980 nm半导体激光照射对种植体表面结构和温度影响的实验研究

目的:980 nm半导体激光可用于种植体周围软组织处理和种植体周围炎治疗,但其对种植体的作用尚未完全被了解。本文研究980 nm半导体激光照射对种植体表面结构和温度变化的影响,以期为临床使用参数设置和操作方法提供依据。方法:980 nm半导体激光照射纯钛圆盘试件,扫描电镜观察钛盘表面结构变化,热电偶检测钛盘温度变化范围和到达骨组织损伤温度升高阈值(10℃)的照射时长。结果:实验全部参数设置下980 nm半导体激光照射,钛盘表面结构均无明显改变。脉冲模式输出功率1 W,移动照射20 s钛盘温度上升19.8℃;经过10.2 s钛盘温度升高可达10℃。连续输出模式或增加输出功率,钛盘温度显著上升,并可在数秒内超过10℃。结论:980 nm半导体激光照射对种植体表面结构无损伤,但存在种植体温度升高造成周围组织热损伤的风险。     

不同烧结温度的羟基磷灰石涂层种植体——骨界面的定量组织学研究

本项研究以成年杂种犬为实验对象,以纯钛种植体为对照,用计算机辅助图象处理的定理组织学方法研究不同烧结温度下HA涂层种植体植入狗股骨后,界面的骨性结合水平。结果表明:涂层种植体在植入一个月后,就能达到较高的骨性结合水平,三个月内继续增高,并稳定在较高的水平,其中以HA2涂层(烧结温度1050℃)种植体优于HAl涂层(烧结为温度为850℃)。纯钛种植体亦能形成较好的骨性结合,但其形成骨性结合的速度和程度均低于涂层种植体。     

Ti2448合金种植体表面不同纳米管径生物活性膜对成骨细胞早期黏附的影响

观察Ti2448合金表面不同纳米管径生物活性膜对成骨细胞早期黏附的影响,筛选出能够抵抗细菌在种植体表面黏附和定植的最适管径,为研制开发具有抗菌性能的种植体提供实验基础。采用阳极氧化法,设定不同氧化电压,在Ti2448合金表面生成具有不同纳米管径的生物活性膜。通过MTT实验检测不同管径纳米管表面在不同时间点黏附的MG-63细胞数量差异;通过免疫荧光法观察接种24h后不同管径纳米管表面黏附细胞形态学差异。结果显示,细胞接种3h和24h后,30nm组的OD值显著高于对照组及其他实验组(P0.05);随着纳米管径的增加,细胞骨架蛋白的铺展范围逐渐减小,细胞由多角形渐变为梭形、椭圆形、圆形。研究结果表明,30nm管径生物活性膜更有利于成骨细胞的早期黏附,可能更能抵抗细菌在钛种植体上的黏附与定植。     

不同处理方法对钛瓷结合强度影响的研究

目的:观察微弧氧化表面处理方法在钛瓷结合方面的作用。方法:将试样分为三组,微弧氧化组,预氧化组,光滑组:对各组钛试件表面进行瓷粉烧结。根据ISO 9693标准,对钛瓷间的三点弯曲结合强度进行测试,并对钛瓷结合界面和瓷剥脱面进行SEM和EDX观察与分析。结果:处理方法不同,钛表面形貌及相结构也不同。钛试样微弧氧化组,预氧化组,光滑组与瓷的结合强度分别是:42.40±4.35Mpa,34.28±2.84Mpa和28.58±2.74Mpa,微弧氧化组的钛瓷结合强度与不进行微弧氧化处理组的钛瓷结合强度相比在统计学上有显著差异(P<0.05);预氧化组的钛瓷结合强度大于光滑组(P<0.05),但明显小于微弧氧化组(P<0.01);未进行微弧氧化处理组的钛瓷界面间可见有明显裂隙;而微弧氧化组的钛瓷界面瓷与钛基体结合紧密,无任何气泡、孔隙存在。结论:钛表面微弧氧化处理后可有效提高钛瓷的结合强度。     

牙龈组织与钛骨内牙种植体的附着

牙龈与牙种植体颈部的附着是影响牙种植体长期功效的重要因素之一,钛是一种非常理想的牙种植生物材料,在临床上得到较广泛的应用。本文就牙龈组织与种植体钛面的附着、附着机理、种植体钛面的性质、结构对龈附着的影响等研究现状作一综述。     

