Demineralized Dentin Matrix Promotes Bone Regeneration Through IDO1-Mediated Th17/Treg Cell Balance Modulation

Demineralised dentin matrix (DDM) is an effective scaffold material for bone tissue engineering. However, the osteoimmunological mechanism of DDM remains unexplored. Th17/Treg cell balance has been noticed as a crucial factor in bone regeneration. The aim of this research is to elucidate the influence of DDM on the balance of Th17/Treg and its underlying molecular mechanisms.

The study examined DDM's effects on Th17/Treg cell balance by co-incubating DDM extract with peripheral blood mononuclear cells (PBMCs). Transcriptomics and bioinformatics identified DDM-related differentially expressed genes. A naïve CD4⁺ T cell-bone marrow-derived mesenchymal stem cells (BMSCs) co-culture model assessed DDM's immunomodulatory effects on Th17/Treg differentiation and their impact on BMSC viability and osteogenic differentiation. In vivo, a rabbit mandibular defect model with DDM implantation evaluated bone repair and Th17/Treg regulation.

DDM promoted Treg cell differentiation in PBMCs and increased anti-inflammatory cytokines. Transcriptomic analysis shows that indoleamine 2,3-dioxygenase 1 (IDO1) is a key regulator of Th17/Treg balance. In naïve CD4⁺ T cells, DDM enhanced Treg differentiation, which was blocked by IDO1 inhibition. This immunomodulation improved BMSCs viability and osteogenic differentiation which was attenuated by IDO1 inhibition. Co-culture with naïve CD4⁺ T cells further amplified DDM's pro-osteogenic effects. In rabbit mandibular defects, DDM enhanced bone repair and was suppressed with IDO1 inhibition.

This study demonstrates that DDM regulates the Th17/Treg cell balance by promoting Treg cell differentiation through IDO1 upregulation, with minimal impact on Th17 cells. This immunomodulatory mechanism is a key factor underlying DDM's significant role in bone repair.