Inhibitory Effects of Antimicrobial Photodynamic Therapy with Curcumin on Biofilm-Associated Gene Expression Profile of Aggregatibacter actinomycetemcomitans.

Maryam Pourhajibagher, Nasim Chiniforush, Abbas Monzavi, Hamidreza Barikani, Mohammad Moein Monzavi, Shaghayegh Sobhani, Sima Shahabi, Abbas Bahador
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Abstract

Objectives: Periodontitis is an inflammation of periodontal tissues that is caused by the biofilm of periodontal pathogens. Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans) is an opportunistic periodontopathogen that can be the cause of periodontal diseases via fimbriae as a virulence factor. In this study, we aimed to determine the expression level of A. actinomycetemcomitans rcpA gene as a virulence factor associated with biofilm formation after antimicrobial photodynamic therapy (aPDT) as a relatively new therapeutic modality.

Materials and methods: To determine sub-lethal doses of aPDT against A. actinomycetemcomitans ATCC 33384 strain, we used curcumin (CUR) as a photosensitizer at a final concentration of 40 μmol/ml, which was excited with a light-emitting diode (LED) at the wavelength of 450 nm. Quantitative real-time polymerase chain reaction (qRT-PCR) was then applied to monitor rcpA gene expression in A. actinomycetemcomitans.

Results: 10-40 μmol/ml of CUR caused a significant reduction in the growth of A. actinomycetemcomitans compared to control group (P<0.05). Also, the cell viability of A. actinomycetemcomitans was significantly decreased after more than four minutes of LED irradiation. Therefore, the sub-lethal dose of aPDT against A. actinomycetemcomitans was 5 μmol/ml of CUR with three minutes of LED irradiation at a fluency of 180-240 J/cm2, which reduced the expression of the rcpA gene by approximately 8.5-fold.

Conclusions: aPDT with CUR leads to decreased cell survival and virulence of A. actinomycetemcomitans. Thus, CUR-aPDT can be used as an alternative approach for the successful treatment of periodontitis in vivo.

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姜黄素抗菌素光动力疗法对放线菌群生物膜相关基因表达谱的抑制作用。
目的:牙周炎是由牙周病原体的生物膜引起的牙周组织炎症。放线菌聚集菌(a .放线菌)是一种机会性牙周病病原体,可作为毒力因子通过菌膜引起牙周病。在这项研究中,我们旨在确定放线菌rcpA基因作为一种与生物膜形成相关的毒力因子,在抗菌光动力治疗(aPDT)作为一种相对较新的治疗方式后的表达水平。材料与方法:以姜黄素(curcumin, CUR)为光敏剂,终浓度为40 μmol/ml,用发光二极管(LED)在波长450 nm处激发,测定aPDT对放线菌ATCC 33384的亚致死剂量。采用实时定量聚合酶链反应(qRT-PCR)技术检测放线菌中rcpA基因的表达。结果:与对照组相比,10-40 μmol/ml CUR可显著抑制放线菌的生长。LED照射4分钟以上,放线菌数量明显减少。因此,aPDT对放线菌的亚致死剂量为5 μmol/ml CUR,在180-240 J/cm2的流畅度下照射3分钟,rcpA基因的表达减少了约8.5倍。结论:aPDT联合CUR可降低放线菌的细胞存活率和毒力。因此,curr - apdt可以作为一种成功治疗体内牙周炎的替代方法。
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