Morphological behavior of osteoblast-like cells on surface-modified titanium in vitro.

In recent papers, we reported the results of a study on the graded porous titanium coatings on titanium by plasma spraying and amino-group ion implantation. The paper is to preliminarily evaluate the biocompatibility of surface-modified titanium through 2, 5 and 7 days cell culture in vitro. Cell morphology was observed by a scanning electron microscope. Cell proliferation and type I collagen synthesis were measured by 3(4.5-dimethyl-thiazole-2-yl)2,5-diphenyl tetrazolium bromide (MTT) and enzyme-linked immunosorbent assay (ELISA), respectively. Our experimental results showed that osteoblast-like cells attached and spread well on surface-modified titanium. Cells were observed to grow into the pores and form extracellular matrix. MTT and ELISA results showed no detrimental effect on the development of cell. These studies support the biocompatibility of surface-modified titanium.     

Biomimetic coating of compound titania and hydroxyapatite on titanium

The modification on the titanium implant surface is an effective method to improve the biocompatibility of titanium. This article describes efforts to improve implant biocompatibility by applying titania and hydroxyapatite to form a three-layer coating on the titanium surface. This three-layer coating is made up of HA as the top layer (formed by hydrothermal treatment), porous TiO2 as the middle layer (formed by micro-arc oxidation) and a dense TiO2 film as the inner layer (formed by preanodic oxidation). The physicochemical characteristics, cell behavior and in vivo studies were assessed. The physicochemical characteristics were investigated using scanning electron micoscopy observation, fibronectin and laminin adsorption, corrosion test and X-ray diffraction analysis. Cell behavior included morphology observation with scanning electron microscopy (SEM), number count with methylthiazol tetrazolium (MTT) assay and Alkaline phosphatase (ALP, a representative enzyme of osteoblastic differentiation) activity of osteoblast-like MC3T3-E1 cells. In study in vivo the specimens were embedded in skull wound for repair. By the analysis of experiments, the titanium coated with this three-layer coating has been proved to have excellent corrosion resistance and good biocompatibility, which can promote cell proliferation and bone formation. So this modified titanium is an improved alternative to untreated titanium for bone repair applications.     

Biocompatibility of supercritical CO2-treated titanium implants in a rat model

Supercritical phase CO2 is a promising method for sterilizing implantable devices and tissue grafts. The goal of this study is to evaluate the biocompatibility of titanium implants sterilized by supercritical phase CO2 in a rat subcutaneous implantation model. At 5 weeks post implantation titanium implants sterilized by supercritical phase CO2 produce a soft tissue reaction that is comparable to other methods of sterilization (steam autoclave, ultraviolet light radiation, ethylene oxide gas, and radio-frequency glow-discharge), as indicated by the thickness and density of the foreign body capsule, although there were some differences on the capillary density. Overall the soft tissue response to the implants was similar among all methods of sterilization, indicating supercritical phase CO2 treatment did not compromise the biocompatibility of the titanium implant.     

Surface engineering of titanium thin films with silk fibroin via layer-by-layer technique and its effects on osteoblast growth behavior

The objective of the present study was to surface modify the titanium thin films to improve its biocompatibility. A layer-by-layer (LBL) self-assembly technique, based on the electrostatic interactions mediated adsorption of chitosan (Chi) and silk fibroin (SF), was used leading to the formation of multilayers on the titanium thin film surfaces. The surface chemistry and wettability of LBL films were investigated by X-ray photoelectron spectroscopy (XPS), water contact angle measurement, and atomic force microscopy, respectively. XPS and contact angle measurement results indicated that a full SF/Chi pair film was formed after the deposition layers of PEI/(SF/Chi)(2) on the titanium film surfaces. The topographies of multilayered films were directly related to the corresponding outmost layer components. The build-up of such SF/Chi pair films on titanium films may in turn affect the biocompatibility of the modified titanium films. Therefore, an in vitro investigation was performed to confirm this hypothesis. Cell proliferation, cell viability, DNA synthesis as well as differentiation function (alkaline phosphatase) of osteoblasts on LBL-modified titanium films and control samples were investigated, respectively. Osteoblasts cultured on modified titanium films was found to be higher proliferation tendency than that on control (p < 0.05). Cell viability, alkaline phosphatase as well as DNA synthesis measurement indicated that osteoblasts on LBL-modified films were greater (p < 0.05 or p < 0.01) than the control, respectively. These results suggest that surface engineering of titanium was successfully achieved via LBL deposition of Chi/SF pairs, and enhanced its cell biocompatibility. The approach presented in the study may be exploited as an efficient alternative for surface engineering of titanium-based implants.     

