应用IBAD方法制备纯钛表面多孔TCP／HA涂层材料的微观分析

目的:为了改善钛种植体的生物相容性,对纯钛表面沉积多孔磷酸三钙/羟基磷灰石(Tricalciumphosphate/hydroxyapatite,TCP/HA)复合涂层材料的表面结构和化学成分进行分析,并与沉积羟基磷灰石(Hydroxyapatite,HA)的钛表面进行对比。方法:用离子束辅助沉积方法(Ionbeamassisteddeposition,IBAD)在纯钛表面沉积HA和TCP/HA涂层材料,通过扫描电镜(Scanningelectronmicroscope,SEM)、原子力显微镜(Atomicforcemicroscopy,AFM)、X射线能谱分析(EnergydispersiveX-rayanalysis,EDX)以及X射线衍射(X-raydiffraction,XRD)技术,检测两种涂层材料表面的微观形态和化学成分,并进行比较。结果:SEM和AFM显示TCP/HA涂层材料表面存在多孔结构,表面化学成分分析显示TCP/HA涂层的钙磷比低于HA,XRD证实TCP/HA涂层内同时存在TCP和HA两种化合物。结论:用IBAD方法在纯钛表面成功地沉积了具有多孔结构的TCP/HA复合涂层材料,该涂层材料和基体材料的结合牢固,改善了基体材料的生物相容性,是一种有应用前景的种植体表面涂层材料。     

HA涂层-多孔TiO2-钛基体梯度涂层对MG63的影响

目的:评价微弧氧化(micro-arc oxidation,MAO)与离子束辅助沉积(ion beam assisted deposition,IBAD)相结合的处理方法对钛种植材料表面微形态及成骨活性的影响。方法:利用IBAD技术,在MAO钛材料表面制备羟基磷灰石(hydroxyapatite,HA)涂层,将钛种植材料按照不同表面处理方法分为机械抛光组(S0)、MAO组(S1)和MAO与IBAD相结合组(S2),以S0作为对照组。利用表面形貌仪分析3组材料表面形貌及粗糙度,利用激光共聚焦显微镜观察成骨细胞在材料表面黏附铺展状态;采用MTT法和碱性磷酸酶(ALP)活性测定,检测不同处理方法对材料成骨活性的影响。所得数据利用SPSS11.0软件包进行单因素方差分析。结果:S2组材料表面粗糙度略小于S1组;复合培养5d后,S2组细胞增殖水平显著高于其他2组;10d后,2粗糙组ALP活性显著高于S0组。结论:MAO与IBAD技术相结合,在MAO表面制备HA活性层,能够显著促进成骨细胞的黏附增殖,促进成骨细胞的矿化。     

羟基磷灰石涂层处理的多孔镍钛合金溶血性及细胞黏附性评价

目的通过研究经羟基磷灰石(HA)涂层处理后多孔镍钛(NiTi)合金的溶血率及成骨细胞在其表面的附着、增殖及分化情况,评价其体外生物相容性。方法测定经HA涂层处理的圆盘状多孔NiTi合金(NiTi-HA组;直径10mm,厚2mm)的生物相容性,未经涂层处理的多孔NiTi合金(NiTi组)及致密纯钛(Ti组)试样设为对照组。采用分光光度法分析其溶血性能;将成骨细胞接种于试样表面,用扫描电镜(SEM)观察成骨细胞黏附形态,MTS法及碱性磷酸酶(ALP)试剂检测细胞附着、增殖情况及ALP活性,对数据进行重复测量方差分析。结果 NiTi-HA组、NiTi组及Ti组试样的溶血率分别为(0.30±0.11)%、(0.51±0.07)%及(0.27±0.06)%,均低于国家标准(YY/T0127.1)规定的5%。SEM观察显示,NiTi-HA组及NiTi组试样细胞黏附形态良好。NiTi-HA组试样表面细胞附着及增殖数量均高于NiTi组试样(P值分别为0.000与0.001)。NiTi-HA组及NiTi组试样表面细胞ALP活性无差异(P=1),但均高于Ti组试样(P值分别为0.001与0.0004)。结论 NiTi-HA组、NiTi组及Ti组试样均无溶血作用;NiTi-HA组较NiTi组更有利于成骨细胞附着和增殖,且ALP活性均高于纯钛。     

