Dataset to: Low-Intensity Pulsed Ultrasound as a BioPhysical Modality to Assess and Modulate Mucosal Activity in experimental Colitis

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Dataset to: Low-Intensity Pulsed Ultrasound as a BioPhysical Modality to Assess and Modulate Mucosal Activity in experimental Colitis Abstract Background: Low-Intensity Pulsed Ultrasound (LIPUS) is a non-thermal mechanical energy modality that has garnered considerable interest, not only for its applications in neuromodulation and musculoskeletal contexts, but also for its notable anti-inflammatory properties reported in various preclinical models. However, its potential impact on inflamed mucosal tissue remains poorly explored. This study aims to investigate the use of LIPUS as an innovative biophysical therapeutic approach for the treatment of inflammatory bowel disease (IBD). Methods: A single, precisely targeted LIPUS stimulation (38 kHz, 150 mW/cm², 180 s) was applied to the colonic area of mice with Dextran Sodium Sulfate (DSS)-induced colitis. Biological effects were assessed through clinical, histological, and molecular analyses at 2, 24, and 96 hours post-stimulation. To validate the translational relevance of the observed mechanisms, an ex-vivo human co-culture model was established using Lamina Propria Mononuclear Cells (LPMCs) from Ulcerative Colitis patients and human intestinal organoids. For the statistical analysis the distribution is evaluated through Shapiro test, then the significance was evaluated trough t-test or One-way ANOVA. Results: A single LIPUS stimulation significantly improved clinical recovery (body weight, Disease Activity Index) compared to controls. While systemic pro-inflammatory cytokines (TNF-α, IL-1β, IFN-γ) were reduced at 24 and 96 hours, the decisive immunomodulatory event occurred earlier. At 2 hours post-stimulation, LIPUS triggered a rapid local clearance of immune infiltration (CD45+), induced a macrophage phenotype switch toward the restorative M2 profile (CD163+) and promoted both mucosal remodelling and epithelial barrier restoration. In the human co-culture model, LIPUS-treated LPMCs downregulated pathological signaling (NF-κB, STAT3) and, when co-cultured with organoids, actively preserved epithelial tight junctions (ZO-1) and stemness markers (LGR5) against inflammatory damage Conclusions: Overall, our findings demonstrate that LIPUS is a safe and effective biophysical approach capable of modulating mucosal inflammation through a precise, time-dependent reprogramming of the immune microenvironment. The conservation of this immune-epithelial crosstalk in human tissue highlights the potential of LIPUS as a novel, gut-targeted "mechanomedicine" strategy for inducing remission and tissue repair in IBD.

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