Spray-assisted layer-by-layer assembly on hyaluronic acid scaffolds for skin tissue engineering

Abstract

Tissue engineering approaches for the development of a single epidermal-dermal scaffold to treat full-thickness skin defects have been limited by difficulties in the fabrication of a bilayer scaffold combining the specific properties of the epidermis and the dermis. Here we present an innovative approach to developing a scaffold that holds promise for skin tissue engineering. We utilize the spray-assisted layer-by-layer assembly technique to deposit a polyelectrolyte multilayer film composed of hyaluronic acid and poly-L-lysine (the epidermal component) on a porous hyaluronic acid scaffold (the dermal component), in a rapid and controlled manner. The multilayer film promotes cell adhesion, contributing to regeneration of the epidermal barrier functions of skin. While human keratinocytes attached and proliferated on the coated porous scaffolds, they did not invade the porous dermal component, thus leaving room for seeding of relevant fibroblast cell types in this scaffold. This scaffold therefore holds promise for co-culture of different cells, which may be useful for treatment of full-thickness skin defects as well as other tissue engineering applications.

Influence of poly‐l‐lysine molecular weight on antibacterial efficacy in polymer multilayer films

“…Hepatoyctes, fibroblasts, and embryonic stem cells have been successfully co‐cultured on PLL and HA patterned surfaces . Keratinocytes have been cultured successfully on (PLL/HA) 150.5 films . HA containing hydrogels and coated surfaces are nonadhesive due to their high hydrophilicity, but incorporating PLL as the last layer has been shown to aid cells in adhering .…”

“…Keratinocytes have been cultured successfully on (PLL/HA) 150.5 films . HA containing hydrogels and coated surfaces are nonadhesive due to their high hydrophilicity, but incorporating PLL as the last layer has been shown to aid cells in adhering . Despite their uses in cell culture, it is well known that polycations, including PLL, can be toxic to mammalian cells at high MW and concentrations .…”

“…93 HA containing hydrogels and coated surfaces are nonadhesive due to their high hydrophilicity, but incorporating PLL as the last layer has been shown to aid cells in adhering. 61,93 Despite their uses in cell culture, it is well known that polycations, including PLL, can be toxic to mammalian cells at high MW 114 and concentrations. [115][116][117] Our results indicated that while NIH 3T3 fibroblasts incubated in media exposed to PLL 30 and PLL 90 coatings remained similarly viable to untreated controls, cells exposed to media incubated in PLL 400 coatings over 24 h lost viability (Fig.…”

Mussel-Inspired Multifunctional Hydrogel Coating for Prevention of Infections and Enhanced Osteogenesis

“…In particular, photo-cross-linkable hydrogels provide the option to first spray the prepolymer solution onto the implants, followed by in situ photo-cross-linking to cure the coating layer. 12,20,21 However, to achieve long-term stability, hydrogels should possess strong adhesion to Ti surfaces, especially under wet environments. Inspired by the unusual ability of mussels to adhere to essentially any surfaces under water, extensive studies in the past few decades suggest that the presence of catechol motifs contributes to improved adhesion.…”

Biomaterials for Skin Substitutes

“…Keratinocytes were later seeded on this scaffold to make a bilayer skin graft . HA has been used to make mono‐ and bilayer scaffolds. Its negatively charged nature makes it an interesting option to be combined with positively charged polymers like poly‐ l ‐lysine (PLL) and chitosan .…”