Transgenic high Expression of EPHB4 in Canine Periodontal Ligament Stem Cells （cPDLSCs）Modulates Osteogenic Differentiation and migration in cPDLSCs

Abstract Stem cells and bioactive molecules are fundamental components of tissue regeneration. Particularly from oral tissues, including periodontal ligament stem cells (PDLSCs), dental pulp stem cells (DPSCs), and stem cells from the apical papilla (SCAPs), adult stem cells have great potential to develop into mesodermal lineages. Multilineage potential—that which spans bone/teeth-forming, fat-forming, and nerve-forming cells—defines these mesenchymal stem cells. Emphasizing its relevance, DPSCs and PDLSCs have been effectively stimulated to produce bone-forming cells in both in vitro and in vivo environments. Human adipose-derived stem cells (ASCs) and Bone Marrow Mesenchymal Stem Cells (BMMSCs) also show this. The EphB4 receptor, a vital member of the Eph family of receptor tyrosine kinases, plays a pivotal role in guiding mesenchymal stem cells towards the osteoblast lineage. The bone remodeling process relies significantly on the EphrinB2/EphB4 axis. EphrinB2's reverse signaling specifically reduces osteoclast activity by lowering the activity of c-Fos and NFATC1, therefore lowering the bone resorption. By reducing RhoA activity, EphB4's forward signaling improves osteoblast development and increases bone production. EphrinB2 can also bind to EphB4, therefore guiding the differentiation of BMMSCs, ASCs, PDLSCs, and SHEDs into osteoblast cells. Here we want to examine the effects of EphB4 gene transfection on canine periodontal ligament stem cells (cPDLSCs) and assess their osteogenic ability by means of osteogenic markers (RunX2, OCN, COL1) and phosphorylation levels of EphB4 in cPDLSCs.This approach aims to unveil the potential molecular mechanisms involved in this process.