A Novel Self-assembling Peptides Nanofiberscaffolds For 3-dimensional Real-time Monitor Cell Cultures And Anti-aging Study

Objective: Using a new synthesis chiral self-assembling peptide called GFS-15 to simulate extracellular matrix growth environment, and studying the feasibility of the peptide as a biological material. The special sequence structure and physical and chemical properties of the self-assembled peptide are applied to be connected to the gold electrode sensor, exploring the effect of this kind of nano material be used as real time monitoring for 3D cell culture, and analyze the process and mechanism of resveratrol anti-aging under the condition of 3D micro environment.Method: The two stage structure, nano morphology and macroscopic morphology of the chiral self-assembled short peptide GFS-15 were mainly characterized by circular chromatograph, atomic force microscope and Congo red staining. Cyclic voltammetry curve, circular dichroism to detect self-assembly process influenced by the factors of time, concentration, and salt ions of; three-dimensional cell culture, cell cycle distribution, apoptosis staining, qRT-PCR to study the short peptide of anti-aging cell biological function.Results: The short peptide GFS-15 is a typical beta-sheet structure, and the atomic force microscope(AFM) shows that it can self-assemble in ionic condition to form nano-fiber structure; Congo red staining showed that the peptide hydrogel with transparent by eyes, can be used as three-dimensional culture cells; Electrochemical study results show that the short peptide can be combined on the surface of the gold electrode, stop of the electron transfer and the peak-to-peak separation(Ep) decreased; the 3D real-time trial shows that the peptide can combine with the gold electrode and form nano-fiber to attach cell and support cell growth. After2 hours, the peptide can combine with the gold electrode and the modified gold-surface support cell growth; Time to 24 hours, the nano-fiber still exist, and also can support cell growth. The study of resveratrol anti-aging cultured in 3D microenvironment results show that the cell cycle of 3D anti-aging group arrest in G2 was 11.89%. Compared to 2D anti-aging group, it’s decreased about 7.84%.Conclusions: Novel chiral self-assembling GFS-15 has a stable secondary structure, forms an ordered nano-fibrous scaffolds, can be used as an ideal material for 3D cell culture; Binding Au electrode(Au)to modified the surface and form stable nanofiber-scaffold, as 3D cell culture materials of real-time monitoring are obtained; Compared to cell culture in 2D Petri dish and 3D microenvironment model, resveratrol anti-aging effect is tested, 3D cell culture process is obviously better to 2D process.