类金刚石膜的生物医学性能与应用

类金刚石膜具有高硬度、高光洁度、低摩擦系数、高耐磨性及耐腐蚀性等很多优异性能,并且易于掺杂而改变性能以适应不同需要.良好的生物医学性能也使其在体内植入物及医疗器械的表面改性方面有广阔的应用前景.主要就类金刚石膜在生物医学方面的性能与应用进行综述.     

The human micro-vascular endothelial cells in vitro interaction with atomic-nitrogen-doped diamond-like carbon thin films

This paper reports the initial response of atomic nitrogen doped diamond like carbon (DLC) to endothelial cells in vitro. The introduction of nitrogen atoms/molecules to the diamond like carbon structures leads to an atomic structural change favorable to the attachment of human micro-vascular endothelial cells. Whilst the semi-conductivity induced by nitrogen in DLC is thought to play a part, the increase in the non-bonded N atoms and N(2) molecules in the atomic doped species (with the exclusion of the charged species) seems to contribute to the improved attachment of human microvascular endothelial cells. The increased endothelial attachment is associated with a lower work function and slightly higher water contact angle in the atomic doped films, where the heavy charged particles are excluded. The films used in the study were synthesized by the RF PECVD technique followed by post deposition doping with nitrogen, and afterwards the films were characterized by XPS, Raman spectroscopy, SIMS and Kelvin probe. The water contact angles were measured, and the counts of the adherent endothelial cells on the samples were carried out. This study is relevant and contributory to improving biocompatibility of surgical implants and prostheses.     

Incorporation of Si and SiOx into diamond-like carbon films: Impact on surface properties and osteoblast adhesion

The interaction of human osteoblast cells with diamond-like carbon films incorporating silicon and silicon oxide (SiO_x, 1 ? x ? 1.5) and synthesized using the direct-current plasma-activated chemical vapour deposition method was investigated. Cell culture studies were performed for films with Si contents ranging from ～4 at.% to 15 at.%. Substantial differences between Si-incorporated and SiO_x-incorporated films were found for the bonding environments of Si atoms and the hybridization of underlying carbon structures. However, osteoblast-attachment studies did not show statistically significant trends in properties of cell growth (count, area and morphology) that can be attributed either to the Si content of the films or to the chemical structure of the films. The surface energy decreased by 40% as the Si content of the SiO_x incorporated DLC films increased to 13 at.%. The cell adhesion properties however did not change in response to lowering of the surface energy. The incorporation of both Si and SiO_x leads to a beneficial reduction in the residual stress of the films. The average roughness of the films increases and the hardness decreases when Si and SiO_x are added to DLC films. The impact of these changes for load-bearing biomedical applications can be determined only by carefully controlled experiments using anatomic simulators.     

A rapid, quantitative method for measuring leukocyte adhesion to normal and balloon-injured arteries in vitro

Many of the currently available techniques for quantifying leukocyte adhesion require monolayers of cells and are therefore unsuitable for use in ex vivo arterial tissue. Here we describe a rapid method to measure adhesion of leukocytes to intact artery strips and to determine the effect of artery injury on adhesiveness of leukocytes with and without activation. Leukocytes were isolated from rabbit blood, labelled with 51Cr, and added to the luminal face of the left and right subclavian arteries derived from the same animal. In some experiments the endothelium was removed before addition of leukocytes and in another series of experiments the artery was injured by inflating a balloon catheter within the lumen in vitro before leukocyte addition. After washing, the adhesion of labelled leukocytes was quantified by gamma counting. To determine localization of the leukocytes, some arteries were fixed in situ and examined microscopically, with confirmation of leukocyte identification by enzyme cytochemistry. The adhesion of leukocytes increased progressively during 60 min and was inhibited by reducing the temperature to 4 degrees C. Adhesion was increased by the nitric oxide synthase inhibitor L-NAME. Stretching the artery wall in vitro using a balloon catheter increased leukocyte adhesion within 1 h after injury. In contrast, this did not occur following simple arterial denudation. Histological examination of stained en face preparations and transverse sections of the subclavian arteries revealed loosely adherent granulocytic leukocytes on the endothelial surface. This technique is straightforward and allows accurate and rapid measurement of autologous leukocyte adhesion to normal and pathologically altered arteries ex vivo.     

