Intestinal epithelial cell (IEC) homeostasis during IBD can be facilitated by increased expression of cytokines, such as tumor necrosis factor-α (TNF-α), and by toll-like receptor (TLR) ligands derived from microbiota. However, the mechanisms of immune system-mediated tissue repair in IBD remain elusive. We have previously found that anti-TNF-α therapy induced IEC restitution is mediated by IL 22-driven IEC proliferation. Next, we revealed that antibiotic-induced changes in microbiota distinctly contributed to the restoration of the epithelial barrier via a TLR4-dependent mechanism. Thus, we aim to dissect the significance of the microbiota-TLR4-TNFα axis for IEC layer restoration during IBD.