喷砂酸蚀混合碱热处理后纯钛表面生物活性的研究

目的研究喷砂酸蚀混合碱热处理后的纯钛表面其生物活性。方法将经抛光(M)、喷砂酸蚀(SLA)、喷砂酸蚀混合碱热(AHH)处理后的纯钛试件分为3组。扫描电子显微镜(SEM)及能谱分析(EDS)对3组表面结构和表面化学元素及其含量进行分析;荧光显微镜下观察成纤维细胞在样品表面的粘附及铺展情况;并将各组试件浸泡模拟体液(SBF)中观察羟基磷灰石沉积情况。结果纯钛表面微纳复合结构中,微米孔直径3~5μm,纳米孔直径100~200 nm,同时在试件表面引入钙钠元素。成纤维细胞粘附:DAPI染色后细胞核呈蓝色荧光,罗丹明B染细胞骨架红色,SLA组、AHH组细胞铺展较M组好,呈空间铺展;且AHH组表面细胞数量明显多于其他两组。第1周时,AHH组表面沉积磷灰石明显可见,而未在M、SLA组检测到羟基磷灰石。结论喷砂酸蚀混合碱处理后的纯钛表面,表面活性好,有助于促进成纤维细胞早期粘附及其在空间上的铺展,同时又促进羟基磷灰石的沉积,进而体现了良好的生物性能,可以为种植体表面处理方法提供参考。     

牙周膜细胞在碱处理后的多孔钛片表面的活性

目的 以碱处理和仿生溶液在多孔钛表面制备类骨磷灰石涂层,观察人牙周膜细胞(PDLC)在碱处理前后多孔钛表面的黏附生长,评价多孔钛类骨磷灰石表面的生物相容性.方法 松装烧结法制备多孔钛片,60℃下行碱处理,在模拟体液中沉积类骨磷灰石涂层.扫描电镜和X射线衍射仪分析涂层表面形貌、物相和结构,能量散射光谱测定仪分析其表面成分...     

钛表面激光快速成形CaTiO3涂层成骨细胞相容性的研究

目的：探讨激光快速成形技术制备的纯钛表面CaTiO3涂层的成骨细胞相容性。方法：分别制备纯钛表面激光快速成形CaTiO3涂层试件和经表面处理的纯钛试件各6个,采用成骨细胞与材料共培养技术,利用MTT法分别检测成骨细胞在两种材料表面的黏附和增殖数量,并进行统计学分析;利用扫描电镜观察法分别观察成骨细胞在两种材料表面的铺展状态。结果：纯钛表面激光快速成形CaTiO3涂层材料表面成骨细胞的黏附和增殖数量明显高于经表面处理的纯钛（P〈0．05）;成骨细胞在经表面处理纯钛表面呈板状平铺,而在CaTiO3涂层表面呈多极性、立体伸展,并表现出更加旺盛的功能状态。结论：激光快速成形技术在纯钛表面制备的CaTiO3涂层能够促进成骨细胞在材料表面的黏附、铺展和增殖。     

