口腔生物医学 ›› 2022, Vol. 13 ›› Issue (4): 208-213.

• 论著 • 上一篇    下一篇

自产氧抗菌光动力纳米体系对炎症调节和成骨分化的影响

博梦1,王林2,孙晓琳1   

  1. 1. 吉林大学口腔医院种植科
    2. 吉林大学口腔医学院
  • 收稿日期:2022-07-25 修回日期:2022-11-17 出版日期:2022-12-25 发布日期:2022-12-24
  • 通讯作者: 孙晓琳 E-mail:sxl2673366@126.com
  • 基金资助:
    国家自然科学基金面上项目

Effect of oxygen self-sufficient nanoplatform on the anti-inflammation and osteogenesis of periodontitis

1, 1,   

  • Received:2022-07-25 Revised:2022-11-17 Online:2022-12-25 Published:2022-12-24

摘要: 目的:探讨自产氧的抗菌光动力纳米体系——F@Ce6-M对炎症调节和成骨分化的影响。方法:构建F@Ce6-M并对其进行表征、生物安全性的评价。使用630 nm光照射牙龈卟啉单胞菌(P. gingivalis)菌液处理小鼠成纤维细胞L929,建立炎症模型,分别加入F@Ce6和F@Ce6-M并设立相应对照组。实时荧光定量PCR检测促炎因子和抗炎因子的基因表达水平,评价F@Ce6-M介导的抗菌光动力治疗(aPDT)对炎症调节的影响;通过免疫荧光检测NF-κB/p65亚基易位情况,分析F@Ce6-M的免疫调节机制;通过实时荧光定量PCR检测成骨相关基因表达,并采用碱性磷酸酶(ALP)进行染色,评价F@Ce6-M在炎症微环境中对前成骨细胞成骨分化的调控作用。结果:实时荧光定量PCR结果显示,与阴性对照组相比,F@Ce6-M介导aPDT能明显降低白细胞介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α基因表达,并且效果明显优于F@Ce6-M光照组(P<0.05);而各组间转化生长因子(TGF)-β1、精氨酸(Arg)-1和IL-10的基因表达差异无统计学意义(P>0.05)。在巨噬细胞中NF-κB/p65易位的免疫荧光实验中,F@Ce6-M光照组可抑制NF-κB/p65亚基转位(P<0.05)。对于成骨标志物基因表达水平,与阴性对照组相比,光照组ALP、Runt相关转录因子2(RUNX2)、Ⅰ型胶原蛋白(COL-1)、骨钙蛋白(OCN)基因表达量显著上升,F@Ce6-M光照组的基因表达量升高最为显著(P<0.05)。在ALP染色实验中,光照组的ALP活性水平显著上升,F@Ce6-M光照组ALP活性水平升高最为显著(P<0.05)。结论:自产氧的抗菌光动力纳米体系能够减少促炎因子的释放,并对成骨细胞的成骨潜能有显著的恢复作用,从而增强aPDT治疗牙周炎的效果。

关键词: 抗菌光动力疗法, 牙周炎, 炎症因子, 炎症调节, 成骨分化

Abstract: Objective:To investigate the effect of self-oxygenated antibacterial photodynamic nanoplatform--F@Ce6-M on inflammation regulation and osteogenic differentiation. Methods:The mouse fibroblast L929 was treated with Porphyromonas gingivalis (P. gingivalis) bacterial solution by light irradiation, and the inflammation model was established. F@Ce6 and F@Ce6-M were respectively added, and the corresponding control group was set up. The gene expression levels of pro-inflammatory and anti-inflammatory factors were detected by qRT-PCR to evaluate the effect of F@Ce6-M mediated antimicrobial photodynamic therapy (aPDT) on inflammation regulation. NF-κB/p65 subunit translocation was detected by immunofluorescence to analyze the immunomodulatory mechanism of F@Ce6-M. The expression of osteoblast-related genes was detected by qRT-PCR and stained with ALP to evaluate the regulatory effect of F@Ce6-M on osteogenic differentiation of preosteoblasts. Results:qRT-PCR results showed that F@Ce6-M mediated aPDT significantly decreased the expression of interleukin (IL-6), IL-1β, tumor necrosis factor (TNF)-α gene compared with negative control group, and the effect was significantly better than that of F@Ce6-M no light group (P<0.05); There were no significant differences in the gene expressions of transforming growth factor (TGF)-β1, arginine (Arg)-1 and IL-10 among all groups (P>0.05). F@Ce6-M light group inhibited NF-κB/p65 subunit translocation (P<0.05). The osteogenic marker expressions of ALP, runt-related transcription factor 2 (RUNX2), collagen type Ⅰ (COL-1) and osteocalcin (OCN) in light group were significantly increased compared with negative control group, and the gene expression levels in F@Ce6-M group were the most significantly increased (P<0.05). The activity level of ALP in the light group was significantly increased, the most significantly increased in the F@Ce6-M light group (P<0.05). Conclusions:F@Ce6-M could reduce the release of pro-inflammatory factors, and significantly restore the osteogenic potential of osteoblasts, thereby enhancing the effect of aPDT on periodontitis.

Key words: antibacterial photodynamic therapy, Periodontitis, inflammatory factors, immunomodulation, osteogenesis differentiation