Investigating the functionality of diamond-like carbon films on an artificial heart diaphragm

In this study, the authors used diamond-like carbon film to coat the ellipsoidal diaphragm (polyurethane elastomer) of artificial hearts. The purpose of such coatings is to prevent the penetration of hydraulic silicone oil and blood through the diaphragm. To attach diamond-like carbon film uniformly on the diaphragm, the authors developed a special electrode. In estimating the uniformity of the diamond-like carbon film, the thickness was measured using a scanning electron microscope, and the characteristics of the diamond-like carbon film was investigated using infrared spectroscopy, Ar-laser Raman spectrophotometer, and x-ray photoelectron spectrometer. Also, to estimate the penetration of silicone oil through the diaphragm, in vitro testing was operated by alternating the pressure of silicone oil for 20 days. The authors were able to successfully attach uniform deposition of diamond-like carbon film on the ellipsoidal diaphragm. In this in vitro test, diamond-like carbon film was proven to have good stability. The amount of silicone oil penetration was improved by one-third using the diamond-like carbon film coating compared with an uncoated diaphragm. It is expected that through the use of the diamond-like carbon film, the dynamic compatibility of an artificial heart diaphragm will increase.     

新型生物医学材料——类金刚石膜的研究进展

类金刚石因具有良好的细胞相容性、血液相容性及高耐磨性高硬度等特点 ,而成为一种很有发展前景的生物膜材料。本文综述了 2 0世纪 90年代以来类金刚石的理化性质、生物学方面的研究进展 ,并对未来的发展进行了展望。     

Development of a new dynamic method for quantitative evaluation of in vitro hemocompatibility of biomedical materials

In this study a new dynamic method is introduced allowing the estimation of blood cell adhesion on flat test surfaces by measuring the cell loss in the bulk phase of surface contacting test blood under defined rheological conditions. This was achieved by constructing a novel test chamber permitting the contact of small amounts of blood with a large geometrical test surface. The construction consists of a spiral-shaped flow channel of 0.3 cm width, 0.02 cm height and 78 cm length covered with the biomaterials to be tested from both sides. Laminarity of blood flow in the conduit was confirmed theoretically by the calculation of an equivalent to the Reynolds number for curved systems the so-called Dean number. Furthermore, flow laminarity was proved experimentally finding that the flow rate of blood with different hematocrit values was proportional to the hydrostatic pressure applied. The applicability of the novel 'spiral method' for the estimation of hemocompatibility was demonstrated by evaluation of platelet adhesion onto different polymers in comparison to siliconized and fibrinogen coated glass as reference surfaces. Additionally, it was possible under distinct conditions to determine the adhesion of leucocytes and the detachment of platelet aggregates. Therefore, it was concluded that the spiral method can be used for the assessment of the hemocompatibility of flat biomedical polymers. As main advantages of the new method can be considered the high time efficiency and accuracy without labelling or optical detection of adherent cells.     

Concurrent improvement in biocompatibility and bioinertness of diamond-like carbon films with nitrogen doping

The surfaces of implantable biomaterials improving biocompatibility and bioinertness are critical for new application of bioimplantable devices. Diamond-like carbon (DLC) film is a promising biomaterial with use for coating bioimplantable devices because of its good biocompatibility, bioinertness, and mechanical properties. In this study, concurrent improvement in biocompatibility and bioinertness of DLC films has been achieved using N-incorporation technique. The N doping degree was found to play an important role in affecting the biocompatibility and bioinertness of N-doped DLC films. The results indicated that the N-doped DLC films deposited at N(2) concentration of 5% could help to create suitable condition of surface/structure/adhesion combination of DLC films in the both affinity of the L929 mouse fibroblasts and electrochemical inertness in the Hank's balanced salt solutions (simulating human body fluids). N doping supports the attachment and proliferation of cells and prevents the permeation of electrolyte solutions, thereby simultaneity improved the biocompatibility and bioinertness of DLC films. This finding is useful for the fabrication and encapsulation of in vivo devices without induced immune response in the human body. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 100A:3151-3156, 2012.     

