Aerosolizable Formulations of Porcine Extracellular Matrix with Antibacterial and Immunomodulatory Effects

Controlling bacterial presence and inflammation in the larynx remains a significant challenge, especially in contexts where infection and immune responses can affect tissue health, prompting growing interest in developing bioactive materials that can be delivered locally and noninvasively to modulate microbial load and inflammatory responses. In this study, we developed and characterized aerosolizable formulations derived from decellularized extracellular matrix (dECMx) of porcine vocal fold lamina propria (VFLPx) and thyroid cartilage (Cx). These formulations were evaluated for their antibacterial and immunomodulatory effects. The dECMx formulations exhibited heterogeneous microstructures and aggregate sizes smaller than 5 μm, which is consistent with requirements for aerosol delivery. Antibacterial activity against Staphylococcus aureus was assessed through colony-forming unit (CFU) quantification and bacterial growth kinetics, comparing the liquid and aerosol forms. In liquid form, VFLPx reduced CFU counts by ∼66% and Cx by ∼16%, both showing statistically significant reductions compared to controls. In aerosolized form, both formulations demonstrated comparable antibacterial efficacy, with approximately ∼47% reductions in CFU for VFLPx and Cx. Immunomodulatory potential was evaluated using a THP-1 ascGFP reporter cell line, which was stimulated with LPS to activate NF-κB signaling. Exposure to both dECMx formulations significantly attenuated the LPS-induced GFP fluorescence, indicating a suppression of NF-κB-mediated activation. These in vitro results demonstrate that liquid dECM-derived formulations maintain significant antibacterial activity after processing and aerosolization while also inhibiting inflammatory pathways in human macrophages. This highlights their potential as localized treatments to effectively control bacterial infections without causing inflammation.