Characterisation of the stability and bio-functionality of tethered proteins on bioengineered scaffolds: Implications for stem cell biology and tissue repair

Ting Yi Wang, Kiara A.F. Bruggeman, Rebecca K. Sheean, David R. Nisbet, Clare L. Parish

Research output: Chapter in Book/Report/Conference proceedingConference PaperOtherpeer-review

1 Citation (Scopus)

Abstract

In the context of biological applications, the timely delivery of molecules can be critical for cellular and organ function. As such, previous studies have demonstrated the superior long-term protein delivery, by way of protein tethering onto bioengineered scaffolds, compared to conventional delivery of soluble protein in vitro and in vivo. Despite such benefits little knowledge exists regarding the stability, release kinetics, longevity, activation of intracellular pathway and functionality of these proteins over time. By way of example, here we examined the stability, degradation and functionality of a protein, glial derived neurotrophic factor (GDNF), which is known to influence neuronal survival, differentiation and neurite morphogenesis. Enzyme-linked immuno-sorbent assays revealed that GDNF, covalently tethered onto polycaprolactone electrospun nanofibrous scaffolds, remained present on the scaffold surface for 120 days, with no evidence of protein leaching or degradation. The tethered GDNF protein remained functional and capable of activating downstream signalling cascades, as revealed by its capacity to phosphorylate intracellular Erk in a neural cell line. Furthermore, immobilisation of GDNF protein promoted cell survival and differentiation in culture at both 3 and 7 days. This study provides important evidence of the stability and functionality kinetics of tethered molecules.

Original languageEnglish
Title of host publication1st Global Conference on Biomedical Engineering and 9th Asian-Pacific Conference on Medical and Biological Engineering
PublisherSpringer-Verlag London Ltd.
Pages46-49
Number of pages4
Volume47
ISBN (Electronic)9783319122618
DOIs
Publication statusPublished - 2015
Externally publishedYes
EventGlobal Conference on Biomedical Engineering (GCBME) / Asian-Pacific Conference on Medical and Biological Engineering (APCMBE) 2014 - Tainan, Taiwan
Duration: 9 Oct 201412 Oct 2014

Conference

ConferenceGlobal Conference on Biomedical Engineering (GCBME) / Asian-Pacific Conference on Medical and Biological Engineering (APCMBE) 2014
Abbreviated titleGCBME 2014
CountryTaiwan
CityTainan
Period9/10/1412/10/14
Other1st Global Conference on Biomedical Engineering, GCBME 2014 and 9th Asian-Pacific Conference on Medical and Biological Engineering, APCMBE 2014

Keywords

  • Neurons
  • Protein tethering
  • Scaffolds
  • Stability

Cite this

Wang, T. Y., Bruggeman, K. A. F., Sheean, R. K., Nisbet, D. R., & Parish, C. L. (2015). Characterisation of the stability and bio-functionality of tethered proteins on bioengineered scaffolds: Implications for stem cell biology and tissue repair. In 1st Global Conference on Biomedical Engineering and 9th Asian-Pacific Conference on Medical and Biological Engineering (Vol. 47, pp. 46-49). Springer-Verlag London Ltd.. https://doi.org/10.1007/978-3-319-12262-5_14