Physical Chemistry I and Research Center of Micro- and Nanochemistry and (Bio)Technology (Cμ), Department of Chemistry and Biology, University of Siegen, Siegen, Germany.
INSERM, Univ Brest, EFS, UMR 1078, GGB-GTCA, Brest, France.
Front Cell Infect Microbiol. 2022 Jul 13;12:941014. doi: 10.3389/fcimb.2022.941014. eCollection 2022.
Women with cystic fibrosis (CF) have a significantly lower life expectancy compared to men, which is indicated by an earlier impairment of lung function due to chronic colonization with biofilm formed by . There is growing evidence that blood serum concentrations of the steroid sex hormone estradiol (E) correlate with the occurrence of pulmonary exacerbations in CF but also play a role in the mucoid switch of . This study aims to shed light on possible microbiological reasons for sexual dimorphism in CF by investigating the influence of E on biofilm formation of . CF isolates. For this purpose, 10 CF isolates of the respiratory tract derived from different CF patients have been treated with E in a microtiter plate biofilm model. Biofilms have been examined by crystal violet assays, field emission scanning electron microscopy (FE-SEM), 3D laser scanning microscopy (LSM), and quorum sensing (QS) reporter assays of the supernatants taken from biofilms. This allowed us to simultaneously investigate the effects of E on attached biofilm mass, biofilm ultrastructure, and QS activity. Upon E treatment, six out of 10 investigated CF isolates showed an increase of attached biofilm mass, whereas biofilms from two tested non-CF laboratory strains (PAO1 and ATCC19660) did not. Moreover, FE-SEM and 3D LSM analyses of the E responsive CF biofilms revealed ultrastructural remodeling of biofilm structure at different scales with increased formation of prominent biofilm spots, enhanced coverage with extracellular polymeric substance (EPS), and extended average surface roughness. QS activity measurements performed in biofilm supernatants luminescence acyl homoserine lactone (AHL) reporter assays further showed that E treatment may also modulate QS signaling, as shown in an E sensitive CF isolate. Together, our results suggest the biofilm modulating effects of E on various clinical CF isolates that are documented by both biomass and ultrastructural changes of biofilms. The gained new insight into the influence of steroid hormones on biofilm phenotypes might pave the way for novel future approaches in personalized medicine based on the patients' sex and hormonal status.
患有囊性纤维化 (CF) 的女性与男性相比，预期寿命明显较低，这是由于慢性生物膜定植导致肺功能受损所致。越来越多的证据表明，类固醇性激素雌二醇 (E) 的血清浓度与 CF 中肺部恶化的发生有关，但也在 CF 生物膜的粘液开关中发挥作用。这项研究旨在通过研究 E 对 CF 生物膜形成的影响，揭示 CF 中性别二态性的可能微生物学原因。为此，我们使用雌二醇 (E) 在微滴定板生物膜模型中处理了来自不同 CF 患者的 10 种呼吸道 CF 分离株。通过结晶紫测定、场发射扫描电子显微镜 (FE-SEM)、三维激光扫描显微镜 (LSM) 和生物膜上清液中的群体感应 (QS) 报告物测定来检查生物膜。这使我们能够同时研究 E 对附着生物膜质量、生物膜超微结构和 QS 活性的影响。在 E 处理后，10 种研究的 CF 分离株中有 6 种显示附着生物膜质量增加，而来自两种测试的非 CF 实验室菌株 (PAO1 和 ATCC19660) 的生物膜则没有。此外，对 E 反应性 CF 生物膜的 FE-SEM 和 3D LSM 分析表明，生物膜结构在不同尺度上的超微结构重塑，生物膜斑点形成增加，细胞外聚合物 (EPS) 覆盖增强，平均表面粗糙度增加。生物膜上清液中的 QS 活性测量通过 luminescence 酰基高丝氨酸内酯 (AHL) 报告物测定进一步表明，E 处理也可能调节 QS 信号，如在 E 敏感的 CF 分离株中所示。总之，我们的结果表明，E 对各种临床 CF 分离株具有生物膜调节作用，这通过生物膜的生物量和超微结构变化得到证实。我们对类固醇激素对生物膜表型的影响有了新的认识，这可能为基于患者性别和激素状态的个性化医学的新方法铺平道路。
