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A comparative study of in vitro apatite deposition on heat-, H(2)O(2)-, and NaOH-treated titanium surfaces. 体外磷灰石沉积在热处理、H(2)O(2)-和naoh处理钛表面的比较研究。
Pub Date : 2001-02-01 DOI: 10.1002/1097-4636(200102)54:2<172::aid-jbm3>3.0.co;2-#
X X Wang, S Hayakawa, K Tsuru, A Osaka

Commercially pure titanium specimens are subjected to three different treatments, and their bioactivity are evaluated by immersing the specimens in a simulated body fluid (SBF, Kokubo's recipe) for various periods up to 7 days, with particular attention being paid to the differences in apatite deposition between surfaces open to SBF and surfaces in contact with the container's bottom. The treatment with a H(2)O(2)/HCl solution at 80 degrees C for 30 min followed by heating at 400 degrees C for 1 h produces an anatase titania gel layer on the specimen surface. This gel layer deposits apatite both on the contact and on open surfaces, and apatite deposition ability does not change with pre-staking in distilled water. The treatment with a NaOH solution at 60 degrees C for 3 days produces a sodium titanate gel layer. This gel layer can deposit apatite only on the contact surface, and the apatite deposition ability is completely lost after 1 day of pre-staking in distilled water. It is concluded, therefore, that the bioactivity of the titania gel originates from the favorable structure of the gel itself while the bioactivity of the sodium titanate gel depends heavily on ion release from the gel. The third treatment, a simple heat treatment at 400 degrees C for 1 h, produces a dense (not porous) oxide layer on the specimen surface. The specimens can deposit apatite on the contact surface after only 3 days of staking in SBF, but they cannot deposit apatite on the open surface for up to 2 months of staking. The implications of such apatite deposition behavior have been discussed in relation to the environments of titanium implants in bone as well as to the methodology of the SBF staking experiment.

对商业纯钛标本进行三种不同的处理,并通过将标本浸泡在模拟体液(SBF, Kokubo的配方)中长达7天的不同时期来评估其生物活性,特别注意向SBF开放的表面和与容器底部接触的表面之间磷灰石沉积的差异。用H(2)O(2)/HCl溶液在80℃下处理30分钟,然后在400℃下加热1小时,在样品表面产生锐钛矿型二氧化钛凝胶层。该凝胶层在接触表面和开放表面沉积磷灰石,并且在蒸馏水中预桩不会改变磷灰石的沉积能力。用60℃的NaOH溶液处理3天,形成钛酸钠凝胶层。该凝胶层只能在接触面沉积磷灰石,在蒸馏水中预桩1天后,磷灰石沉积能力完全丧失。因此,二氧化钛凝胶的生物活性源于凝胶本身的良好结构,而钛酸钠凝胶的生物活性很大程度上取决于凝胶中的离子释放。第三种处理,在400℃下进行1小时的简单热处理,在试样表面产生致密的(不多孔的)氧化层。试件在SBF中桩敷3天后就能在接触表面沉积磷灰石,但在开放表面长达2个月的桩敷中不能沉积磷灰石。讨论了这种磷灰石沉积行为与骨内钛植入物环境的关系以及SBF桩接实验的方法。
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引用次数: 170
Effects of geometry of hydroxyapatite as a cell substratum in BMP-induced ectopic bone formation. 羟基磷灰石作为细胞基质在bmp诱导异位骨形成中的作用。
Q M Jin, H Takita, T Kohgo, K Atsumi, H Itoh, Y Kuboki
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引用次数: 0
Prophylaxis of implant-related staphylococcal infections using tobramycin-containing bone cement. 含妥布霉素骨水泥预防种植体相关葡萄球菌感染。
Pub Date : 2000-12-15 DOI: 10.1002/1097-4636(20001215)52:4<754::aid-jbm21>3.0.co;2-#
M W Nijhof, W J Dhert, A Fleer, H C Vogely, A J Verbout

