阳极氧化伴水热处理制备纯钛羟基磷灰石薄涂层

为了解纯钛阳极氧化伴水热处理的技术路线及羟基磷灰石薄涂层在体内的成骨效应，以钛片为阳极，β－磷酸甘油钠和醋酸钙为电解质，经200－400V直流电解15min，维持电流密度≤50mA/cm^2，而后经280℃－300℃水热处理2h，取光滑，喷砂表面作对照，分别植入12只兔股骨内，枯术后4，8，16周取出带种植体骨块制作磨片，观察新骨生成情况及术后8周光滑和涂层种植体的界面超微结构，并作表面能谱分析，结果发现水热处理后，钛片表面出现了羟基磷灰石涂层，动物实验显示涂层种植体8周后编织骨的转化和成熟较快，未见剥脱的羟基磷灰石碎片，其表面的钙，磷含量在种植后增加明显，这说明阳极氧化伴水热处理可以制备出纯钛表面羟基磷灰石薄涂层，其早期促编织骨形成作用明显，可以加快编织骨转化成板层骨。     

纯钛种植体的表面预备

钛的优良特性早已被公认为首选牙用种植材料，其表面氧化膜是形成骨融合的关键。为了进一步增加其耐腐蚀性和生物相容性，人们采取了多种方式进行表面处理，最终达到在植入体的不同部位产生不同的粗糙表面，以利于不同细胞在不同地方的附着，尽量提高其表面能，增加其生物活性，同时，达到避免各种污染物对纯钛种植体影响的目的。另外，由于无法控制种植体的表面电荷，因此，可以忽略其对种植体的影响作用     

钛种植体表面改性策略及对骨整合的影响

背景：纯钛因具有优良的生物相容性、机械性能和与骨组织相近的弹性模量等被广泛应用于口腔种植领域。 目的：综述近年来钛金属种植体材料表面物理改性、化学改性和生物化学改性策略的研究进展。 方法：以“titanium,implant,surface modification,osseointegration”为检索词,检索PubMed数据库,以“钛,种植体,表面改性,骨整合”为检索词,检索中国知网数据库,限定时间范围为2007年1月至2013年2月。文献检索语种为英文和中文。纳入内容与钛种植体表面改性方法及其对骨整合影响密切相关的文献,排除重复文献。 结果与结论：计算机初检得到199篇文献,根据纳入排除标准,对其中76篇文献进行分析。钛种植体本身是生物惰性材料,通过表面改性对钛金属表面进行活化处理,使之具有生物功能性,与骨组织形成早期骨整合,是国内外口腔种植材料研究的热点问题。物理改性、化学改性和生物化学改性可缩短钛种植体种植周期,获得早期骨整合和更高的结合强度。今后的发展趋势是将多种改性方法有机结合,从分子水平深入研究钛金属材料表面与机体细胞和组织之间的界面分子作用机制,完善种植体的表面改性技术,实现种植体和骨组织的早期和更加稳定的骨整合。     

微弧氧化/电化学沉积钙磷涂层纯钛种植体的骨内植入

背景：近年来已有对微弧氧化/电化学沉积技术制备涂层在材料性能方面的相关报道,但对这种材料植入体内的性能研究较少见。 目的：观察纯钛种植体经微弧氧化/电化学沉积处理后的骨结合和新骨形成情况。 方法：通过微弧氧化/电化学沉积方法在纯钛上制备含钙磷元素的涂层,然后将该种植体和纯钛种植体分别植入羊两侧胫骨种植窝内,于动物处死前15,5 d分别进行注射四环素进行四环素标记。术后4,12周分别进行X射线、扫描电镜及激光共聚焦观察。 结果与结论：两侧X射线表现相似,种植体周围均无明显阴影,骨小梁排列和骨质密度与宿主骨基本一致。术后4周时,在电镜下可观察到两组种植体和骨组织之间均有间隙,部分见骨性结合;术后12周时,微弧氧化/电化学沉积种植体组可形成新骨,并且新骨与种植体和原来骨组织结合紧密,涂层与钛基体没有明显间隙,纯钛种植体组也可见新骨生成,但可看到明显裂隙。激光共聚焦观察显示,微弧氧化/电化学沉积种植体组双标记带间距离及骨矿化沉积率均高于纯钛种植体组(P < 0.05)。表明微弧氧化/电化学沉积处理可增强纯钛种植体的骨结合能力及新骨形成。     

