Pro-inflammatory Analysis of Macrophages in Contact with Titanium Particles and Porphyromonas gingivalis.
Pro-inflammatory Analysis of Macrophages in Contact with Titanium Particles and Porphyromonas gingivalis.
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DOI：
10.1590/0103-6440201701382
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发表时间：
2017-07
影响因子：
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通讯作者：
Cury AADB
中科院分区：
文献类型：
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作者：
Dodo CG;Meirelles L;Aviles-Reyes A;Ruiz KGS;Abranches J;Cury AADB
在钛牙种植体的插入过程中，颗粒可能会从种植体剪切到种植体周围区域，导致骨溶解，并且它们与细菌的结合会加剧炎症反应。然而，口腔中的一种高侵袭性细菌牙龈卟啉单胞菌 (Pg) 与钛颗粒之间的关联仍不清楚。本研究评估了人类巨噬细胞与牙龈卟啉单胞菌脂多糖 (PgLPS) 相关的钛微粒和纳米颗粒接触时的促炎反应。使用THP-1细胞处理12、24和48小时，分6组：对照(C)、PgLPS(L)；微粒子（M）；纳米颗粒（N）； PgLPS 和微粒 (LM)； PgLPS 和纳米颗粒 (LN)。进行了以下测定：i) 使用 MTS 检测细胞活力，ii) 使用 SEM 检测细胞形态，以及 iii) 通过 qRT-PCR 和 ELISA 检测肿瘤坏死因子 α (TNF-α)、白细胞介素 1 β (IL-1 β) 和白细胞介素 6 (IL-6) 的表达。对于统计，使用了双向方差分析和 Tukey 检验（p<0.05）。治疗后，细胞表现出相似的活力和形态，表明治疗无法诱导细胞死亡。在 N 组和 LN 组中，12 小时后 TNF-α 和 IL1-β 的基因表达显着升高，24 小时后 IL-6 的基因表达显着升高。随着时间的推移，细胞因子的产生呈上升曲线，TNF-α在48小时达到峰值，IL1-β和IL-6在时间点之间呈直线，尽管N组细胞在48小时出现显着的IL-6产生。总之，这些结果表明钛纳米粒子刺激巨噬细胞中更强的促炎反应，与它们与牙龈卟啉单胞菌LPS的关联无关。
During insertion of titanium dental implants, particles may shear from the implant to the periimplant region causing osteolysis, and their association with bacteria can exacerbate the inflammatory reaction. However, the association of a high invasive bacterium from the oral cavity, Porphyromonas gingivalis (Pg), and titanium particles remains unknown. This study evaluated pro-inflammatory reaction of human macrophages in contact with micro and nanoparticles of titanium associated with Porphyromonas gingivalis lipopolysaccharide (PgLPS). THP-1 cell were used and treated for 12, 24 and 48 h following 6 groups: Control(C), PgLPS (L); Microparticles (M); Nanoparticles (N); PgLPS and microparticles (LM); PgLPS and nanoparticles (LN). The following assays were carried out: i) cell viability using MTS, ii) cell morphology by SEM and iii) expression of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1 β) and interleukin-6 (IL-6) by qRT-PCR and ELISA. For statistics two-way ANOVA followed by Tukey’s test was used (p<0.05). After treatment, cells presented similar viability and morphology demonstrating that the treatments were not able to induce cell death. Gene expression was significantly higher for TNF-α and IL1-β after 12 h, and for IL-6 after 24 h in the N and LN groups. Cytokine production over time was an ascending curve for TNF-α with the peak at 48 h and IL1-β and IL-6 had a straight line among the time points, although cells from N group presented a significant production of IL-6 at 48 h. In conclusion, these results suggest that titanium nanoparticles stimulate stronger pro-inflammatory response in macrophages, independent of their association with LPS from P.gingivalis.