In a rabbit model, premixed tobramycin-containing bone cement was studied for its efficacy to prevent infections with two frequently encountered staphylococcal species in arthroplasty surgery. After intramedullary inoculation with staphylococci, either standard or premixed tobramycin-containing Simplex-P bone cement was injected in the right femur of 120 rabbits. Development of infection was examined by culture of femoral bone after 7 or 28 days. Loss of body weight and elevated erythrocyte sedimentation rate in the control rabbits inoculated with Staphylococcus aureus were seen in the first postoperative week, returning to normal in 28 days. Inoculation with Staphylococcus epidermidis resulted only in a low-grade infection. All rabbits receiving premixed tobramycin-containing bone cement were free of signs of infection, and all their cultures were negative. Culture yield from Staphylococcus aureus controls increased with time and inoculum dose. Staphylococcus epidermidis controls needed higher inoculum doses to establish an infection, while culture yield decreased in time. These differences in mode of prosthesis-related infection are explained by differences in virulence factors.

在兔模型中,研究了含妥布霉素的预混合骨水泥预防关节置换术中常见的两种葡萄球菌感染的效果。120只家兔右股骨经葡萄球菌髓内接种后,分别注射含妥布霉素的标准型或预混型Simplex-P骨水泥。7天或28天后用股骨培养检查感染的发展情况。接种金黄色葡萄球菌的对照兔术后第一周体重减轻,红细胞沉降率升高,28天后恢复正常。接种表皮葡萄球菌只导致低度感染。所有接受含妥布霉素骨水泥预混剂的家兔均无感染迹象,培养结果均为阴性。金黄色葡萄球菌对照的培养产量随时间和接种量的增加而增加。表皮葡萄球菌对照需要较高的接种剂量来建立感染,而培养产量随着时间的推移而下降。这些假体相关感染模式的差异是由毒力因素的差异来解释的。
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引用次数: 48
Well-defined sulfobetaine-based statistical copolymers as potential antibioadherent coatings. 良好定义的基于磺基甜菜碱的统计共聚物作为潜在的抗生素粘附涂层。
Pub Date : 2000-10-01 DOI: 10.1002/1097-4636(200010)52:1<88::aid-jbm11>3.0.co;2-#
A B Lowe, M Vamvakaki, M A Wassall, L Wong, N C Billingham, S P Armes, A W Lloyd

The potential use of novel poly(sulfobetaine) copolymers as antibioadherent coatings was investigated using Pseudomonas aeruginosa as a model microorganism and human macrophages and 3T3 mouse embryonic fibroblasts. Two well-defined statistical copolymers with narrow molecular weight distributions were prepared by group transfer copolymerization of n-butyl methacrylate (nBuMA) with either 10 or 30 mol % 2-(dimethylamino)ethyl methacrylate (DMAEMA). Sulfobetainized nBuMA-DMAEMA copolymers (poly[sulfobetaine-stat-nBuMA]) were obtained by treating these precursor polymers with 1,3-propanesultone under mild conditions. Both proton NMR spectroscopy and elemental microanalyses indicated that essentially all the DMAEMA residues were derivatized in both copolymers. Poly(methyl methacrylate) (PMMA) discs were coated with the sulfobetainized nBuMA-DMAEMA copolymers and the bioadherent properties of these coated materials were compared with those of PMMA. Statistically significantly fewer (p<.05) bacteria, macrophages, and fibroblasts adhered to the poly(sulfobetaine-stat-nBuMA)-coated PMMA than to the uncoated PMMA. The poly(sulfobetaine-stat-nBuMA) copolymer containing the higher proportion (30 mol %) sulfobetainized DMAEMA residues proved to be the more effective antibioadherent coating. The antibioadherent properties of these coating materials may allow the cost-effective production of dirt-resistant, easy to clean work surfaces, bioinert coatings for medical devices, and antifouling coatings for marine, agricultural, and industrial applications.

