Recombinant human fibronectin segment (rhFN) hydrogel carried hPDLSCs to repair diabetic trauma by activated NF-κB signaling pathway.

The accumulation of advanced glycation end products (AGEs) plays a crucial role in chronic inflammation and delayed wound healing in individuals with diabetes. In this context, fibronectin has been identified as a crucial protein that promotes the differentiation of human periodontal ligament stem cells (hPDLSCs) into myofibroblasts, which play a vital role in the repair of diabetic skin ulcers. This process is intimately associated with the integrin β1 receptor and the NF-κB signaling pathway, both crucial for cellular responses to fibronectin. To validate our hypothesis, we expressed rhFN, a recombinant protein containing the integrin β1 affinity-binding domain from human fibronectin segments 12-14. This protein was used to formulate a hydrogel for hPDLSCs. rhFN's binding affinity to integrin β1 was confirmed by molecular docking and the cellular thermal shift assay (CETSA). We developed -hPDLSCs with stable ITGB1 knockdown using and compared their proliferation, migration and adhesion to wild-type hPDLSCs. Morphological changes were observed via SEM, and α-SMA expression levels were measured in AGEs-damaged hPDLSCs. We created full-thickness wound models in diabetic mice to assess pharmacodynamics. The study showed that rhFN stimulated hPDLSCs differentiation into myofibroblasts by boosting ITGB1 expression. rhFN also reduced AGEs' negative effects on hPDLSCs, as seen through SEM analysis and α-SMA levels. In full-thickness wound models, hPDLSCs and rhFN accelerated re-epithelialization and collagen synthesis. rhFN is proposed to interact with the ITGB1 receptor on hPDLSCs, activating the NF-κB pathway to neutralize AGEs-induced pro-inflammatory cytokines. This study suggests rhFN as a potential biomedical material for tissue repair.