRNA levels for osteocalcin were found at day 22. These results indicate that collagen-coating alone is not sufficient to accelerate differentiation of rat calvarial osteoblasts on Ti6Al4V.     

The effects of pulsed electromagnetic field on the functions of osteoblasts on implant surfaces with different topographies

The use of pulsed electromagnetic fields (PEMFs) is a promising approach to promote osteogenesis. However, few studies have reported the effects of this technique on the osseointegration of endosseous implants, especially with regard to different implant topographies. We focused on how the initial interaction between cells and the titanium surface is enhanced by a PEMF and the possible regulatory mechanisms in this study. Rat osteoblasts were cultured on three types of titanium surfaces (Flat, Micro and Nano) under PEMF stimulation or control conditions. Protein adsorption was significantly increased by the PEMF. The number of osteoblasts attached to the surfaces in the PEMF group was substantially greater than that in the control group after 1.5 h incubation. PEMF stimulation oriented the osteoblasts perpendicular to the electromagnetic field lines and increased the number of microfilaments and pseudopodia formed by the osteoblasts. The cell proliferation on the implant surfaces was significantly promoted by the PEMF. Significantly increased extracellular matrix mineralization nodules were observed under PEMF stimulation. The expression of osteogenesis-related genes, including BMP-2, OCN, Col-1,ALP, Runx2 and OSX, were up-regulated on all the surfaces by PEMF stimulation. Our findings suggest that PEMFs enhance the osteoblast compatibility on titanium surfaces but to different extents with regard to implant surface topographies. The use of PEMFs might be a potential adjuvant treatment for improving the osseointegration process.