Mucus-penetrating archaeolipid nanocarriers delivering Thymus vulgaris essential oil and tobramycin: A multifunctional approach against Pseudomonas aeruginosa biofilms and inflammation.

Cystic fibrosis (CF) is characterized by thick mucus obstruction, persistent Pseudomonas aeruginosa biofilms, and chronic airway inflammation, leading to progressive lung damage. Inhaled tobramycin (TB) is the standard antibiotic therapy but has limited efficacy due to poor biofilm penetration and bacterial tolerance, while Thymus vulgaris essential oil (EOT) provides antioxidant and anti-inflammatory effects but is hindered by low solubility and instability. In this study, a comprehensive in vitro evaluation was conducted on EOT and TB co-loaded into nanostructured archaeolipid carriers (NAL-EOT/TB), composed of a compritol-miglyol lipid matrix with a surface coating of Tween 80 and archaeolipids from the archaea Halorubrum tebenquichense. NAL-EOT/TB interacted minimally with mucins and diffused efficiently through artificial mucus. Under mucus-covered conditions, macrophage uptake was higher for NAL-EOT/TB than for non-archaeolipid nanocarriers (NLC-EOT/TB), likely due to improved mucus permeation and scavenger receptor recognition. In epithelial cell-biofilm co-cultures, NAL-EOT/TB significantly decreased epithelial cell damage compared to both free EOT/TB and NLC-EOT/TB. Moreover, NAL-EOT/TB lowered ROS in neutrophil-like cells and achieved greater IL-8 reduction than dexamethasone. These findings support NAL-EOT/TB as a multifunctional nanotherapeutic platform to address mucus penetration, biofilm disruption, and airway inflammation in CF.