钛银合金及其表面纳米管化的生物相容性

目的分析纳米钛银合金纳米管复合物的生物相容性。方法首先采用熔融法制备钛银合金(3%的银含量),然后通过阳极氧化技术在钛银合金表面制备纳米管。采用扫描电镜实验方法观察复合物的表面形貌。采用钛银合金及钛银合金纳米管和商业钛金属分别与成骨细胞共同培养方法,扫描电镜下观察细胞在支架上的黏附、增殖及凋亡。采用细胞毒性实验(MTT)实验方法检测金属的相容性。结果通过改变阳极氧化条件能制备不同表面形貌的钛银合金纳米管,将钛银合金及钛银合金纳米管和商业钛金属分别与成骨细胞共同培养3天后,倒置显微镜观察金属材料周围的成骨细胞生长情况,发现钛银合金纳米管及钛银合金周围细胞增殖与商业钛金属无明显差异。同时通过扫面电镜观察细胞在金属材料上的生长增殖情况,细胞在钛银合金纳米管上生长良好,可见大量细胞伪足附着于其上。同时通过细胞毒性实验(MTT),发现钛银合金纳米管及钛银合金与商业钛金属之间无明显差异。结论钛银合金纳米管具有良好的生物相容性。     

飞秒激光处理促进人牙周成纤维细胞在纯钛材料表面的增殖和分化

背景：目前对种植体表面优化处理促进人工牙种植体骨整合的形成是近年来这一领域的研究热点,而通过飞秒激光这种表面处理钛金属表面改性促进成纤维细胞的增殖和分化的相关性研究报道较少。目的：观察飞秒激光处理对于人牙周膜成纤维细胞在钛金属表面增殖和分化的影响。方法：将人牙周膜成纤维细胞接种在分别经过飞秒激光处理、TiO₂颗粒喷砂和砂纸顺向抛光的纯钛片表面(飞秒激光组,TiO₂颗粒喷砂组和对照组),观察细胞增殖和分化情况。结果与结论：与对照组相比,飞秒激光组和TiO₂颗粒喷砂组钛片表面粗糙度明显增加,表面细胞密度明显增加,RUNX2和OSX mRNA表达水平增加;且飞秒激光组上述指标高于TiO₂颗粒喷砂组。提示飞秒激光处理的纯钛表面更加粗糙,更有利于人牙周膜成纤维细胞的增殖和分化。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

纯钛表面阳极氧化增强成骨细胞生物活性及护骨素基因的表达

背景:纯钛阳极氧化改性后形成的纳米结构与骨组织具有良好的生物相容性。目的:观察纯钛表面纳米孔结构的形貌和物相构成,以及其对MC3T3-E1小鼠前成骨细胞增殖、黏附等生物学行为和促成骨基因护骨素表达的影响。方法:取纯钛片24份,其中12份仅进行机械抛光,作为对照组;另外12份进行机械抛光后,应用阳极氧化技术在纯钛表面制备纳米孔结构,作为实验组。将小鼠前成骨细胞MC3T3-E1分别接种于两组试件表面,接种7 d后扫描电镜下观察细胞形态,采用MTT法检测细胞增殖情况,绘制生长曲线;同时检测细胞促成骨基因护骨素的表达。结果与结论:阳极氧化后钛片表面形成规格统一的纳米孔结构,但是物相构成并未发生变化。与接种于对照组试件上的成骨细胞相比,实验组试件表面的细胞密度变大,覆盖金属的面积更多,呈现多边形结构,突触向周围移行,可见板状伪足向周围材料伸出;接种第7天时,实验组细胞数目约为对照组的1.4倍,同时纳米孔表面成骨细胞护骨素基因的表达高于对照组(P0.01)。结果表明阳极氧化后形成纳米孔结构的钛片更有利于成骨细胞的黏附、增殖和护骨素基因的表达,进而促进成骨细胞生长,具有良好的生物相容性。     

阳极氧化产品表面元素定性分析探讨

目的对表面经阳极氧化的钛及钛合金产品进行表面元素定性分析,并对其数据进行探讨。方法采用扫描电镜及能谱仪对表面元素进行测定。结果阳极氧化产品表面经扫描电镜及能谱仪分析,所包含元素除基体元素外,还可包含少量Na、Si、Ca、P等元素。结论纯钛、钛合金类产品植入物表面形成的氧化膜具有良好的生物相容性,通过对钛及钛合金表面阳极氧化工艺和成分控制,可以有效降低产品生物安全风险。     

新型经皮植入材料的生物学性能评价研究

参照ISO 10993系列标准,通过细胞毒性试验、刺激试验和致敏试验对阳极氧化表面改性纯钛作为经皮植入材料进行了生物学性能评价研究.试验结果表明,此种改性纯钛无细胞毒性,皮肤刺激反应属于极微弱类型,不存在致敏潜能,是生物学性能优良的经皮植入材料.     

