纯钛表面阳极氧化电压对成骨细胞早期黏附及伸展的影响

目的:通过成骨细胞离体培养试验,观察不同阳极氧化电压对成骨细胞早期黏附及伸展的影响。方法:分别在电压为140V、200V及260V,电流密度为70A/m2等条件下,在0.03mol/L甘油磷酸钙(Ca-GP)和0.15mol/L醋酸钙混合电解液中,对纯钛样本表面进行阳极氧化处理。对阳极氧化后样本的平均粗糙度进行测量,并在其表面进行人成骨细胞培养,对早期细胞黏附及伸展等情况进行研究。采用SPSS13.0forWindows统计分析软件中的单因素方差分析进行统计学处理。结果:纯钛表面平均粗糙度为0.17μm,经阳极氧化后,随电压升高,平均粗糙度分别为0.23μm、0.26μm及0.33μm,统计学分析表明有显著性差异(P<0.05)。经过2h细胞培养后,阳极氧化处理后,纯钛表面细胞骨架发生形态学改变,且随阳极氧化电压的增高,细胞形态不规则呈现升高的趋势。统计分析结果表明,260V电压阳极氧化组,细胞黏附数显著高于对照组(P<0.05)。结论:阳极氧化处理可改善纯钛表面特性,成骨细胞在其表面的黏附及伸展等早期细胞行为受阳极氧化电压的影响。     

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。     

人牙周膜成纤维细胞在微弧氧化陶瓷膜表面的生物相容性研究

目的　观察通过微弧氧化（MAO）和水热处理形成的陶瓷膜表面人牙周膜成纤维细胞（hPDLF细胞）的粘附和增殖情况,评价微弧氧化陶瓷膜表面的生物相容性及MAO工艺应用于钛植入材料表面处理的可行性。方法　纯钛试件分为3组,微弧氧化处理组（M组）、微弧氧化-水热处理组（M-H组）和纯钛对照组,每组各20枚。对不同时间点hPDLF细胞在材料表面的附着、生长、增殖情况以及ALP活性进行检测,并对实验结果进行统计分析。结果　陶瓷膜具有良好的生物相容性,培养5 d后,改性两组材料的表面细胞增殖比纯钛组及空白对照组明显增多,各组间细胞增殖情况存在统计学差异。同时培养4 d后,M组和M-H组细胞ALP活性大幅提高,各组间比较均有统计学差异。结论　MAO处理后的纯钛表面对hPDLF的生物相容性优于未处理组,增加水热处理工艺较一般MAO工艺可以更有效提高hPDLF细胞的早期粘附、增殖及ALP活性。     

纯钛微弧氧化-壳聚糖-海藻酸钠涂层的细胞相容性研究

目的:通过对不同涂层表面进行成骨细胞生物相容性检测,评价其生物相容性。方法:纯钛微弧氧化组为对照A组、纯钛微弧氧化-碱处理-壳聚糖复合处理为实验B组、纯钛微弧氧化-碱处理-壳聚糖-海藻酸钠复合处理为C组。通过CCK-8实验检测不同时间细胞黏附情况以及增殖情况,激光共聚焦检测早期细胞骨架形态,扫描电镜观察试件表面形貌,ALP检测其不同时间碱性磷酸酶活性。结果:激光共聚焦显示B组和C组表面细胞的生长和黏附均优于A组,C组优于B组。CCK-8和ALP的检测结果显示3组材料表面细胞的增殖、活性的顺序为C组>B组>A组,3组的比较差异均具有统计学意义。结论:3组材料均具有良好的生物相容性,纯钛微弧氧化--碱处理-壳聚糖-海藻酸钠复合处理涂层细胞相容性最佳。[关键词]纯钛成骨细胞壳聚糖海藻酸钠生物相容性     