钛- 载药纳米复合材料的组装及其性质的研究

目的:构建一种能结合钛表面的载药纳米粒及钛-载药纳米复合材料的组装和性质研究。方法:(1)多巴胺修饰的非离子表面活性剂多巴胺-泊洛沙姆188(Dop-Poloxamer188)的合成和检测;(2)Dop-Poloxamer188作为表面活性剂、PLGA作为油相基质,制备纳米粒及纳米粒载药和表征;(3)钛片的预处理及钛片与修饰后的纳米粒的结合;(4)纳米粒修饰后的钛表面的表征。结果:新合成的Dop-Poloxamer188在285 nm左右有紫外吸收峰,说明多巴胺成功的修饰在Poloxamer188的两端;Dop-Poloxamer188能和PLGA制备出很好的纳米粒,平均粒径在110 nm左右,PDI小于0.1;多巴胺修饰的纳米粒与钛片通过简单的浸渍过程结合后,通过水接触角、场发射扫面电镜(Fe-SEM)、荧光显微镜、X射线光电子能谱(XPS)等仪器检测都显示多巴胺修饰的纳米粒成功且牢固的修饰在钛片表面。结论:成功达到钛表面的载药纳米粒修饰的目的,为钛种植体的载药系统提供了新的思路和方法。     

微钛钉种植体治疗上前牙前突的临床研究

目的应用微钛钉种植体治疗上前牙前突,评价微钛钉种植体增强磨牙支抗的安全性、有效性。方法选择20例患者,上牙列前牙区牙齿覆盖在Ⅱ°以上,平均覆盖为8.3 mm。应用微钛钉种植体,植入上颌第一恒磨牙和第二前磨牙颊侧根尖之间的牙槽间隔,以每侧200g力滑动内收前牙。通过比较矫治前后头颅侧位片分析其治疗的有效性。结果 20例牙弓突度均得到明显改善,切牙切缘平均内收6.8mm,支抗磨牙平均前移0.6 mm,磨牙在前后向和垂直向均未发生显著性移动,均无显著性扭转,种植体保持稳定。结论微钛钉种植体能有效治疗上颌前牙前突,起到磨牙强支抗的效能。     

口腔种植的新型钛材料生物安全性研究

目的:在动物体内进行新型钛材料的生物安全性研究。方法:采用TAI纯钛板材(Ti≥99.9%,宝鸡英耐特有色金属有限公司),通过对钛材料的喷砂、酸蚀、碱热等处理后,构建微纳双级骨仿生结构。对处理材料进行扫描电镜观察后,再经过兔皮下实验、小鼠急性溶血性实验确定此种材料的体内安全性。结果:1.扫描电镜结果显示:经过喷砂酸蚀碱热处理后的实验组具有微纳复合结构,与骨组织表面形态更为接近,从仿生学出发,更利于骨组织愈合与抑制骨组织周围炎的发生。2.皮下植入实验结果:经过喷砂酸蚀碱热处理后的实验组与只经过喷砂酸蚀的对照组对生物组织无明显刺激,未见明显的周围组织炎。3.急性毒性实验:未见经过喷砂酸蚀碱热处理后的实验组与只经过喷砂酸蚀的对照组出现明显的生物学反应,未见明显体重下降。结论:新型表面处理钛材料在生物安全性的动物体外实验方面无异常,可以进行下一步实验。     

口腔种植材料表面特性与生物膜形成研究进展

人类口腔环境为微生物提供了适宜生存的条件,多种微生物在牙齿表面形成了由基质包裹相互粘附的口腔生物膜,口腔生物膜是口腔微生物生存、代谢和致病的基础。随着1965年Brnemark种植体在临床上的成功应用,种植相关材料周围致病菌导致的种植体周围炎成为种植修复最常见的并发症之一,影响种植修复的远期效果。种植体周围炎引起了许多关注,并且口腔种植材料表面的特性和口腔生物膜的形成密切相关。本文就种植材料及天然牙齿表面的生物膜形成、种植材料表面特性对口腔生物膜及细菌粘附的影响因素、增强种植材料抗菌性能的方法以及未来的研究方向等作一综述。     