Easy assessment of the biocompatibility of Ni-Ti alloys by in vitro cell culture experiments on a functionally graded Ni-NiTi-Ti material

The biocompatibility of nickel-titanium alloys was investigated by single-culture experiments on functionally graded samples with a stepwise change in composition from pure nickel to pure titanium, including an Ni-Ti shape memory alloy for a 50:50 mixture. This approach permitted a considerable decrease of experimental resources by simultaneously studying a full variation of composition. The results indicate a good biocompatibility for a nickel content up to about 50%. The cells used in the biocompatibility studies comprised osteoblast-like osteosarcoma cells (SAOS-2, MG-63), primary human osteoblasts (HOB), and murine fibroblasts (3T3).     

Development of the DexAide right ventricular assist device inflow cannula

Cannula design and cannulation site can pose major limitations to chronic pump implantations in animal studies. The aim of this study was to evaluate the biocompatibility of various inflow cannula designs for the DexAide right ventricular assist device (RVAD). The DexAide RVAD was implanted for intended durations of 14, 30, or 90 days in 19 animals (mean 20 +/- 11 days). Seven inflow cannula designs were evaluated: angled titanium conduit with caged tip (two cases); flexible polyurethane coated polyvinyl chloride (PVC) tube (one case); open ended titanium (one case); a titanium cannula with a flange (six cases); a cannula with a gelatin coated flange (five cases); a cannula with an angled flange (one case); and open ended titanium with two side holes (three cases). The open ended titanium inflow cannula with two side holes positioned through the diaphragmatic surface of the right ventricle (RV) via a right thoracotomy showed good biocompatibility for the chronic animal study. Other cannulae inserted into the infundibular portion of the RV via a left thoracotomy showed significant depositions. Gelatin coated inflow cannula had the advantage to prevent tissue growth around the inflow cannula. The DexAide RVAD pump itself showed good biocompatibility, although nonadherent depositions originating from the inflow cannulae were captured onto the primary impeller blades.     

Effect of calcium and phosphorus ion implantation on the corrosion resistance and biocompatibility of titanium

This paper is concerned with the corrosion resistance and biocompatibility of titanium after surface modification by the ion implantation of calcium or phosphorus or calcium + phosphorus. Calcium and phosphorus ions were implanted in a dose of 10(17) ions/cm(2). The ion beam energy was 25 keV. The microstructure of the implanted layers was examined by TEM. The chemical composition of the surface layers was determined by XPS and SIMS. The corrosion resistance was examined by electrochemical methods in a simulated body fluid (SBF) at a temperature of 37 degrees C. The biocompatibility was evaluated in vitro. As shown by TEM results, the surface layers formed during calcium, phosphorus and calcium + phosphorus implantation were amorphous. The results of the electrochemical examinations (Stern's method) indicate that the calcium, phosphorus and calcium + phosphorus implantation into the surface of titanium increases its corrosion resistance in stationary conditions after short- and long-term exposures in SBF. Potentiodynamic tests show that the calcium-implanted samples undergo pitting corrosion during anodic polarisation. The breakdown potentials measured are high (2.5 to 3 V). The good biocompatibility of all the investigated materials was confirmed under the specific conditions of the applied examination, although, in the case of calcium implanted titanium it was not as good as that of non-implanted titanium.     

