Burn Injury Reprograms Human Adipose SVF-Derived Organoids Toward an IL-6/IL-1β–Dominant Inflammatory Phenotype

Authors:
Filip Vlavcheski, Marc Jeschke

Body of Abstract:
Background: Severe burn injury triggers profound systemic inflammation and metabolic dysregulation, yet the contribution of adipose-resident stromal vascular fraction (SVF) cells to this response remains poorly defined. Since adipose tissue is a major endocrine organ and a large immune cell reservoir, injury-induced reprogramming of its stromal compartment may significantly shape host inflammatory trajectories. This study examines whether SVF-derived adipose organoids generated from burn patients exhibit altered cytokine secretion and inflammatory signaling compared with non-burn controls.

Methods: Subcutaneous adipose tissue was collected during surgery from adult burn patients and from non-burn donors undergoing elective procedures. SVF was isolated enzymatically, embedded in 3D matrices, and differentiated into adipose organoids under identical conditions. Conditioned media were analyzed for IL-6, IL-1β, and TNFα by ELISA. Organoid lysates were examined for NF-κB activation markers (phospho-p65, IκBα degradation) and nuclear localization of NF-κB by immunofluorescence. Data represent n≥3 donors per group and were analyzed using unpaired t tests.

Results: Burn-derived SVF organoids exhibited a pronounced inflammatory profile, showing significantly higher IL-6 (p<0.01) and IL-1β (p<0.05) secretion relative to non-burn controls. TNFα remained low in both groups without significant differences. Burn organoids showed clear biochemical evidence of NF-κB activation, including elevated p65 phosphorylation and accelerated IκBα loss. Confocal imaging confirmed increased nuclear p65 accumulation in burn organoids, whereas control organoids displayed primarily cytosolic localization indicating activation of canonical pro-inflammatory NF-κB signaling. Conclusions: Human SVF-derived adipose organoids from burn patients exhibit selective IL-6/IL-1β hypersecretion coupled with canonical NF-κB activation, despite minimal TNFα induction. These findings indicate that burn injury reprograms adipose stromal cells toward a distinct pro-inflammatory state that may contribute to systemic inflammation, immune dysregulation, and metabolic stress in surgical critical illness. This organoid platform provides a translational human model for dissecting adipose-driven inflammatory mechanisms after burn injury and may support future therapeutic development.