Creating a stem cell niche in the inner ear using self-assembling peptide amphiphiles

Abstract

The use of human embryonic stem cells (hESCs) for regeneration of the spiral ganglion will require techniques for promoting otic neuronal progenitor (ONP) differentiation, anchoring of cells to anatomically appropriate and specific niches, and long-term cell survival after transplantation. In this study, we used self-assembling peptide amphiphile (PA) molecules that display an IKVAV epitope (IKVAV-PA) to create a niche for hESC-derived ONPs that supported neuronal differentiation and survival both in vitro and in vivo after transplantation into rodent inner ears. A feature of the IKVAV-PA gel is its ability to form organized nanofibers that promote directed neurite growth. Culture of hESC-derived ONPs in IKVAV-PA gels did not alter cell proliferation or viability. However, the presence of IKVAV-PA gels increased the number of cells expressing the neuronal marker beta-III tubulin and improved neurite extension. The self-assembly properties of the IKVAV-PA gel allowed it to be injected as a liquid into the inner ear to create a biophysical niche for transplanted cells after gelation in vivo. Injection of ONPs combined with IKVAV-PA into the modiolus of X-SCID rats increased survival and localization of the cells around the injection site compared to controls. Human cadaveric temporal bone studies demonstrated the technical feasibility of a transmastoid surgical approach for clinical intracochlear injection of the IKVAV-PA/ONP combination. Combining stem cell transplantation with injection of self-assembling PA gels to create a supportive niche may improve clinical approaches to spiral ganglion regeneration.

Ile‐Lys‐Val‐ala‐Val (IKVAV) peptide for neuronal tissue engineering

“…Self‐assembled IKVAV‐PA gels facilitated ependymal stem cell–derived neuronal progenitor cell to neuronal differentiation and cell survival both in vitro and in vivo. Increased β3 tubulin expression and improved neurite extension of the cells were observed in vitro, whereas the cell survival rate increased in vivo with the combination of IKVAV‐PA hydrogel . The adhesion of sensory neurons was also improved by the IKVAV‐PA nanofiber gels .…”

“…Increased β3 tubulin expression and improved neurite extension of the cells were observed in vitro, whereas the cell survival rate increased in vivo with the combination of IKVAV-PA hydrogel. 37 The adhesion of sensory neurons was also improved by the IKVAV-PA nanofiber gels. 38 A hybrid matrix of neuro-active IKVAV-PA and collagen scaffold was designed for combinatorial control of the matrix bioactivity by adjusting laminin epitope concentration and collagen.…”

Extracellular Matrix and Cellular Plasticity in Musculoskeletal Development

“…For example, scaffolds such as the self-assembling peptide amphiphiles ( Sato et al, 2018 ) developed in recent years is an example of how these scaffolds can be specifically designed to suit regeneration of different tissues such as muscle ( Sleep et al, 2017 ) and nerves ( Cheng et al, 2013 ; Matsuoka et al, 2017 ). In these studies, peptides mimicking various ECM proteins, such as collagen ( Gore et al, 2001 ; Luo and Tong, 2011 ), laminin ( Cheng et al, 2013 ), heparin ( Mammadov et al, 2011 ), and TNC ( Sever et al, 2014 ), were incorporated into peptide amphiphiles.…”

Self‐Assembling Peptide Gels for 3D Prostate Cancer Spheroid Culture

“…Several β-sheet fibrillizing peptides have been available commercially, notably Puramatrix from Corning, HydroMatrix from Sigma-Aldrich, and PGD-HydroGels from PeptiGelDesign. Considering additional designs beyond linear fibrillizing peptides such as β-hairpins 57,58 ; aromatic peptide amphiphiles 59–63 ; peptide amphiphiles 64–66 , and others 67,68 , supramolecular peptide matrices for 3D cell culture represent a diverse family.…”