Recombinant Spider Silk Functionalized Silkworm Silk Matrices as Potential Bioactive Wound Dressings and Skin Grafts

Abstract

Silk is considered to be a potential biomaterial for a wide number of biomedical applications. Silk fibroin (SF) can be retrieved in sufficient quantities from the cocoons produced by silkworms. While it is easy to formulate into scaffolds with favorable mechanical properties, the natural SF does not contain bioactive functions. Spider silk proteins, on the contrary, can be produced in fusion with bioactive protein domains, but the recombinant procedures are expensive, and large-scale production is challenging. We combine the two types of silk to fabricate affordable, functional tissue-engineered constructs for wound-healing applications. Nanofibrous mats and microporous scaffolds made of natural silkworm SF are used as a bulk material that are top-coated with the recombinant spider silk protein (4RepCT) in fusion with a cell-binding motif, antimicrobial peptides, and a growth factor. For this, the inherent silk properties are utilized to form interactions between the two silk types by self-assembly. The intended function, that is, improved cell adhesion, antimicrobial activity, and growth factor stimulation, could be demonstrated for the obtained functionalized silk mats. As a skin prototype, SF scaffolds coated with functionalized silk are cocultured with multiple cell types to demonstrate formation of a bilayered tissue construct with a keratinized epidermal layer under in vitro conditions. The encouraging results support this strategy of fabrication of an affordable bioactive SF-spider silk-based biomaterial for wound dressings and skin substitutes.

In Situ Forming Injectable Silk Fibroin Hydrogel Promotes Skin Regeneration in Full Thickness Burn Wounds

“…Management of burn wounds thus poses challenging clinical and economical problems. This has encouraged the exploration and development of affordable novel matrices with ideal physical and biological properties . Developing a matrix with capacity to innately guide cell migration and proliferation while also holding suitable mechanical properties, remains a big challenge in tissue engineering.…”

Hydrogel Preparation Methods and Biomaterials for Wound Dressing

“…The functionalized recombinant spider silk proteins expressed in bacteria hold great potential for the development of advanced biomaterials in the field of tissue engineering [ 115 , 127 ]. The wound dressing made by Chouhan et al using recombinant spider silk protein (4RepCT) not only stimulated cell proliferation but also provided antibacterial effects, thereby improving the wound healing process [ 128 ].…”

Spider Silk for Tissue Engineering Applications

“…The spider silk was functionalized with fused binding motifs from fibronectin containing the RGD sequence (Blue), a growth factor (basic fibroblast growth factor, FGF2) (red), or cationic peptides with antimicrobial properties (AMP) (gray), to enhance the cell-binding activity and cellular growth. Adopted and modified with permission [84]. Copyright 2018, American Chemical Society.…”