微弧氧化钛基种植材料对成骨细胞早期黏附的影响

目的：检测纯钛种植体材料微弧氧化（microarc oxidation，MAO）表面改性后的成骨细胞生物相容性，探讨微弧氧化技术在钛种植体表面改性中的价值。方法：在纯钛种植体材料表面用微弧氧化法制备羟基磷灰石陶瓷薄膜，将MAO改性钛种植体材料作为实验组Ⅰ，将纯钛表面阳极氧化改性处理的种植体材料作为实验组Ⅱ．并设立对照组Ⅰ（纯钛种植体材料）和对照组Ⅱ（即细胞直接生长在培养板上），分别进行扫描电镜（SEM）、能谱分析（EDS）、X射线衍射图谱分析（XRD）等检测，比较成骨细胞的黏附水平，对数据采用SPSS11．0统计软件包进行单因素方差分析。结果：MAO改性后生成粗糙、多孔的陶瓷薄膜层，与处理前电解液成分相比，钙磷比无显著改变。MAO改性钛组黏附的细胞密度显著高于其他组（P〈0．01），而对照组Ⅰ、Ⅱ的细胞密度无显著差异（P〉0．05）。结论：与阳极氧化表面改性材料相比，该钛基微弧氧化薄膜层能够显著促进成骨细胞的附着，具有良好的细胞相容性。     

超细晶纯钛种植体表面微弧氧化处理的实验研究

目的:探讨微弧氧化技术用于超细晶纯钛表面处理的可行性。方法:通过微弧氧化方法在超细晶纯钛和普通纯钛(对照组)表面制备氧化涂层,然后测定各组材料表面氧化涂层的组成、厚度、粗糙度,并分析其表面形貌和涂层的结合强度。结果:通过微弧氧化在超细晶纯钛和普通纯钛表面形成的氧化涂层主要由O、Ti、P、Si、C组成,两组的各元素比例均无明显差异(P>0.05),两组涂层的表面形貌均为疏松多孔结构,超细晶纯钛组的涂层厚度为(7.833±0.771)μm,普通纯钛组(4.775±0.558)μm(P<0.05);超细晶纯钛实验组和普通纯钛对照组粗糙度值分别为(1.391±0.143)μm和(1.346±0.091)μm(P>0.05),涂层的结合强度分别为(23.900±1.267)N和(23.867±1.134)N(P>0.05)。结论:超细晶纯钛经微弧氧化处理后可在其表面形成疏松而多孔的氧化涂层;涂层表面的元素组成未发生改变,结合强度能满足临床中种植体使用的要求。     

微纳复合结构对成纤维细胞黏附增殖的影响

目的：探讨经不同方法处理后的纯钛表面对成纤维细胞黏附增殖的影响。方法：将36个试件分平均为3组：机械抛光组（A组）;喷砂酸蚀组（B组）;喷砂酸蚀碱热组（C组）,每组均12个试件。通过扫描电子显微镜（SEM）观察分析3组试件表面微观结构和细胞在试件表面的铺展情况,激光共聚焦显微镜（CLSM）检测各试件表面的粗糙度;运用CCK-8试剂盒在450 nm波长下检测各试件对成纤维细胞（ L929）黏附与增殖的吸光度值（ OD值）。结果：A组表面光滑,试件表面成纤维细胞骨架大多呈梭形铺展,伸展较差;B组和C组表面粗糙,且C组表面可见微纳复合结构,试件表面成纤维细胞骨架呈三维空间向铺展,表面黏附成纤维细胞数量明显多于A组和B组。观察第1,3,5 d试件表面细胞增殖情况,可见粗糙表面较光滑表面更利于成纤维细胞的增殖。结论：喷砂酸蚀碱热方法处理后的纯钛表面形成微米-纳米复合孔洞,表面活性好,促进成纤维细胞早期黏附及表面铺展,且不抑制细胞的增殖。     