钛合金内固定材料修复脊柱结核：生物相容性及力学性能

BACKGROUND: Currently, surgical implant fixation is mainly applied for spinal tuberculosis. How to choose implant materials, however, is still under discussion. OBJECTIVE: To compare the biocompatibility and mechanical properties of titanium alloy and stainless steel for rabbit spinal tuberculosis. METHODS: Thirty rabbits were chosen to prepare spinal tuberculosis models. Then, the rabbits were equivalently randomized into two groups, which underwent implant fixation with stainless steel or titanium alloy, respectively. At 30 days after implantation, biocompatibility and biomechanical properties of the two materials in the repaired region of spine were observed and detected, respectively. RESULTS AND CONCLUSION: In view of the biocompatibility, infection and immunological rejection could not been found in the titanium alloy group; in contrast, infection appeared in three rabbits of the stainless steel group. Flexion, extension and lateral bending displacements under the spinal loading in the titanium alloy group were significantly less than those in the stainless steel group (P < 0.05); axial pull-out strength in the titanium alloy group was significantly higher than that in the stainless steel group (P < 0.05); flexion, extension, lateral bending and axial compression in the titanium alloy group were significantly greater than those in the stainless steel group (P < 0.05). In conclusion, titanium alloy material has good biocompatibility that can be used to restore and maintain the spinal stability.     

两种钛合金成品表面特性表征及血液相容性的对比研究

目的对两种不同加工工艺生产的钛合金成品(白色和黑色)进行表面特性鉴定,并评价其血液相容性,为选择合适的工艺流程提供理论依据。方法使用扫描电子显微镜(scanning electron microscopy,SEM)、能谱分析(energy dispersive spectroscopy,EDS)、原子力显微镜(atomic force microscopy,AFM)、X射线衍射仪(X-ray diffraction,XRD)、X射线光电子能谱仪(X-ray photoelectron spectrometer,XPS)对两种钛合金的表面结构、表面物理和化学性能进行表征。通过体外纤维蛋白原吸附、血小板黏附与激活、凝血时间的测定等实验,以及植入狗右心房10 d观察血栓形成情况,系统评价两种钛合金的血液相容性。结果白色钛合金表面为粗且深的犁沟,成分为Ti、Al、V,黑色钛合金表面呈孔样层状叠加分布,且成分除了Ti、Al、V外,还有Si、Na、Ca、Fe等杂质。XRD结果显示两种钛合金物相结构均为钛。XPS结果显示两种钛合金表层都形成极薄的TiO_2薄膜,但黑色钛合金形成的TiO_2薄膜略厚于白色钛合金。体外纤维蛋白原吸附、血小板黏附与激活及凝血时间结果均显示两种钛合金血液相容性无明显差异。体内植入实验结果显示两种钛合金表面均形成了血栓,但黑色钛合金有明显异物反应,形成了增生的肉芽组织,且电镜下显示,白色钛合金表面黏附的有形成分少于黑色钛合金。结论结合体内、体外实验,白色钛合金的生物相容性优于黑色钛合金。     

钛合金与不锈钢内固定材料置入修复脊柱结核的生物相容性

背景：钛合金和不锈钢是两种较为常用的内固定植入材料,但两种材料的治疗效果和生物相容性存在一定的差异。 目的：分析钛合金和不锈钢内固定材料置入治疗脊柱结核的效果及生物相容性。 方法：纳入脊柱结核患者71例,其中男35例,女36例,年龄17-81岁,35例进行钛合金内固定材料置入治疗,36例进行不锈钢内固定材料置入治疗。随访12个月,分析两组脊柱后凸畸形角度变化、治疗效果、Frankel脊髓损伤分级及材料生物相容性。 结果与结论：两组治疗前的脊柱后凸畸形角度、Frankel脊髓损伤分级比较差异无显著性意义,两组治疗后末次随访的脊柱后凸畸形角度、Frankel脊髓损伤分级均较治疗前明显改善(P < 0.05),但两组间比较差异无显著性意义;钛合金内固定组治愈34例(97%),不锈钢内固定组治愈33例(92%),两组间治愈率比较差异无显著性意义。两种材料均具有良好的生物相容性,未发生感染等不良反应。表明在脊柱结核治疗过程中,置入钛合金或不锈钢内固定材料均能获得良好的效果,并具有良好的生物相容性。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

Effects of TiO2-containing phosphate glasses on solubility and in vitro biocompatibility

