三维多孔碳纤维/聚乳酸/壳聚糖复合支架材料的体外细胞相容性评价

利用溶液共混法以及冷冻干燥法制备了三维多孔碳纤维/聚乳酸/壳聚糖(CF/PLA/CS)复合生物支架材料,通过相差显微镜和扫描电子显微镜检测了鼠骨髓基质细胞（BMSCs）与该材料的生物相容性,得到了MTT生长曲线,评价了材料的毒性。 结果表明,以实验组材料的浸提液培养细胞,8 d后细胞开始连片生长,没有观察到细胞变形、坏死现象;在实验组材料上培养4 d后细胞的SEM图像显示,细胞形貌正常,并已开始向孔隙深部生长;MTT法绘制的增值曲线表明,培养4 d后实验组的细胞增殖速度高出空白对照组30%。 以上细胞形态学观察法和细胞增殖法评价结果表明,三维多孔 CF/PLA/CS复合材料没有细胞毒性,并对细胞有良好的粘附、增殖能力,有望成为骨修复材料。     

油页岩干馏瓦斯中的丙烯组分在不同耐热合金上的积炭特性

采用自制的连续流动反应装置,分别以2520及316L不锈钢管为反应器,对不同气体流量(20、30、40 m L/min)、不同壁温(700、750、800℃)、恒定反应时间(20min)条件下干馏瓦斯中丙烯组分在不同反应器内的积炭特性的实验结果表明,两种管壁上初始积炭温度范围分别为630~650℃及670~690℃;扫描电镜对积炭微观形态观察表明,管内积炭都以丝状炭和无定形炭为主;程序升温氧化方法对不同工况下两种反应管内积炭量分析得出,316L耐热合金具有抗金属粉化特性;热重及差示扫描量热法对管内剥离炭样活性的表征结果证实,316L耐热合金内积炭活性比2520耐热合金内的高。     

聚二甲基硅氧烷-纸复合微流控芯片上的肝癌细胞三维培养

研发了一种聚二甲基硅氧烷-纸复合型微流控芯片用于肝癌细胞三维培养.芯片使用明胶处理硝酸纤维素薄膜作为细胞培养基底,以水凝胶网格作为三维培养支撑.结合微通道主动灌流与水凝胶中的被动扩散,模拟体内的流体运输形式实现细胞与外界物质交换.实验结果显示,芯片上的液滴生成以及细胞定位种植简便可靠.连续监测显示肝癌HcpG2细胞在水凝胶微球中增殖形成类似组织的三维结构.细胞增殖动力学分析以及生化检测结果显示了芯片三维培养与二维培养的差别.这种芯片三维细胞培养方法操作简便可靠,仿真度高,适合于肿瘤细胞研究.     

Gd3+对原代培养的小鼠骨髓基质细胞成骨分化和成脂分化的影响

本文采用MTT法、碱性磷酸酶活性测定、矿化功能的测定以及油红O的染色和定量测定等手段研究了Gd3+对原代培养的小鼠骨髓基质细胞成骨分化和成脂分化的影响。研究结果表明,浓度为1×10-10和1×10-8 mol.L-1的Gd3+对小鼠骨髓基质细胞的增殖没有影响,其他测试浓度下的Gd3+则抑制小鼠骨髓基质细胞的增殖。当Gd3+与小鼠骨髓基质细胞作用7 d时,其对小鼠骨髓基质细胞成骨分化的影响与作用浓度有关,当Gd3+与小鼠骨髓基质细胞作用14 d时,在全部测试浓度范围内,抑制小鼠骨髓基质细胞成骨分化。除1×10-8和1×10-5 mol.L-1外,其他测试浓度下的Gd3+促进小鼠骨髓基质细胞的矿化功能。当Gd3+与小鼠骨髓基质细胞作用10 d时,其抑制小鼠骨髓基质细胞的成脂分化,当Gd3+与小鼠骨髓基质细胞作用16 d时,除1×10-9mol.L-1外,其他浓度的Gd3+也抑制小鼠骨髓基质细胞的成脂分化。实验结果提示,Gd3+可能通过促进骨髓基质细胞的成骨分化、抑制其成脂分化途径起到对骨的保护作用。Gd3+对原代培养的小鼠骨髓基质细胞成骨分化和成脂分化的影响与作用浓度和时间有关,而且,它们是影响Gd3+对骨是损伤还是保护作用转变的关键因素。     