成骨细胞在微弧氧化处理后纯钛表面的附着、增殖及ALP活性

目的 :对表面微弧氧化 (microarcoxidation ,MAO)处理后的纯钛材料进行成骨细胞生物相容性检测 ,评价改进的MAO工艺应用于钛植入材料表面处理的可能性。方法 :纯钛材料经过 2种MAO处理后 (MAO 1和MAO 2工艺 ) ,采用MC 3T3细胞系对不同时间点成骨细胞在材料表面的附着率、生长增殖情况以及ALP活性进行检测 ,以未经处理光滑纯钛表面作为对照 ,以SPSS对实验结果进行统计分析。结果 :早期 (0 .5h、1h)细胞附着率差异有统计学意义 ,MAO 1组 >MAO 2组 >纯钛组 ;2h后 ,MAO 1组与MAO 2组无差异 ,2组细胞附着率都显著高于纯钛组。细胞增殖及ALP活性测试中 ,MAO 1处理组在各时间点都显著高于另外 2组。结论 :MAO处理后的纯钛对成骨细胞的生物相容性优于未处理组 ,改进的MAO 1处理工艺较一般工艺可以更有效提高成骨细胞的早期粘附、增殖及ALP活性。     

种植体表面TiO2/HA梯度涂层的生物相容性研究

目的：通过观察大鼠成骨细胞（osteoblast,OB）在微弧氧化（micro-arc oxidation,MAO）和水热处理形成的TiO2/HA梯度涂层表面的粘附和增殖等情况,评价微弧氧化陶瓷膜表面的生物相容性及微弧氧化-水热处理工艺应用于钛植入材料表面处理的可行性。方法：将纯钛试件共60枚分为微弧氧化处理组（M组）、微弧氧化-水热处理组（M-H组）和纯钛对照组。采用扫描电镜（scanning electron microscopy,SEM）观察表面形貌并用X线衍射（X-ray diffraction,XRD）对其进行成分分析。然后对不同时间点OB细胞在各组材料表面的附着、生长、增殖情况以及碱性磷酸酶（alkaline phosphatase,ALP）活性进行检测,并对实验结果进行统计分析。结果：微弧氧化处理后试件表面呈现多孔状,主要为锐钛矿型TiO2,也包含金红石型TiO2。经过后续水热处理,从扫描电镜照片可以看到试件表面析出一层白色柱形结晶体,同时XRD谱线出现了羟基磷灰石的衍射峰。试件与OB细胞共培养后可见细胞在材料周围生长良好,培养5d后,改性两组的材料表面细胞增殖比纯钛组及空白对照组明显增多。培养7d后,M组和M-H组细胞ALP活性大幅提高,各组间比较均有统计学差异。结论：MAO及水热处理后的TiO2/HA梯度涂层表面能有效提高OB细胞的早期粘附、增殖及ALP活性。     

纯镁复合硅烷/植酸载锌膜层的细胞相容性实验研究

目的:制备纯镁复合硅烷/植酸载锌膜层,希望可以改善纯镁的耐腐蚀性及其细胞生物相容性。方法:制备纯镁阳极氧化、阳极氧化-硅烷处理、阳极氧化-硅烷/植酸双复合处理及硅烷/植酸双复合载Zn试样,采用扫描电镜观察涂层表面形貌,并进行分析元素,通过CCK-8检测细胞增殖和黏附能力,激光共聚焦检测细胞黏附形态。结果:纯镁阳极氧化表面存在微孔,复合处理起到封孔的作用,载不同浓度锌增加了细胞增殖和黏附能力,其中阳极氧化-硅烷/植酸双复合载Zn 20 g/L可防止细胞液浸入,生物相容性最佳。结论:纯镁阳极氧化-硅烷/植酸双复合载Zn 20 g/L膜层,细胞增长和形态铺展最好,明显促进了成骨细胞在材料表面的增殖、黏附能力及其生物功能表达。     

微弧氧化处理钛铌锆锡合金对成骨细胞生长影响的体外研究

目的:研究新型钛铌锆锡合金(Ti-24Nb-4Zr-7.9Sn,TNZS)表面经过微弧氧化(micro-arc oxidation,MAO)改性后对成骨细胞在其表面的附着计数,增殖特性及碱性磷酸酶(ALP)活性的影响.方法:以光滑纯钛表面作为对照,实验组为未作处理的钛铌锆锡合金和经微弧氧化处理的钛铌锆锡合金,然后对其粗糙度和表面能进行分析,并观察原代培养的成骨细胞在其表面的生长情况.结果:经MAO处理的TNZS表面粗糙度和表面能明显高于未处理TNZS和纯钛组;早期(0.5、1 h)细胞附着计数3组之间无明显差异;2h后,细胞在MAO- TNZS表面的附着数高于其他2组;细胞增殖及ALP活性测试中,MAO-TNZS处理组在培养7 d和10 d后显著高于未处理的TNZS组,而Ti组最低.结论:钛铌锆锡合金表面采用微弧氧化法处理后表面形貌及表面能发生改变,这种改变促进了细胞在其表面的早期附着及细胞的生长和功能表达.     