不同基台材料对人牙龈上皮细胞半桥粒黏附相关基因及蛋白表达的影响

目的探讨聚醚醚酮、氧化锆、纯钛基台材料对人牙龈上皮细胞半桥粒黏附相关基因及蛋白表达的影响, 以期筛选出更适合上皮附着的基台材料。方法制备聚醚醚酮、氧化锆、纯钛试件各48枚, 用扫描电镜观察各组表面形貌, 用白光干涉仪测量表面粗糙度, 光学接触角测量仪测量接触角。采用扫描电镜观察人牙龈上皮细胞在各组试件表面的早期黏附状态, 并使用细胞计数试剂盒评估各组人牙龈上皮细胞的增殖能力, 实时荧光定量PCR和蛋白质印迹法分别检测各组人牙龈上皮细胞半桥粒黏附相关基因及蛋白的表达水平。结果 3组试件均呈平坦、光滑的表面形貌;聚醚醚酮组、氧化锆组和纯钛组平均粗糙度(Ra值)分别为(95.63±2.06)、(37.93±3.56)、(134.2±4.62)nm(F=368.16, P<0.001), 平均最大高度(Rz值)分别为(2.42±0.22)、(0.87±0.10)、(3.77±0.28)nm(F=91.95, P<0.001), 组间差异均有统计学意义(P<0.05);接触角总体差异无统计学意义(F=0.45, P=0.658)。人牙龈上皮细胞在3组试件表面均呈扁平伸展的多边形...     

氟离子注入钛表面改性对成骨细胞黏着斑形成的影响

目的探讨氟离子注入纯钛进行表面改性后,钛表面改性层化学组成的变化及其对人成骨细胞样细胞系MG- 63黏着斑形成的影响。方法应用等离子体浸没离子注入装置对钛试件进行氟离子注入,通过X线光电子能谱（XPS）分析研究其表面化学组成和各元素的化学结合价态。将人MG- 63细胞接种于氟离子注入组和纯钛组钛试件表面,用激光共聚焦显微镜观察两组钛试件表面对人MG- 63细胞黏着斑形成的影响。结果XPS全谱图显示氟离子注入后试件由钛、氧、氟和碳元素组成,拟合结果显示钛表面改性层的元素以二氧化钛和三氟化钛的形式存在。MG-63细胞培养6 h后,氟离子注入组钛试件表面的黏着斑形成数量比纯钛组钛试件表面形成得多,二者间差异具有统计学意义（P<0.05）。结论氟离子注入可成功地将氟元素引入纯钛表面形成含有二氧化钛和三氟化钛的表面改性层,该层可促进成骨细胞黏附初期黏着斑的形成,提示其具有良好的生物相容性。     

In vivo characterization of titanium implants coated with synthetic hydroxyapatite by electrophoresis

This study compared in vivo the performances of commercially pure titanium (cp Ti) screw dental implants either uncoated or coated with synthetic hydroxyapatite (HA) by electrophoresis. The HA coating was characterized by scanning electron microscopy, energy dispersive spectroscopy (EDS) and Fourier-transform infrared (FT-IR) spectroscopy. Well-adhered carbonated-hydroxyapatite layers (4- to-8-microm-thick) were obtained. In vivo tests were carried out by insertion of both uncoated and HA-coated implants into rabbit tibiae for 8 or 12 weeks. Histomorphometric analysis was performed by scanning electron microscopy with the aid of image-processing software. Results showed significantly greater bone-implant contact for HA-coated implants (p<0.05) than cp Ti implants. Comparison of bone content inside the screw implants showed no significant differences (p>0.05) between both types of implants, although cp Ti had numerically higher percentage of bone content than HA-coated implants. In conclusion, the HA-coated implants had better performance regarding the bone-implant contact area than the uncoated implants; coating by electrophoresis proved to be a valuable process to coat metallic implants with an osteoconductive material such as hydroxyapatite.     

