Effect of Osteoclast Co-culture on the Differentiation of Human Mesenchymal Stem Cells Grown on Bone Graft Granules

Abstract

Traditional approaches to bone repair are currently being integrated with innovative tissue-engineering techniques, as researchers and clinicians shift their treatment focus toward regenerating functional tissue rather than just filling a defect to provide structural support. Cells are expanded and incorporated into implantable systems in hopes of enhancing the bone-forming capabilities of traditional bone graft substitutes. The present study examined how osteoclasts might be used to stimulate the differentiation of human mesenchymal stem cells (hMSCs) into bone forming cells. The two cell types were co-cultured on a resorbable, three-dimensional bone graft substitute. Osteoclasts were seeded prior to the addition of hMSCs, as well as simultaneously, to determine if resorption of the scaffold would have any bearing on observed response by hMSCs. When seeded directly with hMSCs on the 3-D substrates, the osteoclasts had an increase in TRAP expression over time if seeded simultaneously. The co-culture setup had a positive influence on the proliferation of hMSCs. Late stage osteoblast differentiation markers (bone sialoprotein) were positively affected by direct co-culture with osteoclasts. The addition of RANKL to the culture medium for osteoclastogenesis appears to be a factor in the observed responses by hMSCS, but is not the only factor influencing the MSCs. Osteoclasts were shown to have an influence on the development of mesenchymal stem cells into osteoblasts when cultured in vitro. Findings from this study, coupled with the knowledge obtained from our previous work, will aid in the development of a clinically viable mesenchymal stem cell based bone graft system.

Keywords:

• BONE GRAFT
• MESENCHYMAL STEM CELL
• OSTEOCLASTS
• TISSUE ENGINEERING