miR-210-3p suppresses osteogenic differentiation of MC3T3-E1 by targeting brain derived neurotrophic factor (BDNF)

Abstract

Background and objective As an important mediator of intercellular interaction and formation of extracellular bone matrix, porous scaffolds are widely used for bone regeneration. Accumulating evidences demonstrate that microRNA are involved in the regulation of scaffolds-induced bone regeneration. Recently, we revealed that miR-210-3p was highly expressed during osteogenesis induced by HAG. In present study, we further explored the molecular mechanism underlying the effect of miR-210-3p on osteogenic differentiation. Materials and methods In this study, miR-210-3p mimics and inhibitors were synthesized and transfected into MC3T3-E1 cells to explore their effects on osteogenic differentiation. The expression of osteogenic marker (Alp and Runx2) were detected by real-time quantitative PCR (qRT-PCR) and western blotting. After osteogenesis induction for 7 days, Alp staining were used to detected osteoblast differentiation of MC3T3-E1 cells. CCK8 and Transwell assays were performed to detected cell proliferation and migration. Then, top ranking list of target genes of miR-210-3p obtained from TargetScan and the expression of BDNF were detected by qRT-PCR and ELISA. The relationship between miR-210-3p and BDNF was verified by luciferase report assay. Furthermore, the effect of BDNF on osteoblast differentiation was verified by transfecting siRNA or adding BDNF to the culture medium. Results MiR-210-3p mimics markedly suppress osteogenic differentiation, cell migration and cell proliferation of MC3T3-E; nevertheless, silencing of miR-210-3p dramatically enhanced MC3T3-E1 osteogenesis, cell migration and proliferation. Furthermore, luciferase reporter assay verified that brain derived neurotrophic factor (BDNF) is a directly target of miR-210-3p. Moreover, BDNF siRNA significantly decreased the expression levels of ALP and cell migration. The addition of BDNF partially rescued the inhibition of osteogenesis by miR-210-3p. Conclusion miR-210-3p inhibited the osteogenic differentiation via targeting BDNF. Our Results provide a promising target for regulating osteogenic differentiation.

MicroRNA expression in JAG1/Notch-activated periodontal ligament stem cells

“…Thus, Li et al [ 35 ] found overexpression of mhesiR-133a-3p markedly inhibited the expression of osteogenesis-associated genes such as RUNX2 and OSX , reduced ALP activity and mineralization by targeting ankyrin repeat domain 44 (ANKRD44) in BMMSCs. Similarly, Deng et al [ 36 ] observed significant inhibition of osteogenic differentiation, cell migration, and cell proliferation in MC3T3-E1 cells due to miR-210-3p. This suppression was attributed to the direct targeting of brain-derived neurotrophic factor (BDNF) by miR-210-3p.…”

Three-Dimensional Mechanical Microenvironment Rescued the Decline of Osteogenic Differentiation of Old Human Jaw Bone Marrow Mesenchymal Stem Cells

“…54 Another study also found that miR-210-3p suppresses osteogenic differentiation of MC3T3-E1 by targeting brain derived neurotrophic factor. 61 No study has reported the relationship between miR-210 and osteogenic differentiation of hJBMSCs. This experiment will continue to investigate the role of miR-210 in osteogenic differentiation of hJBMSCs and create a new research direction for miR-210.…”

“…A study by Li et al found that MiR-210-3p inhibits osteogenic differentiation and promotes adipogenic differentiation correlated with Wnt signaling in ERalpha-deficient rBMSCs . Another study also found that miR-210-3p suppresses osteogenic differentiation of MC3T3-E1 by targeting brain derived neurotrophic factor . No study has reported the relationship between miR-210 and osteogenic differentiation of hJBMSCs.…”

The effect of Staphylococcus aureus on innate and adaptive immunity and potential immunotherapy for S. aureus-induced osteomyelitis

“…Its effect on osteoblasts depends on the dosage. Under physiological conditions, TNF-a promotes the migration and proliferation of osteoblasts by activating the p38/MAPK pathway (49). In contrast, under acute inflammatory conditions, excessive TNF-a inhibits osteoblastogenesis by inhibiting the Wnt pathway by regulating GSK3b (50).…”