选择性激光烧结多孔钛种植体及其性能研究

目的 分析选择性激光烧结（SLS）多孔钛种植体的机械性能及生物相容性,探讨其与壳聚糖（CS）/羟磷灰石（HA）复合涂层结合后的促骨结合作用。方法 制备Ti6Al4V试件,部分试件表面进行CS/HA涂层处理;对试件进行扫描电子显微镜观察和机械性能检测;体外培养MC3T3-E1细胞,进行活/死细胞染色、甲基噻唑基四唑（MTT）及碱性磷酸酶（ALP）水平检测;将柱状螺纹种植体植入兔股骨髁部,分析其体内生物学性能。结果 试件准弹性梯度随孔隙率增大而减小,孔隙率为30%时与皮质骨接近,70%时与松质骨接近;试件具有良好的生物相容性。复合CS/HA涂层后可促进MC3T3-E1细胞增殖、分化,有利于骨组织长入孔隙,形成良好的骨结合。结论 多孔钛种植体具有良好的机械性能和生物相容性,与CS/HA涂层结合后具有骨诱导性,利于形成稳定的骨结合。     

Hydroxyapatite coating for titanium fibre mesh scaffold enhances osteoblast activity and bone tissue formation

This study investigated the bone regeneration properties of titanium fibre mesh as a tissue engineering material. A thin hydroxyapatite (HA) coating on the titanium fibre web was created using the developed molecular precursor method without losing the complex interior structure. HA-coated titanium fibre mesh showed apatite crystal formation in vitro in a human osteoblast culture. Titanium fibre mesh discs with or without a thin HA coating were implanted into rat cranial bone defects, and the animals were killed at 2 and 4 weeks. The in vivo experience revealed that the amount of newly formed bone was significantly higher in the HA-coated titanium fibre mesh than in the non-coated titanium fibre mesh 2 weeks after implantation. These results suggest that thin HA coating enhances osteoblast activity and bone regeneration in the titanium fibre mesh scaffold. Thin HA-coating improved the ability of titanium fibre mesh to act as a bone regeneration scaffold.     

Photocoupling of fibronectin to titanium surfaces influences keratinocyte adhesion, pellicle formation and thrombogenicity.

OBJECTIVES: Coating of implant surfaces with biomolecules can influence basic host responses and enhance subsequent tissue integration. The biological factors have to be immobilized on the implant material. Human fibronectin (Fn) was used as a model protein and covalently coupled to titanium (Ti) surfaces via silanization and an anthraquinone linker. The impact on several aspects of initial host/biomaterial interactions (keratinocyte adhesion, platelet interactions and pellicle formation) was studied. METHODS: Coupling efficiency was characterized by immunological techniques. The effects of coupled Fn on initial host/biomaterial interactions were assessed. Cell adhesion and spreading were investigated by fluorescent staining, pellicle formation by an acoustic sensor system (quartz crystal microbalance with dissipation, QCM-D), and platelet adhesion as one parameter mediating the inflammatory response by scanning electron microscopy (SEM) and immunological assays. RESULTS: Coupling efficiency was related to irradiation time used for photochemical coupling of the UV-activated anthraquinone to the silanized Ti surface. With an optimized protocol, the amount of Fn coupled to the surface could be almost doubled compared to standard dip-coating methods. On the anthraquinone-coupled Fn coatings, cell adhesion and spreading of human keratinocytes was significantly enhanced. Online detection of pellicle formation revealed strong reversibility of saliva protein adhesion on Fn coated surfaces compared to the pure Ti surface. Furthermore, the Fn coated Ti showed a low thrombogenicity. SIGNIFICANCE: This study suggests that anthraquinone-coupled biological coatings may be useful for biofunctionalization of Ti dental implants by enhancement of soft tissue re-integration (restoration of the epithelial seal) combined with diminished pellicle formation.     