Development of a new dynamic method for quantitative evaluation of in vitro hemocompatibility of biomedical materials.

In this study a new dynamic method is introduced allowing the estimation of blood cell adhesion on flat test surfaces by measuring the cell loss in the bulk phase of surface contacting test blood under defined rheological conditions. This was achieved by constructing a novel test chamber permitting the contact of small amounts of blood with a large geometrical test surface. The construction consists of a spiral-shaped flow channel of 0.3 cm width, 0.02 cm height and 78 cm length covered with the biomaterials to be tested from both sides. Laminarity of blood flow in the conduit was confirmed theoretically by the calculation of an equivalent to the Reynolds number for curved systems the so-called Dean number. Furthermore, flow laminarity was proved experimentally finding that the flow rate of blood with different hematocrit values was proportional to the hydrostatic pressure applied. The applicability of the novel 'spiral method' for the estimation of hemocompatibility was demonstrated by evaluation of platelet adhesion onto different polymers in comparison to siliconized and fibrinogen coated glass as reference surfaces. Additionally, it was possible under distinct conditions to determine the adhesion of leucocytes and the detachment of platelet aggregates. Therefore, it was concluded that the spiral method can be used for the assessment of the hemocompatibility of flat biomedical polymers. As main advantages of the new method can be considered the high time efficiency and accuracy without labelling or optical detection of adherent cells.     

An in vitro investigation of diamond-like carbon as a femoral head coating

Wear of polyethylene acetabular components of hip implants is a significant clinical problem. In prosthetic hip surgery, polyethylene wear is identified as a factor that limits the life of the implant; it is known that the production of debris can cause adverse tissue reactions that may lead to extensive bone loss around the implant, and consequently loosening of the fixation. A new class of so-called Diamond-Like Carbon coatings, applied to titanium femoral heads was compared to ceramic and metallic heads in terms of wear behavior against UHMWPE using a hip joint simulator with a bovine calf serum lubricant. A thin film of Diamond-Like Carbon was deposited directly onto titanium (Ti6Al4V) head using chemical vapor deposition. The wear of polyethylene coupled with Diamond-Like Carbon coated femoral heads was comparable to that obtained with the polyethylene coupled with commercial alumina femoral heads.     

A combinatorial partitioning method to identify multilocus genotypic partitions that predict quantitative trait variation

Recent advances in genome research have accelerated the process of locating candidate genes and the variable sites within them and have simplified the task of genotype measurement. The development of statistical and computational strategies to utilize information on hundreds -- soon thousands -- of variable loci to investigate the relationships between genome variation and phenotypic variation has not kept pace, particularly for quantitative traits that do not follow simple Mendelian patterns of inheritance. We present here the combinatorial partitioning method (CPM) that examines multiple genes, each containing multiple variable loci, to identify partitions of multilocus genotypes that predict interindividual variation in quantitative trait levels. We illustrate this method with an application to plasma triglyceride levels collected on 188 males, ages 20--60 yr, ascertained without regard to health status, from Rochester, Minnesota. Genotype information included measurements at 18 diallelic loci in six coronary heart disease--candidate susceptibility gene regions: APOA1--C3--A4, APOB, APOE, LDLR, LPL, and PON1. To illustrate the CPM, we evaluated all possible partitions of two-locus genotypes into two to nine partitions (approximately 10(6) evaluations). We found that many combinations of loci are involved in sets of genotypic partitions that predict triglyceride variability and that the most predictive sets show nonadditivity. These results suggest that traditional methods of building multilocus models that rely on statistically significant marginal, single-locus effects, may fail to identify combinations of loci that best predict trait variability. The CPM offers a strategy for exploring the high-dimensional genotype state space so as to predict the quantitative trait variation in the population at large that does not require the conditioning of the analysis on a prespecified genetic model.     

