3D Living Dressing Improves Healing and Modulates Immune Response in a Thermal Injury Model

Abstract

Thermal injury trauma can induce a state of immunosuppression, causing wounds to become chronic in nature. Stem cell-based therapies represent a promising new approach to treat such wounds due to their capacity to self-renew and their multi-lineage potential. Mesenchymal stem cells (MSCs) are known to secrete endogenous factors that stimulate wound healing by promoting angiogenesis, extracellular matrix remodeling, skin regeneration, and by dampening down inflammation. MSC delivery in a biomaterial construct can augment their wound-healing capacity by concentrating cells at the burn site and upregulating trophic factor secretion. The work presented is the first to evaluate repair in an in vitro raft thermal injury model using a regenerative, dual cell delivery three-dimensional (3D) core/shell (c/s) "living dressing"construct. This previously characterized 3D c/s bioprinted construct, which delivers both MSCs and endothelial cells, was used to treat an in vitro 3D raft skin thermal injury wound model. The mesenchymal stromal cell line (T0523) was encapsulated within a gelatin-based shell bioink, and human umbilical vein endothelial cells within a chitosan-based core bioink to biofabricate a living dressing for enhanced thermal injury repair and regeneration. We hypothesized that the cell-laden c/s tissue engineered construct (TEC) would strengthen the wound's proangiogenic, anti-inflammatory, and skin regeneration potential. An in vitro thermal injury in a 3D raft skin model showed a slight delay in wound closure in the presence of the c/s TEC but was augmented by corresponding increases in the release of wound-healing factors, epidermal growth factor, matrix metalloproteinases-9, transforming growth factor-α, platelet-derived growth factor; a decrease in pro-inflammatory factor interleukin-6, and evidence of neovascularization. Core/shell three-dimensional (3D) bioprinted living dressings were used to heal an in vitro thermal injury wound in a 3D raft skin model. The mesenchymal stem cells/human umbilical vein endothelial cell-laden 3D tissue engineered constructs when compared with the untreated control resulted in a slight delay in wound closure along with corresponding increases in the secretion of wound-healing factors epidermal growth factor, matrix metalloproteinases-9, transforming growth factor-α, and platelet-derive growth factor; a decrease in pro-inflammatory factor interleukin-6; and promotion of neovascularization. The present work is the first to evaluate repair and corresponding cytokine response in an in vitro 3D raft thermal injury model using a "living dressing"construct.