Time-Dependent Inflammatory Cytokine Responses in a Human Fetal Membrane Dual-Compartment Model

Preterm birth is a leading cause of neonatal morbidity and mortality, with inflammation at the feto-maternal interface playing a central role in its pathogenesis. Human fetal membrane models provide a physiologically relevant platform to investigate inflammatory mechanisms and evaluate targeted interventions.

Human fetal membrane explants from term cesarean deliveries were cultured in a dual-compartment system enabling side-specific stimulation and compartment-resolved analysis. Inflammation was induced by amniotic application of lipopolysaccharide (LPS; 1,000-10,000 ng/ml), and interleukin (IL)-1β and IL-6 concentrations were quantified over 1-48 h. Rytvela (0.1-10 μM), an allosteric IL-1 receptor modulator, was evaluated in a subset of experiments using preceding, simultaneous, and delayed administration relative to LPS exposure.

LPS induced time- and dose-dependent increases in IL-1β and IL-6. IL-6 responses were detectable from 6 h onward, whereas IL-1β increased at later time points (24-48 h). Rytvela alone did not induce cytokine secretion. Modulation of IL-1β responses differed between treatment regimens: while IL-1β levels were increased across all LPS-containing conditions, preceding rytvela was associated with comparatively lower IL-1β levels than simultaneous or delayed administration. In contrast, IL-6 levels were not consistently affected by rytvela across experimental conditions.

The dual-compartment fetal membrane model enables time-resolved analysis of inflammatory responses under physiologically relevant conditions. Preceding administration of rytvela was associated with lower IL-1β levels compared to simultaneous or delayed treatment, whereas IL-6 responses were not consistently affected. These findings suggest that timing and pathway-specific regulation may influence inflammatory responses in fetal membranes.