Phosphate-based glasses with different amounts of titanium dioxide (TiO(2)), having the following molar composition 50P(2)O(5)-30CaO-9Na(2)O-3SiO(2)-3MgO-(5-x)K(2)O-xTiO(2), (where x = 0, 2.5, 5 mol %), were synthesised and characterized in terms of solubility (according to ISO 10993-14), and in vitro biocompatibility using human MG-63 osteoblast cells. Dissolution tests were carried out in Tris-HCl (pH 7.4) to simulate the physiological pH and in citric acid (pH 3.0) to simulate an acidic environment. The weight loss decreased with increasing TiO(2) content, a process further enhanced in acidic medium. TiO(2) reduced the pH changes usually caused by the dissolution products released. Cellular tests showed that all the glasses studied (0-5 mol % TiO(2)) and TiCl(4), used to investigate the biocompatibility of titanium ions, did not produce cytotoxic effects on human MG-63 osteoblasts for up to 5 days in culture. On the basis of these results, we suggest that TiO(2)-containing phosphate glasses could be promising substrates for bone tissue engineering applications.     

Titanate biomaterials with enhanced antiinflammatory properties

While titanium implants are generally recognized as having excellent biocompatibility, the mechanistic basis for this has yet to be established. We previously demonstrated that TiO2, found on surfaces of titanium, has antioxidant properties that degrade the reactive oxygen species (ROS) which mediate the inflammatory response. We hypothesized that the antioxidant mechanism was similar to that known to mediate photocatalysis by titanium oxides. Specifically, we investigated whether the electronic or valence state of the surface titanium atoms mediates the catalytic degradation of ROS. Surface Ti(IV) atoms in TiO2 and SrTiO3 single crystal substrates were converted into Ti(III) while maintaining the bulk crystalline structure by vacuum annealing or Niobium doping. The degradation of both chemically-induced and neutrophil-derived ROS were significantly increased by changing the valence state of surface titanium. These results suggest that titanium-mediated degradation of ROS is through a catalytic mechanism. Furthermore, we describe a series of novel biomaterials that have antioxidant properties superior to those of titanium.     

Development of a new system for evaluating the biocompatibility of implant materials using an osteogenic cell line (MC3T3-E1)

A new culture system was developed to clarify the biocompatibility of implant materials with bone tissue using the MC3T3-E1 osteogenic cell line. The cells were inoculated onto specimens such as aluminium oxide, titanium, dental casting silver-palladium alloy (PD), and a plastic coverslip. To study the effects of these materials on cell growth, differentiation, and calcification, DNA and protein content, alkaline phosphatase activity, and calcium content, respectively, were determined. The results from biochemical analysis suggest titanium and aluminum oxide to have adequate biocompatibility, while PD has an irritant effect on cell metabolism. It is clear that an objective view of the differentiation and calcification processes of osteogenic cells can be understood through such analysis. From the results of this study, our culture system appears suitable for evaluating the biocompatibility of implant materials with bone tissue.     

不同材料修复颅骨缺损的比较

BACKGROUND: Currently, different methods materials used for craniocerebral injury and skull defect repair have their own clinical efficacy and adverse reactions. OBJECTIVE: To compare the clinical effects of autologous bone, allograft bone, porous tricalcium phosphate and titanium mesh for skull defects. METHODS: 200 patients with skull defects were selected, including 120 males and 80 females, aged from 18-76 years old, and then were allotted into four groups by random number table (n=50 per group), undergoing repair with autologous bone, allograft bone, porous tricalcium phosphate and titanium mesh, respectively. The postoperative complications were compared among groups, and osseous healing properties were observed through the follow-up. RESULTS AND CONCLUSION:The healing property of autologous bone, allograft bone and titanium mesh was significantly superior to that of porous tricalcium phosphate (P < 0.05), but there were no significant differences amorg the autologous bone, allograft bone and titanium mesh groups. In addition, the incidence of complications of autologous bone and titanium mesh goup was significantly less than that of porous tricalcium phosphate and allograft bone groups (P < 0.05), and comparison between the latter two groups showed no significant difference. In conclusion, autologous bone and titanium mesh for skull defect repair have good biocompatibility, and can promote bone healing.     