以铜绿假单胞菌为模型微生物,以人巨噬细胞和3T3小鼠胚胎成纤维细胞为实验对象,研究了新型聚磺基甜菜碱共聚物作为抗生素粘附膜的应用前景。用甲基丙烯酸正丁酯(nBuMA)与10或30 mol % 2-(二甲氨基)甲基丙烯酸乙酯(DMAEMA)进行基团转移共聚,制备了两种分子量分布较窄、性能良好的统计共聚物。用1,3-丙磺酸在温和条件下处理这些前驱体聚合物,得到了磺胺甜菜碱化nBuMA-DMAEMA共聚物(聚[磺胺甜菜碱-stat- nbuma])。质子核磁共振谱和元素微量分析表明,DMAEMA残基在两种共聚物中都发生了衍生化。将磺化的nBuMA-DMAEMA共聚物包覆在聚甲基丙烯酸甲酯(PMMA)圆盘上,并与PMMA的生物粘附性能进行了比较。统计学上显著减少(p
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引用次数: 82
Platelet adhesion and procoagulant activity induced by contact with radiofrequency glow discharge polymers: roles of adsorbed fibrinogen and vWF. 接触射频辉光放电聚合物诱导血小板粘附和促凝活性:吸附纤维蛋白原和vWF的作用。
Pub Date : 2000-09-15 DOI: 10.1002/1097-4636(20000915)51:4<669::aid-jbm15>3.0.co;2-#
J M Grunkemeier, W B Tsai, M R Alexander, D G Castner, T A Horbett

The potential hemocompatibility of radiofrequency glow discharge (RFGD) polymers made by copolymerization of mixtures of hexafluoropropene and ethylene (C(3)F(6)/C(2)H(4)) or acrylic acid and 1,7-octadiene was investigated using in vitro assays for platelet adhesion and platelet catalyzed thrombin generation. Thrombin generation rate normalized to platelet number was used as a measurement of platelet activation (procoagulant activity). RFGD polymers produced by copolymerization of acrylic acid and 1, 7-octadiene contained varying amounts of carboxylic acid species as determined by electron spectroscopy for chemical analysis (ESCA). These polymers induced little variation in platelet adhesion, thrombin generation, or platelet activation. RFGD polymerization of C(3)F(6) and C(2)H(4) resulted in polymers with varying proportions of fluorinated species, as determined by ESCA. Fibrinogen adsorption from plasma was maximal on a polymer made with 25% C(3)F(6) (75% C(2)H(4)) in the feed. However von Willebrand factor (vWF) adsorption was greater on polymers made with increased %C(3)F(6) in the feed. Platelet adhesion decreased with increasing %C(3)F(6) in the feed. Thrombin generation was lowest for platelets adherent to polymers made from both C(3)F(6) and C(2)H(4). Therefore, procoagulant activity of platelets increased for polymers made with increased %C(3)F(6) in the feed, similar to the trend in vWF adsorption. These findings suggest that increased incorporation of fluorinated species into RFGD polymers leads to decreased platelet adhesion and increased platelet activation (which is possibly due to increased vWF adsorption).

采用体外血小板粘附和血小板催化凝血酶生成实验研究了六氟丙烯和乙烯(C(3)F(6)/C(2)H(4))或丙烯酸和1,7-辛二烯共聚制备的射频辉光放电(RFGD)聚合物的潜在血液相容性。凝血酶生成率归一化血小板数作为血小板活化(促凝活性)的测量。由丙烯酸和1,7 -辛二烯共聚制备的RFGD聚合物含有不同数量的羧酸,这是通过电子能谱(ESCA)进行化学分析的结果。这些聚合物在血小板粘附、凝血酶生成或血小板活化方面几乎没有引起变化。经ESCA测定,C(3)F(6)和C(2)H(4)的RFGD聚合产生的聚合物含不同比例的氟化物质。在饲料中添加25% C(3)F(6) (75% C(2)H(4))的聚合物上,血浆中纤维蛋白原的吸附量最大。然而,饲料中增加%C(3)F(6)的聚合物对冯氏血友病因子(vWF)的吸附更大。血小板粘附随着饲料中%C(3)F(6)的增加而降低。血小板粘附在由C(3)F(6)和C(2)H(4)制成的聚合物上凝血酶生成最低。因此,饲料中增加%C(3)F(6)的聚合物提高了血小板的促凝活性,与vWF吸附的趋势相似。这些发现表明,氟化物质在RFGD聚合物中的掺入增加导致血小板粘附减少和血小板活化增加(这可能是由于vWF吸附增加)。
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引用次数: 49
Biphasic calcium phosphates: influence of three synthesis parameters on the HA/beta-TCP ratio. 双相磷酸钙:三个合成参数对HA/ β - tcp比的影响。
Pub Date : 2000-09-15 DOI: 10.1002/1097-4636(20000915)51:4<680::aid-jbm16>3.0.co;2-#
J M Bouler, R Z LeGeros, G Daculsi

