Effects of specialized proresolving mediators on gut epithelial barrier in early life

Damage to the intestinal epithelial barrier is a hallmark of inflammatory diseases such as necrotizing enterocolitis. Specialized proresolving mediators (SPMs), such as lipoxin A4, resolvin D1, and resolvin E1, which are derived from essential fatty acids, have been shown to aid in resolving inflammation and promote mucosal healing. This study aimed to explore the effects of specific SPMs on intestinal inflammatory response in an early life in vitro model. We established 3-dimensional and 3-dimensional organoid cultures from fetal and pediatric intestines and investigated the effect of an SPM cocktail (lipoxin A4, resolvin D1, and resolvin E1) on gut epithelial maturation and barrier function. An inflammatory response of the gut barrier was provoked by lipopolysaccharide and flagellin stimulations combined with proinflammatory cytokines, tumor necrosis factor-α, and interferon gamma. Additionally, repetitive mechanical wounding was developed to test the effects of the SPM cocktail on 2-dimensional organoid monolayers. Under physiological conditions, we observed no effect of SPM cocktail treatment on gut epithelial maturation. Upon cytokine challenge, there was no modulation of the inflammatory tone of the gut barrier by the SPM cocktail. However, during the repetitive wounding and recovery assay, SPM cocktail treatment accelerated barrier recovery and maintained barrier integrity for 24 hours after repeated injuries. Our findings suggest that the SPM cocktail does not affect bacterial product- or cytokine-induced epithelial inflammation, although it may accelerate epithelial barrier recovery in mechanically wounded monolayers. These results provide valuable insights into the therapeutic potential of SPMs in neonatal intestinal inflammation. SIGNIFICANCE STATEMENT: Using early life intestinal organoid models, we found that although specialized proresolving mediators did not alter cytokine- or bacterial product-induced inflammation, they significantly enhanced epithelial barrier recovery following repeated mechanical injury.