Abstract
BackgroundImproved understanding of the interactions between bone cells and endothelial cells involved in osteogenesis should aid the development of new strategies for bone tissue engineering. The aim of the present study was to determine whether direct communication between bone marrow stromal cells (MSC) and human umbilical vein endothelial cells (EC) could influence the osteogenic potential of MSC in osteogenic factor-free medium.MethodsAfter adding EC to MSC in a direct-contact system, cell viability and morphology were investigated with the WST assay and immnostaining. The effects on osteogenic differentiation of adding EC to MSC was systematically tested by the using Superarray assay and results were confirmed with real-time PCR.ResultsFive days after the addition of EC to MSC in a ratio of 1:5 (EC/MSC) significant increases in cell proliferation and cellular bridges between the two cell types were detected, as well as increased mRNA expression of alkaline phosphatase (ALP). This effect was greater than that seen with addition of osteogenic factors such as dexamethasone, ascorbic acid and β-glycerophosphate to the culture medium. The expression of transcription factor Runx2 was enhanced in MSC incubated with osteogenic stimulatory medium, but was not influenced by induction with EC. The expression of Collagen type I was not influenced by EC but the cells grown in the osteogenic factor-free medium exhibited higher expression than those cultured with osteogenic stimulatory medium.ConclusionThese results show that co-culturing of EC and MSC for 5 days influences osteogenic differentiation of MSC, an effect that might be independent of Runx2, and enhances the production of ALP by MSC.
Highlights
The goal of bone tissue engineering is the generation of new bone from osteogenic cells, supported with biocompatible, biodegradable three-dimensional scaffolds [1,2]
In most of the co-culture studies to date, cells have been treated with osteogenic stimulatory medium [14,19,20], whereas the present study was undertaken without the addition of osteogenic supplements to marrow stromal cells (MSC) or to the coculture group
Under osteogenic factor-free conditions, the addition of endothelial cells (EC) induced the expression of alkaline phosphatase (ALP) by MSC in a short incubation time (5 days)
Summary
The goal of bone tissue engineering is the generation of new bone from osteogenic cells, supported with biocompatible, biodegradable three-dimensional scaffolds [1,2]. The type of osteogenic cell most appropriate for bone tissue engineering has yet to be determined. As MSC can be isolated from adult bone marrow and expanded in vitro without loss of differentiation potential, they are suitable candidates for bone tissue engineering [1,5,6,7]. Improved understanding of the interactions between bone cells and endothelial cells involved in osteogenesis should aid the development of new strategies for bone tissue engineering. The aim of the present study was to determine whether direct communication between bone marrow stromal cells (MSC) and human umbilical vein endothelial cells (EC) could influence the osteogenic potential of MSC in osteogenic factor-free medium
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