Self-Assembling Peptide Scaffold Carrying Neural-Cell Adhesion Molecule-Derived Mimetic-Peptide Transplantation Promotes Proliferation and Stimulates Neurite Extension by Modulating Tau Phosphorylation and Calpain/Glycogen Synthase Kinase 3 beta (GSK-3β) in Neurons

Ann Transplant. 2020 Jul 7:25:e924093. doi: 10.12659/AOT.924093.

Abstract

BACKGROUND Self-assembling peptide scaffolds have been extensively applied in tissue engineering. Many investigations have modified self-assembling peptide scaffolds by integrating functional motifs, with promising applications. This study aimed to generate a novel RADA16 self-assembling peptide scaffold integrating a neural-cell adhesion molecule-derived mimetic-peptide (SIDRVEPYSSTAQ) and evaluated the effects on neuron proliferation. MATERIAL AND METHODS A 37-amino-acids peptide of RADA16-activation motif containing neural-cell adhesion molecule-derived mimetic-peptide (SIDRVEPYSSTAQ) was synthesized and self-assembled into a scaffold. Dorsal root ganglion (DRG) and spinal cord motor neurons (SCMN) were primarily isolated and identified. Neurons (DRG and SCMN) were divided into FRM, FRM-MP, and FRM-MP-LiCl groups. The adherence ability of neurons was evaluated using toluidine blue staining. Proliferation and apoptosis of neurons were assessed using CCK-8 and flow cytometry assay, respectively. Immunofluorescence assay was used to measure neurite extension. Western blot assay was used to assess GSK-3ß/p-GSK-3ß, Tau/p-Tau, and calpain expression in neurons. RESULTS FRM-MP-LiCl released multiple-peptide with higher efficiency. FRM-MP-LiCl significantly enhanced proliferation and inhibited apoptosis compared to FRM and FRM-MP groups (p.

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Cell Proliferation / drug effects*
  • Ganglia, Spinal / cytology
  • Ganglia, Spinal / drug effects
  • Ganglia, Spinal / metabolism
  • Glycogen Synthase Kinase 3 beta / metabolism*
  • Neural Cell Adhesion Molecules / pharmacology*
  • Neurites / drug effects
  • Neurites / metabolism
  • Neuronal Outgrowth / drug effects*
  • Neurons / cytology
  • Neurons / drug effects*
  • Neurons / metabolism
  • Phosphorylation / drug effects
  • Rats
  • Rats, Wistar
  • tau Proteins / metabolism*

Substances

  • Neural Cell Adhesion Molecules
  • tau Proteins
  • Glycogen Synthase Kinase 3 beta