Catelas I, Huk O L, Petit A, Zukor D J, Marchand R, Yahia L
Institute of Biomedical Engineering, Ecole Polytechnique de Montréal, Biomechanics/Biomaterials Research Group, Québec, Canada.
J Biomed Mater Res. 1998 Sep 15;41(4):600-7. doi: 10.1002/(sici)1097-4636(19980915)41:4<600::aid-jbm12>3.0.co;2-i.
Using the J774 macrophage cell line, we designed an in vitro model to analyze by flow cytometry the effects of size, concentration, and composition of ceramic (Al2O3 and ZrO2) and high density polyethylene (HDP) particles on phagocytosis and cell mortality. Inflammatory mediator (TNF-alpha) also was measured by ELISA. Kinetic studies revealed that phagocytosis of the particles begins very early after cell exposure, increasing with time and particle concentration and reaching a plateau after 15 h. This implies that the optimum period to evaluate cellular response to particulate debris is between 15 and 24 h of incubation. Results also showed that phagocytosis increases with concentration for particles up to 2 microns. For larger particles (up to 4.5 microns), phagocytosis seems to reach a plateau independent of size and concentration, which suggests a saturation of phagocytosis that is most likely dependent on overall particle volume ingested. We did not detect any significant difference in phagocytosis between Al2O3 and ZrO2 at 0.6 microns. Al2O3 seems to be more easily phagocytosed than HDP at the same size (4.5 microns) and concentrations. Cytotoxicity studies revealed that macrophage mortality increases with particle size and concentration for sizes greater than 2 microns. Smaller particles (0.6 microns) cause cell mortality only at higher concentrations (from 1,250 particles per cell), but the mortality is still very low (10%). No significant difference in cell mortality and TNF-alpha release was found between Al2O3 and ZrO2. Effects of Al2O3 and HDP at 4.5 microns were compared by measuring TNF-alpha release. Results showed that TNF-alpha release increases with particle concentrations and is higher with HDP than with Al2O3.
我们使用J774巨噬细胞系设计了一个体外模型，通过流式细胞术分析陶瓷（Al2O3和ZrO2）及高密度聚乙烯（HDP）颗粒的大小、浓度和组成对吞噬作用及细胞死亡率的影响。还通过酶联免疫吸附测定法检测了炎性介质（肿瘤坏死因子-α）。动力学研究表明，细胞接触颗粒后，吞噬作用很早就开始了，随时间和颗粒浓度增加，15小时后达到平稳状态。这意味着评估细胞对颗粒碎片反应的最佳时期是孵育15至24小时之间。结果还表明，对于直径达2微米的颗粒，吞噬作用随浓度增加。对于更大的颗粒（达4.5微米），吞噬作用似乎达到一个与大小和浓度无关的平稳状态，这表明吞噬作用的饱和很可能取决于摄入的颗粒总体积。在0.6微米时，我们未检测到Al2O3和ZrO2之间吞噬作用有任何显著差异。在相同大小（4.5微米）和浓度下，Al2O3似乎比HDP更易被吞噬。细胞毒性研究表明，对于直径大于2微米的颗粒而言，巨噬细胞死亡率随颗粒大小和浓度增加。较小的颗粒（0.6微米）仅在较高浓度（每个细胞1250个颗粒起）时导致细胞死亡，但死亡率仍然很低（10%）。未发现Al2O3和ZrO2之间细胞死亡率及肿瘤坏死因子-α释放有显著差异。通过检测肿瘤坏死因子-α释放，比较了4.5微米的Al2O3和HDP的作用。结果表明，肿瘤坏死因子-α释放随颗粒浓度增加，且HDP比Al2O3更高。