基于甲基三苯乙炔基硅烷的新型聚三唑树脂的制备与性能

首先采用格氏试剂法合成了甲基三苯乙炔基硅烷(MTPES),通过傅里叶变换红外光谱(FT-IR)、核磁共振氢谱(1 H-NMR)对其结构进行了表征。然后以MTPES和4,4’-二叠氮甲基联苯(BAMBP)为原料制备了新型聚三唑树脂(MPTA)。利用FT-IR和差示扫描量热(DSC)研究了MPTA树脂的固化行为,通过动态力学热分析(DMA)和热重分析(TG)研究了炔基与叠氮基配比对树脂热性能的影响,并通过测试凝胶时间随贮存时间的变化研究了树脂及其四氢呋喃(THF)溶液的贮存稳定性。结果表明,固化后的树脂玻璃化转变温度(Tg)达到236℃,在氮气中的5%热失重温度(T_(d5))在320℃左右。MPTA树脂在35℃和25℃下分别贮存7d和20d后,100℃下树脂的凝胶时间分别为40min和25min,MPTA树脂的THF溶液在同样条件下贮存28d后,凝胶时间分别为54min和61min,具有比现有聚三唑树脂更好的贮存稳定性。单向T700碳纤维-MPTA复合材料常温下的弯曲强度为1 660 MPa,弯曲模量为129 GPa,150℃下的弯曲强度保留率为70%。     

酸水解法从菊芋中提取果糖

采用热水浸提法提取菊芋中的菊糖,并用磷酸水解为还原糖(大部分为果糖)。从固液比、提取时间、温度、水解时间、温度和菊糖液pH值等方面试验了对还原糖得率的影响,通过正交实验确定了最佳条件为:固液比1∶30,浸提温度80℃,浸提时间30min,水解温度80℃,水解时间30min,pH值为2,可以得到较令人满意的结果。     

聚乙烯醇固定化的微球菌AD3对除草剂阿特拉津的生物降解

从农药厂的工业废水和污泥的混合物中分离到高效降解除草剂阿特拉津的藤黄微球菌(Micrococcusluteus)AD3菌株。以聚乙烯醇(PVA)为包埋材料进行AD3菌株的固定化,对影响固定化细胞降解阿特拉津的因素(如阿特拉津浓度、温度、pH)和操作稳定性进行了研究。结果表明,在最适条件下(30℃、pH7.2、阿特拉津浓度500mg/L)固定化细胞的阿特拉津降解速度明显高于游离细胞。固定化细胞在低温、低pH和高阿特拉津浓度条件下的阿特拉津降解率,以及贮存和操作稳定性,也明显好于游离细胞。固定化细胞在15℃、pH7.2、阿特拉津浓度为500mg/L的条件下培养72h以后,阿特拉津降解率为90%,在相同条件下游离细胞的降解率仅为35%。固定化细胞在pH5.0、30℃、阿特拉津浓度为500mg/L的条件下培养72h以后,降解率为96%,在相同条件下游离细胞的降解率仅为40%。固定化细胞能够快速降解1000mg/L的阿特拉津,而游离细胞则不能。稳定性实验表明,固定化细胞重复使用30次以后其降解活力没有明显降低。     

动力学因素对热诱导相分离法制备亲水性乙烯-丙烯酸共聚物微孔膜结构的影响

选择 3种不同丙烯酸含量的乙烯 丙烯酸共聚物 (EAA)为原材料 ,二苯醚 (DPE)为稀释剂 ,研究了淬冷温度、粗化时间等影响液滴生长的动力学因素对热诱导相分离法 (TIPS)制备EAA DPE亲水性高分子微孔膜结构的影响 .淬冷温度的高低决定了EAA DPE体系是发生液 液相分离还是固 液相分离 ,而产生相分离的机理不同将影响稀释剂液滴的生长 ,最终影响微孔膜的孔径 .实验结果表明 ,在相同粗化时间的条件下 ,随着EAA1 41 0 DPE、EAA3 0 0 2 DPE、EAA3 0 0 3 DPE三体系冷却温度的逐渐升高 ,孔径逐渐变大 .在结晶温度以下 ( 0℃、3 0℃、60℃ )粗化时间相同时 ,温度对微孔膜的孔径影响较小 ,例如 0℃和 3 0℃的恒温条件粗化 1 0min,微孔膜的孔径在 1～ 3 μm之间 ;在 60℃的恒温条件粗化 1 0min ,微孔膜的孔径在 3～ 5 μm之间 .而在 90℃的恒温条件粗化相同的时间 ,由于体系始终处于结晶温度线以上 ,体系始终处在液 液相分离区域 ,最终得到微孔膜的孔径达到了 6～8μm .在结晶温度以下 ( 3 0℃ )进行恒温粗化 ,由于体系的过冷程度很大 ,液滴相的粗化过程被抑制住 ,所以粗化时间对微孔膜的孔径影响不大 ;而在结晶温度以上 ( 90℃ )进行恒温粗化时 ,则是随着粗化时间的延长 ,微孔膜的孔径逐渐变大     

