The mechanical stimulation of cells in 3D culture within a self-assembling peptide hydrogel

Abstract

The aim of this present study was to provide a scaffold as a tool for the investigation of the effect of mechanical stimulation on three-dimensionally cultured cells. For this purpose, we developed an artificial self-assembling peptide (SPG-178) hydrogel scaffold. The structural properties of the SPG-178 peptide were confirmed by attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) and transmission electron microscopy (TEM). The mechanical properties of the SPG-178 hydrogel were studied using rheology measurements. The SPG-178 peptide was able to form a stable, transparent hydrogel in a neutral pH environment. In the SPG-178 hydrogel, mouse skeletal muscle cells proliferated successfully (increased by 12.4 ± 1.5 times during 8 days of incubation; mean ± SEM). When the scaffold was statically stretched, a rapid phosphorylation of ERK was observed (increased by 2.8 ± 0.2 times; mean ± SEM). These results demonstrated that the developed self-assembling peptide gel is non-cytotoxic and is a suitable tool for the investigation of the effect of mechanical stimulation on three-dimensional cell culture.

Hierarchical Structure of the Fibrillar Hydrogel Network of a Self‐Assembled Synthetic Peptide Revealed by X‐ray Scattering and Atmospheric Scanning Electron Microscopy

“…A sharp diffraction peak corresponding to spacing of 0.480 nm (q ¼ 13.08 nm À1 ) was observed in the WAXS profile of the SPG-178 hydrogel ( Figure 6) at room temperature. It is assigned to the b-sheet structure formed in the hydrogel, and was consistent with our previous findings [13] in which ATR FT-IR spectroscopy measurements of the SPG-178 peptide hydrogel (in deuterium water) revealed the formation of an antiparallel b-sheet structure in the gel.…”

“…A molecular model of the antiparallel b-sheet structure and fibril formation of SPG-178 were previously obtained using Facio, a 3D-graphics program, and by employing Tinker with a charm 22 force field parameter. [24,25] The dimension of the peptide monomer was calculated using Swiss-PdbViewer [13,26] ( Figure 1). …”

“…[13] The peptide was expected to form a fibrous structure by self-assembly in water, as illustrated in Figure 1, and the formation of a hydrogel composed of peptide fibrils may have been caused by network formation. Figure 4 shows TEM micrographs of the self-assembled peptides.…”

“…The use of synthetic peptide hydrogels may minimize the risk of biological contamination. Previous studies examined synthetic self-assembling peptide-based gels [5,[6][7][8][9] and their applications, such as 3D cell culture scaffolds, [10][11][12][13] hemostats, [14,15] lubricants for osteoarthritis, [16] regenerative medicine, [17] wound healing, [18] and drug delivery systems. [19,20] The efficacy of these self-assembled materials may be affected by various factors, including pH, temperature, and the concentrations of salt and peptides.…”

Design of Tetra-arm PEG-crosslinked Thermoresponsive Hydrogel for 3D Cell Culture

“…PNIPAAm hydrogel expresses reversible swelling and shrinking behaviors in aqueous solution when the temperature is changed to be above or below the LCST of PNIPAAm, which is called volume phase transition temperature (VPTT). 13 Because activity and function of cells change when they are physically stretched and deformed, 14,15 PNIPAAm gel is promising as a novel material for a 3D cell culture method that can actively control cell fate. We developed the molecular design of a gel that achieves cell biocompatibility and maintains transparency even at high temperature.…”