选择性激光熔融打印钛种植体的制备和表面优化处理及其成骨性能的研究

目的　通过表征和体外相关实验分析选择性激光熔融（SLM）3D打印钛种植体试样表面的生物相容性能，为SLM打印种植体最终应用于临床提供理论基础和实验依据。 方法　使用SLM方法制备得圆片形钛种植体试样60枚，经不同处理后分为三组：喷砂酸蚀组（S组）、阳极氧化组（SA组）、阳极氧化+rhBMP-2表面组（SAB组）；场发射扫描电镜（FE-SEM）和原子力显微镜（AFM）观察各组试样的表面形貌和粗糙度，接触角测量仪检测亲水性；SEM观察骨髓间充质干细胞（BMMSCs）在各组试样表面的粘附状态；CCK-8检测BMMSCs在各组试样表面的增殖水平。 结果　SEM观察结果显示阳极氧化后的试样SA组表面可见一层均匀密布的二氧化钛纳米管，直径约为100纳米；AFM观察试样表面粗糙度约为300纳米；接触角测量仪测得经表面处理和改性的SA组，SAB组试样具有超亲水性；BMMSCs粘附和增殖实验显示各组试样表面的促细胞粘附和增殖能力SAB组>SA组>S组。 结论　SLM打印种植体试样未处理组和处理组都具有较好的生物相容性能，阳极氧化与加载BMP-2生长因子处理结合的方法能有效促进BMMSCs的粘附和增殖。     

The influence of surface bioactivated modification on titanium percutaneous implants anchored in bone

In order to achieve biological sealing and resist mechanical damage of load-bearing percutaneous devices, Ti with excellent mechanical properties was anodic-oxidized to be endowed with bioactivity, with plasma-sprayed hydroxyapatite coated Ti as control. Similar to previous works, hydroxyapatite coating could bond tightly with living tissues, resulting in implant stability for whole implantation periods. Meanwhile, when anodic-oxidized bioactivated Ti was implanted percutaneously in vivo, it could induce a layer of calcium phosphate at the interface of tissues/implant. This layer of Ca-P not only induced the fibrous tissue or collagen ingrowth in its structure, but also improved the osteointegration between the bone and the implant. There was no significant biological response difference for the anodic-oxidized Ti and HA coated Ti at different implantation period with histological statistical analysis (p>0.05). Accordingly, suitable bioactivated modified surface of Ti by anodic-oxidized method could not only obtain the same results as the HA coating, but also might avoid some drawbacks of plasma-sprayed HA coatings to achieve biological sealing for a long period in vivo.     

Preliminary study on HA coating percutaneously implanted in bone

A comparative investigation on the possibility of hydroxyapatite (HA) coating and pure Ti column to form biological sealing with skin tissue was completed in this study. HA coating and pure Ti column were percutaneously implanted in the tibia of rabbits. Compared with titanium (Ti) implant, HA coating forms epithelial sealing with skin tissue at 6 weeks postoperatively, while the Ti implant may loosen from the implanted site and be lost. The Ti column loosing rate at this time was 50%. However, once the Ti implant becomes fixed with the bone tissue, it can form epithelial sealing with skin tissue just like the HA coating, at 8 weeks postoperatively. At 8 weeks postoperatively, the epithelial sealing is not destroyed in spite of the fact that the HA coating is biodegraded. Our results show that the HA coating can become fixed with the bone faster than the Ti, which is beneficial for epithelial sealing formation. The main role of HA coating for epithelial sealing is beneficial for sealing at the initial period after it is implanted.     

