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Related Experiment Video

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Bone Conditioned Medium: Preparation and Bioassay

Bone Conditioned Medium: Preparation and Bioassay

Published on: July 8, 2015

CCN3 increases BMP-4 expression and bone mineralization in osteoblasts.

Tzu-Wei Tan¹, Yuan-Lin Huang, Jung-Tzu Chang

¹Department of Pharmacology, School of Medicine, China Medical University and Hospital, Taichung, Taiwan.

Journal of Cellular Physiology

|September 8, 2011

Summary

Nephroblastoma overexpressed (NOV) gene, also known as CCN3, promotes bone formation by increasing BMP-4 expression in osteoblasts. This process involves integrins, ILK, p38, JNK, and AP-1 signaling pathways.

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Isolation of Mesenchymal Stem Cells from Human Alveolar Periosteum and Effects of Vitamin D on Osteogenic Activity of Periosteum-derived Cells

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Published on: May 4, 2018

Area of Science:

Cell Biology
Molecular Biology
Biochemistry

Background:

The nephroblastoma overexpressed (NOV) gene, or CCN3, is known to regulate skeletal mesenchymal cell differentiation.
Bone morphogenetic proteins (BMPs) are critical for osteoblast differentiation and bone formation.
The specific role of CCN3 in regulating BMP expression and bone formation in osteoblasts remains largely uncharacterized.

Purpose of the Study:

To investigate the effects of CCN3 on BMP expression and bone nodule formation in cultured osteoblasts.
To elucidate the signaling pathways involved in CCN3-mediated osteoblast differentiation.

Main Methods:

Cultured osteoblasts were treated with CCN3.
Expression of BMP-4 was measured.
Bone nodule formation was assessed.
Involvement of integrins, integrin-linked kinase (ILK), p38, JNK, and activator protein-1 (AP-1) signaling pathways was examined using antibodies and inhibitors.
Kinase activity, protein phosphorylation, and nuclear translocation of c-Jun were analyzed.

Main Results:

CCN3 significantly increased BMP-4 expression and bone nodule formation in osteoblasts.
Inhibition of α5β1 and αvβ5 integrins, ILK, p38, and JNK abolished CCN3-induced effects.
CCN3 enhanced ILK kinase activity and phosphorylation of p38 and JNK.
AP-1 inhibitors suppressed CCN3-enhanced bone nodule formation and BMP-4 expression.
CCN3 promoted c-Jun nuclear translocation and binding to the BMP-4 promoter, which was blocked by ILK, p38, and JNK inhibitors.

Conclusions:

CCN3 enhances BMP-4 expression and bone nodule formation in osteoblasts.
The integrin receptor, ILK, p38, JNK, and AP-1 signaling pathways are crucial mediators of CCN3's effects on osteoblast differentiation and bone formation.