Hydroxyapatite (HA) contents measurements were conducted on eight biphasic calcium phosphate (BCP) samples obtained by sintering calcium-deficient apatite formed previously by hydrolyzing a dicalcium phosphate dihydrate (DCPD) powder. We evaluated the influences and interactions of three synthesis factors: alkalinity, process duration, and concentration of the water suspension in DCPD. Those parameters were varied simultaneously between two limit levels. Experiments used a factorial design method (FDM) allowing optimization of the number of samples as well as statistical analysis of results. FDM showed that HA content, in a defined experimental area, can be described by a first-order polynomial equation in which the initial alcalinity and the DCPD/water ratio are the major influences. Experiment prove that pH measured at the end of the hydrolysis was predictive of the HA content in the final BCP. This study leads up to an isoresponse line diagram which will allow the synthesis of some BCP with fitted HA/beta-tricalcium phosphate ratios.

对8个双相磷酸钙(BCP)样品进行了羟基磷灰石(HA)含量测定,该样品是由磷酸二氢钙(DCPD)粉末水解形成的缺钙磷灰石烧结而成的。我们评估了三个合成因素的影响和相互作用:碱度、工艺时间和DCPD中水悬浮液的浓度。这些参数在两个极限水平之间同时变化。实验使用因子设计方法(FDM),允许优化样本数量以及结果的统计分析。FDM表明,在确定的实验区域内,HA含量可以用一阶多项式方程来描述,其中初始碱度和DCPD/水比是主要影响因素。实验证明,水解结束时测定的pH值可以预测最终BCP中HA的含量。这项研究导致了一个等响应线图,这将允许合成一些具有合适的HA/ β -磷酸三钙比例的BCP。
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引用次数: 146
Anodic oxidation and hydrothermal treatment of titanium results in a surface that causes increased attachment and altered cytoskeletal morphology of rat bone marrow stromal cells in vitro. 钛的阳极氧化和水热处理导致体外大鼠骨髓基质细胞的附着增加和细胞骨架形态改变。
Pub Date : 2000-09-05 DOI: 10.1002/1097-4636(20000905)51:3<398::aid-jbm14>3.0.co;2-#
J Takebe, S Itoh, J Okada, K Ishibashi

Previous studies have suggested the usefulness of a new coating method-namely, the forming of a thin hydroxyapatite (HA) layer on commercially pure titanium (cpTi) by anodization and hydrothermal treatment-for use as a dental root implant material. In vivo and in vitro studies confirmed that an HA layer on cpTi (HA/cpTi) implants showed good compatibility with bone tissue, rat bone marrow stromal (RBM) cells, and immune cells. The aim of the present investigation was to further characterize the in vitro early cellular behavior of RBM cells on HA/cpTi implants. Therefore, in this study we performed surface analysis, analysis of cell initial attachment, and analysis of cell morphology and the cytoskeleton. Drops of distilled water or cell culture medium showed smaller contact angles with HA/cpTi than with cpTi. RBM cells were cultured for 30, 60, and 120 min on HA/cpTi and cpTi, and the level of cell adhesion was shown to increase with time on both substrates. However, cell adhesion on HA/cpTi was significantly higher than on cpTi at 60 and 120 min. Especially at 120 min, when compared with cpTi, the cell morphology on the surface of HA/cpTi not only adopted a flattened and spreading form, but also extended filopodium-like processes with irregular edges that were intimately adapted to the surface of the HA microcrystals. The cytoskeleton on HA/cpTi showed well-formed actin filaments that were parallel to each other and the long axis of RBM cells. The actin filaments of RBM cells on the HA/cpTi surface were localized to the periphery (corresponding to the edge of the filopodium-like processes) well after 120 min. This suggests that actin filaments of RBM cells need to be anchored at the HA/cpTi surface and the numerous HA microcrystals precipitated on the HA/cpTi surface. These findings were similar to the scanning electron microscopic morphology. The peripheral anchorage provide sufficient strength of attachment to allow recognization of actin filaments upon HA/cpTi. The surface of HA/cpTi was more hydrophilic and exhibited markedly improved wettability compared to untreated cpTi, and higher levels of early cell attachment were observed on surfaces after anodization and hydrothermal treatment than on surfaces with untreated cpTi. The results of in vitro experiments suggest that this new method for forming a thin HA layer on the surface of cpTi could be useful to ensure excellent cellular behavior on implant surfaces. The characterization of cell morphology on the thin HA layer formed by anodization and hydrothermal treatment on cpTi implant material suggests that physicochemical or biological conditioning of the implant surface involves implant surface topography.

