Characterization of adipose-derived mesenchymal stem cell combinations for vascularized bone engineering.
Since bone repair and regeneration depend on vasculogenesis and osteogenesis, both of these processes are essential for successful vascularized bone engineering. Using adipose-derived stem cells (ASCs), we investigated temporal gene expression profiles, as well as bone nodule and endothelial tubule formation capacities, during osteogenic and vasculogenic ASC lineage commitment. Osteoprogenitor-enriched cell populations were found to express RUNX2, MSX2, SP7 (osterix), BGLAP (osteocalcin), SPARC (osteonectin), and SPP1 (osteopontin) in a temporally specific sequence. Irreversible commitment of ASCs to the osteogenic lineage occurred between days 6 and 9 of differentiation. Endothelioprogenitor-enriched cell populations expressed CD34, PECAM1 (CD31), ENG (CD105), FLT1 (Vascular endothelial growth factor [VEGFR1]), and KDR (VEGFR2). Capacity for microtubule formation was evident in as early as 3 days. Functional capacity was assessed in eight coculture combinations for both bone nodule and endothelial tubule formation, and the greatest expression of these end-differentiation phenotypes was observed in the combination of well-differentiated endothelial cells with less-differentiated osteoblastic cells. Taken together, our results demonstrate vascularized bone engineering utilizing ASCs is a promising enterprise, and that coculture strategies should focus on developing a more mature vascular network in combination with a less mature osteoblastic stromal cell.
Institute of Reconstructive Plastic Surgery Laboratories, New York University Langone Medical Center, New York, New York 10016, USA.
Cristian D
CD
Valenzuela
Alexander C
AC
Allori
Derek D
DD
Reformat
Alexander M
AM
Sailon
Robert J
RJ
Allen
Edward H
EH
Davidson
Mani
M
Alikhani
Timothy G
TG
Bromage
John L
JL
Ricci
Stephen M
SM
Warren
D000273:Adipose Tissue / Q000166:cytology*; D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D001842:Bone and Bones / Q000098:blood supply* / Q000502:physiology*; D002454:Cell Differentiation; D019070:Cell Lineage; D018920:Coculture Techniques; D005260:Female; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D008870:Microtubules / Q000378:metabolism; D008875:Middle Aged; D018919:Neovascularization, Physiologic; D010011:Osteocytes / Q000166:cytology / Q000378:metabolism; D010012:Osteogenesis; D023822:Tissue Engineering / Q000379:methods*; D055815:Young Adult
D015415:Biomarkers