摘要
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 integrinβ1 receptor and the NF-κB signaling pathway,both crucial for cellular responses to fibronectin.To validate our hypothesis,we expressed rhFN1024,a recombinant protein containing the integrinβ1 affinity-binding domain from human fibronectin segments 12–14.This protein was used to formulate a hydrogel for hPDLSCs.rhFN1024's binding affinity to integrinβ1 was confirmed by molecular docking and the cellular thermal shift assay(CETSA).We developed sh-ITGB1-hPDLSCs with stable ITGB1 knockdown using shRNA-ITGB1 and compared their proliferation,migration and adhesion to wildtype hPDLSCs.Morphological changes were observed via SEM,andα-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 rhFN1024 stimulated hPDLSCs differentiation into myofibroblasts by boosting ITGB1 expression.rhFN1024 also reduced AGEs'negative effects on hPDLSCs,as seen through SEM analysis andα-SMA levels.In full-thickness wound models,hPDLSCs and rhFN1024 accelerated re-epithelialization and collagen synthesis.rhFN1024 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 rhFN1024 as a potential biomedical material for tissue repair.
基金
funded by the National Key Research and Development Program of China(2022YFC2403102)
Kea-Area Research and Development Program of Guangdong Province,China(2022B1111080007)
Guangzhou Research and Development Plan in Key Fields,China(202103030003).
