Calcified Tissue International

, Volume 74, Issue 4, pp 388–397 | Cite as

Quantification, Localization, and Expression of IGF-I and TGF-β1 During Growth Factor-Stimulated Fracture Healing

  • B. Wildemann
  • G. Schmidmaier
  • N. Brenner
  • M. Hüning
  • R. Stange
  • N. P. Haas
  • M. Raschke
Article

Abstract

Because of the increasing interest on stimulating fracture healing, knowledge about the role and chronology of growth factors during the healing process is important. The purpose of this study was to quantify the protein concentration of IGF-I and TGF-β1 during rat tibial fracture healing 5, 10, and 15 days after fracture using ELISA methods and to analyze the distribution of the proteins and the related mRNA expression in the fracture callus by immunohistochemistry and in situ hybridization. The following three groups were analyzed: Fractured tibiae intramedullary stabilized with K-wires coated with IGF-I and TGF-β1 compared with fractures stabilized with uncoated K-wires and unfractured tibiae. The weight of the callus increased during the healing process in both experimental groups. The protein concentration of IGF-I and TGF-β1 in the fracture callus showed significant changes between the investigated time points and treatment groups compared with the unfractured tibia. IGF-I increased with healing time whereas TGF-β1 revealed a constantly elevated level at the investigated time points. Mesenchymal cells, osteoblasts, osteocytes, proliferating and immature chondrocytes, and osteoclasts expressed both growth factors. No differences in the expression and localization pattern of the growth factors were detectable among the groups. Using the different methods for quantification and visualization of the growth factors, no differences (except the increased IGF-I concentration at day 15 in the growth factor group) were seen between the normal and the growth factor-stimulated fracture healing as an indication for physiological healing after exogenous growth factor treatment.

Keywords

Growth healing ELISA In situ Hybridization Immunohistochemistry Fracture healing 

Notes

Acknowledgements

This work was supported by a grant to Dr. G. Schmidmaier by the Deutsche Forschungsgemeinschaft (DFG Schm 1436 1-2) and to Dr. Britt Wildemann by the Berliner Sparkassen Stiftung. We thank Ms. C. Bergmann for her excellent technical help with the in situ hybridization.

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Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • B. Wildemann
    • 1
  • G. Schmidmaier
    • 1
  • N. Brenner
    • 1
  • M. Hüning
    • 1
  • R. Stange
    • 1
  • N. P. Haas
    • 1
  • M. Raschke
    • 2
  1. 1.Department of Trauma and Reconstructive SurgeryCharité, Humboldt-University of Berlin, BerlinGermany
  2. 2.Department of Trauma and Reconstructive SurgeryUniversity Hospital MuensterMuenster, Germany

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