Chignier E, Guidollet J, Freyria A M, Ardail D, McGregor J L, Louisot P
INSERM U 331, A. Carrel Medical School, Lyon, France.
J Biomed Mater Res. 1993 Aug;27(8):1087-94. doi: 10.1002/jbm.820270814.
Biomaterials induce an inflammatory reaction characterized by a rapid recruitment at the implantation site of polymorphonuclear cells and macrophages. In the course of the inflammatory response, the cellular activation triggers expression of a number of enzymes, such as 5'-nucleotidase, which is widely distributed in animal cell membranes as an ectoenzyme. It is now well established that 5'-nucleotidase activity decreases following the contact of inflammatory cells with foreign particles. In this paper we investigate a possible correlation between the enzymatic activities and the dynamic properties of the cell membrane bilayer. Dacron pieces were introduced into rats' peritoneal cavities for a period of 6 h, after which the peritoneal cells were harvested, and various enzyme assays performed, including those for cytoplasmic, lysosomal, and ectoenzymes. In parallel, we studied cell membrane fluidity, using fluorescence polarization of 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH), and cellular ultrastructural alteration resulting from the cell-biomaterial interactions using scanning and transmission electron microscopy. Our results show that: 1) macrophages spread around the Dacron fibers with cytoplasmic finger-like projections, but no phagolysosomes, 2) 5'-nucleotidase levels decrease with surgical trauma in comparison with the resident cell exudate, 3) implantation of biomaterials slightly modify the 5'-nucleotidase levels observed in the sham animal, 4) no differences in the anisotropy values indicating that membrane lipid order within the cells could not account for the observed decrease of 5'-nucleotidase activity. Thus, we can suggest that 5'-nucleotidase expression may reflect a particular feature of cell activation without a phagocytic process.
生物材料会引发炎症反应，其特征是多形核细胞和巨噬细胞迅速聚集到植入部位。在炎症反应过程中，细胞活化会触发多种酶的表达，比如5'-核苷酸酶，它作为一种外切酶广泛分布于动物细胞膜中。现在已经明确，炎症细胞与异物颗粒接触后，5'-核苷酸酶的活性会降低。在本文中，我们研究了酶活性与细胞膜双层动态特性之间可能存在的相关性。将涤纶片植入大鼠腹腔6小时，之后收集腹腔细胞，并进行各种酶检测，包括对细胞质酶、溶酶体酶和外切酶的检测。同时，我们使用1-(4-三甲基铵苯基)-6-苯基-1,3,5-己三烯（TMA-DPH）的荧光偏振研究细胞膜流动性，并使用扫描电子显微镜和透射电子显微镜研究细胞-生物材料相互作用导致的细胞超微结构改变。我们的结果表明：1）巨噬细胞围绕涤纶纤维扩散，有细胞质指状突起，但没有吞噬溶酶体；2）与驻留细胞渗出液相比，5'-核苷酸酶水平随手术创伤而降低；3）生物材料的植入对假手术动物中观察到的5'-核苷酸酶水平有轻微影响；4）各向异性值没有差异，这表明细胞内的膜脂有序性无法解释观察到的5'-核苷酸酶活性降低。因此，我们可以认为5'-核苷酸酶的表达可能反映了细胞活化的一个特殊特征，而无需吞噬过程。
