Influence of bone morphogenetic protein-2 on spiral ganglion neurite growth in vitro

Eur Arch Otorhinolaryngol. 2009 Sep;266(9):1381-9. doi: 10.1007/s00405-009-0930-y. Epub 2009 Mar 6.

Abstract

Recombinant human bone morphogenetic protein-2 (rhBMP-2) is a growth factor of the transforming growth factor-beta superfamily. Members of this protein family are involved in the development of various mammalian tissues, including the inner ear. As their notations indicate, they also have well-known effects on bone formation and regeneration. In this study, we examined the influence of rhBMP-2 on spiral ganglion (SG) neurite growth in vitro and showed the presence of its most preferred receptor BMPR-IB in spiral ganglion cells both in vitro and in vivo. SG explants of postnatal day 4 rats were analysed for neurite length and number after organotypical cell culture for 72 h, fixation and immunolabeling. Different concentrations of rhBMP-2 were used in a serum-free culture media. Neurite growth was compared with control groups that lacked stimulative effects; with neutrophin-3 (NT-3), which is a well-established positive stimulus on neurite length and number; and with combinations of these parameters. The results display that neurite number and total neurite length per explant in particular concentrations of rhBMP-2 increased by a maximum factor of two, while the mean neurite length was not affected. NT-3 demonstrated a much more potent effect, delivering a maximum increase of a factor of five. Furthermore, a combination of both growth factors shows a predominant effect on NT-3. Immunohistological detection of BMPR-IB was successful both in cell culture explants and in paraffin-embedded sections of animals of different ages. The results show that rhBMP-2 is, among other growth factors, a positive stimulus for SG neurite growth in vitro. Most growth factors are unstable and cannot be attached to surfaces without loss of their biological function. In contrast, rhBMP-2 can be attached to metal surfaces without loss of activity. Our findings suggest in vivo studies and a future clinical application of rhBMP-2 in cochlear implant technology to improve the tissue/electrode interface.

MeSH terms

  • Animals
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Protein Receptors, Type I / metabolism
  • Bone Morphogenetic Proteins / pharmacology*
  • Culture Media, Serum-Free
  • Neurites / drug effects
  • Neurites / physiology*
  • Neurogenesis / drug effects*
  • Neurotrophin 3 / pharmacology
  • Organ Culture Techniques
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Proteins / pharmacology*
  • Spiral Ganglion / drug effects
  • Spiral Ganglion / metabolism
  • Spiral Ganglion / physiology*
  • Transforming Growth Factor beta / pharmacology*

Substances

  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins
  • Culture Media, Serum-Free
  • Neurotrophin 3
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2
  • Bone Morphogenetic Protein Receptors, Type I