Corrosion resistance and biocompatibility of a new porous surface for titanium implants

Alterations of the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study was to investigate the biocompatibility of cpTi when submitted to a new, porous titanium, surface treatment (porous Ti). Five types of surface treatments, namely sintered microspheres porous titanium (porous Ti), titanium plasma spray (TPS), hydroxyapatite (HA), sandblasted and acid etched (SBAE), and resorbable blast medium, sandblasted with hydroxyapatite (RBM) were made. In the experimental methods, the corrosion potentials were measured over time, and then a linear sweep voltammetric analysis measured the polarization resistances and corrosion currents. For biocompatibility evaluation, MG63 osteoblast-like cells were used. Cell morphology, cell proliferation, total protein content, and alkaline phosphatase (ALP) activity were evaluated after 2 h, and after 2, 4 and 7 d. Porous Ti and SBAE showed a better corrosion resistance, with a weak corrosion current and a high polarization resistance, than the other surfaces. Cell attachment, cell morphology, cell proliferation, and ALP synthesis were influenced by the surface treatments, with a significant increase observed of the activity of osteoblast cells on the porous coating (porous Ti). Based on these results, it is suggested that the porous Ti surface has a significantly better biocompatibility than the other surface treatments and an excellent electrochemical performance.     

Torsional stability of HA-coated and grit-blasted titanium dental implants.

Hydroxylapatite (HA)-coated and grit-blasted (non-HA-coated) titanium dental implants were inserted into healed extraction sites of canine mandibles. After six weeks, the animals were killed and the implants mechanically tested in torsion to failure. Interface attachment strength, implant/tissue compatibility, integrity of the HA coating, and the location of interface failure were evaluated. Mechanical testing demonstrated an interface torsional strength of 3.98 +/- 0.93 MPa for the HA-coated implants and 2.25 +/- 0.65 MPa for the grit-blasted implants. This represents a 76.9% improvement in the maximum torsional interface strength, and is statistically-significant (p = 0.0004). On qualitative histologic analysis, interface failure was seen to occur primarily at the HA/implant interface, although failure through the HA coating and regions of bone/HA interface failure were observed. The HA-coated implants had bone in direct apposition to their surface with no fibrous tissue interposition. The grit-blasted implants also had regions of direct bone-implant apposition, but these areas were limited to a smaller proportion of the total interface area. There was no evidence of breakdown or change in thickness of the HA coating.     

Evaluation of the titanium Ti-6Al-7Nb alloy with and without plasma-sprayed hydroxyapatite coating on growth and viability of cultured osteoblast-like cells

BACKGROUND: Osseointegrated dental implants are currently recognized as a standard treatment method in dentistry. Titanium (Ti) and its alloys are the metals of choice for endosseous parts of currently available dental implants. Ti-6Al-4V is the most used Ti alloy, however; an improved version, Ti-6Al-7Nb, has been recently developed. METHODS: Rat osteoblast-like cells (osteo- 1 culture) were used to analyze the biocompatibility of Ti-6Al-7Nb alloy with and without hydroxyapatite (HA) coating. The cells were grown on culture Petri dishes on the top of either plain Ti-6Al-7Nb or HA-coated Ti-6Al-7Nb disks. Osteo-1 cells grown on plain culture dishes were used as controls. Growth and cell viability curves were obtained by scanning electron microscopy. For the growth and viability curves, 10(4) cells were seeded on 35 mm dishes. Cells from each group were counted, in triplicate at 3, 7, 11, and 15 days after seeding using the Trypan blue dye exclusion assay. RESULTS: The cells grew as multiple layers on both Ti-6Al-7Nb substrates, showing extracellular matrix only when grown on HA-coated Ti-6Al-7Nb disks. The cells grown on HA-coated Ti-6Al-7Nb grew more slowly than the other 2 groups, with significantly smaller cell numbers than control cultures at the end of the experimental time. Additionally, the HA coated Ti-6Al-7Nb group presented smaller percentage of cell viability when compared to the control group. However, no significant differences were observed between the Ti groups. CONCLUSIONS: The presence of HA on the Ti-6Al-7Nb surface impaired the cell growth and viability of osteo-1 cells. However, this coating improved the extracellular matrix formation. Thus, our cell viability and structural studies showed that Ti-6Al-7Nb with or without HA coating has relevant physical and biological properties as an implant material.