PHBHHx膜表面亲水改性及其与神经干细胞生物相容性

对羟基丁酸-羟基己酸共聚酯(PHBHHx)膜进行表面改性,研究神经干细胞(NSCs)在改性后的PHBHHx膜表面的贴附、增殖及分化情况,为开发新型脑组织工程支架材料奠定基础。采用溶剂挥发法制备PHBHHx膜,扫描电镜观察其表面性状;分别通过脂肪酶处理,NaOH处理的方法对PHBHHx膜进行表面改性,测量接触角以检测膜表面亲水性。分离培养孕14.5 d大鼠胚胎大脑皮质NSCs,接种在表面改性后的PHBHHx膜表面进行体外培养,扫描电镜观察膜表面细胞形态,MTT法检测细胞活力,免疫细胞化学染色观察NSCs存活和分化情况。结果显示,与未处理的PHBHHx膜相比,脂肪酶、NaOH处理能够显著提高PHBHHx膜表面亲水性,增加NSCs在PHBHHx膜表面贴附数量;NSCs在改性后的PHBHHx膜表面能够良好地存活并分化为神经元和胶质细胞。结果提示PHBHHx膜表面碱处理通过提高材料表面亲水性和粗糙程度,增加其与NSCs的生物相容性,改性后的PHBHHx材料是一种非常有潜力的新型脑组织工程支架材料,有望在NSCs移植修复脑损伤中发挥作用。     

Characterization of the properties of differentially modified titanium surfaces for dental implantology. 2: In vitro biocompatibility studies]

The aim of the present study was to determine whether specific surface modifications are capable of improving the biocompatibility of a titanium surface, and whether there is a correlation between the physico-chemical properties of the implant material and its biocompatibility. To this end, the properties of titanium surfaces were modified using various methods or the latter were coated with various materials. Plasma treatments under different atmospheric conditions (N2-plasma, SO2-plasma, acetylene plasma) as well as plasma polymerization were used to affect the biological response. Characterization of the physico-chemical surface properties by means of X-ray photoelectron spectroscopy (XPS), contact angle measurements and the calculation of surface tensions or surface energy provided important information on the interactions at the interface between the implant material and the aqueous environment. The influence of the respective surface modification on cell proliferation, cell viability and the activity of mitochondrial dehydrogenases was evaluated in specific in vitro tests with human gingiva fibroblasts. It was show that different modifications of the titanium samples induce different biological responses of the gingiva fibroblasts. The results confirm the existence of correlations between thermodynamic surface properties and cellular reactions under in vitro conditions.     

In vitro assays for adhesion and migration of osteoblastic cells (Saos-2) on titanium surfaces

The first event occurring at the boundary between a metal implant and living tissue is the attachment of cells onto the metal surface of the implant. The attachment characteristics of the metal in this situation are critical in determining its biocompatibility and usefulness as artificial bone and tooth implants. Using the human osteosarcoma cell line Saos-2, we attempted to establish simple and reliable methods for evaluating the attachment of cultured osteoblastic cells onto titanium samples that had been subjected to various surface treatments. Fluorescence actin imaging showed that cells cultured on titanium with hydrofluoric acid etching (HF-Ti) exhibited delayed spreading of their cytoplasm, as compared to cells cultured for the same length of time on nitrided titanium or physically polished titanium. The HF-Ti-cultured cells also exhibited poor assembly of focal contacts, as visualized by vinculin immunofluorescence. Furthermore, in motility assays based on an in vitro wound model, cells cultured on HF-Ti migrated more slowly than cells cultured on other titanium surfaces. These data suggest that Saos-2 cells attach less effectively to the HF-Ti surface. The methods described in this study should be useful for assessing the initial interactions of cultured cells with various materials, including metals.S.G. is the recipient of a grant awarded to foreign students by the government of Japan. This study was supported by the Integrated Center for Science (INCS) at Ehime University.     

Review: the potential impact of surface crystalline states of titanium for biomedical applications

In many biomedical applications, titanium forms an interface with tissues, which is crucial to ensure its long-term stability and safety. In order to exert control over this process, titanium implants have been treated with various methods that induce physicochemical changes at nano and microscales. In the past 20?years, most of the studies have been conducted to see the effect of topographical and physicochemical changes of titanium surface after surface treatments on cells behavior and bacteria adhesion. In this review, we will first briefly present some of these surface treatments either chemical or physical and we explain the biological responses to titanium with a specific focus on adverse immune reactions. More recently, a new trend has emerged in titanium surface science with a focus on the crystalline phase of titanium dioxide and the associated biological responses. In these recent studies, rutile and anatase are the major two polymorphs used for biomedical applications. In the second part of this review, we consider this emerging topic of the control of the crystalline phase of titanium and discuss its potential biological impacts. More in-depth analysis of treatment-related surface crystalline changes can significantly improve the control over titanium/host tissue interface and can result in considerable decreases in implant-related complications, which is currently a big burden on the healthcare system.     