Protein adsorption on thin films of carbon and carbon nitride monitored with in situ ellipsometry

Thin films of amorphous carbon and amorphous, graphitic and fullerene-like carbon nitride were deposited by reactive magnetron sputtering and optically characterized with spectroscopic ellipsometry. Complementary studies using scanning electron microscopy and atomic force microscopy were performed. The films were exposed to human serum albumin (HSA) and the adsorption was monitored in situ using dynamic ellipsometry. From the ellipsometric data the adsorbed amount of proteins was quantified in terms of surface mass density using de Feijter's model. The results indicate larger adsorption of proteins onto the amorphous films compared to the films with a more textured structure. Complementary studies with 125I-labeled HSA showed an apparent protein adsorption up to six times larger compared to the ellipsometry measurement. In addition, the four types of films were incubated in blood plasma followed by exposure to anti-fibrinogen, anti-HMWK or anti-C3c, revealing the materials' response to complement and contact activation. The amorphous and graphitic carbon nitride exhibit rather high immune activity compared to a titanium reference, whereas the amorphous carbon and the fullerene-like CNx show less immune complement deposition. Compared to the reference, all films exhibit indications of a stronger ability to initiate the intrinsic pathway of coagulation. Finally, the surfaces' bone-bonding ability was investigated by examination of their ability to form calcium phosphate crystals in a simulated body fluid, with a-CNx depositing most calcium phosphate after 21 days of incubation.     

A new quantitative method to evaluate the in vitro bioactivity of melt and sol-gel-derived silicate glasses

Two melt-derived glasses (45S5 and 60S) and four sol-gel glasses (58S, 68S, 77S, and 91S) have been synthesized. The activation energy for the silicon release was determined, and a very close correlation was observed between this value and published results of the bioactive behavior of the glasses. This relationship can be explained in terms of the influence of chemical composition, textural properties, and structural density on the silanol group formation and silicon dissolution. These measurements provide a quantitative method to evaluate the in vitro bioactivity of SiO(2)-based glasses. Preliminary studies suggest an activation energy gap (Ea) of 0.35-0.5 eV as a boundary between bioactive and nonbioactive glasses.     

The effects of amorphous carbon films deposited on polyethylene terephthalate on bacterial adhesion

There is an increasing interest in developing new methods to reduce bacteria adhesion onto polymeric materials that are used in biomedical implants. The antibacterial behavior on polyethylene terephthalate (PET) treated by acetylene (C2H2) plasma immersion ion implantation-deposition (PIII-D) is investigated. The surface structure of the treated PET is determined by laser Raman spectroscopy, X-ray photoelectron spectroscopy (XPS) and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). The results show that a thin amorphous polymer-like carbon (PLC) layer is formed on the PET surface. Atomic force micrographs (AFM) show that C2H2 PIII-D significantly changes the surface morphology of PET. The capacities of Staphylococcus aureus (SA) and Staphylococcus epidermidis (SE) to adhere onto PET are quantitatively determined by plate counting and Gamma-ray counting of 125I radio labeled bacteria in vitro. The results indicate that the adhesion of the two kinds of bacteria to PET is suppressed by PLC. The adhesion efficiency of SE on the coated surface is only about 14% of that of the untreated PET surface, and that of SA is about 35% of that of the virgin surface. The electrokinetic potentials of the bacterial cells and substrates are determined by zeta potential measurement. All the substrates as well as the bacterial strain have negative zeta potentials, and it means that bacterial adhesion is not mediated by electrostatic interactions. The surface energy components of the various substrates and bacteria are calculated based on measurements in water, formamide and diiodomethane. The surface free energies obtained are used to calculate the interfacial free energies of adhesion ( deltaFAdh ) of SA and SE onto various substrates, and it is found that bacterial adhesion is energetically unfavorable on the PLC deposited on PET by C2H2 PIII-D.     

钛基植人物生物陶瓷涂层的研究进展

医用钛合金材料属于生物惰性材料,广泛应用于硬组织的替换与修复领域.采用表面改性技术在钛基材料表面制成生物陶瓷涂层可改善钛基材料的生物活性和生物相容性.羟基磷灰石涂层已在临床上获得应用,但使用效果仍然受其较低的结合强度和结晶度所制约.为了获得综合性能更好的植入材料,制备了多种新型生物陶瓷涂层,其具有良好的生物活性、较好的...     