纯钛阳极氧化及表面薄层羟基磷灰石形成技术的研究 Ⅱ．电解质及电压对氧化膜性能的影响

目的：了解电解质和电压因素对氧化膜成分、形态和稳定性能的影响。方法：将纯钛片分别在六种不同浓度电解质中（β－磷酸甘油钠：醋酸钙＝0.01M:0.15M,0.02M:0.20M,0.03M:0.25M,0.04M:0.25M,0.05M:0.25M,0.06M:0.30M)和4种不同电压条件下（200V,250V,300V,350V）进行阳极氧化,观察表面氧化膜性能的改变。结果：随着电解质和电压的增加,钛表面氧化膜突起高度增加,孔隙,边缘逐渐不平滑,形状逐渐不规则。能谱分析表明,在形成近HA化学当量的6组电解质中,Ca/Ti比随电解质浓度的增加而增加。结论：根据HA的析出量和氧化膜的稳定性,确定β－磷酸甘油钠：醋酸钙=0.04M:0.25M为较佳电解质浓度比（Ca/Ti比为3.6);根据Ca/P比值,确定300V为最佳电解条件（Ca/P比为1．8）。     

纯钛微弧氧化-多巴胺-环型多肽涂层的细胞相容性研究

目的:探讨纯钛种植体表面加载环型多肽对成骨细胞生物行为的影响。方法:微弧氧化组为M组,微弧氧化加载多巴胺组为P组,微弧氧化加载多巴胺接枝环环肽组为R组。扫描电镜对3组钛片进行表面形貌分析,能谱仪分析3组试件元素成分。CCK-8检测成骨细胞不同时间点在3组的增殖黏附能力,激光共聚焦观察成骨细胞在3组涂层表面的铺展情况。结果:扫描电镜及能谱显示涂层加载成功,CCK-8显示R组的A值在各个时间点上都高于M组和P组,且各组比较具有统计学意义(P<0.05)。激光共聚焦观察R组细胞数目最多,伪足较多,细胞联系最为紧密。结论:纯钛表面加载环肽后能有效的促进成骨细胞早期的黏附及增殖。     

Improvement of in vitro titanium bioactivity by three different surface treatments.

OBJECTIVE: Dental implants are usually made from commercially pure titanium or titanium alloys. The aim of this investigation was to determine the influence of surface treatments of commercially pure titanium samples on in vitro bioactivity. METHODS: Commercially pure (cp) titanium (Ti) sheets were submitted to three different surface treatments, including, for all samples, etching with an HCl/H(2)SO(4) solution. Part of each etched sample was further submitted either to anodic oxidation by using an H(3)PO(4) solution or to thermal oxidation. Treated and non-treated samples were analyzed by using scanning electron microscopy (SEM), profilometry and photoelectron X-ray spectroscopy (XPS). The in vitro assessment was carried out through the immersion of samples in simulated body fluid (SBF). In vitro testing was carried out by SEM and by the determination of calcium (Ca) content in solution by atomic absorption spectrometry (AAS). The non-treated titanium samples were used as the control group. RESULTS: This study has shown that, after up to 7-day exposure, a calcium phosphate layer precipitated only on samples submitted to at least one of the three treatments used. This result, based on SEM images, is in good agreement with Ca content and XPS analysis, in which remarkable effects of surface modifications on Ti samples are highlighted. SIGNIFICANCE: These results suggest that suitable surface treatments, such as employed here, may improve in vitro titanium bioactivity in a SBF solution at 37 degrees C. This behavior suggests a possibility of a further favorable in vivo response.     

