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.
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.
Department of Orthopedic Surgery, Stanford University, Stanford, CA 94305, USA.
Yunqing
Y
Kang
Sungwoo
S
Kim
Monica
M
Fahrenholtz
Ali
A
Khademhosseini
Yunzhi
Y
Yang
D000469:Alkaline Phosphatase / Q000378:metabolism; D001483:Base Sequence; D001846:Bone Development; D001854:Bone Marrow Cells / Q000166:cytology* / Q000201:enzymology; D002130:Calcium Phosphates; D002478:Cells, Cultured; D018920:Coculture Techniques; D017931:DNA Primers; D004730:Endothelium, Vascular / Q000166:cytology* / Q000201:enzymology; D005455:Fluorescent Antibody Technique; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology; D008855:Microscopy, Electron, Scanning; D018919:Neovascularization, Physiologic; D060888:Real-Time Polymerase Chain Reaction; D054457:Tissue Scaffolds; D014471:Umbilical Veins / Q000166:cytology* / Q000201:enzymology
D002130:Calcium Phosphates; D017931:DNA Primers; C485817:beta-tricalcium phosphate; D000469:Alkaline Phosphatase