Contact profilometry and correspondence analysis to correlate surface properties and cell adhesion in vitro of uncoated and coated Ti and Ti6Al4V disks

A fundamental goal in the field of implantology is the design of specific devices able to induce a controlled and rapid "osseointegration". This result has been achieved by means of surface modifications aimed at optimizing implant-to-bone contact; furthermore, bone cell adhesion on implant surface has been directly improved by the application of biomolecules that stimulate new tissue formation, thus controlling interactions between biological environment and implanted materials. Actually, methods for biochemical factor delivery at the interface between implant surface and biological tissues are under investigation; a reliable technique is represented by the inclusion of biologically active molecules into biocompatible and biodegradable materials used for coating implant surface. This paper focuses the application of three polymeric materials already acknowledged in the clinical practice, i.e. poly-L-lactic acid (PLLA), poly-DL-lactic acid (PDLA), and sodium alginate hydrogel. They have been used to coat Ti (Ti2) and Ti6Al4V (Ti5) disks; their characteristics have been determined and their performances compared, with specific regard to the ability in allowing osteoblast adhesion in vitro. Moreover, profilometry data analysis permitted to identify a specific roughness parameter (peak density) which mainly controls the amount of osteoblast adhesion.     

钛表面纳米仿生磷灰石涂层对成骨样细胞活性的生物增强效应

目的：观察钛表面纳米仿生磷灰石涂层对成骨样细胞行为的影响，为骨科常用钛植入体的表面改性及其生物效应提供实验依据。方法: 商业用纯钛经过物理、化学和生物处理，表面生成均匀薄层仿生的纳米磷灰石涂层，将仿生涂层的钛金属板与成骨样细胞复合培养，以纯钛和只经磨砂、酸蚀处理的钛板作为对照，采用MTT法检测细胞活力和增殖变化、扫描电镜和激光共聚焦荧光显微镜观察细胞形态、RT-PCR检测碱性磷酸酶基因表达。结果: 纳米仿生磷灰石涂层比非涂层钛金属表面细胞的增殖数量明显增高，细胞的形态和分布也优于对照组；培养12 d，涂层对细胞ALP基因表达的量明显高于对照组。结论: 钛金属表面纳米仿生磷灰石涂层可以增强细胞的生物效应，提高钛植入体的骨界面早期结合，具有很好的应用前景。     

树脂水门汀口腔黏结材料SuperBond C&B的体外细胞毒性

BACKGROUND: Because of long-term contact with tooth tissues, gingival tissues and saliva in the mouth, oral bonding materials must have the characteristics of biological safety and low or no cytotoxicity, so as to ensure no irritation response to the oral mucous and no toxicity and allergic reactions when contacting with human tissues. OBJECTIVE: To observe the in vitro cytotoxicity of an oral bonding material, SuperBond C&B. METHODS: The logarithmic human periodontal ligament fibroblasts were obtained and inoculated in 96-well plates with a cell concentration of 1.0×108/L, 100 μL/hole. Supernatant was removed after 24 hours of routine culture, and the specimens were grouped and cultured. 90 μL DMEM culture liquid+10 μL SuperBond C&B were added in the experimental group. 100 μL medium was added in the negative control group. 100 μL phenol solution was added in the positive control group. After 72 hours of culture, the cell survival rate in each group was calculated by trypan blue exclusion test. RESULTS AND CONCLUSION: The cell survival rates of the experimental group and the negative control group were higher than that of the positive control group (P < 0.05), and there was no significant difference between the experimental group and the negative control group (P > 0.05). These results show that SuperBond C&B has no cytotoxicity.        

脂肪干细胞与外源性血管内皮生长因子及纤维蛋白胶复合物体内构建血管化组织工程脂肪

背景：血运重建机制是组织工程化脂肪组织成功构建的决定性因素。 目的：观察脂肪干细胞与外源性血管内皮生长因子和纤维蛋白胶复合物在体内构建血管化组织工程脂肪的可行性。 方法：从健康成年人吸脂术后的脂肪组织中分离脂肪干细胞并行原代及传代培养,将第3代经BrdU标记的脂肪干细胞向脂肪细胞定向诱导2周后,制成5×10¹⁰ L^-1细胞悬液。由0.5 mL细胞悬液、100 μL的血管内皮生长因子工作液或DMEM培养基与0.5 mL纤维蛋白胶组成实验组和对照组移植物,分别植入裸鼠背部皮下。 结果与结论：术后8周取材时：①实验组可见血管增生并长入材料,呈轻度纤维包裹;对照组有少量血管长入材料中,也有轻度纤维包裹现象。实验组新生组织湿质量大于对照组(P < 0.01)。②苏木精-伊红染色均可见移植物中有新生脂肪组织形成和不同程度的微血管长入;实验组微血管数多于对照组(P < 0.01)。③新生组织免疫荧光染色示,两组新生脂肪细胞的胞核及部分微血管内皮细胞的胞核呈现绿色荧光。结果说明脂肪干细胞与外源性血管内皮生长因子和纤维蛋白胶复合物在体内可构建血管化组织工程脂肪,脂肪干细胞与外源性血管内皮生长因子共同参与新生脂肪组织的血管化过程。     