Modification of the Micro Arc-oxidized Ti Surface for Implant Applications

Surface treatments applied to titanium and its alloys for implant applications are important for the development of bio properties.In this study,first an oxide layer was formed on the surface of the titanium plate by micro arc oxidation,and then both calcium phosphate and calcium phosphate/chitosan accumulation were performed for different samples by the sol-gel method.FE-SEM/EDS examinations,XRD,FTIR and thermal analysis were performed for these micro arc-oxidized,cal-cium phosphate-coated and calcium phosphate/chitosan-coated surfaces.The surface roughnesses for these surfaces were measured between 10 μm and 100 μm,suitable for bone development on the surface.The effect of chitosan addition on the calcium phosphate-coated surface on apatite formation ability and antibacterial properties was investigated.Although the addition of chitosan slows down the formation of apatite,it ensured that the coating had antibacterial properties.The calcium phosphate/chitosan biocomposite obtained can be recommended for dental and orthopedic implants.     

Cell cultivation on porous titanium implants with various structures

The paper presents data on the cultivation of human dermal fibroblasts and rabbit mesenchymal stromal cells on two types of porous titanium implants, i.e., those with irregular pores formed by pressed titanium particles and those with regular pores formed by the cohesion of one-size titanium particles inside the implant. The goal of this study was to determine what type of titanium implant porosity ensured its strongest interaction with cells. Cells were cultivated on implants for 7 days. During this period, they formed a confluent monolayer on the implant surface. Cells grown on titanium implants were monitored by scanning electron microscopy. Fibroblasts interaction with implants depended on the implant porosity structure. On implants with irregular pores cells were more spread. On implants with regular pores fibroblasts enveloped particles and were only occasionally bound with neighboring particles by small outgrowths. There was no tight interaction of particles inside the implant. In implants formed by pressed particles, cells grow not only on surface, but also in the depth of the implant. Thus, we suppose that a tighter interaction of cells with the titanium implant and, supposedly, tissues with the implant in the organism will take place in the variant when the implant structure is formed by pressed titanium particles, i.e., cellular interaction was observed inside the implant. In implants with irregular pores, cells grew both on the surface and in the depth. Thus, cells exhibited more adequate interactions with irregular pore titanium implants in vitro and hopefully the same interaction will be true in tissues after the implantation of the prosthesis into the organism.     

Matrix-assisted laser desorption ionization mass spectrometry: a new tool for probing interactions between proteins and metal surfaces. Use in dental implantology.

The fixation in the bone of an artificial titanium tooth root is believed to be initiated by the rapid adsorption of the proteins present in the surgical cavity on the titanium surface. The study of this adsorption should make it possible to predict the osseointegration capacities of new implant surface treatments. We describe here a new method, based on matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS), for quantifying proteins adsorbed on titanium surfaces fully identical to these designed for implantology. The key step of this method is a new MALDI-MS sample preparation allowing the adsorbed proteins to be removed from the surface and to be homogeneously dispersed in the matrix crystals. The adsorption of a model protein (lysozyme) on two titanium surfaces (polished and sandblasted) was studied in order to evaluate the method. The absolute MALDI-MS intensity was shown to vary linearly with the amount of adsorbed lysozyme. After dipping the titanium surfaces for different times in lysozyme solutions at different concentrations, the maximum amount of adsorbed lysozyme was measured by MALDI-MS and was shown to correspond to a lysozyme monolayer, which is consistent with results described in the literature.     

Micro-plasma textured Ti-implant surfaces

The surface state of titanium implants modulates bone response and implant anchorage. This evidence brought implant manufacturers to switch from the standard surface refinements and implement new surface treatments for more bone apposition and enhanced interfacial strength measured by removal torque or push-out tests. Anodic plasma-chemical treatment of implant surfaces is a cost-effective process to modify surface topography and chemistry. This technique is used for structuring connected with a coating of implant surfaces. The aim of our investigations, here, is to texture the implant surface in the nanoscale without coating. Ti disks with different mechanical pre-treatment (grinded, glass blasted) were used as substrate. Micro-plasma texturing was carried out in an aqueous electrolyte. By applying a pulsed DC voltage to the specimen, micro-plasma discharge was generated in the thin steam film between immersed specimen and electrolyte. The electrical process parameter current density was varied. The micro-plasma textured Ti surfaces were characterised optically by SEM and electrochemically by CV- (for testing the corrosion parameters), CA- (to give the enlargement of the real surface) and EIS-measurement in range of 100 kHz-100 microHz. We found that the initial structure of the material surface has small or no influence on the results of the micro-plasma treatment. The properties of the thick oxide layer resulting from the plasma process are influenced by electrical process parameters. After removal of the thick oxide layer a fine, micro- and nanoscaled surface structure of the titanium remains.     