极限稀释法微孔池培养测定造血前体细胞

根据极限稀释法(Limiting dilution)原理,应用微孔池培养无增殖细胞生长的“阴性池”及有增殖细胞生长的“阳性池”分析,测定了小鼠造血前体细胞Haemopoietic progenitor cells(HPC)在骨髓中的含量。以形态学及~3H-TdR参入率为指标所测定的Balb/c小鼠HPC的含量分别为33及31/10~5骨髓细胞。此外,在培养后4天及7天,按不同浓度接种的细胞数与收获细胞数之间呈现良好的线性关系,反映了在微孔池中HPC的增殖。本方法除用以测定在骨髓中的HPC含量以外,尚可用于分析各种培养条件、造血因子以及细胞毒药物等对其增殖和分化的影响。     

顶空-毛细管气相色谱法测定印刷品中15种痕量挥发性有机物

采用顶空-气相色谱法对印刷品中的挥发性有机物(苯、甲苯、乙苯、二甲苯、乙醇等15种)进行分析,研究了不同升温程序、顶空平衡温度、平衡时间等对分析结果的影响,优化的实验参数为:进样口温度:200℃;检测器温度:250℃。升温程序:初始温度40℃,保持5 min;以20℃/min速率升温至200℃,保持3 min。以高纯氮气作为载气,流速为8.0 mL/min保持5 min,再以0.2 mL/min的速率降至5.0 mL/min(分流比5:1);顶空条件:进样瓶平衡温度为90℃,平衡时间30 min,顶空进样体积1.0 mL。本方法线性关系良好,15种挥发性有机物的相关系数均大于0.9998,高、中、低3个浓度的回收率为77.8%~100.6%;相对标准偏差均低于6.0%(n=6)。方法可用于印刷品中挥发性有机物的分析。     

Spatiotemporal control of cell adhesion on a self-assembled monolayer having a photocleavable protecting group

Control of cell adhesion is a key technology for cell-based drug screening and for analyses of cellular processes. We developed a method to spatiotemporally control cell adhesion using a photochemical reaction. We prepared a cell-culturing substrate by modifying the surface of a glass coverslip with a self-assembled monolayer of an alkylsiloxane having a photocleavable 2-nitrobenzyl group. Bovine serum albumin (BSA) was adsorbed onto the substrate to make the surface inert to cell adhesion. When exposed to UV light, the alkylsiloxane underwent a photocleavage reaction, leading to the release of BSA from the surface. Fibronectin, a protein promoting cell adhesion, was added to cover the irradiated regions and made them cell-adhesive. Seeding of cells on this substrate resulted in their selective adhesion to the illuminated regions. By controlling the sizes of the illuminated regions, we formed cell-adhesive spots smaller than single cells and located focal adhesions of the cells. Moreover, by subsequently illuminating the region alongside the cells patterned on the substrate in advance, we released their geometrical confinements and induced migration and proliferation. These manipulations were conducted under a conventional fluorescence microscope without any additional instruments. The present method of cell manipulation will be useful for cell biological studies as well as for the formation of cell arrays.     

