Nerve growth factor promotes osteogenic differentiation of MC3T3-E1 cells via BMP-2/Smads pathway.

BACKGROUND: Exogenous nerve growth factor (NGF) can induce osteogenic precursor cell differentiation and promote fracture healing. However, the molecular mechanism by which NGF induces osteogenesis is not well understood. BMP-2 has good osteogenic efficacy and is one of the most osteogenic-inducing growth factors known. Therefore, this study aimed to determine whether NGF induces osteogenic differentiation of mouse embryonic osteogenic precursor cell line MC3T3-E1 by BMP-2 and search further mechanisms of NGF on BMP-2. METHODS: MC3T3-E1 cells were treated with NGF at a concentration gradient for indicated times, after which the cell viability was measured by CCK-8 kit. Osteogenic differentiation was detected with quantification of alkaline phosphatase (ALP) activity also visualized with ALP staining. The transcription and expression of relevant genes were detected by qPCR and western blotting, respectively. NGF's effect on BMP2 was studied with qPCR and luciferase reporter assay. The phosphorylation of Smads was probed with specific antibodies by western blotting, and the location of Smads was observed through immunofluorescence. RESULTS: We found that NGF promoted proliferation and osteogenic differentiation of MC3T3-E1, increased the expression level of BMP-2, as well as the phosphorylation and nuclear translocation of Smad1/5/8. However, neutralization of BMP-2 with si-BMP-2 or BMP-2 signal inhibitors reversed NGF induced phosphorylation and nuclear translocation of Smad1/5/8, as well as the expression of Runx2, type I collagen, osteocalcin and osteopontin. In addition, si-BMP-2 abrogated NGF-induced ALP activity. CONCLUSION: NGF induced osteogenic differentiation of MC3T3-E1 cells through BMP-2/Smads pathway and induction of Runx2. Our study would provide a theoretical basis for clinical treatment of fractures using NGF.

Intra-articular Injection of Chitosan-Based Supramolecular Hydrogel for Osteoarthritis Treatment.

BACKGROUND: Pain and cartilage destruction caused by osteoarthritis (OA) is a major challenge in clinical treatment. Traditional intra-articular injection of hyaluronic acid (HA) can relieve the disease, but limited by the difficulty of long-term maintenance of efficacy. METHODS: In this study, an injectable and self-healing hydrogel was synthesized by in situ crosslinking of N-carboxyethyl chitosan (N-chitosan), adipic acid dihydrazide (ADH), and hyaluronic acid-aldehyde (HA-ALD). RESULTS: This supramolecular hydrogel sustains good biocompatibility for chondrocytes. Intra-articular injection of this novel hydrogel can significantly alleviate the local inflammation microenvironment in knee joints, through inhibiting the inflammatory cytokines (such as TNF-α, IL-1ß, IL-6 and IL-17) in the synovial fluid and cartilage at 2- and even 12-weeks post-injection. Histological and behavioral test indicated that hydrogel injection protected cartilage destruction and relieved pain in OA rats, in comparison to HA injection. CONCLUSION: This kind of novel hydrogel, which is superior to the traditional HA injection, reveals a great potential for the treatment of OA.