Osterix-mCherry Expression Allows for Early Bone Detection in a Calvarial Defect Model

Adv Biosyst. 2019 Dec;3(12):e1900184. doi: 10.1002/adbi.201900184. Epub 2019 Nov 4.

Abstract

The process of new bone formation following trauma requires the temporal recruitment of cells to the site, including mesenchymal stem cells, preosteoblasts, and osteoblasts, the latter of which deposit minerals. Hence, bone repair, a process that is assessed by the extent of mineralization within the defect, can take months before it is possible to determine if a treatment is successful. Here, a fluorescently tagged Osterix, an early key gene in the bone formation cascade, is used as a predictive measure of bone formation. Using a calvarial defect model in mice, the ability to noninvasively track the Osterix transcription factor in an Osterix-mCherry mouse model is evaluated as a measure for bone formation following treatment with recombinant human Bone-Morphogenetic-Protein 2 (rhBMP-2). Two distinct delivery materials are utilized, an injectable nanocomposite hydrogel and a collagen sponge, that afford distinct release kinetics and it is found that cherry-fluorescent protein can be detected as early as 2 weeks following treatment. Osterix intensity correlates with subsequent bone formation and hence can serve as a rapid screening tool for osteogenic drugs or for the evaluation and optimization of delivery platforms.

Keywords: BMP-2; Osterix; bone regeneration; imaging; nanocomposite; scaffold.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 2 / pharmacology
  • Cells, Cultured
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Osteoblasts / cytology
  • Osteoblasts / metabolism
  • Osteogenesis / drug effects
  • Osteogenesis / physiology*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / pharmacology
  • Red Fluorescent Protein
  • Regeneration / drug effects
  • Skull / metabolism*
  • Sp7 Transcription Factor / genetics
  • Sp7 Transcription Factor / metabolism*
  • Tissue Scaffolds / chemistry
  • Transforming Growth Factor beta / pharmacology

Substances

  • Bone Morphogenetic Protein 2
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Sp7 Transcription Factor
  • Sp7 protein, mouse
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2