The effects of FOXC2-gene-manipulated human periodontal ligament stem cells on bone regeneration of craniofacial bone defect

The progress and development of bone tissue engineering technology has brought new hope for the repair of oral and maxillofacial bone defects. As one of the ideal seed cells, the induction of human periodontal ligament stem cells (hPDLSCs) into osteoblasts has become a focus in current research. Forkhead box C2 (FOXC2) may be a good candidate gene for bone regeneration. Therefore, the aim of this study was to verify the role of FOXC2 in hPDLSCs osteogenesis and genetically-manipulate seed cells based on FOXC2 gene for enhancing their bone regeneration potentials. The hPDLSCs models with down-regulated and up-regulated expression of FOXC2 were constructed by lentivirus transfections. ALP staining, alizarin red staining, qRT-PCR, WB analysis and histological staining were used to verify the function of FOXC2 in regulating osteogenic differentiation of hPDLSCs. Finally, through transcriptome sequencing and qRT-PCR verification, the downstream core genes, and possible mechanism of FOXC2 were speculated. All data were presented as means ± standard deviations (SDs). P values less than 0.05 were considered statistically significant. GraphPad PRISM software (version 9.0; La Jolla, CA, USA) was used. The research results suggest that FOXC2 gene is involved in the regulation of osteogenic differentiation of hPDLSCs which may be related to promoting extracellular matrix interaction, cell adhesion and collagen formation. hPDLSCs with FOXC2 up-regulation are expected to be used as gene-enhanced seed cells in bone tissue engineering.