Decellularised matrices from force loaded periodontal ligament stem cells support osteogenic differentiation

Periodontal ligament stem cells (hPDLSCs) are mechanosensing cells responding to mechanical forces. This study investigates the impact of decellularised extracellular matrix (dECM) derived from intermittent compressive force (ICF)-treated hPDLSCs on osteogenic differentiation. hPDSCLs were subjected to ICF loading at 1.5 g/cm² for 24 h and then maintained with normal medium (N) or osteogenic induction medium (OM) followed by a decellularisation process. dECMs derived from ICF (dECM-ICF) were characterised using a scanning electron microscope, energy-dispersive X-ray spectroscopy, and proteomic analysis. hPDLSCs were re-seeded on dECM-ICF. Cell proliferation and viability were examined by resazurin and LIVE/DEAD assays. Mineralisation was determined by Alizarin Red S staining. Results demonstrated that dECM-ICF-derived from OM (dECM-ICF-OM) exhibited a fibrillar network structure and showed no cellular component while preserving fibronectin and type I collagen. dECM-ICF exhibited biocompatibility, as indicated by the absence of cytotoxic effects and the ability of hPDLSCs to attach, spread, and proliferate. dECM-ICF-OM significantly enhanced mineral deposition compared to dECM-ICF from normal conditions (dECM-ICF-N). Proteomic analysis of dECM-ICF demonstrated the upregulated proteins in the PI3K-Akt, Ras, MAPK, mTOR, ErbB, TNF, and VEGF signallings. In conclusion, dECM-ICF supports hPDLSCs growth and osteogenic differentiation. dECM-ICF is a promising cell-free natural scaffold to promote periodontal regeneration.