TY - JOUR
T1 - Osteogenesis by human mesenchymal stem cells cultured on silk biomaterials
T2 - Comparison of adenovirus mediated gene transfer and protein delivery of BMP-2
AU - Meinel, Lorenz
AU - Hofmann, Sandra
AU - Betz, Oliver
AU - Fajardo, Robert
AU - Merkle, Hans P.
AU - Langer, Robert
AU - Evans, Christopher H.
AU - Vunjak-Novakovic, Gordana
AU - Kaplan, David L.
N1 - Funding Information:
We thank Elvire Gouze for support with the FACS analysis. This work was supported by the German Alexander von Humboldt Foundation (Feodor-Lynen fellowship to LM), the Association for Orthopaedic Research, and the National Institutes of Health (DE13405-04, NIH AR46563-01A2 to DK, NIH P41 EB002520 to DK, and NIH AR050243-01 to CE). The authors thank Trudel Inc. (Zurich, Switzerland) for providing B. mori cocoons.
PY - 2006/10
Y1 - 2006/10
N2 - Bone tissue engineering, gene therapy based on human mesenchymal stem cells (MSCs) and silk fibroin biomaterials were combined to study the impact of viral transfection on MSC osteogenic performance in vitro. MSCs were transduced with adenovirus containing a human BMP-2 (Ad-BMP-2) gene at clinically reasonable viral concentrations and cultured for 4 weeks. Controls with nontransfected MSCs, but exposed to exogenous BMP-2 concentrations on an analogous time profile as that secreted by the Ad-BMP-2 group, were compared. Both the Ad-BMP-2 MSC group and the exogenous protein BMP-2 group strongly expressed osteopontin and bone sialoprotein. Cells secreted a matrix that underwent mineralization on the silk fibroin scaffolds, forming clusters of osseous material, as determined by micro-computed tomography. The expression of osteogenic marker proteins and alkaline phosphatase was significantly higher in the Ad-BMP-2 MSC group than in the exogenous protein BMP-2 group, and no significant differences in mineralization were observed in two of the three MSC sources tested. The results demonstrate that transfection resulted in higher levels of expression of osteogenic marker genes, no change in proliferation rate and did not impact the capacity of the cells to calcify tissues on these protein scaffolds. These findings suggest additional options to control differentiation where exogenous additions of growth factors or morphogens can be replaced with transfected MSCs.
AB - Bone tissue engineering, gene therapy based on human mesenchymal stem cells (MSCs) and silk fibroin biomaterials were combined to study the impact of viral transfection on MSC osteogenic performance in vitro. MSCs were transduced with adenovirus containing a human BMP-2 (Ad-BMP-2) gene at clinically reasonable viral concentrations and cultured for 4 weeks. Controls with nontransfected MSCs, but exposed to exogenous BMP-2 concentrations on an analogous time profile as that secreted by the Ad-BMP-2 group, were compared. Both the Ad-BMP-2 MSC group and the exogenous protein BMP-2 group strongly expressed osteopontin and bone sialoprotein. Cells secreted a matrix that underwent mineralization on the silk fibroin scaffolds, forming clusters of osseous material, as determined by micro-computed tomography. The expression of osteogenic marker proteins and alkaline phosphatase was significantly higher in the Ad-BMP-2 MSC group than in the exogenous protein BMP-2 group, and no significant differences in mineralization were observed in two of the three MSC sources tested. The results demonstrate that transfection resulted in higher levels of expression of osteogenic marker genes, no change in proliferation rate and did not impact the capacity of the cells to calcify tissues on these protein scaffolds. These findings suggest additional options to control differentiation where exogenous additions of growth factors or morphogens can be replaced with transfected MSCs.
KW - Adenoviral gene transfer
KW - Mesenchymal stem cells
KW - Micro-computed tomography
KW - Silk
KW - Tissue engineering
KW - bone morphogenetic protein (BMP-2)
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U2 - 10.1016/j.biomaterials.2006.05.021
DO - 10.1016/j.biomaterials.2006.05.021
M3 - Article
C2 - 16765437
AN - SCOPUS:33744966656
SN - 0142-9612
VL - 27
SP - 4993
EP - 5002
JO - Biomaterials
JF - Biomaterials
IS - 28
ER -