Early Osteogenic Differentiation of Mouse Preosteoblasts Induced by Collagen-Derived DGEA-Peptide on Nanofibrous Phage Tissue Matrices

Abstract

Specific biochemical and physical cues in tissue extracellular matrices play a critical role in regulating cellular growth processes and their fate. We report initial responses of bone stem cells induced by collagen-derived DGEA-peptides on nanofibrous M13 phage tissue matrices. We constructed genetically engineered M13 phage with DGEA-peptide displayed in high density on the major coat proteins and biomimetic nanofibrous tissue-like matrices in two and three dimensions. We investigated the effects of biochemical cues, specifically DGEA-peptides on preosteoblast (MC3T3) morphologies. The preosteoblasts grown on the top of the DGEA-incorporated phage matrices exhibited significant outgrown morphology with early bone cell marker protein expression. Through soluble peptide competition assays and control experiments, we verified that the observed cellular morphologies and osteogenic protein marker expression were specifically caused by the DGEA-peptides. We confirmed that the outgrown morphologies are linked with the early phase of osteogenic protein expression through mRNA quantification and bone cell protein marker expression. Additionally, we demonstrated that the phage-based tissue matrix systems could work as a good cell culture platform to investigate the specific effect of biochemical cues, which can be tuned precisely at a single amino acid level with little change in other physical and chemical properties of the environment. Our study advances the understanding of osteogenic differentiation and our phage-based tissue matrices have the potential for future bone regeneration therapy and systemic investigation of specific cellular responses to biochemical ligand stimulation.