Biocompatibility of electrophoretical deposition of nanostructured hydroxyapatite coating on roughen titanium surface: in vitro evaluation using mesenchymal stem cells

A nano hydroxyapatite (HAp) layer was coated on a roughen titanium surface by means of electrophoretic deposition with an acetic anhydride solvent system. The objectives of this current study are to investigate whether nano-HAp can improve mechanical strength at a lower sintering temperature and biocompatibility. Densification temperature was lowered from usual 1000 to 800 degrees C. The coating interfacial bonding strength, phase purity, microstructure, and biocompatibility were investigated. Degradation of HA phase was not detected in XRD. A porous TiO2 layer acts as a gradient coating layer with an intermediate thermal expansion coefficient between hydroxyapatite and titanium that reduces the thermal stress. From SEM image, the coating does not contain any crack. Mesenchymal stem cell (MSC) is the progenitor cell for various tissues in mature animals, which can improve integration of bone tissue into implant. In this in vitro study, rabbit MSCs culture indicated that the HAp/Ti nanocomposite biomaterial had good biocompatibility and bioactivity. Around materials and on its surface cell grew well with good morphology. Proliferation of the MSCs on the nano-HAp coating was higher than its micron counterpart in XTT assay. These properties show potential for the orthopaedic and dental applications.     

Polysaccharide-protein surface modification of titanium via a layer-by-layer technique: characterization and cell behaviour aspects

To improve the surface biocompatibility of titanium films, a layer-by-layer (LBL) self-assembly technique, based on the polyelectrolyte-mediated electrostatic adsorption of chitosan (Chi) and gelatin (Gel), was used leading to the formation of multilayers on the titanium thin film surfaces. The film growth was initialized by deposition of one layer of positively charged poly(ethylene imine) (PEI). Then the thin film was formed by the alternate deposition of negatively charged Gel and positively charged Chi utilizing electrostatic interactions. The LBL film growth was monitored by several techniques. The chemical composition, surface topography as well as wettability were investigated by using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), confocal laser scanning microscopy (CLSM) and water contact angle measurement, respectively. Quantitative XPS analysis showed the alternative change of C/N ratio after four sequential cycles coating of Ti/PEI/Gel/Chi/Gel, which indicated the discrete layer structure of coatings. Uncoated titanium (control sample) displayed a smooth surface morphology (root mean square (RMS) roughness was around 2.5 nm). A full coverage of coating with Gel/Chi layers was achieved on the titanium surface only after the deposition layers of PEI/(Gel/Chi)2. The PEI/Gel/(Chi/Gel)3 layer displayed a rough surface morphology with a tree-like structure (RMS roughness is around 82 nm). These results showed that titanium films could be modified with Chi/Gel which may affect the biocompatibility of the modified titanium films. To confirm this hypothesis, cell proliferation and cell viability of osteoblasts on LBL-modified titanium films as well as control samples were investigated in vitro. The proliferation of osteoblasts on modified titanium films was found to be greater than that on control (p<0.05) after 1 and 7 days culture, respectively. Cell viability measurement showed that the Chi/Gel-modified films have higher cell viability (p<0.05) than the control. These data suggest that Chi/Gel were successfully employed to surface engineer titanium via LBL technique, and enhanced its cell biocompatibility. The approach presented here may be exploited for fabrication of titanium-based implant surfaces.     

Effect of dual ion implantation of calcium and phosphorus on the properties of titanium

This study is concerned with the effect of dual implantation of calcium and phosphorus upon the structure, corrosion resistance and biocompatibility of titanium. The ions were implanted in sequence, first Ca and then P, both at a dose of 10(17) ions/cm2 at a beam energy of 25 keV. Transmission electron microscopy was used to investigate the microstructure of the implanted layer. The chemical composition of the implanted layer was examined by XPS and SIMS. The corrosion resistance was determined by electrochemical methods in a simulated body fluid (SBF) at a temperature of 37 degrees C. The biocompatibility tests were performed in vitro in a culture of human-derived bone cells (HDBC) in contact with the tested materials. The viability of the cells was determined by an XTT assay and their activity by the measurements of the alkaline phosphatase activity in contact with implanted and non-implanted titanium samples. The in vitro examinations confirmed that, under the conditions prevailing during the experiments, the biocompatibility of Ca + P ion-implanted titanium was satisfactory. TEM results show that the surface layer formed by the Ca + P implantation is amorphous. The corrosion resistance of titanium, examined by the electrochemical methods, appeared to be increased after the Ca + P ion implantation.     

