Osteoprotegerin Enhances Osteogenesis of Human Mesenchymal Stem Cells

Abstract

Mesenchymal stem cells (MSCs) can differentiate into osteoblasts and hold promise for applications of bone regeneration such as bone tissue engineering. However, current approaches for in vitro osteogenesis cannot effectively induce osteogenesis and need to be modified to produce quality bone for clinical applications. Previous studies have shown that the conditioned medium (CM) from osteoblast culture enhances osteogenesis of MSCs, and soluble osteogenic factors in the CM may be involved in the regulation. However, these factors are not fully identified. In this study, we profiled soluble factors secreted from MG-63 cells using a comparative protein array and found that osteoprotegerin (OPG), known as a potent anti-osteoclastogenic protein, was at the highest relative level among 507 soluble molecules detected by the array. Furthermore, treating hMSCs with OPG before osteogenic induction significantly increased the expression of osteocalcin mRNA transcript and the production of calcium deposits compared to the untreated control cells, suggesting that OPG is capable of priming undifferentiated hMSCs for the enhancement of subsequent osteogenesis. Furthermore, we showed that the nuclear factor-kappaB (NF-κB) was activated by OPG in undifferentiated hMSCs and that blocking NF-κB activation before osteogenic induction decreased osteogenesis of OPG-pretreated cells upon receiving osteogenic stimuli. Taken together, our results suggest that OPG is a pro-osteogenic factor that can be used as an osteogenic supplement in a growth medium to prime the osteogenic capacity of undifferentiated hMSCs to enhance osteogenesis for bone tissue engineering.