Synergistic effect of Indian hedgehog and bone morphogenetic protein-2 gene transfer to increase the osteogenic potential of human mesenchymal stem cells.

Synergistic effect of Indian hedgehog and bone morphogenetic protein-2 gene transfer to increase the osteogenic potential of human mesenchymal stem cells. Reichert, J. C., Schmalzl, J., Prager, P., Gilbert, F., Quent, V. M. C., Steinert, A. F., Rudert, M., & Noth, U. Stem cell research & therapy, 2013.
doi abstract bibtex

INTRODUCTION: To stimulate healing of large bone defects research has concentrated on the application of mesenchymal stem cells (MSCs). METHODS: In the present study, we induced the overexpression of the growth factors bone morphogenetic protein 2 (BMP-2) and/or Indian hedgehog (IHH) in human MSCs by adenoviral transduction to increase their osteogenic potential. GFP and nontransduced MSCs served as controls. The influence of the respective genetic modification on cell metabolic activity, proliferation, alkaline phosphatase (ALP) activity, mineralization in cell culture, and osteogenic marker gene expression was investigated. RESULTS: Transduction had no negative influence on cell metabolic activity or proliferation. ALP activity showed a typical rise-and-fall pattern with a maximal activity at day 14 and 21 after osteogenic induction. Enzyme activity was significantly higher in groups cultured with osteogenic media. The overexpression of BMP-2 and especially IHH + BMP-2 resulted in a significantly higher mineralization after 28 days. This was in line with obtained quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analyses, which showed a significant increase in osteopontin and osteocalcin expression for osteogenically induced BMP-2 and IHH + BMP-2 transduced cells when compared with the other groups. Moreover, an increase in runx2 expression was observed in all osteogenic groups toward day 21. It was again more pronounced for

@article{reichert_synergistic_2013,
	title = {Synergistic effect of {Indian} hedgehog and bone morphogenetic protein-2 gene transfer to increase the osteogenic potential of human mesenchymal stem cells.},
	volume = {4},
	doi = {10.1186/scrt316},
	abstract = {INTRODUCTION: To stimulate healing of large bone defects research has concentrated on the application of mesenchymal stem cells (MSCs). METHODS: In the present study, we induced the overexpression of the growth factors bone morphogenetic protein 2 (BMP-2) and/or Indian hedgehog (IHH) in human MSCs by adenoviral transduction to increase their osteogenic potential. GFP and nontransduced MSCs served as controls. The influence of the respective genetic modification on cell metabolic activity, proliferation, alkaline phosphatase (ALP) activity, mineralization in cell culture, and osteogenic marker gene expression was investigated. RESULTS: Transduction had no negative influence on cell metabolic activity or proliferation. ALP activity showed a typical rise-and-fall pattern with a maximal activity at day 14 and 21 after osteogenic induction. Enzyme activity was significantly higher in groups cultured with osteogenic media. The overexpression of BMP-2 and especially IHH + BMP-2 resulted in a significantly higher mineralization after 28 days. This was in line with obtained quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analyses, which showed a significant increase in osteopontin and osteocalcin expression for osteogenically induced BMP-2 and IHH + BMP-2 transduced cells when compared with the other groups. Moreover, an increase in runx2 expression was observed in all osteogenic groups toward day 21. It was again more pronounced for},
	language = {eng},
	number = {5},
	journal = {Stem cell research \& therapy},
	author = {Reichert, Johannes C. and Schmalzl, Jonas and Prager, Patrick and Gilbert, Fabian and Quent, Verena M. C. and Steinert, Andre F. and Rudert, Maximilian and Noth, Ulrich},
	year = {2013},
	pmid = {24004723},
	pmcid = {PMC3854715},
	keywords = {*Osteogenesis, Adenoviridae/genetics, Bone Marrow Cells/cytology, Bone Morphogenetic Protein 2/genetics/*metabolism, Cell Differentiation, Cells, Cultured, Core Binding Factor Alpha 1 Subunit/genetics/metabolism, Extracellular Matrix/metabolism, Genetic Vectors/genetics/metabolism, Hedgehog Proteins/genetics/*metabolism, Humans, Mesenchymal Stromal Cells/*cytology/metabolism, Osteocalcin/genetics/metabolism, Osteopontin/genetics/metabolism, Time Factors, Transfection},
}

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METHODS: In the present study, we induced the overexpression of the growth factors bone morphogenetic protein 2 (BMP-2) and/or Indian hedgehog (IHH) in human MSCs by adenoviral transduction to increase their osteogenic potential. GFP and nontransduced MSCs served as controls. The influence of the respective genetic modification on cell metabolic activity, proliferation, alkaline phosphatase (ALP) activity, mineralization in cell culture, and osteogenic marker gene expression was investigated. RESULTS: Transduction had no negative influence on cell metabolic activity or proliferation. ALP activity showed a typical rise-and-fall pattern with a maximal activity at day 14 and 21 after osteogenic induction. Enzyme activity was significantly higher in groups cultured with osteogenic media. The overexpression of BMP-2 and especially IHH + BMP-2 resulted in a significantly higher mineralization after 28 days. This was in line with obtained quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analyses, which showed a significant increase in osteopontin and osteocalcin expression for osteogenically induced BMP-2 and IHH + BMP-2 transduced cells when compared with the other groups. Moreover, an increase in runx2 expression was observed in all osteogenic groups toward day 21. 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