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In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts.

Publication ,  Journal Article
Gersbach, CA; Guldberg, RE; García, AJ
Published in: Journal of cellular biochemistry
April 2007

Genetic engineering with osteogenic factors is a promising approach for cell-based therapeutics and orthopedic regeneration. However, the relative efficacy of different strategies for inducing osteoblastic differentiation remains unclear and is further complicated by varied delivery vehicles, cell types, and evaluation criteria. In order to elucidate the effects of distinct gene-based strategies, we quantitatively evaluated osteoblastic differentiation and mineralization of primary skeletal myoblasts overexpressing either the BMP-2 growth factor or Runx2 transcription factor. Retroviral delivery of BMP-2 or Runx2 stimulated differentiation into an osteoblastic phenotype, as demonstrated by the induction of osteogenic gene expression, alkaline phosphatase activity, and matrix mineralization in monolayer culture and on collagen scaffolds both in vitro and in an intramuscular site in vivo. In general, BMP-2 stimulated osteoblastic markers faster and to a greater extent than Runx2, although we also identified experimental conditions under which these two factors produced similar effects. Additionally, Runx2-engineered cells did not utilize paracrine signaling via secreted osteogenic factors, in contrast to cells overexpressing BMP-2, as demonstrated by conditioned media studies and activation of Smad signaling. These results emphasize the complexity of gene therapy-based orthopedic therapeutics as an integrated relationship of differentiation state, construct maturation, and paracrine signaling of osteogenic cells. This study is significant in evaluating proposed therapeutic systems and defining a successful strategy for integrating gene medicine and orthopedic regeneration.

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Published In

Journal of cellular biochemistry

DOI

EISSN

1097-4644

ISSN

0730-2312

Publication Date

April 2007

Volume

100

Issue

5

Start / End Page

1324 / 1336

Related Subject Headings

  • Transforming Growth Factor beta1
  • Transforming Growth Factor beta
  • Smad Proteins
  • Signal Transduction
  • Retroviridae
  • RNA, Messenger
  • Paracrine Communication
  • Osteogenesis
  • Osteoblasts
  • Myoblasts, Skeletal
 

Citation

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Gersbach, C. A., Guldberg, R. E., & García, A. J. (2007). In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts. Journal of Cellular Biochemistry, 100(5), 1324–1336. https://doi.org/10.1002/jcb.21118
Gersbach, Charles A., Robert E. Guldberg, and Andrés J. García. “In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts.Journal of Cellular Biochemistry 100, no. 5 (April 2007): 1324–36. https://doi.org/10.1002/jcb.21118.
Gersbach CA, Guldberg RE, García AJ. In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts. Journal of cellular biochemistry. 2007 Apr;100(5):1324–36.
Gersbach, Charles A., et al. “In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts.Journal of Cellular Biochemistry, vol. 100, no. 5, Apr. 2007, pp. 1324–36. Epmc, doi:10.1002/jcb.21118.
Gersbach CA, Guldberg RE, García AJ. In vitro and in vivo osteoblastic differentiation of BMP-2- and Runx2-engineered skeletal myoblasts. Journal of cellular biochemistry. 2007 Apr;100(5):1324–1336.
Journal cover image

Published In

Journal of cellular biochemistry

DOI

EISSN

1097-4644

ISSN

0730-2312

Publication Date

April 2007

Volume

100

Issue

5

Start / End Page

1324 / 1336

Related Subject Headings

  • Transforming Growth Factor beta1
  • Transforming Growth Factor beta
  • Smad Proteins
  • Signal Transduction
  • Retroviridae
  • RNA, Messenger
  • Paracrine Communication
  • Osteogenesis
  • Osteoblasts
  • Myoblasts, Skeletal