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首页> 外文期刊>Biomaterials >Quantitative analysis of macrophage apoptosis vs. necrosis induced by cobalt and chromium ions in vitro.
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Quantitative analysis of macrophage apoptosis vs. necrosis induced by cobalt and chromium ions in vitro.

机译：定量分析钴和铬离子诱导的巨噬细胞凋亡与坏死的关系。

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The potential toxicity of metal ions in tissues surrounding metal-metal hip replacements is a cause for concern. Previous studies conducted in our laboratory demonstrated that Co(2+) and Cr(3+) induce TNF-alpha secretion in macrophages, as well as cell mortality. However, the degree of apoptosis and necrosis remained to be investigated. The aim of the present study was to quantify the rate of macrophage mortality by apoptosis vs. necrosis induced by Co(2+) and Cr(3+). J774 mouse macrophages were incubated in growth medium containing 0-10ppm Co(2+) and 0-500ppm Cr(3+) for 24 and 48h under conventional cell culture conditions. Transmission electron microscopy, flow cytometry (Annexin-V fluorescein isothiocyanate/propidium iodide assay) and a specific cell death detection ELISA were used to illustrate cell death and differentiate between apoptotic and necrotic cells. Cell culture exposed to low concentrations of Co(2+) (0-6ppm) revealed a low degree of mortality. In contrast, at the highest concentrations(8-10ppm), late apoptosis occurred within 24h. After 48h, however, there was a clear evidence for an increase in the rate of necrosis while apoptosis occurred at much lower rate. Macrophages exposed to Cr(3+) demonstrated a predominance of apoptosis after 24h. At concentrations lower than 250ppm, early and late apoptosis occurred at the same rate. At higher concentrations (250-500ppm), the number of early apoptotic cells decreased in favor of late apoptosis. After 48h, lower concentrations of Cr(3+) (150ppm) induced a higher degree of early apoptosis than after 24h, and some necrosis. At higher concentrations, the percentage of early apoptotic cells decreased, while necrosis became predominant over late apoptosis. In conclusion, this study demonstrates that macrophage mortality induced by metal ions depends on the type and concentration of metal ions as well as the duration of their exposure. Overall, apoptosis was predominant after 24h with both Co(2+) and Cr(3+) ions, but high concentrations induced mainly necrosis at 48h. These results point to the potential for these ions of inducing tissue damage by necrosis if present in large concentrations in vivo.
机译：金属离子对金属髋关节置换物周围组织中金属离子的潜在毒性令人担忧。在我们实验室中进行的先前研究表明，Co（2+）和Cr（3+）诱导巨噬细胞中的TNF-α分泌以及细胞死亡。然而，细胞凋亡和坏死的程度仍有待研究。本研究的目的是量化由凋亡与坏死由Co（2+）和Cr（3+）诱导的巨噬细胞死亡率的比率。在常规细胞培养条件下，将J774小鼠巨噬细胞在含有0-10ppm Co（2+）和0-500ppm Cr（3+）的生长培养基中孵育24和48h。透射电镜，流式细胞仪（Annexin-V异硫氰酸荧光素/碘化丙啶含量测定）和特异性细胞死亡检测ELISA用于说明细胞死亡并区分凋亡细胞和坏死细胞。暴露于低浓度的Co（2+）（0-6ppm）的细胞培养显示出较低的死亡率。相反，在最高浓度（8-10ppm）下，细胞凋亡在24h内发生。但是，在48小时后，有明显的证据表明坏死率增加，而凋亡发生率要低得多。暴露于Cr（3+）的巨噬细胞在24小时后显示出凋亡的优势。在低于250ppm的浓度下，早期和晚期凋亡发生的速率相同。在较高浓度（250-500ppm）下，早期凋亡细胞的数量减少，有利于晚期凋亡。 48小时后，较低浓度的Cr（3+）（150ppm）诱导的早期细胞凋亡程度高于24小时后，并且出现一些坏死。在较高的浓度下，早期凋亡细胞的百分比降低，而坏死则占晚期细胞凋亡的主导。总之，这项研究表明，由金属离子引起的巨噬细胞死亡率取决于金属离子的类型和浓度以及其暴露时间。总体而言，细胞凋亡在24h后以Co（2+）和Cr（3+）离子为主，但高浓度在48h时主要引起坏死。这些结果表明，如果这些离子以高浓度存在于体内，那么这些离子就有可能通过坏死而引起组织损伤。

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