微弧氧化法在纯钛表面生成含羟基磷灰石陶瓷膜的研究

目的：利用微弧氧化法在纯钛表面直接生成含羟基磷灰石的陶瓷膜。方法：通过微弧氧化技术,采用双相脉冲电源,以0．4mol／L醋酸钙和0．2mol／L磷酸二氢钠为电解液体系,设定脉冲频率为60Hz,正相电流密度为20A／dm^2,处理时间分别为5rain、10min、20min、30min、60min,在纯钛表面制备陶瓷膜。利用扫描电镜观察膜层的表面形貌,能谱仪分析氧化膜的元素组成,X射线衍射仪分析膜层的相组成,电涡流涂层测厚仪测量膜层的厚度。结果：以0．4mol／L醋酸钙和0．2mol／L磷酸二氢钠为电解液体系,经微弧氧化处理后,在纯钛表面形成含有Ti、O、Ca和P元素的微孔结构的氧化膜,该膜由锐钛矿型TiO2、金红石型TiO2及羟基磷灰石组成。随微弧氧化反应时间的延长,膜层增厚,膜层表面微孔孔径增大,数量减少,粗糙度增大。钛元素的相对含量减少,Ca、P元素相对含量增加,O元素变化不明显。钛、锐钛矿型TiO2和金红石型TiO2的衍射峰逐渐减弱,羟基磷灰石的衍射峰逐渐增强。结论：应用微弧氧化技术,以0．4mol／L醋酸钙和0．2mol／L磷酸二氢钠为电解液体系,在纯钛表面能直接生成含有羟基磷灰石成份的微孔结构的陶瓷膜。     

不同粗化处理对超细晶纯钛表面性能的影响

目的:研究不同粗化处理对超细晶纯钛表面性能及成骨细胞黏附和增殖的影响.方法:将超细晶纯钛棒切割为直径7 mm、厚度2mm的试件,按不同喷砂压力(0.2～0.8 MPa)分组,对其表面进行喷砂酸蚀处理,对照组为普通纯钛.通过表面形貌、粗糙度、亲水性研究材料的表面性能,然后将大鼠胚胎成骨细胞(MC3T3-E1)接种到各组钛片表面,观察细胞初期黏附形态,测定其增殖密度.结果:超细晶纯钛粗化处理后,表面呈现出由喷砂和酸蚀所形成的大小不同的弹坑状双层结构.随着喷砂压力增大,超细晶纯钛表面坑孔直径和粗糙度逐渐增大,但二者均小于普通纯钛对照组(P＜0.05).超细晶纯钛亲水性也随喷砂压力变化而改变,当喷砂压力为0.6 MPa时表现出最佳表面亲水性能.接种细胞后,实验组细胞初期黏附形态优于对照组,当喷砂压力为0.6 MPa时细胞增殖密度最大.结论:对超细晶纯钛喷砂酸蚀处理,喷砂压力为0.6 MPa时,材料表面形貌优于普通纯钛,粗糙度适宜,亲水性良好,更有利于细胞黏附和增殖.     

Preparation and characterization of anodized titanium surfaces and their effect on osteoblast responses

In this study, titanium (Ti) surface was modified by anodizing with a mixture of beta-glycerophosphate sodium and calcium (Ca) acetate, and the anodized surfaces were characterized by scanning electron microscopy, X-ray diffraction, and electron probe microanalysis. In vitro osteoblast response to anodized oxide was also evaluated. The anodic oxide produced was observed to have interconnected pores (0.5-2 microm in diameter) and intermediate roughness (0.60-1.00 microm). In addition, anodic oxide was observed to have amorphous and anatase oxide. Calcium and phosphorus ions were deposited on the Ti oxide during anodization. Osteoblast differentiation, as indicated by alkaline phosphatase production, was enhanced on anodized surfaces. It was thus concluded from this study that Ca phosphate can be deposited on Ti surfaces by anodization. It was also concluded that the phenotypic expression of osteoblast was enhanced by the presence of Ca phosphate and higher roughness on anodized Ti surfaces.     