Microtensile strain on the corrosion performance of diamond-like carbon coating

Hydrogenated diamond-like carbon films (a-C:H DLC) were deposited on STS 304 substrates for the fabrication of vascular stents by means of the r.f. plasma-assisted chemical vapor deposition technique. This study provides reliable and quantitative data for the assessment of the effect of strain on the corrosion performance of DLC-coated systems in the simulated body fluid obtained through electrochemical techniques (potentiodynamic polarization test and electrochemical impedance spectroscopy) and surface analysis (scanning electron microscopy). The electrolyte used in this test was 0.89% NaCl solution at pH 7.4 and 37 degrees C. It was found that the corrosion resistance of the plastically deformed DLC coating was insufficient for use as a protective film in a corrosive body environment. This is due to the increase in the delamination area and degradation of the substrate's corrosion properties with increasing tensile deformation.     

In vitro study of drug loading on polymer-free oxide films of metallic implants

Traditionally, a drug that is loaded onto a metallic surface has to use various polymer bondings as its platform. Unfortunately, polymer coatings on a metallic surface cause numerous problems after implantation, such as late thrombosis, inflammation, and restenosis. This research was conducted to investigate whether an oxide layer can be used as a polymer-free platform for drug loading, especially for cardiovascular stents. The interaction and loading of heparin onto different oxide films on 316LVM stainless steel wire was confirmed in vitro by experimental studies using linear voltammetry, electrochemical impedance spectroscopy, and electron spectroscopy for chemical analysis. The eluting of heparin from heparinized surface was studied by using high-performance liquid chromatography, and activated clotting time in addition to linear voltammetry and electron spectroscopy for chemical analysis analyses. Experimental results show that amorphous oxide could be a potential substitute for the polymer coating of drug-loaded stents for minimizing metallic corrosion, inflammation, late thrombosis, and restenosis.     

A quantitative model to predict pathogenicity of missense variants in the TP53 gene

Germline pathogenic variants in the TP53 gene cause Li‐Fraumeni syndrome, a condition that predisposes individuals to a wide range of cancer types. Identification of individuals carrying a TP53 pathogenic variant is linked to clinical management decisions, such as the avoidance of radiotherapy and use of high‐intensity screening programs. The aim of this study was to develop an evidence‐based quantitative model that integrates independent in silico data (Align‐GVGD and BayesDel) and somatic to germline ratio (SGR), to assign pathogenicity to every possible missense variant in the TP53 gene. To do this, a likelihood ratio for pathogenicity (LR) was derived from each component calibrated using reference sets of assumed pathogenic and benign missense variants. A posterior probability of pathogenicity was generated by combining LRs, and algorithm outputs were validated using different approaches. A total of 730 TP53 missense variants could be assigned to a clinically interpretable class. The outputs of the model correlated well with existing clinical information, functional data, and ClinVar classifications. In conclusion, these quantitative outputs provide the basis for individualized assessment of cancer risk useful for clinical interpretation. In addition, we propose the value of the novel SGR approach for use within the ACMG/AMP guidelines for variant classification.     

一种新的沙门氏菌成簇重复序列的预测方法

目的建立一个新的、直观的鉴定和显示细菌基因组成簇重复序列的方法,了解沙门氏菌基因组成簇重复序列的组成特征。方法直接将细菌的基因组用滑动窗口法切割成重叠片段,每一个片段自体比对,利用Pip Maker生成共线性图。通过共线性图特征识别成簇重复序列区。结果用所设计方案-成簇重复序列预测器(CRpred)对鼠伤寒沙门氏菌LT2菌株基因组进行扫描,总共鉴定出151个成簇重复区。特征分析表明,这些成簇重复区包含低拷贝简单串联重复、高拷贝简单串联重复、间隔串联重复、反向回文重复和反向间隔重复等5种类型。此外,沙门氏菌基因组中有9个复杂重复区域无法使用上述模式进行归类。结论提出了一个新的、简便直观的基因组成簇重复序列的鉴定方法,为搜寻成簇的、规律间隔的短回文重复序列(CRISPR)、数目可变串联重复序列(VNTR)以及其他重复序列相关研究提供支持。