Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio 44106, USA.
J Biomed Mater Res A. 2010 Sep 1;94(3):683-7. doi: 10.1002/jbm.a.32682.
The effect of polymorphonuclear leukocytes (PMNs) on the subsequent chronic phase macrophage-mediated foreign body reaction has not been previously investigated. Furthermore, while monocyte/macrophage-produced cytokines such as GM-CSF, G-CSF, or IL-1beta have been shown to increase PMN survival in vitro, few studies have examined the impact of directly cocultured monocytes/macrophages on PMN viability. To this end, we used our established in vitro system of interleukin (IL)-4-induced monocyte-derived macrophage fusion to examine the role of PMNs in the subsequent foreign body reaction. Monocytes were directly cultured with PMNs for 3 days before the addition of IL-4 to induce monocyte-derived macrophage fusion to facilitate foreign body giant cell (FBGC) formation by days 7 and 10 of culture. Optical microscopy was used to quantitatively determine adherent monocyte density, percent macrophage fusion, and FBGC density. A colorimetric MTT assay was used to assess PMN viability for direct cocultures of monocytes/macrophages and PMNs. Our results strongly suggest that the presence of PMNs inhibit IL-4-induced macrophage fusion and FBGC formation. Additionally, our findings demonstrate that cocultures containing PMNs and monocytes/macrophages increases PMN survival with respect to PMN-only cultures in vitro.
多形核白细胞 (PMN) 对随后的慢性期巨噬细胞介导的异物反应的影响尚未被研究过。此外，虽然单核细胞/巨噬细胞产生的细胞因子，如 GM-CSF、G-CSF 或 IL-1β，已被证明可以增加体外 PMN 的存活率，但很少有研究检查直接共培养的单核细胞/巨噬细胞对 PMN 活力的影响。为此，我们使用我们已建立的白细胞介素 (IL)-4 诱导的单核细胞衍生的巨噬细胞融合的体外系统，来研究 PMN 在随后的异物反应中的作用。在添加 IL-4 以诱导单核细胞衍生的巨噬细胞融合从而促进培养第 7 天和第 10 天的异物巨细胞 (FBGC) 形成之前，将单核细胞直接与 PMN 共培养 3 天。光学显微镜用于定量测定粘附的单核细胞密度、巨噬细胞融合百分比和 FBGC 密度。比色 MTT 测定法用于评估单核细胞/巨噬细胞和 PMN 的直接共培养物中的 PMN 活力。我们的研究结果强烈表明 PMN 的存在抑制了 IL-4 诱导的巨噬细胞融合和 FBGC 形成。此外，我们的发现表明，与仅含 PMN 的培养物相比，含有 PMN 和单核细胞/巨噬细胞的共培养物增加了 PMN 的体外存活率。
