Gene-activated tissue grafts for sustained bone morphogenetic protein-2 delivery and bone engineering: Is muscle with fascia superior to muscle and fat?

J Tissue Eng Regen Med. 2018 Apr;12(4):1002-1011. doi: 10.1002/term.2575. Epub 2017 Nov 22.

Abstract

Previously, we have presented an expedited strategy for sustained delivery of bone morphogenetic protein-2 (BMP-2) to bone lesions based on the implantation of gene-activated fat and muscle fragments. The aim of the present in vitro experiments was to evaluate the potential of muscle with fascia as a BMP-2 delivering osteo-regenerative implant in comparison to fat tissue and muscle alone. Subcutaneous fat, muscle, and muscle with fascia were harvested from Fischer 344 rats. The tissues were cut into small pieces and cultured for up to 90 days after direct transduction with adenoviral BMP-2 or green fluorescence protein vectors. Different vector doses were applied, and proliferation, long-term BMP-2 production, and osteogenic differentiation of the 3 different tissues were investigated in vitro. Muscle with fascia produced the largest amounts of BMP-2. Expression of the transgene was detected for up to 90 days. Proliferation was reduced with increased vector doses. Muscle with fascia showed a higher potential for osteogenic differentiation than fat, but it was not improved as compared to muscle alone. A dose of 4 × 108 plaque forming units of the adenoviral BMP-2 vector appeared to be the optimal dose for transduction of muscle with fascia. Because muscle with fascia produced higher amounts of BMP-2 as compared to muscle alone or fat tissue grafts, showing a high potential for osteogenic differentiation, it might represent an improved osteo-regenerative implant facilitating endogenous repair. Future studies should investigate the effect of muscle with fascia transduced with 4 × 108 plaque forming units on bone healing in vivo.

Keywords: bone regeneration; fascia; fat tissue; gene therapy; muscle tissue; osteogenic differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 2 / biosynthesis*
  • Bone Morphogenetic Protein 2 / genetics
  • Bone and Bones / cytology
  • Cell Differentiation
  • Cell Proliferation
  • Genetic Vectors*
  • Muscle, Skeletal / cytology
  • Muscle, Skeletal / metabolism*
  • Osteogenesis
  • Rats
  • Rats, Inbred F344
  • Subcutaneous Fat / cytology
  • Subcutaneous Fat / metabolism*
  • Tissue Engineering*
  • Transduction, Genetic*

Substances

  • Bmp2 protein, rat
  • Bone Morphogenetic Protein 2