Release Systems Based on Self-Assembling RADA16-I Hydrogels with a Signal Sequence Which Improves Wound Healing Processes

May 9, 2023

Title

Release Systems Based on Self-Assembling RADA16-I Hydrogels with a Signal Sequence Which Improves Wound Healing Processes

Author

Dzierżyńska, Maria, Justyna Sawicka, Milena Deptuła, Paweł Sosnowski, Piotr Sass, Barbara Peplińska, Zuzanna Pietralik-Molińska, Martyna Fularczyk, Franciszek Kasprzykowski, Jacek Zieliński, Maciej Kozak, Paweł Sachadyn, Michał Pikuła, and Sylwia Rodziewicz-Motowidło

Year

2023

Journal

Scientific Reports

Abstract

Self-assembling peptides can be used for the regeneration of severely damaged skin. They can act as scaffolds for skin cells and as a reservoir of active compounds, to accelerate scarless wound healing. To overcome repeated administration of peptides which accelerate healing, we report development of three new peptide biomaterials based on the RADA16-I hydrogel functionalized with a sequence (AAPV) cleaved by human neutrophil elastase and short biologically active peptide motifs, namely GHK, KGHK and RDKVYR. The peptide hybrids were investigated for their structural aspects using circular dichroism, thioflavin T assay, transmission electron microscopy, and atomic force microscopy, as well as their rheological properties and stability in different fluids such as water or plasma, and their susceptibility to digestion by enzymes present in the wound environment. In addition, the morphology of the RADA-peptide hydrogels was examined with a unique technique called scanning electron cryomicroscopy. These experiments enabled us to verify if the designed peptides increased the bioactivity of the gel without disturbing its gelling processes. We demonstrate that the physicochemical properties of the designed hybrids were similar to those of the original RADA16-I. The materials behaved as expected, leaving the active motif free when treated with elastase. XTT and LDH tests on fibroblasts and keratinocytes were performed to assess the cytotoxicity of the RADA16-I hybrids, while the viability of cells treated with RADA16-I hybrids was evaluated in a model of human dermal fibroblasts. The hybrid peptides revealed no cytotoxicity  the cells grew and proliferated better than after treatment with RADA16-I alone. Improved wound healing following topical delivery of RADA-GHK and RADA-KGHK was demonstrated using a model of dorsal skin injury in mice and histological analyses. The presented results indicate further research is warranted into the engineered peptides as scaffolds for wound healing and tissue engineering.

Instrument

J-815

Keywords

the regeneration of severely damaged skin