Enhancing Cutaneous Wound Healing Based on Human Induced Neural Stem Cell-derived Exosomes

Abstract

Background: Wound healing of skin is a complicated process. Cutaneous innervation and neurotrophic factors could participate in multiple stages of wound healing. Neurotrophic factors are mainly produced and released by neurons and neural stem cells (NSCs) which could be obtained in large quantities from human-induced pluripotent stem cells (iPSCs) in vitro. However, the potential wound healing effects of NSC secretions, such as exosomes, are unexplored yet. Methods: NSCs-derived exosomes (NSC-exo) and iPSCs-derived exosomes (iPSC-exo) were isolated from the cell culture supernatants by centrifugation, and then quantified and characterized. The effects of these exosomes on the migration of human dermal fibroblasts (HDF) cells and the tube formation of human umbilical vein endothelial cells (HUVECs) were investigated in vitro. And the in vivo wound healing effect of these exosomes were tested on the mouse skin trauma model. Therefore, a dipeptide/hyaluronic acid (Nap-FF/HA) composite hydrogel was used to encapsulate the exosomes as a sustained release carrier. Histological observations were performed to evaluate the wound healing effect of exosomes. Furthermore, the non-labeling proteomic analysis was performed to explore the possible mechanisms of NSC-exo on wound healing. Results: We obtained extracellular vesicles in a bowl-like structure with membranes which meet the general standards of exosomes. NSC-exo showed promotion effect on the migration of HDF cells and the tube formation of HUVECs in vitro. In a mouse skin injury model, NSC-exo enhanced the wound healing and the Nap-FF/HA hydrogel that contained exosomes did so with less drug frequency by sustaining release of exosomes. Further proteomic analysis demonstrated that the carried neurotrophic factors and immunity-related proteins in NSC-exo may play a functional role in wound healing. Conclusion: NSC-exo may enhance wound healing via neurotrophic factors and immunomodulation.

Biomaterial‐Facilitated Local Delivery of Stem Cell‐Derived Small Extracellular Vesicles: Perspectives in Surgical Therapy

“…The SC-sEV delivered by their dipeptide/HA composite HG improved wound healing through exosomal neurotrophic factors and immunity-related proteins. [129] In addition to wound healing, other plastic surgery-related diseases, such as alopecia, have been proven to be suitable targets for local SC-sEV therapy. [130] Shi et al applied a separable antibacterial chitosan lactate microneedle patch comprised of a HA substrate and PVA needles for sustained release of sEVs derived from adipose MSCs.…”

“…The SC‐sEV delivered by their dipeptide/HA composite HG improved wound healing through exosomal neurotrophic factors and immunity‐related proteins. [ 129 ]…”

Neural stem cell-derived extracellular vesicles: mini players with key roles in neurogenesis, immunomodulation, neuroprotection and aging

“…Therefore, they do respond to factors not only present locally, but also present in other brain regions or in the systemic macroenvironment. Increasing evidence proposed that EVs confer immunomodulatory and anti-inflammatory effects in various inflammatory disorders and tissue injury by suppressing activation of immune cells and reducing pro-inflammatory mediators production ( Akbar et al, 2022 ; Li et al, 2022 ). EVs have been found to regulate immunity and restore function through a variety of pathways.…”

Human pluripotent stem cell-derived extracellular vesicles: From now to the future

“…PSC-EVs have been described as having: Protective effects in in vitro and in vivo models of ischemia-reperfusion kidney injury[ 64 ]; neural protective abilities[ 60 ]; the capacity to modulate neuroinflammation and protect against ischemic stroke through Treg cell expansion[ 73 ]; antifibrotic effects in vivo and in in vivo models of liver injury[ 61 , 72 ]; and reduced cartilage degradation in an osteoarthritis model[ 77 ]. They have shown improvements in wound closure, angiogenesis, and increased nerve fiber density in a wound-healing diabetic mouse model[ 54 , 79 ]; and improved recovery of ovarian function in a premature ovarian failure mouse model[ 67 ]. EVs were also associated with acellular nerve grafts, demonstrating their potential to repair peripheral nerve defects[ 80 ].…”