Molecular Comparison of Macrophage Foreign Body Response

巨噬细胞异物反应的分子比较

基本信息

批准号：
7387029
负责人：
DAVID W GRAINGER
金额：
$ 25.99万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-01-01 至 2008-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7387029
关键词：
apoptosis bioengineering /biomedical engineering biomaterial evaluation biotechnology cell line collagen cytokine foreign body reaction gene expression profiling helper T lymphocyte immune response inflammation laboratory mouse macrophage microarray technology phenotype surface property

项目摘要

DESCRIPTION (provided by applicant): Current use of human- or animal-derived phagocytic cell sources required for biomaterials inflammation and foreign body assay is inconvenient, costly, and potentially unnecessary. Equivalence of secondary macrophage-lineage cell lines in these in vitro assays is largely unproven, although many unpublished anecdotes exist regarding their utility in biomaterials toxicity or inflammatory assays. This proposal's overall objective is to establish, using quantitative modem molecular and cell biological methods, the validity of certain macrophage-derived cell line cultures in accurately duplicating phenotypic behavior of human- and animal-derived phagocytes commonly used to model aspects of the biomaterial implant-generated foreign body response. Our working hypothesis is that phenotypic response and foreign body giant cell formation on several model biomaterials using select secondary macrophage-derived cell lines in cultures designed to monitor cell-mediated inflammatory responses on biomaterials will be measurably similar to that observed using host-derived primary macrophages. Studies seek answers to important, long-standing questions surrounding the appropriate use and limitations of macrophage culture systems to analyze the foreign-body response ubiquitous for all implanted biomaterials. New data will be compared against well-developed primary monocyte/macrophage culture assays now common for biomaterials inflammatory assessments with foreign body giant cells. The following Specific Aims are proposed: (1) Determine selected cell-produced cytokines representing the Th1 and Th2 phenotypic responses in cultured secondary macrophage-derived cell lines on several model biomaterials of controlled surface chemistry; (2) Identify short and long term phenotypic stability in culture through changes in cytokine production and responses in macrophage derived cell cultures as a function of surface chemistry; (3) Quantify several distinct known phenotypic features of foreign body giant cell formation as a function of surface chemistry for macrophage-derived cell lines and primary macrophages over time in culture; (4) Establish reliable, quantitative microarray-based gene analysis to accelerate profiling of specific known markers of macrophage activation and inflammatory behavior, specifically for cytokine expression and apoptosis markers; and (5) Exploit an in vitro co culture system containing cultured macrophages and fibroblasts to study affects of macrophage-derived cytokine production on fibroblast production of collagen and fibrous precursors relevant to encapsulation reactions common to foreign bodies in vivo.

描述（由申请人提供）：生物材料炎症和异物测定所需的人类或动物衍生的吞噬细胞来源的当前使用是不便，昂贵的，并且可能是不必要的。在这些体外测定中，继发性巨噬细胞细胞系的等效性在很大程度上尚未得到证实，尽管许多未发表的轶事在生物材料毒性或炎症测定中存在。该提案的总体目标是使用定量的调制解调器分子和细胞生物学方法建立某些巨噬细胞衍生的细胞系培养物的有效性，以精确地复制人类和动物衍生的吞噬细胞的表型行为，用于模拟生物材料植入物生成的异物反应的方面。我们的工作假设是，在培养物中使用精选的二次巨噬细胞衍生的细胞系上的几种模型生物材料的表型反应和异物巨细胞形成，旨在监测生物材料对细胞介导的炎症反应的培养物，类似于使用宿主衍生的主要巨噬细胞观察到的生物材料的炎症反应。研究寻求回答有关巨噬细胞培养系统适当使用和局限性的重要，长期存在的问题的答案，以分析所有植入的生物材料无处不在的外国体内反应。将新数据与发达的原发性单核细胞/巨噬细胞培养测定法进行比较，目前使用异物巨细胞进行生物材料炎症评估常见。提出了以下特定目的：（1）确定所选的细胞生产的细胞因子，代表培养的次级巨噬细胞衍生的二次巨噬细胞衍生的细胞系中的Th1和Th2表型反应，这些细胞系在受控表面化学的几种模型生物材料上；（2）通过细胞因子产生的变化和巨噬细胞衍生的细胞培养物的反应来确定培养中的短期和长期表型稳定性，这是表面化学的函数；（3）量化了外国体巨细胞形成的几种已知的已知表型特征，这是培养物中巨噬细胞衍生的细胞系和原代巨噬细胞表面化学的函数；（4）建立可靠的，定量的基于微阵列的基因分析，以加快巨噬细胞激活和炎症行为的特定已知标记物的分析，特别是用于细胞因子表达和凋亡标记物的特定标记；（5）利用含有培养的巨噬细胞和成纤维细胞的体外CO培养系统来研究巨噬细胞衍生的细胞因子产生对与与外国体内相关的封装反应相关的胶原蛋白和纤维细胞产生的胶原蛋白和纤维细胞产生的影响。