Bidirectional ephrinB2‑EphB4 signaling regulates the osteogenic differentiation of canine periodontal ligament stem cells

Abstract

The aim of the present study was to evaluate the effect of ephrinB2 gene-transfected canine periodontal ligament stem cells (cPdLScs) on the regulation of osteogenic differentiation. cPdLScs were transfected with a transgenic null-control green fluorescent protein (GFP) vector (termed Vector-cPDLSCs) or with NFNB2 GFP-Blasticidin (termed EfnB2-cPdLScs). Subsequently, the osteogenic differentiation of Vector-cPdLScs and EfnB2-cPdLScs was assessed by reverse transcription-quantitative polymerase chain reaction (RT-qPcR), alkaline phosphatase (ALP) assay and Alizarin Red S staining. The migratory abilities of cPdLScs, Vector-cPdLScs and EfnB2-cPdLScs were also assessed. Following osteogenic induction of Vector-cPdLScs and EfnB2-cPdLScs, the protein expression levels of collagen I, Runt-related transcription factor 2, osteocalcin, ephrin type-B receptor 4 (EphB4), phospho-EphB4, ephrinB2 and phospho-ephrinB2 were analyzed by western blot assays. Following gene transfection, the RT-qPcR and western blotting results revealed that the mRNA and protein expression levels of ephrinB2, respectively, were significantly increased in EfnB2-cPdLScs compared with that in Vector-cPdLScs (P<0.05). ALP and Alizarin Red S staining assays revealed increased ALP activity and mineralization nodules, respectively, in EfnB2-cPdLScs. cell proliferation and migration assays revealed that EfnB2-cPdLScs exhibited enhanced proliferation and migration compared with Vector-cPdLScs (P<0.05). In conclusion, the findings of the current study indicated that ephrinB2 gene-modified cPDLSCs exhibited enhanced osteogenic differentiation, with the ephrinB2 reverse signaling and EphB4 forward signaling pathways serving a key role in this process. Furthermore, ephrinB2 gene modification was observed to promote the migration and proliferation of cPdLScs.

Intermittent parathyroid hormone promotes cementogenesis via ephrinB2‐EPHB4 forward signaling

“…Meanwhile, it is interesting to note that apart from cellular cementum, the PDL areas were also stained positively with ephrinB2 and EPHB4 during the root regeneration process. Previously, it was reported that the ephrinB2/EPHB4 signaling was able to regulate the osteogenic differentiation of PDL stem cells and cocultured preosteoblasts (S. Zhu et al, 2020;S. Y. Zhu et al, 2017).…”

“…Meanwhile, it is interesting to note that apart from cellular cementum, the PDL areas were also stained positively with ephrinB2 and EPHB4 during the root regeneration process. Previously, it was reported that the ephrinB2/EPHB4 signaling was able to regulate the osteogenic differentiation of PDL stem cells and cocultured preosteoblasts (S. Zhu et al, 2020; S. Y. Zhu et al, 2017). Similarly, it could be speculated that there might be ephrinB2/EPHB4 signaling transduction between PDL cells and cementoblasts in our study as well, since intercellular communication was easy within the dense periodontium.…”

Pathological and Therapeutic Approach to Endotoxin-Secreting Bacteria Involved in Periodontal Disease

“…Induced by LPS, TLR4 activates the pro-inflammatory transcription factor NF-κB that enters the nucleus and initiates pro-inflammatory gene transcription that encode the pro-inflammatory cytokines, IL-1 β , IL-6 and IL-8, leading to periodontal tissue destruction [ 54 ]. It increases the expression of Ephrin type-B receptor 4 (EphB4) and inhibits the expression of the protein EphrinB2 [ 74 ]. It also inhibits alkaline phosphatase activity, collagen type 1 Alpha 1, and the osteocalcin production and mineralization in the periodontal ligament stem cells proliferation.…”

Erythropoietin induces odontoblastic differentiation of human‐derived pulp stem cells via EphB4‐Mediated MAPK signaling pathway

“…Recent assays have reported the activation of EphrinB2/EphB4 increased osteogenic/odontogenic differentiation of hDPSCs and canine periodontal ligament stem cells (cPDLSCs) (Heng et al, 2018;Zhu et al, 2020). Moreover, EPO has been shown to regulate bone formation by activating EphrinB2/EphB4 signaling (Li, Shi, et al, 2015).…”

“…The Eph receptors belong to the receptor tyrosine kinase superfamily, and the interaction between ephrin ligands and their receptors contributes to bi‐directional signaling (Lisabeth et al, 2013). Recent assays have reported the activation of EphrinB2/EphB4 increased osteogenic/odontogenic differentiation of hDPSCs and canine periodontal ligament stem cells (cPDLSCs) (Heng et al, 2018; Zhu et al, 2020). Moreover, EPO has been shown to regulate bone formation by activating EphrinB2/EphB4 signaling (Li, Shi, et al, 2015).…”