Godek M L, Sampson J A, Duchsherer N L, McElwee Q, Grainger D W
J Biomater Sci Polym Ed. 2006;17(10):1141-1158. doi: 10.1163/156856206778530731.
The Rho GTPase cellular signaling cascade was investigated in pro-monocyte and (monocyte-)macrophage cells by examining GTPase expression and activation in serum-containing cultures on model biomaterials. Abundance of Rho GDI and the Rho GTPase proteins RhoA, Cdc42 and Rac1 was determined in cells grown on tissue culture polystyrene, polystyrene, poly-l-lactide and Teflon(®) AF surfaces. Protein expression was compared based on cell maturity (pro-monocyte to monocyte to macrophage lineages) and by model surface chemistry: Rho proteins were present in the majority of macrophage cells tested on model surfaces suggesting that a pool of Rho proteins is readily available for signaling events in response to numerous activating cues, including biomaterials surface encounter. Rho GTPase activation profiles in these cell lines indicate active Cdc42 and Rho proteins in RAW 264.7, Rac1 and Rho in J774A.1, and Cdc42 and Rac1 in IC-21 cell lines, respectively. Collectively, these proteins are known to play critical roles in all actin-based cytoskeletal rearrangement necessary for cell adhesion, spreading and motility, and remain important to establishing cellular responses required for foreign body reactions in vivo. Differences in Rho GTPase protein expression levels based on cell sourcing (primary versus secondary-derived cell source), or as a function of surface chemistry were insignificant. Rho GTPase expression profiles varied between pro-monocytic non-adherent precursor cells and mature adherent monocyte/macrophage cells. The active GTP-bound forms of the Rho GTPase proteins were detected from monocyte-macrophage cell lines RAW 264.7 and J774A.1 on all polymer surfaces, suggesting that while these proteins are central to cell adhesive behavior, differences in surface chemistry are insufficient to differentially regulate GTPase activation in these cell types. Active Cdc42 was detected from cells cultured on the more-polar tissue culture polystyrene and poly-l-lactide surfaces after several days, but absent from those grown on apolar polystyrene and Teflon(®) AF, indicating some surface influence on this GTPase in serum-containing cultures.
通过检测含血清培养的模型生物材料上的GTP酶表达和激活情况，研究了原单核细胞和（单核细胞 -）巨噬细胞中的Rho GTP酶细胞信号转导级联反应。在组织培养聚苯乙烯、聚苯乙烯、聚左旋乳酸和特氟龙（®）AF表面生长的细胞中，测定了Rho GDI以及Rho GTP酶蛋白RhoA、Cdc42和Rac1的丰度。基于细胞成熟度（原单核细胞到单核细胞再到巨噬细胞谱系）和模型表面化学性质比较了蛋白质表达：在模型表面测试的大多数巨噬细胞中都存在Rho蛋白，这表明存在大量Rho蛋白，可随时用于响应包括生物材料表面接触在内的众多激活信号的信号转导事件。这些细胞系中的Rho GTP酶激活谱分别表明RAW 264.7细胞系中Cdc42和Rho呈活性状态，J774A.1细胞系中Rac1和Rho呈活性状态，IC - 21细胞系中Cdc42和Rac1呈活性状态。总的来说，已知这些蛋白质在细胞粘附、铺展和运动所需的所有基于肌动蛋白的细胞骨架重排中起关键作用，并且对于建立体内异物反应所需的细胞反应仍然很重要。基于细胞来源（原代细胞与二代细胞来源）或作为表面化学性质的函数，Rho GTP酶蛋白表达水平的差异不显著。Rho GTP酶表达谱在原单核非粘附前体细胞和成熟粘附单核细胞/巨噬细胞之间有所不同。在所有聚合物表面上，均从单核巨噬细胞系RAW 264.7和J774A.1中检测到了Rho GTP酶蛋白的活性GTP结合形式，这表明虽然这些蛋白质对于细胞粘附行为至关重要，但表面化学性质的差异不足以在这些细胞类型中差异调节GTP酶的激活。几天后，在极性更强的组织培养聚苯乙烯和聚左旋乳酸表面培养的细胞中检测到了活性Cdc42，但在非极性聚苯乙烯和特氟龙（®）AF表面生长的细胞中未检测到，这表明在含血清培养中该GTP酶受到一定的表面影响。