In vitro and in vivo biological responses of plasma-sprayed hydroxyapatite coatings with posthydrothermal treatment

This study was undertaken to evaluate the effect of post-hydrothermal treatment on the biological responses of the plasma-sprayed hydroxyapatite (HA)-coated Ti-6Al-4V implant system both in vitro and in vivo. After hydrothermal treatment, the HA coating (HAC) shows the high mechanical strength and indices-of-crystallinity, denser microstructure, lower concentrations of amorphous and impurity phases, when compared with the as-sprayed HAC. The in vitro cell-culture studies, using UMR106 osteoblast-like cell, demonstrated no signifiacnt cell growth on both surface of as-sprayed and hydrothermal-treated HACs during 10-day culture. The in vivo studies, using the transcortical implant model in the femora of goats, evaluated the histological responses of two coatings. After 6 week of implantation, using backscattered electron images, no substantial histological variations in the extents of new bone apposition and new bone healing between the two HACs were observed. However, the as-sprayed HAC, owing to the dissolution induced the granular particles dissociated from the HAC, showed the statically lower extent of new bone apposition than hydrothermal-treated HAC at 12 weeks. The results suggest that hydrothermal treatment could be used to improve the mechanical strength, crystallinity, and phase composition of HAC, which are important factors of long-term fixation and stability of implant. Besides, the treated HAC could also achieve the initial fixation of implant in clinical use.     

Biological response on a titanium implant-grade surface functionalized with modular peptides

Titanium (Ti) and its alloys are among the most successful implantable materials for dental and orthopedic applications. The combination of excellent mechanical and corrosion resistance properties makes them highly desirable as endosseous implants that can withstand a demanding biomechanical environment. Yet, the success of the implant depends on its osteointegration, which is modulated by the biological reactions occurring at the interface of the implant. A recent development for improving biological responses on the Ti-implant surface has been the realization that bifunctional peptides can impart material binding specificity not only because of their molecular recognition of the inorganic material surface, but also through their self-assembly and ease of biological conjugation properties. To assess peptide-based functionalization on bioactivity, the present authors generated a set of peptides for implant-grade Ti, using cell surface display methods. Out of 60 unique peptides selected by this method, two of the strongest titanium binding peptides, TiBP1 and TiBP2, were further characterized for molecular structure and adsorption properties. These two peptides demonstrated unique, but similar molecular conformations different from that of a weak binder peptide, TiBP60. Adsorption measurements on a Ti surface revealed that their disassociation constants were 15-fold less than TiBP60. Their flexible and modular use in biological surface functionalization were demonstrated by conjugating them with an integrin recognizing peptide motif, RGDS. The functionalization of the Ti surface by the selected peptides significantly enhanced the bioactivity of osteoblast and fibroblast cells on implant-grade materials.     

Enhanced mechanical properties and in vitro cell response of surface mechanical attrition treated pure titanium

Surface mechanical attrition treatment (SMAT) was used to fabricate nanocrystalline surface layers on the commercial purity titanium. X-ray diffraction and transmission electron microscopy results indicate that the top layer contained nanograins. Enhanced strength and microhardness were achieved due to the surface nanostructure. Cell culture tests have shown a greater adhered cell density and more extensively spreading morphologies of Saos-2 cells on the SMAT substrates compared to those on the as-received Ti counterparts. Enhanced cell viability and cell cycle were also achieved on the SMAT Ti substrates. These could be attributed to the nanostructure grains with the increased surface hydrophilicity and roughness on the SMAT Ti.     

Improved biological performance of Ti implants due to surface modification by micro-arc oxidation

The surface of a titanium (Ti) implant was modified by micro-arc oxidation (MAO) treatment. A porous layer was formed on the Ti surface after the oxidation treatment. The phase and morphology of the oxide layer were dependent on the voltage applied during the oxidation treatment. With increasing voltage, the roughness and thickness of the film increased and the TiO(2) phase changed from anatase to rutile. During the MAO treatment, Ca and P ions were incorporated into the oxide layer. The in vitro cell responses of the specimen were also dependant on the oxidation conditions. With increasing voltage, the ALP activity increased, while the cell proliferation rate decreased. Preliminary in vivo tests of the MAO-treated specimens on rabbits showed a considerable improvement in their osseointegration capability as compared to the pure titanium implant.