先前的研究表明,一种新的涂层方法是有用的,即通过阳极氧化和水热处理在商业纯钛(cpTi)上形成一层薄薄的羟基磷灰石(HA)层,作为牙根种植材料。体内和体外研究证实,cpTi (HA/cpTi)植入物上的HA层与骨组织、大鼠骨髓基质(RBM)细胞和免疫细胞具有良好的相容性。本研究的目的是进一步表征RBM细胞在HA/cpTi植入物上的体外早期细胞行为。因此,在本研究中,我们进行了表面分析、细胞初始附着分析、细胞形态和细胞骨架分析。液滴蒸馏水或细胞培养基与HA/cpTi的接触角小于与cpTi的接触角。RBM细胞分别在HA/cpTi和cpTi上培养30、60和120分钟,细胞粘附水平随时间的增加而增加。而在60 min和120 min时,细胞在HA/cpTi上的粘附力明显高于在cpTi上的粘附力。特别是在120 min时,与cpTi相比,HA/cpTi表面的细胞形态不仅呈扁平、铺展的形式,而且呈延伸的丝状突起,边缘不规则,与HA微晶表面密切适应。HA/cpTi上的细胞骨架显示形成良好的肌动蛋白丝,与RBM细胞的长轴平行。RBM细胞在HA/cpTi表面的肌动蛋白丝在120分钟后很好地定位到周围(对应于丝状突起的边缘)。这表明RBM细胞的肌动蛋白丝需要锚定在HA/cpTi表面,并且在HA/cpTi表面沉淀了大量的HA微晶。这些发现与扫描电镜形貌相似。外周锚定提供了足够的附着强度,允许在HA/cpTi上识别肌动蛋白丝。与未经处理的cpTi相比,HA/cpTi的表面具有更强的亲水性和明显的润湿性,并且在阳极氧化和水热处理后的表面上观察到比未经处理的cpTi表面更高的早期细胞附着水平。体外实验结果表明,这种在cpTi表面形成薄HA层的新方法可用于确保植入物表面的良好细胞行为。通过阳极氧化和水热处理在cpTi植入材料上形成的薄HA层上的细胞形态表征表明,植入物表面的物理化学或生物调节涉及到植入物表面形貌。
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引用次数: 149
Effects of geometry of hydroxyapatite as a cell substratum in BMP-induced ectopic bone formation. 羟基磷灰石作为细胞基质在bmp诱导异位骨形成中的作用。
Q M Jin, H Takita, T Kohgo, K Atsumi, H Itoh, Y Kuboki

Three different types of porous hydroxyapatite with pore sizes of 100-200 micrometer in diameter-porous particles of hydroxyapatite (PPHAP), porous blocks of hydroxyapatite (PBHAP), and honeycomb-shaped hydroxyapatite (HCHAP)-were compared in terms of their abilities to induce osteogenesis when implanted subcutaneously with recombinant human BMP-2 into rats and extracted at 1, 2, 3, and 4 weeks. Histologically, direct bone formation occurred in PPHAP and PBHAP while only endochondral ossification took place in HCHAP. Interestingly, cartilage in the central zones and bone in the orifice zones of the tunnels of the HCHAP were observed at 2 weeks. After 3 weeks, the cartilage disappeared and bone formation occurred throughout the inner surface of the tunnels of the HCHAP, always leaving space for capillaries within the tunnels. Alkaline phosphatase activity and osteocalcin content were the highest in HCHAP among the three hydroxyapatite implants. These results clearly indicate that BMP-induced bone formation is highly dependent on the geometry of the carrier, which provides feasible structural factors for vascularization.