Evaluation of a new laser surface implant: scanning electron microscopy/energy dispersive X-ray and X-ray photoelectron spectroscopy analyses

The roughness and the purity of implant surfaces are key points in the osteointegration process. The surfaces obtained by classic methods present irregular non-reproducible patterns and furthermore contaminate the implant surface with materials other than titanium which interfere with the process of osteointegration. The aim of the present study is to evaluate, by SEM/EDX and XPS analyses, the surface microstructure and the purity of new laser-treated implant surfaces. The laser treatment of the surface allows to set parameters to determine the roughness in order to obtain a regular and repeatable surface. Furthermore, there being no contact between the implant and the machine, there is no surface contamination with elements other than titanium. In this study we used a diode-pumped solid state laser (DPSS) with Nd:YAG source operating in Q-Switching mode on titanium samples. The resulting samples were analysed by SEM/EDX and XPS to evaluate morphology and purity of the surface. The results show surfaces with very regular roughness and a total absence of contamination.     

Cells,growth factors and bioactive surface properties in a mechanobiological model of implant healing

Interface conditions are of prime importance for implant fixation in the early post-operative period and modelling of specific biochemical interactions at implant surface is still missing. We hypothesized that updating osteoblast adhesion properties and growth factor source in an active zone located at the implant surface was relevant to model biochemical interactions of implant with its environment. We proposed an innovative set of diffusive–convective–reactive equations which relevant parameters were the cell decay factor, the cell motility and the growth factor balance.Initial comparison with histomorphometic results from a stable PMMA canine implant model provided an encouraging base to implement a numerical sensitivity analysis to evaluate the role of three types of bioactive surfaces: acid-etched titanium, coarse grit-blasted acid-etched titanium and coarse grit-blasted acid-etched titanium with RGDS peptide. We found that cell diffusion decrease (acid-etched+RGDS peptide vs. PMMA), and increase of local growth factor fraction (PMMA vs. acid-etched+RGDS peptide), significantly improved the amount of mineralized tissue on the implant surface. When the variation of structural fraction to cell motility and growth factor synthesis was investigated, an envelope pattern with an optimum was obtained but this could be exceeded for strong surface modifications and/or for high growth factor concentrations. The model also confirmed that implant bioactive properties should play a limited role to reduce heterogeneity of new-formed tissue. In conclusion, we suggested that our innovative theoretical approach was relevant to investigate implant fixation and could potentially help in reduction of implant revision.     

Surface Functionalization of Orthopedic Titanium Implants with Bone Sialoprotein

Orthopedic implant failure due to aseptic loosening and mechanical instability remains a major problem in total joint replacement. Improving osseointegration at the bone-implant interface may reduce micromotion and loosening. Bone sialoprotein (BSP) has been shown to enhance bone formation when coated onto titanium femoral implants and in rat calvarial defect models. However, the most appropriate method of BSP coating, the necessary level of BSP coating, and the effect of BSP coating on cell behavior remain largely unknown. In this study, BSP was covalently coupled to titanium surfaces via an aminosilane linker (APTES), and its properties were compared to BSP applied to titanium via physisorption and untreated titanium. Cell functions were examined using primary human osteoblasts (hOBs) and L929 mouse fibroblasts. Gene expression of specific bone turnover markers at the RNA level was detected at different intervals. Cell adhesion to titanium surfaces treated with BSP via physisorption was not significantly different from that of untreated titanium at any time point, whereas BSP application via covalent coupling caused reduced cell adhesion during the first few hours in culture. Cell migration was increased on titanium disks that were treated with higher concentrations of BSP solution, independent of the coating method. During the early phases of hOB proliferation, a suppressive effect of BSP was observed independent of its concentration, particularly when BSP was applied to the titanium surface via physisorption. Although alkaline phosphatase activity was reduced in the BSP-coated titanium groups after 4 days in culture, increased calcium deposition was observed after 21 days. In particular, the gene expression level of RUNX2 was upregulated by BSP. The increase in calcium deposition and the stimulation of cell differentiation induced by BSP highlight its potential as a surface modifier that could enhance the osseointegration of orthopedic implants. Both physisorption and covalent coupling of BSP are similarly effective, feasible methods, although a higher BSP concentration is recommended.