Synthesis and Evaluation of Peptide-based Hybrid Scaffold for Tissue Engineering

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In tissue engineering and wound-healing application, scaffold materials are utilized to provide a mechanical support for cell growth and tissue formation. These scaffold can be modified such that they also provide specific biologic signals to cell in order to control or facilitate tissue formation or regeneration. Cell adhesion peptides Arg-Gly-Asp (RGD) have been incorporated into scaffolds to enhance cell adhesion or to allow biospecific cell adhesion.In this work, for improving the cell-attachment activity of the Arg-Gly-Asp-Ser peptides to tissue engineering scaffold, we tried to stabilize the folded conformations of such peptides by designing the ss-strands peptides, which have the Arg-Gly-Asp-Ser sequence at the C-terminal. These sequence, (Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys)_2 (EAK16) or (Arg-Ala-Arg-Ala-Asp-Ala-Asp-Ala)_2 (RAD16) of the peptides is selected as a typical sequence for having high propensity to form ss-strands.RGDS containing ss-strands model peptides were synthesize … More d using liquid and solid phase procedures. Fibroblast cells attaching to the peptide-immobilized cell culture dishes, indicating that difference in the cell-attachment activity were found for these peptides-immobilized cell culture dishes. EAK16RGDS and RAD16RGDS immobilized polystyrene-dish has the highest spreading activity among the peptide-immobilized ones.The conformational constraint caused by the formation of ss-strands structure is too strong to stabilize the desired conformation at the Arg-Gly-Asp-Ser portion for the peptides in which the Arg-Gly-Asp-Ser sequence is directly linked to the sequences designed for forming ss-strands structure. It is also suggested that designing a spacer sequence is desirable for improving the activity between the Arg-Gly-Asp-Ser sequence and the sequence forming ss-strand structure. Solutions of EAK16RGDS and RAD16RGDS become gelatinous and then form fibers. EAK16RGDS and RAD16RGDS peptides interact with each other and become more structured upon fiber formation. Such peptides may find utility as in vivo tissue engineering 3D scaffolds. Less