In silico pharmacology and 3D-bioprinting reveal Dicranum scoparium as an inhibitor of NF-κB-induced inflammation in the 3D4/31 alveolar macrophage cell line.

Despite the growing interest in medicinal plants, the advancement of phytomedicines has been hindered by their limited clinical efficacy. In this study, we aimed to identify the medicinal properties of Dicranum scoparium aqueous extract (DSAE) using in silico analysis and validate them in an in vitro assay. We then utilize 3D-bioprinting to replace experimental animals for an integrated approach to rapidly identify potential clinical applications. Dicranum scoparium, also known as broom forkmoss, is distributed worldwide and has anti-inflammatory properties. However, its mechanism of action and effects on lung immunity remain unknown. The chemical composition of DSAE was identified using gas chromatography-mass spectrometry. The pharmacological properties of DSAE constituents were predicted using in silico pharmacology and gene ontology. The anti-inflammatory activity of DSAE was assessed using the 3D4/31 alveolar macrophage cell line (3D4/31-AMs) and 3D-bioprinting in vitro. In silico pharmacology showed that DSAE constituents exhibit high physiochemical druglikeness. Target genes identified through in silico prediction were enriched in lung-type cell signatures and regulation of nuclear factor kappa B (NF-κB). In vitro analysis of 3D4/31-AMs revealed that DSAE inhibited NF-κB-induced cell death and reactive oxygen species production. To improve the clinical applicability of DSAE, 3D4/31-AMs were 3D bioprinted using an alginate-gelatin-collagen bioink that increased NF-κB activation sensitivity. DSAE inhibited NF-κB-induced PTGS2, TNF, and IL1B expression in 3D-bioprinted 3D4/31-AMs. Overall, this study introduces a medicinal plant development strategy that integrates in silico analysis with 3D-bioprinting, and suggests DSAE as an inhibitor of NF-κB-mediated lung injury.