Evaluation of shear bond strength at the interface of two porcelains and pure titanium injected into the casting mold at three different temperatures

STATEMENT OF PROBLEM: Titanium has physical and mechanical properties, which have led to its increased use in dental prostheses despite casting difficulties due to high melting point and formation of oxide layers which affect the metal-ceramic bond strength. PURPOSE: This in vitro study evaluated the shear bond strength of the interface of 2 dental porcelains and pure titanium injected into a mold at 3 different temperatures. MATERIAL AND METHODS: Using commercially pure (cp) titanium bars (Titanium, Grade I) melted at 1668 degrees C and cast at mold temperatures of 430 degrees C, 700 degrees C or 900 degrees C, 60 specimens were machined to 4 x 4 mm, with a base of 5 x 1 mm. The 4-mm surfaces were airborne-particle abraded with 100 microm aluminum oxide before applying and firing the bonding agent and evaluating the 2 porcelains (Triceram/Triline ti and Vita Titankeramik). Ten specimens were prepared for each temperature and porcelain combination Shear bond testing was performed in a universal testing machine, with a 500-kg load cell and crosshead speed of 0.5 mm/min. The specimens were loaded until failure. The interfaces of representative fractured specimens of each temperature were examined with a scanning electron microscope (SEM) and energy dispersive spectrometer (EDS). Data for shear bond strength (MPa) were statistically analyzed by 2-way ANOVA and the Tukey test (alpha=.05). RESULTS: The results showed significant differences for the metal/porcelain interaction effect (P=.0464). There were no significant differences for the 2 porcelains (P=.4250). The Tukey test showed a significant difference between the pair cp Ti 430 degrees C Triceram and cp Ti 900 degrees C Triceram, with respective mean values and SDs of 59.74 +/- 11.62 and 34.03 +/- 10.35 MPa. CONCLUSION: Triceram porcelain showed a bond strength decrease with an increase in the mold temperature for casting titanium. The highest bond strength for Vita porcelain and the best metal-ceramic interface observed with the SEM were found with the mold temperature of 700 degrees C.     

In vitro preliminary study of osteoblast response to surface roughness of titanium discs and topical application of melatonin

Objectives: To observe human osteoblast behavior cultured in vitro on titanium discs (Ti) in relation to surface roughness and melatonin application. Study Design: Human osteoblasts (MG-63) were cultured on 60 Ti6Al4V discs divided into three groups: Group I: discs treated with dual acid etching; Group II dual acid etching and blasting with calcium phosphate particles; Group III (control) machined discs. Surface roughness and topography of the discs were examined with scanning electron microscope (SEM) and confocal laser scanning electron microscope( CLSM). Osteoblast adhesion, proliferation and cell morphology were determined by means of fluorescence microscopy with Image-Pro Plus software and SEM. Results: Group II presented the roughest discs, while the least rough were Group III. Cell adhesion was greatest in Group II. The addition of melatonin improved cell proliferation. Conclusions: 1. Surface treatments (dual acid etching, calcium phosphate impaction) increase surface roughness in comparison with machined titanium. 2. Greater surface roughness tends to favor cell adhesion after 24-hour cell culture. 3. The addition of melatonin tends to favor osteoblast proliferation. Key words:Osteoblasts, titanium, roughness, melatonin, dental implants, osseointegration.     

热循环和机械循环对切削Ti2448和纯钛金瓷结合强度的影响

目的 研究热循环和机械循环对切削Ti2448和纯钛金瓷结合强度的影响。方法 按照ISO 9693的标准分别制作纯钛和Ti2448烤瓷试件各30个,每种试件均随机分为3组,A组为对照组,在37 ℃水浴中保存24 h;B组和C组在5~55 ℃水浴中热循环3 000次,在每一个温度下保持60 s,然后采用载荷50 N、频率4 Hz,机械循环20 000次（B组）或40 000次（C组）。采用三点弯曲实验测试试件的金瓷结合强度,通过扫描电镜和能谱分析金瓷剥脱面和金瓷结合界面的显微结构和成分组成,对结果进行两因素方差分析和Tukey检验。结果 A组中,纯钛试件的金瓷结合强度（29.21 MPa±2.20 MPa）明显小于Ti2448试件（44.86 MPa±1.75 MPa）（P<0.01）;纯钛试件和Ti2448试件中,C组与A组之间均有统计学差异（P<0.01）。扫描电镜和能谱分析结果显示,Ti2448瓷剥脱面有明显的瓷残留,而纯钛瓷剥脱面无明显瓷残留。结论 热循环和机械循环处理降低了Ti2448和纯钛的金瓷结合强度。