铅中毒细胞遗传学效应

对确诊为铅吸收和铅中毒的铅冶炼５９名，进行了外周血淋巴细胞染色体畸变（ＣＡ），姐妹单体互换（ＳＣＥ），微核（ＭＮ）等细胞遗传学指标检测观察，并与血铅、尿铅、ＺＰＰ、ＦＥＰ及尿δ－ＡＬＡ等生化指标进行相关分析，结果表明，铅毒作用能引起细胞染色体畸变率、姐妹染色单体交换率及微核率提高，提示铅能引起ＤＮＡ损伤，但无明显致突变作用。     

Electrochemical detection of cell concentration based on reaction of DNA with molybdate

This article describes a fast and simple electrochemical assay for detecting cell concentration.After cell death,the membrane of cells will be broken,and DNA molecules contained in the cells will be released,but this does not happen in living cells.Sodium molybdate can react with the phosphate backbone of the released DNA molecules to form phosphomolybdate precipitation and produces a corresponding redox current.The higher the concentration of DNA,the stronger the intensity of the current generated.Sodium molybdate solution and centrifuged cell supernatant were added onto the glassy carbon electrode to determine the cell concentration by measuring the current intensity.The cell viability,which means the ratio of living cells to the total cells,can also be determined by this method.This assay has the advantages of high sensitivity,low detection limit,and wide detection range.In addition,this method was successfully applied to the detection of cell concentration in human serum,which has potential clinical applications.     

微流控芯片上细胞培养与分析方法研究进展

微流控芯片以其强大的微流体和微小物质控制能力成为研究单细胞、细胞群落乃至生物组织的重要手段。在本篇综述中,我们将以微流控芯片上细胞体外培养模型的建立为主,对近几年来重要的研究工作加以评述,全面地介绍微流控技术在细胞生命科学研究中应用的优势和未来发展方向,具体包括微流控芯片的细胞操控能力、细胞培养微环境的构建以及芯片联用检测手段,希望为从事这一领域研究工作的读者提供一些新的思路。     

Recent Advances on Surface-modified Biomaterials Promoting Selective Adhesion and Directional Migration of Cells

The surfaces and interfaces of biomaterials interact with the biological systems in multi-scale levels, and thereby influence the biological functions and comprehensive performance in vitro and in vivo. In particular, a surface promoting the selective adhesion and directional migration of desired types of cells in complex environment is extremely important in the repair and regeneration of tissues such as peripheral nerve and blood vessel, and long-term application of intracorporal devices such as intravascular implants. Therefore, surface modification of biomaterials is a facile and effective method to achieve the desired cell-biomaterials interactions. In this short review, recent advances on the surface modification of biomaterials to regulate selective cell adhesion and migration are briefly summarized. In particular, the surface properties of biomaterials are manipulated via the convenient introduction of amino groups to the ester-based polymers, the formation of polyelectrolyte multilayers, and the fabrication of topology and gradient cues, etc., followed by the association of chemical and biological signals such as collagen, heparin, hyaluronic acid, peptides and cell growth factors. The selective adhesion and directional migration of various types of cells such as endothelial cells(ECs), smooth muscle cells(SMCs), hepatocytes and Schwann cells(SCs) are achieved over the competitive counterpart cells by the use of cell-resisting substances and cell-selective motifs on gradient substrates in most cases. Recent advances on cell behaviors in three-dimensional(3D) cell-extracellular matrix(ECM)-mimicking substrates are also reviewed.     

The effects of flow type on aptamer capture in differential mobility cytometry cell separations

In this work, differential mobility cytometry (DMC) was used to monitor cell separation based on aptamer recognition for target cells. In this device, open-tubular capillaries coated with Sgc8 aptamers were used as affinity chromatography columns for separation. After cells were injected into the columns, oscillating flow was generated to allow for long-term cell adhesion studies. This process was monitored by optical microscopy, and differential imaging was used to analyze the cells as they adhered to the affinity surface. We investigated the capture time, capture efficiency, purity of target and control cells, as well as the reusability of the affinity columns. Capture time for both CCRF-CEM cells and Jurkat T cells was 0.4 ± 0.2 s, which demonstrated the high separation affinity between aptamers and target cells. The capture efficiency for CCRF-CEM cells was 95% and purity was 99% in a cell mixture. With the advantage of both high cell capture efficiency and purity, DMC combined with aptamer-based separation emerges as a powerful tool for rare cell enrichment. In addition, aptamer-based DMC channels were found to be more robust than antibody based channels with respect to reuse of the separation device.     

