Osteogenic and angiogenic potentials of monocultured and co-cultured human-bone-marrow-derived mesenchymal stem cells and human-umbilical-vein endothelial cells on three-dimensional porous beta-tricalcium phosphate scaffold

Acta Biomater. 2013 Jan;9(1):4906-15. doi: 10.1016/j.actbio.2012.08.008. Epub 2012 Aug 16.


The use of biodegradable beta-tricalcium phosphate (β-TCP) scaffolds holds great promise for bone tissue engineering. However, the effects of β-TCP on bone and endothelial cells are not fully understood. This study aimed to investigate cell proliferation and differentiation of mono- or co-cultured human-bone-marrow-derived mesenchymal stem cells (hBMSCs) and human-umbilical-vein endothelial cells (HUVECs) on a three-dimensional porous, biodegradable β-TCP scaffold. In co-culture studies, the ratios of hBMSCs:HUVECs were 5:1, 1:1 and 1:5. Cellular morphologies of HUVECs, hBMSCs and co-cultured HUVECs/hBMSCs on the β-TCP scaffolds were monitored using confocal and scanning electron microscopy. Cell proliferation was monitored by measuring the amount of double-stranded DNA (dsDNA) whereas hBMSC and HUVEC differentiation was assessed using the osteogenic and angiogenic markers, alkaline phosphatase (ALP) and PECAM-1 (CD31), respectively. Results show that HUVECs, hBMSCs and hBMSCs/HUVECs adhered to and proliferated well on the β-TCP scaffolds. In monoculture, hBMSCs grew faster than HUVECs on the β-TCP scaffolds after 7 days, but HUVECs reached similar levels of proliferation after 14 days. In monoculture, β-TCP scaffolds promoted ALP activities of both hBMSCs and HUVECs when compared to those grown on tissue culture well plates. ALP activity of cells in co-culture was higher than that of hBMSCs in monoculture. Real-time polymerase chain reaction results indicate that runx2 and alp gene expression in monocultured hBMSCs remained unchanged at days 7 and 14, but alp gene expression was significantly increased in hBMSC co-cultures when the contribution of individual cell types was not distinguished.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Base Sequence
  • Bone Development*
  • Bone Marrow Cells / cytology*
  • Bone Marrow Cells / enzymology
  • Calcium Phosphates*
  • Cells, Cultured
  • Coculture Techniques
  • DNA Primers
  • Endothelium, Vascular / cytology*
  • Endothelium, Vascular / enzymology
  • Fluorescent Antibody Technique
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / enzymology
  • Microscopy, Electron, Scanning
  • Neovascularization, Physiologic*
  • Real-Time Polymerase Chain Reaction
  • Tissue Scaffolds*
  • Umbilical Veins / cytology*
  • Umbilical Veins / enzymology


  • Calcium Phosphates
  • DNA Primers
  • beta-tricalcium phosphate
  • Alkaline Phosphatase