Abstract：
In our previous studies, Chlorin-e6 (Ce6) demonstrated a significant reduction of microorganisms\' viability against multi-species biofilm related to periodontitis while irradiated with blue light. However, the conjugation of Ce6 and antimicrobial peptides, and the incorporation of this photosensitizer in a nanocarrier, is still poorly explored. We hypothesized that chlorin-e6 conjugated to the antimicrobial peptide LL-37 loaded nanoemulsion could inhibit a multi-species biofilm related to periodontitis during photodynamic therapy (PDT), the pre-treatment with hydrogen peroxide was also tested. The nanoemulsion (NE) incorporated with Ce6 was characterized regarding the physiochemical parameters. Images were obtained by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Later, the Ce6 and LL-37 incorporated in NE was submitted to UV-Vis analysis and Reactive Oxygen Species (ROS) assay. Finally, the combined formulation (Ce6+LL-37 in nanoemulsion) was tested against multi-species biofilm related to periodontitis. The formed nanoformulation was kinetically stable, optically transparent with a relatively small droplet diameter (134.2 unloaded and 146.9 loaded), and weak light scattering. The NE system did not impact the standard UV-VIS spectra of Ce6, and the ROS production was improved while Ce6 was incorporated in the NE. The combination of Ce6 and LL-37 in NE was effective to reduce the viability of all bacteria tested. The treatment with hydrogen peroxide previous to PDT significantly impacted bacterial viability. The current aPDT regimen was the best already tested against periodontal biofilm by our research team. Our results suggest that this combined protocol must be exploited for clinical applications in localized infections such as periodontal disease. - Nanoemulsion demonstrated to be an excellent nanocarrier for photodynamic application. - Chlorin-e6 incorporated in nanoemulsion showed great physicochemical and biophotonic parameters. - The combination of chlorin-e6 and LL-37 peptide in nanoemulsion is effective to eliminate periodontal pathogenic bacteria. - The treatment with hydrogen peroxide previous to PDT significantly impacted bacterial viability.
摘要：
在我们之前的研究中,在蓝光照射下，Chlorin-e6（Ce6）显示出微生物对与牙周炎有关的多物种生物膜的生存能力显着降低。然而,Ce6和抗菌肽的结合，并将这种光敏剂掺入纳米载体中，仍然缺乏探索。我们假设在光动力疗法（PDT）期间，与负载有抗菌肽LL-37的纳米乳液缀合的氯苷-e6可以抑制与牙周炎相关的多物种生物膜，还测试了用过氧化氢的预处理。结合Ce6的纳米乳液(NE)在物理化学参数方面进行表征。通过透射电子显微镜(TEM)和扫描电子显微镜(SEM)获得图像。稍后，将掺入NE的Ce6和LL-37进行UV-Vis分析和活性氧（ROS）测定。最后,联合制剂(纳米乳剂中的Ce6+LL-37)针对与牙周炎相关的多物种生物膜进行了试验.形成的纳米制剂是动力学稳定的，光学透明，具有相对较小的液滴直径（卸载134.2和装载146.9），和微弱的光散射。NE系统不影响Ce6的标准UV-VIS光谱,并且当Ce6掺入NE中时ROS产生得到改善。在NE中Ce6和LL-37的组合有效地降低所有测试的细菌的活力。在PDT之前用过氧化氢处理显著影响细菌活力。当前的aPDT方案是我们研究小组已经针对牙周生物膜进行的最佳测试。我们的结果表明，这种联合方案必须用于局部感染如牙周病的临床应用。-纳米乳液被证明是用于光动力应用的优异的纳米载体。-掺入纳米乳液中的氯-e6显示出良好的物理化学和生物光子参数。-在纳米乳剂中的二氢卟啉-e6和LL-37肽的组合有效消除牙周致病菌。-在PDT之前用过氧化氢处理显著影响细菌活力。
