The Recombinant Protein EphB4-Fc Changes the Ti Particle-Mediated Imbalance of OPG/RANKL via EphrinB2/EphB4 Signaling Pathway and Inhibits the Release of Proinflammatory Factors In Vivo

Abstract

Aseptic loosening caused by wear particles is one of the common complications after total hip arthroplasty. We investigated the effect of the recombinant protein ephB4-Fc (erythropoietin-producing human hepatocellular receptor 4) on wear particle-mediated inflammatory response. In vitro, ephrinB2 expression was analyzed using siRNA-NFATc1 (nuclear factor of activated T-cells 1) and siRNA-c-Fos. Additionally, we used Tartrate-resistant acid phosphatase (TRAP) staining, bone pit resorption, Enzyme-linked immunosorbent assay (ELISA), as well as ephrinB2 overexpression and knockdown experiments to verify the effect of ephB4-Fc on osteoclast differentiation and function. In vivo, a mouse skull model was constructed to test whether the ephB4-Fc inhibits osteolysis and inhibits inflammation by micro-CT, H&E staining, immunohistochemistry, and immunofluorescence. The gene expression of ephrinB2 was regulated by c-Fos/NFATc1. Titanium wear particles activated this signaling pathway to the promoted expression of the ephrinB2 gene. However, ephrinB2 protein can be activated by osteoblast membrane receptor ephB4 to inhibit osteoclast differentiation. In in vivo experiments, we found that ephB4 could regulate Ti particle-mediated imbalance of OPG/RANKL, and the most important finding was that ephB4 relieved the release of proinflammatory factors. The ephB4-Fc inhibits wear particle-mediated osteolysis and inflammatory response through the ephrinB2/EphB4 bidirectional signaling pathway, and ephrinB2 ligand is expected to become a new clinical drug therapeutic target.

Low-intensity pulsed ultrasound regulates osteoblast-osteoclast crosstalk via EphrinB2/EphB4 signaling for orthodontic alveolar bone remodeling

“…This suggests that the differentiation and maturation of osteoclasts require the interaction of osteoblasts. However, the result is in contradiction with the finding that EphB4 could inhibits osteoclast differentiation as a reverse signal (Ge et al, 2020). Thus, we speculate that this effect may be through other signaling pathways; for example, the RANKL/OPG pathway of osteoblasts may be increased by LIPUS.…”

Osteoblast Jmjd3 regulates osteoclastogenesis via EphB4 and RANKL signalling

“…Therefore, further studies are needed to investigate the role of Jmjd3 in RANKL expression. Our results and previous studies have shown that RANKL expression increases when the expression of EphB4 decreases (Ge et al, 2020). After overexpressing EphB4 in Jmjd3‐silenced osteoblasts, the formation of osteoclasts and the expression of NFATc1, c‐Fos, TRAP and cathepsin K in RAW264.7 cells was significantly inhibited when co‐cultured with the supernatant of these cells.…”

“…The binding of EphB4 on the membrane of osteoblasts to EphrinB2 on the membrane of osteoclasts activates downstream signalling pathways and suppresses osteoclastogenesis, a process referred to as a ‘reverse signal’. Conversely, when EphrinB2 binds to EphB4 on the osteoblast membrane, the binding promotes the mineralization of osteoblasts, which is known as a ‘forward signal’ (Ge et al, 2020; Mundy & Elefteriou, 2006; Zhao et al, 2006).…”

The Therapeutic Effects of EFNB2-Fc in a Cell Model of Kawasaki Disease

“…Knockdown of EphB4 led to the inhibition of both p-ERK and p-STAT1, but the ultimate effect on tube formation is reduction. A previous study showed that EphB4 relieved the release of proinflammatory factors [32]. In our study, the knockdown of EphB4 showed a tendency to increase inflammation-related factors, such as IL-6, but did not bring a significant difference.…”