Zenni G C, Ellinger J, Lam T M, Greisler H P
Loyola University Medical Center, Maywood, Illinois 60153.
J Invest Surg. 1994 Mar-Apr;7(2):135-41. doi: 10.3109/08941939409015357.
This study quantifies macrophage acid phosphatase release as a marker of cell activation when cultured with or without biomaterials. Peritoneal macrophages were harvested from six New Zealand White rabbits and cultured in minimum essential media with 10% equine serum. After cell identification by morphology, nonspecific esterase, and RAM 11 immunoperoxidase, cells were passaged twice, and second-passage macrophages were seeded in 96-well plates (5,000 cells/well) and grown to confluence. After collection of day zero media, circular disks of polyglactin 910 and two types of commercially available polyethylene terephthalate with different construction and sterilization characteristics were placed on the cell monolayer. Controls without biomaterials were also established. Media was collected and pooled for each group and time point beginning on day 2 and continuing every other day for 22 days. Conditioned media were quantitatively assayed for acid phosphatase colorimetrically at 402 nm using p-nitrophenylphosphate as the substrate. Acid phosphatase activity increased progressively at late time points for each group but no difference was noted between groups at any time point. These data show that the activation of cultured macrophages with time is not altered differentially by the presence of biomaterials. The previously demonstrated monokine release following biomaterial exposure is therefore a specific event and not simply part of the generalized activation phenomenon.
本研究对与生物材料一起培养或不与生物材料一起培养时巨噬细胞酸性磷酸酶的释放进行了量化,以此作为细胞活化的标志物。从6只新西兰白兔中获取腹腔巨噬细胞,并在含有10%马血清的最低必需培养基中培养。通过形态学、非特异性酯酶和RAM 11免疫过氧化物酶对细胞进行鉴定后,细胞传代两次,将第二代巨噬细胞接种到96孔板中(每孔5000个细胞),培养至汇合。收集第0天的培养基后,将聚乙交酯910圆盘以及两种具有不同结构和灭菌特性的市售聚对苯二甲酸乙二酯圆盘放置在细胞单层上。同时也设立了不放置生物材料的对照组。从第2天开始,每隔一天收集并合并每组在各个时间点的培养基,持续22天。使用对硝基苯磷酸酯作为底物,在402 nm处通过比色法对条件培养基中的酸性磷酸酶进行定量测定。每组在后期时间点酸性磷酸酶活性均逐渐增加,但在任何时间点各组之间均未观察到差异。这些数据表明,生物材料的存在并不会使培养的巨噬细胞随时间的活化产生差异。因此,先前证明的生物材料暴露后单核因子的释放是一个特定事件,而不仅仅是一般活化现象的一部分。