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首页> 外文学位 >Growth and behavior of chondrocytes on nano engineered surfaces and construction of micropatterned co-culture platforms using layer-by-layer platforms using layer-by-layer assembly lift-off method.
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Growth and behavior of chondrocytes on nano engineered surfaces and construction of micropatterned co-culture platforms using layer-by-layer platforms using layer-by-layer assembly lift-off method.

机译：纳米工程表面上软骨细胞的生长和行为以及使用逐层组装剥离法的逐层平台构建微模式共培养平台。

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摘要

Several approaches such as self-assembled monolayers and layer-by-layer assembled multilayer films are being used as tools to study the interactions of cells with biomaterials in vitro. In this study, the layer-by-layer assembly approach was used to create monolayer, bilayer, trilayer, five, ten and twenty-bilayer beds of eleven different biomaterials. The various biomaterials used were poly(styrene-sulfonate), fibronectin, poly-L-lysine, poly-D-lysine, laminin, bovine serum albumin, chondroitin sulfate, poly(ethyleneimine), polyethylene glycol amine, collagen and poly(dimethyldiallyl-ammonium chloride) with unmodified tissue-culture polystyrene as standard control. Three different cell lines---primary bovine articular chondrocytes, and two secondary cell lines, human chondrosarcoma cells and canine chondrocytes were used in these studies. Chondrocyte morphology and attachment, viability, proliferation, and functionality were determined using bright field microscopy, the Live/Dead viability assay, MTT assay, and immunocytochemistry, respectively.; Atomic force microscopy of the nanofilms indicated an increase in surface roughness with increasing number of layers. The most important observations from the studies on primary bovine articular chondrocytes were that these cells exhibited increasing viability and cell metabolic activity with increasing number of bilayers. The increase in viability was more pronounced than the increase in cell metabolic activity. Also, bovine chondrocytes on bilayers of poly(dimethyldiallyl-ammonium chloride, poly-L-lysine, poly(styrene-sulfonate), and bovine serum albumin were substantially bigger in size and well-attached when compared to the cells grown on monolayer and trilayers. Lactate dehydrogenase assay performed on chondrosarcoma cells grown on 5- and 10-bilayer multilayer beds indicated that the 10-bilayer beds had reduced cytotoxicity compared to the 5-bilayer beds. MTT assay performed on canine chondrocytes grown on 5-, 10-, and 20-bilayer nanofilm beds revealed increasing cell metabolic activity for BSA with increasing bilayers.; Micropatterned multilayer beds having poly-L-lysine, poly-D-lysine, laminin poly(dimethyldiallyl-ammonium chloride) and poly(ethyleneimine) as the terminating layers were fabricated using the Layer-by-layer Lift-off (LbL-LO) method that combines photolithography and LbL self-assembly. Most importantly, micropatterned co-culture platforms consisting of anti-CD 44 rat monoclonal and anti-rat osteopontin (MPIIIB101) antibodies were constructed using the LbL-LO method for the first time. These co-culture platforms have several applications especially for studies of stem and progenitor cells. Co-culture platforms exhibiting spatiotempora-based differentiation can be built with LbL-LO for the differentiation of stem cells into the desired cell lineage.
机译：诸如自组装单层和逐层组装的多层薄膜等几种方法正被用作研究细胞与生物材料在体外相互作用的工具。在这项研究中，采用逐层组装的方法来创建11种不同生物材料的单层，双层，三层，五层，十层和二十层双层床。使用的各种生物材料是聚（苯乙烯磺酸盐），纤连蛋白，聚-L-赖氨酸，聚-D-赖氨酸，层粘连蛋白，牛血清白蛋白，硫酸软骨素，聚（乙烯亚胺），聚乙二醇胺，胶原蛋白和聚（二甲基二烯丙基-氯化铵）和未改性的组织培养聚苯乙烯作为标准对照。在这些研究中，使用了三种不同的细胞系-原代牛关节软骨细胞和两种次生细胞系，即人软骨肉瘤细胞和犬软骨细胞。分别使用明视野显微镜，活/死活力测定法，MTT测定法和免疫细胞化学测定软骨细胞的形态和附着，活力，增殖和功能。纳米膜的原子力显微镜检查表明，表面粗糙度随层数的增加而增加。对原代牛关节软骨细胞的研究中最重要的观察结果是，随着双层数的增加，这些细胞显示出更高的生存力和细胞代谢活性。活力的增加比细胞代谢活性的增加更为明显。而且，与单层和三层细胞相比，聚二甲基二烯丙基氯化铵，聚L-赖氨酸，聚苯乙烯磺酸盐和牛血清白蛋白双层的牛软骨细胞的大小明显更大，且附着力更好。 。对在5层和10层多层床上生长的软骨肉瘤细胞进行的乳酸脱氢酶测定表明，与5层床相比，10层床具有降低的细胞毒性；对在5、10-， 20层纳米膜床揭示了BSA随双层数的增加而增加的细胞代谢活性；具有聚L-赖氨酸，聚D-赖氨酸，层粘连蛋白聚二甲基二烯丙基氯化铵和聚亚乙基亚胺的微图案化多层床层是使用光刻和LbL自组装相结合的逐层剥离（LbL-LO）方法制造的，最重要的是，由抗CD 44大鼠monocl组成的微模式共培养平台首次使用LbL-LO方法构建了抗大鼠骨桥蛋白（MPIIIB101）抗体。这些共培养平台具有多种应用，特别是用于干细胞和祖细胞的研究。可以使用LbL-LO构建展示基于时空差异的共培养平台，以将干细胞分化为所需的细胞谱系。

著录项

  • 作者

    Shaik, Jameel.;

  • 作者单位

    Louisiana Tech University.;

  • 授予单位 Louisiana Tech University.;
  • 学科 Engineering Biomedical.; Engineering Materials Science.
  • 学位 Ph.D.
  • 年度 2007
  • 页码 140 p.
  • 总页数 140
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 生物医学工程;工程材料学;
  • 关键词

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