Abstract
The apparent rise of bone disorders demands advanced treatment protocols involving tissue engineering. Here, we describe self-assembling tetrapeptide scaffolds for the growth and osteogenic differentiation of human mesenchymal stem cells (hMSCs). The rationally designed peptides are synthetic amphiphilic self-assembling peptides composed of four amino acids that are nontoxic. These tetrapeptides can quickly solidify to nanofibrous hydrogels that resemble the extracellular matrix and provide a three-dimensional (3D) environment for cells with suitable mechanical properties. Furthermore, we can easily tune the stiffness of these peptide hydrogels by just increasing the peptide concentration, thus providing a wide range of peptide hydrogels with different stiffnesses for 3D cell culture applications. Since successful bone regeneration requires both osteogenesis and vascularization, our scaffold was found to be able to promote angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. The results presented suggest that ultrashort peptide hydrogels are promising candidates for applications in bone tissue engineering.
Highlights
In the context of bone tissue engineering, Bone-marrow-derived mesenchymal stem cells (BM-MSCs) have been reported to have the capability to differentiate into the osteogenic lineage in vitro if cultured with media supplemented with appropriate differentiation cocktail.[13]
There are questions about the safety of Matrigel in the possibility to use it for clinical purposes because its components are originated from Engelbreth−Holm−Swarm sarcomas[19] and because it is established that Matrigel and the basement membrane matrix promote tumor growth and tumorigenesis in vivo.[20,21]
The positively charged amine group from the lysine residue and the polarity of the surface have been reported to mediate cell adhesion and spreading.[52,53]. These peptides are composed of a positively charged amino acid (Lys) in the C-terminal and three nonpolar amino acids as a hydrophobic tail. Due to their amphiphilic structure, both peptides are able to self-assemble to form ordered aggregates.[52−54] The aggregation rate of the peptides can be enhanced by alternating the aromatic phenylalanine (Phe, F)
Summary
The mineral produced by the cells cultured in the IVFK hydrogel in the osteogenic medium produced the most intense ARS staining when compared to the other scaffolds tested. The results below (Figure 5b) show the time course of ALP activity in MSCs cultured on 2D and in different scaffolds: collagen, IVFK, and IVZK after 7 and 14 days.
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