Osteogenic Potential of Porous Titanium. An Experimental Study in Sheep

The search for osteoinductive as well as osteoconductive materials has led to the novel idea of using titanium in bone augmentations of the alveolar crest. Due to its excellent biocompatibility and favorable osteogenic properties, highly porous TiO₂ granules has been proposed as a promising material for non-resorbable synthetic bone grafts in the restoration of large bone defects, and for bone augmentation in dental applications.ObjectivesThe aim of this study was to investigate the osteoconductive properties and biological performance of porous titanium granules used in osseous defects adjacent to the maxillary sinus in sheep. The experimental animal study involved 15 yearling sheep with a focus on the osteogenic potential of porous titanium used for subantral augmentation.Material and methodsCalibrated defects were prepared in the subantral region of sheep. The defects were randomized into tests and control group. The test defects were grafted with porous titanium granules (PTG), whereas control defects were left empty (sham). Defects were left for healing for 30, 60, and 90 days. After healing, the grafted areas were removed and finally osteoconductivity was analyzed by an orthopantograph (OPG} and histology.ResultsSignificantly more new bone formed in PTG grafted defects compared with sham. The control group showed significantly less expression of key inflammation cells, but with no significant difference in key inflammation cells compared with the experimental groups.ConclusionPorous titanium can offer as an effective alternative to calcium phosphate and bone collagen-based materials used for subantral augmentation of the maxillary bone in cases of dental implantation.     

Biocompatibility of beta-stabilizing elements of titanium alloys

In comparison to the presently used alpha + beta titanium alloys for biomedical applications, beta-titanium alloys have many advantageous mechanical properties, such as an improved wear resistance, a high elasticity and an excellent cold and hot formability. This will promote their future increased application as materials for orthopaedic joint replacements. Not all elements with beta-stabilizing properties in titanium alloys are suitable for biomaterial applications-corrosion and wear processes cause a release of these alloying elements to the surrounding tissue. In this investigation, the biocompability of alloying elements for beta- and near beta-titanium alloys was tested in order to estimate their suitability for biomaterial components. Titanium (grade 2) and the implant steel X2CrNiMo18153 (AISI 316 L) were tested as reference materials. The investigation included the corrosion properties of the elements, proliferation, mitochondrial activity, cell morphology and the size of MC3T3-E1 cells and GM7373 cells after 7 days incubation in direct contact with polished slices of the metals. The statistical significance was considered by Weir-test and Lord-test (alpha = 0.05). The biocompatibility range of the investigated metals is (decreasing biocompatibility): niobium-tantalum, titanium, zirconium-aluminium-316 L-molybdenum.     

羟基磷灰石涂层钛合金材料生物相容性研究初探

目的探讨一种新型的代骨材料--羟基磷灰石涂层的钛合金材料的生物相容性。方法制备羟基磷灰石涂层钛合金材料浸提液后,采用细胞毒性实验以观察实验样品浸提液对L929小鼠成纤维细胞的毒性反应;通过对小鼠尾静脉及腹腔注射试验样品浸提液后,观察其对小鼠的急性全身毒性反应;Ames实验及迟发型超敏反应实验对其遗传毒性及致敏性进行安全性评价。结果羟基磷灰石涂层钛合金材料浸提液对L929小鼠成纤维细胞的相对增殖率(RGR)为96.9%,细胞毒性反应为1级,无细胞毒性反应;对小鼠亦无明显的急性全身毒性作用,实验样品组与阴性对照组动物体质量差异无统计学意义(P0.05);遗传毒性Ames实验表明,在活化与非活化条件下,该材料浸提液对鼠伤寒沙门氏菌株的回变菌落数与对照组比均未增加2倍,对该菌株无诱变性;迟发型超敏反应实验显示,该材料浸提液无潜在的皮肤接触致敏性。结论羟基磷灰石涂层的钛合金材料具有良好的生物相容性。