背景:这项研究旨在研究牙周膜(PDL)干细胞来源的外泌体负载大黄素(Emo@PDL-Exo)在抗微生物光动力疗法(aPDT)中对作为致龋细菌的变异链球菌和嗜酸乳杆菌的有效性。
方法:在分离和表征PDL-Exo之后,该研究继续准备和验证Emo@PDL-Exo的存在。抗菌作用,抗生物膜活性,和Emo的抗代谢能力,PDL-Exo,和Emo@PDL-Exo然后用和不照射波长为405±10nm的蓝色激光进行评估,输出强度为150mW/cm2,持续时间为60s。该研究使用计算机分子对接评估了大黄素与GtfB和SlpA蛋白的结合亲和力。最终,该研究检查了内源性活性氧(ROS)的产生以及gelE和sprE基因表达水平的变化。
结果:研究发现,使用Emo@PDL-Exo介导的aPDT导致嗜酸乳杆菌和变形链球菌显著减少4.90±0.36和5.07log10CFU/mL,分别为(P<0.05)。研究发现,使用Emo@PDL-Exo进行aPDT可显著降低嗜酸乳杆菌和变形链球菌生物膜的44.7%和50.4%,分别,与对照组未处理的生物膜相比(P<0.05)。此外,嗜酸乳杆菌和变形链球菌的代谢活性分别下降58.3%和71.2%,分别为(P<0.05)。分子对接分析显示大黄素与SlpA和GtfB蛋白具有很强的结合亲和力,对接评分为-7.4和-8.2kcal/mol,分别。该研究还发现,使用Emo@PDL-Exo组的aPDT导致slpA和gtfB基因表达最显著的减少,减少了4.2倍和5.6倍,分别,与对照组相比(P<0.05),可能是由于内源性ROS的产生增加。
结论:研究表明,使用Emo@PDL-Exo的aPDT可有效降低细胞活力,生物膜活性,以及变异链球菌和嗜酸乳杆菌的代谢效力。由于内源性ROS产生增加,aPDT也显著降低gtfB和slpAmRNA的表达水平。研究结果表明,Emo@PDL-Exo介导的aPDT可能是一种有前途的抗龋微生物方法。
BACKGROUND: This study was conducted to investigate the efficiency of periodontal ligament (PDL) stem cell-derived exosome-loaded Emodin (Emo@PDL-Exo) in antimicrobial photodynamic therapy (aPDT) on Streptococcus mutans and Lactobacillus acidophilus as the cariogenic bacteria.
METHODS: After isolating and characterizing PDL-Exo, the study proceeded to prepare and verify the presence of Emo@PDL-Exo. The antimicrobial effect, anti-biofilm activity, and anti-metabolic potency of Emo, PDL-Exo, and Emo@PDL-Exo were then evaluated with and without irradiation of blue laser at a wavelength of 405 ± 10 nm with an output intensity of 150 mW/cm2 for a duration of 60 s. In addition, the study assessed the binding affinity of Emodin with GtfB and SlpA proteins using in silico molecular docking. Eventually, the study examined the generation of endogenous reactive oxygen species (ROS) and changes in the gene expression levels of gelE and sprE.
RESULTS: The study found that using Emo@PDL-Exo-mediated aPDT resulted in a significant decrease in L. acidophilus and S. mutans by 4.90 ± 0.36 and 5.07 log10 CFU/mL, respectively (P < 0.05). The study found that using Emo@PDL-Exo for aPDT significantly reduced L. acidophilus and S. mutans biofilms by 44.7% and 50.4%, respectively, compared to untreated biofilms in the control group (P < 0.05). Additionally, the metabolic activity of L. acidophilus and S. mutans decreased by 58.3% and 71.2%, respectively (P < 0.05). The molecular docking analysis showed strong binding affinities of Emodin with SlpA and GtfB proteins, with docking scores of -7.4 and -8.2 kcal/mol, respectively. The study also found that the aPDT using Emo@PDL-Exo group resulted in the most significant reduction in gene expression of slpA and gtfB, with a decrease of 4.2- and 5.6-folds, respectively, compared to the control group (P < 0.05), likely due to the increased generation of endogenous ROS.
CONCLUSIONS: The study showed that aPDT using Emo@PDL-Exo can effectively reduce the cell viability, biofilm activity, and metabolic potency of S. mutans and L. acidophilus. aPDT also significantly reduced the expression levels of gtfB and slpA mRNA due to the increased endogenous ROS generation. The findings suggest that Emo@PDL-Exo-mediated aPDT could be a promising antimicrobial approach against cariogenic microorganisms.