Study of cell behaviour on a cellulose anti-adhesive substratum

Carboxymethylcellulose (CMC) is used as an anti-adhesive in several biomaterial applications. We have shown that an original bi-layered hydroxypropylmethylcellulose (HPMC)-CMC-coated substratum (CEL) is suitable for studying the molecular mechanisms underlying cell–substratum and cell–cell interactions. The present work was carried out to assess the anti-adhesive properties of a CEL substratum by analysing the behaviour of three types of adherent cells (Swiss 3T3 fibroblasts, MC-3T3-E1 pre-osteoblasts and B16F10 highly metastatic melanoma cells). Tissue culture polystyrene (tPS) was used as an adhesive control and poly(2-hydroxyethyl methacrylate) PolyHEMA as an anti-adhesive control. Cell morphology, growth, cell cycle analysis and apoptosis were compared. The proliferation of cells on CEL was significantly decreased, cells were arrested in the G1 phase and underwent apoptosis (except for melanoma cells) 48 h post-seeding. We conclude that CEL is an effective anti-adhesive cellulose biomaterial that gives reproducible and demonstratable results, especially for apoptosis. Its advantage as a coating for culture devices is its long shelf-live. It may therefore also be a very good candidate for other biomedical applications requiring adhesion-resistant substrata.     

新型杀菌剂氟吗啉对黄瓜疫霉病菌细胞壁主要组分合成及分布的影响

应用同位素示踪和组织特异性荧光染色等生物化学方法研究了新型杀菌剂氟吗啉对黄瓜疫霉病菌细胞壁主要组分合成及分布的影响. 结果表明, 氟吗啉对细胞壁的主要组成物质纤维素和非纤维素葡聚糖的合成没有抑制作用, 但破坏细胞壁物质在菌丝正确位置的形成和分布, 扰乱菌丝的极性生长.     

聚苯乙烯细胞培养板表面的糖化温敏修饰及其对细胞行为的影响

采用紫外光固定化法, 对组织培养用聚苯乙烯板进行半乳糖糖化温敏修饰. 通过红外光谱(ATR-FTIR)和X射线光电子能谱(XPS)对改性表面的化学组成及结构进行了表征, 并采用原子力显微镜(AFM)观察了改性表面形貌, 发现改性表面比未经修饰表面粗糙度增加. 静态接触角测试结果表明, 改性表面具有良好的温度响应性. 对人肝肿瘤(HepG-2)细胞在改性表面的吸/脱附行为的研究结果表明, HepG-2细胞在半乳糖糖化温敏表面表现出比在未经修饰聚苯乙烯细胞培养板表面更好的生长趋势, 当环境温度降低时, 细胞发生自动脱附, 避免了酶解法对细胞功能造成的损伤.     

Subcellular topological effect of particle monolayers on cell shapes and functions

We studied topological effects of subcellular roughness displayed by a closely packed particle monolayer on adhesion and growth of endothelial cells. Poly(styrene-co-acrylamide) (SA) particles were prepared by soap-free emulsion copolymerization. Particle monolayers were prepared by Langmuir–Blodgett deposition using particles, which were 527 (SA053) and 1270 nm (SA127) in diameter. After 24-h incubation, cells tightly adhered on a tissue culture polystyrene dish and randomly spread. On the other hand, cells attached on particle monolayers were stretched into a narrow stalk-like shape. Lamellipodia spread from the leading edge of cells attached on SA053 monolayer to the top of the particles and gradually gathered to form clusters. This shows that cell–cell adhesion became stronger than cell–substrate interaction. Cells attached to SA127 monolayer extended to the reverse side of a particle monolayer and engulfed particles. They remained immobile without migration 24 h after incubation. This shows that the inhibition of extensions on SA127 monolayer could inhibit cell migration and cell proliferation. Cell growth on the particle monolayers was suppressed compared with a flat TCPS dish. The number of cells on SA053 gradually increased, whereas that on SA127 decreased with time. When the cell seeding density was increased to 200,000 cells cm⁻², some adherent cells gradually became into contact with adjacent cells. F-actin condensations were formed at the frame of adherent cells and the thin filaments grew from the edges to connect each other with time. For the cell culture on SA053 monolayer, elongated cells showed a little alignment. Cells showed not arrangement of actin stress fibers but F-actin condensation at the contact regions with neighboring cells. Interestingly, the formed cell monolayer could be readily peeled from the particle monolayer. These results indicate that endothelial cells could recognize the surface roughness displayed by particle monolayers and the response was dependent on the pitch of particle monolayers.