Non-healing chronic wounds are severe complications, which can often eventually lead to amputations. As such, there is a clear clinical need for dressings that promote the healing of chronic wounds. An advanced wound dressing aims to keep wound tissues moist while offering increased healing rates, preventing scar formation, reducing pain, minimizing infection, improving cosmetics, and lowering overall health care costs. We have previously developed tetrameric peptides Ac-IVZK-NH 2 (Ac-Ile-Val-Cha-Lys-NH 2 ) and Ac-IVFK-NH 2 (Ac-Ile-Val-Phe-Lys-NH 2 ) that self-assemble into nanofibrous hydrogels with biomimetic properties resembling those of collagen. In our study, we tested if these nanogels can fulfill the wound healing criteria mentioned above, and found that the nanogels are suitable scaffolds for encapsulating human dermal fibroblasts. We selected peptide nanogels Ac-IVZK-NH 2 and Ac-IVFK-NH 2 and produced silver nanoparticles in situ within the nanogels to assess their efficacy on micropigs with full-thickness excision wounds. The in situ generation of the silver nanoparticles was done solely through UV irradiation, no reducing agent was used. Application of the peptide nanogels on full thickness micropig wounds demonstrated that the scaffolds are biocompatible and did not trigger wound inflammation. This suggests that the scaffolds are safe for topical application. A comparison of the effect of both nanogels-even without the addition of the silver nanoparticles, revealed that the scaffold itself has a high potential to act as an antibacterial agent, which may suppress both the inflammatory reaction and the activity of proteases. Interestingly, the effect on wound closure of the peptide nanogels was comparable to those of standard care hydrogels. Despite our promising results, there is still much to learn about the molecular basis underlying the efficacy of tetrameric peptide nanogels in wound healing. This will support the urgent demand for advanced treatments of diabetic wounds, based on scientifically and clinically validated studies.

The ability of skeletal muscle to self-repair after a traumatic injury, tumor ablation, or muscular disease is slow and limited, and the capacity of skeletal muscle to self-regenerate declines steeply with age. Tissue engineering of functional skeletal muscle using 3D bioprinting technology is promising for creating tissue constructs that repair and promote regeneration of damaged tissue. Hydrogel scaffolds used as biomaterials for skeletal muscle tissue engineering can provide chemical, physical and mechanical cues to the cells in three dimensions thus promoting regeneration. Herein, we have developed two synthetically designed novel tetramer peptide biomaterials. These peptides are self-assembling into a nanofibrous 3D network, entrapping 99.9% water and mimicking the native collagen of an extracellular matrix. Different biocompatibility assays including MTT, 3D cell viability assay, cytotoxicity assay and live-dead assay confirm the biocompatibility of these peptide hydrogels for mouse myoblast cells (C2C12). Immunofluorescence analysis of cell-laden hydrogels revealed that the proliferation of C2C12 cells was well-aligned in the peptide hydrogels compared to the alginate-gelatin control. These results indicate that these peptide hydrogels are suitable for skeletal muscle tissue engineering. Finally, we tested the printability of the peptide bioinks using a commercially available 3D bioprinter. The ability to print these hydrogels will enable future development of 3D bioprinted scaffolds containing skeletal muscle myoblasts for tissue engineering applications.

Injured skeletal muscles which lose more than 20% of their volume, known as volumetric muscle loss, can no longer regenerate cells through self-healing. The traditional solution for recovery is through regenerative therapy. As the technology of three-dimensional (3D) bioprinting continues to advance, a new approach for tissue transplantation is using biocompatible materials arranged in 3D scaffolds for muscle repair. Ultrashort self-assembling peptide hydrogels compete as a potential biomaterial for muscle tissue formation due to their biocompatibility. In this study, two sequences of ultrashort peptides were analyzed with muscle myoblast cells (C2C12) for cell viability, cell proliferation, and differentiation in 3D cell culture. The peptides were then extruded through a custom-designed robotic 3D bioprinter to create cell-laden 3D structures. These constructs were also analyzed for cell viability through live/dead assay. Results showed that 3D bioprinted structures of peptide hydrogels could be used as tissue platforms for myotube formation – a process necessary for muscle repair.

Tissue engineering is a promising alternative to organ transplantation, where the number of waiting patients is not supported by the number of available donors. Tissue engineering aims to fabricate functional tissue using biocompatible scaffolds. Nanogels made from self-assembling ultrashort peptides are promising scaffold biomaterials. We focus on two compounds of a novel class of rationally designed tetrameric peptides for biomedical applications that have the advantage of being natural but synthetic hydrogels. These compounds have an innate tendency to self-assemble into nanofibrous hydrogels, which can be used for the fabrication of three-dimensional (3D) skin grafts, treating full-thickness wounds in minipigs and skeletal muscle tissue proliferation and differentiation. We were able to produce in situ silver nanoparticles within the peptide nanogels, solely through ultraviolet irradiation, with no reducing agent present. Applying the peptide nanogels on full-thickness minipig wounds demonstrated that the scaffolds were biocompatible, with no notable wound inflammation, and comparable to standard care solutions. Interestingly, the peptide scaffolds revealed a high potential to act as antibacterial agents. Microscopic observation demonstrated the ability of human umbilical vein endothelial cells to form tube-like structures within peptide nanogels. Moreover, we successfully produced artificial 3D vascularized skin substitutes using these peptide scaffolds. Additionally, we could demonstrate that both tetrameric peptides support 3D bioprinting, indicating their possible use as future bioinks. We believe that the results described represent an advancement in the context of engineering skin and skeletal muscle tissue, thereby providing the opportunity to rebuild missing, failing, or damaged parts.

Millions of people worldwide suffer from skin injuries, which create significant problems in their lives and are costly to cure. Tissue engineering is a promising approach that aims to fabricate functional organs using biocompatible scaffolds. We designed ultrashort tetrameric peptides with promising properties required for skin tissue engineering. Our work aimed to test the efficacy of these scaffolds for the fabrication of dermal grafts and 3D vascularized skin tissue models. We found that the direct contact of keratinocytes and fibroblasts enhanced the proliferation of the keratinocytes. Moreover, the expression levels of TGF-β1, b-FGF, IL-6, and IL-1α is correlated with the growth of the fibroblasts and keratinocytes in the co-culture. Furthermore, we successfully produced a 3D vascularized skin co-culture model using these peptide scaffolds. We believe that the described results represent an advancement in the fabrication of skin tissue equivalent, thereby providing the opportunity to rebuild missing, failing, or damaged parts. Graphical abstract [Formula: see text]