Bernatchez S F, Parks P J, Gibbons D F
Center for Interfacial Engineering, University of Minnesota, Minneapolis 55455, USA.
Biomaterials. 1996 Nov;17(21):2077-86. doi: 10.1016/0142-9612(96)00014-2.
Fibrous materials have the potential of being used for tissue scaffolding. The interaction of macrophages with fibres of various compositions and sizes was observed in vitro. The following materials were tested: individual gold fibres; woven fibres of polyester and nylon; non-woven fibres of polybutylene/polypropylene 80:20 and polyester. All fibres had diameters between 2 and 40 microns. At the end of the 24 h incubation time, culture media were retrieved for the assay of tumour necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6), two cytokines secreted by activated macrophages. Fibre samples were processed for scanning electron microscopy (SEM), or for immunofluorescence labelling of the MAC-1 and ICAM-1 cell surface markers. Confocal microscopy was used for the latter, which was performed with the woven and non-woven samples. None of the fibre samples induced significant amounts of TNF-alpha or IL-6 secretion in the culture medium, suggesting that the cells did not activate this pathway. SEM on individual gold fibres showed that the fibre diameter had an effect on the morphology of the cells, namely on their extent of spreading. Larger fibres had a higher number of cells, which tended to cluster together without spreading extensively. When comparing woven and non-woven fibres, SEM showed that cells spread extensively on the woven fibres, whereas they tended to maintain their spherical shape on the non-woven fibres. Confocal microscopy demonstrated a difference between materials in the number of MAC-1 and ICAM-1 positive cells. These results demonstrate that a combination of morphological, immune and biochemical markers can be used to distinguish the response of elicited macrophages to various materials. The cells appeared to be only moderately activated on all materials tested, with changes in their morphology but without increased secretion of cytokines. The measured responses imply interactions between nominal fibre composition and fibre diameter.
纤维材料具有用于组织支架的潜力。在体外观察了巨噬细胞与各种组成和尺寸的纤维之间的相互作用。测试了以下材料:单根金纤维;聚酯和尼龙的编织纤维;聚丁烯/聚丙烯80:20和聚酯的非织造纤维。所有纤维的直径在2至40微米之间。在24小时孵育时间结束时,收集培养基用于检测肿瘤坏死因子α(TNF-α)和白细胞介素-6(IL-6),这是两种由活化巨噬细胞分泌的细胞因子。纤维样品经过处理用于扫描电子显微镜(SEM),或用于MAC-1和ICAM-1细胞表面标志物的免疫荧光标记。后者使用共聚焦显微镜进行,对编织和非编织样品进行了该操作。没有一种纤维样品在培养基中诱导大量的TNF-α或IL-6分泌,这表明细胞没有激活该途径。对单根金纤维的SEM显示纤维直径对细胞形态有影响,即对其铺展程度有影响。较大的纤维有更多的细胞,这些细胞倾向于聚集在一起而不广泛铺展。当比较编织纤维和非编织纤维时,SEM显示细胞在编织纤维上广泛铺展,而在非编织纤维上它们倾向于保持球形。共聚焦显微镜显示材料之间MAC-1和ICAM-1阳性细胞数量存在差异。这些结果表明,形态学、免疫和生化标志物的组合可用于区分引发的巨噬细胞对各种材料的反应。在所有测试材料上,细胞似乎仅被适度激活,其形态发生了变化,但细胞因子分泌没有增加。所测得的反应暗示了标称纤维组成和纤维直径之间的相互作用。