Abstract
BackgroundThe main goal of bone tissue engineering has been the generation of healthy bone in order to replace affected tissue. Therefore, optimized biomaterials are needed which allow the survival and growth of mesenchymal stem cells. Until now the key challenge in the clinical application of cell-based tissue engineering bone implants was poor diffusion of oxygen into the tissue, making functional blood vessel networks a necessity. With their ability to evolve into different cell types, to expand extensively in vitro, and to release paracrine soluble factors, bone marrow stromal cells (BMSC) are highly attractive for tissue engineering. During the last years hypoxia became a proven method to control proliferation, differentiation, and pluripotency of BMSC. Here we applied different methods to characterize metabolically conditioned media (MCM) in comparison to hypoxia conditioned media (HCM) and evaluated their ability to attract BMSC in 2-D migration assays.MethodsBMSC and fibroblasts of human origin were isolated and cultivated to obtain HCM and MCM. Both media were characterized by angiogenesis arrays, cytokine arrays, and ELISA for selected factors. 2-D migration tests were performed with Corning Transwell®-96 permeable support chambers with porous polyester membranes with a pore size of 8.0 μm.ResultsCharacterization of HCM and MCM revealed that the concentration of angiogenic factors was higher in MCM than in HCM. However, the chemoattractive capacity of MCM for BMSC was equivalent to that of HCM. HCM and MCM produced by human skin fibroblasts attracted human BMSC as efficiently as HCM and MCM produced by human BMSC.ConclusionsHCM and MCM have a high chemoattractive capacity for BMSC. Both conditioned media harbor high concentrations of angiogenic factors which are important for angiogenesis and cell migration. Both chemoattracting conditioned media can also be derived from skin fibroblasts which can easily be obtained from patients in individualized therapy approaches.
Highlights
The main goal of bone tissue engineering has been the generation of healthy bone in order to replace affected tissue
Knowing that the secretion of vascular endothelial growth factor (VEGF) from these cells can be upregulated by hypoxic conditions in a hypoxia-inducible factor-1 (HIF1)-dependent way [14], we showed in our previous research that hypoxia conditioned media have a speciesspecific chemoattractive capacity for human, ovine, equine, and canine bone marrow stromal cells (BMSC) [15]
Characteristics of hypoxia conditioned media (HCM) and metabolically conditioned media (MCM) In order to determine which angiogenic factors were released from BMSC and foreskin fibroblasts under hypoxic and starving conditions we performed a Human Angiogenesis Array analysis
Summary
The main goal of bone tissue engineering has been the generation of healthy bone in order to replace affected tissue. Until now the key challenge in the clinical application of cell-based tissue engineering bone implants was poor diffusion of oxygen into the tissue, making functional blood vessel networks a necessity With their ability to evolve into different cell types, to expand extensively in vitro, and to release paracrine soluble factors, bone marrow stromal cells (BMSC) are highly attractive for tissue engineering. Optimized biomaterials are needed which allow survival and growth of mesenchymal stem cells, a subset of bone marrow stromal cells (BMSC), with the ability to differentiate into osteoblasts and osteocytes. Another critical point is to attract endothelial cells in order to provide neovascularization. BMSC are able to secrete significant levels of chemoattractive agents like VEGF, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), MIP-1β, and monokine induced by IFN-γ (MIG) [5]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
