The Mechanism Of Cthrc1 In The Osteogentic Differentation Of Hpdlscs

To investigate the role and underlying mechanisms of collagen triple helix repeat containing 1 (CTHRC1) in the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs). 1. Primary hPDLSCs were isolated from premolars of healthy orthodontic patients via enzymatic digestion. 2. The mesenchymal identity of these cells was confirmed using flow cytometry and immunofluorescence. 3. A lentiviral vector was constructed to overexpress CTHRC1, with transfection efficiency verified by Western blot and qRT-PCR. 4. Osteogenic differentiation was assessed via alkaline phosphatase activity and Alizarin red staining. 5. Western blot analyzed ERK1/2 and pERK1/2 expression, then examined osteogenic markers (RUNX2, OCN, and OSTERIX) before and after ERK1/2 pathway inhibition, evaluating CTHRC1′s role in osteogenic differentiation. 1. The isolated hPDLSCs exhibited mesenchymal characteristics, multipotency, and clonal proliferative capacity, meeting the requirements for subsequent experiments. 2. Western Blot and qRT-PCR analyses confirmed CTHRC1 overexpression after transfection. 2. Osteogenic differentiation potential was elevated, with increased alkaline phosphatase activity and mineralized nodule formation in the CTHRC1 overexpression group. And western blot revealed that CTHRC1 overexpression activated the ERK1/2 signaling pathway. 3. CTHRC1 overexpression upregulated osteogenic markers (RUNX2, OCN, and OSTERIX) at the protein levels. However, inhibition of ERK1/2 activation via ERK1/2 inhibitor attenuated this effect, suggesting that CTHRC1 enhances hPDLSC osteogenic differentiation through ERK1/2 pathway activation and phosphorylation. CTHRC1 significantly promotes the osteogenic differentiation of hPDLSCs by activating the ERK1/2 signaling pathway.