我们比较了孔径为100-200微米的三种不同类型的多孔羟基磷灰石——多孔羟基磷灰石颗粒(PPHAP)、多孔羟基磷灰石块(phap)和蜂窝状羟基磷灰石(HCHAP)——在重组人BMP-2皮下植入大鼠并于1、2、3和4周提取时诱导成骨的能力。组织学上,PPHAP和PBHAP发生直接成骨,而HCHAP仅发生软骨内成骨。有趣的是,在2周时观察到HCHAP隧道中心区域的软骨和孔口区域的骨。3周后,软骨消失,整个HCHAP隧道内表面出现骨形成,隧道内毛细血管始终留有空间。三种羟基磷灰石种植体HCHAP的碱性磷酸酶活性和骨钙素含量最高。这些结果清楚地表明bmp诱导的骨形成高度依赖于载体的几何形状,这为血管化提供了可行的结构因素。
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引用次数: 0
NIST workshop on needs for reference biomaterials. NIST关于参考生物材料需求的研讨会。
Pub Date : 2000-08-01 DOI: 10.1002/(sici)1097-4636(200008)51:2<155::aid-jbm2>3.0.co;2-#
J A Tesk
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引用次数: 1
Effect of gas composition on spore mortality and etching during low-pressure plasma sterilization. 低压等离子体灭菌过程中气体成分对孢子死亡率和腐蚀的影响。
Pub Date : 2000-07-01 DOI: 10.1002/(sici)1097-4636(200007)51:1<128::aid-jbm17>3.0.co;2-#
S Lerouge, M R Wertheimer, R Marchand, M Tabrizian, L Yahia

The aim of this work was to investigate possible mechanisms of sterilization by low-temperature gas plasma: spore destruction by plasma is compared with etching of synthetic polymers. Bacillus subtilis spores were inoculated at the bottom of glass vials and subjected to different plasma gas compositions (O(2), O(2)/Ar, O(2)/H(2), CO(2), and O(2)/CF(4)), all known to etch polymers. O(2)/CF(4) plasma exhibited much higher efficacy than all other gases or gas mixtures tested, with a more than 5 log decrease in 7.5 min, compared with a 2 log decrease with pure oxygen. Examination by scanning electron microscopy showed that spores were significantly etched after 30 min of plasma exposure, but not completely. We speculate about their etch resistance compared with that of synthetic polymers on the basis of their morphology and complex coating structure. In contrast to so-called in-house plasma, sterilization by Sterrad(R) tended to increase the observed spores' size; chemical modification (oxidation), rather than etching, is believed to be the sterilization mechanism of Sterrad(R).

这项工作的目的是研究低温气体等离子体灭菌的可能机制:等离子体破坏孢子与合成聚合物的蚀刻进行了比较。将枯草芽孢杆菌孢子接种于玻璃瓶底部,并接受不同的等离子体气体成分(O(2), O(2)/Ar, O(2)/H(2), CO(2)和O(2)/CF(4)),所有这些都已知会腐蚀聚合物。O(2)/CF(4)等离子体表现出比所有其他气体或气体混合物更高的效率,在7.5分钟内降低了5倍以上,而纯氧降低了2倍。扫描电镜检查显示,暴露于等离子体30分钟后,孢子明显蚀刻,但不完全蚀刻。我们根据其形态和复杂的涂层结构推测其与合成聚合物的耐蚀性。与所谓的内部等离子体相比,Sterrad(R)灭菌倾向于增加观察到的孢子的大小;化学修饰(氧化),而不是蚀刻,被认为是Sterrad(R)的杀菌机制。
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引用次数: 166
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