GMSC-Derived Exosomes Combined with a Chitosan/Silk Hydrogel Sponge Accelerates Wound Healing in a Diabetic Rat Skin Defect Model

Abstract

Background: Delayed wound healing in diabetic patients is one of the most challenging complications in clinical medicine, as it poses a greater risk of gangrene, amputation and even death. Therefore, a novel method to promote diabetic wound healing is of considerable interest at present. Previous studies showed that injection of MSC-derived exosomes has beneficial effects on wound healing. In current studies, we aimed to isolate exosomes derived from gingival mesenchymal stem cells (GMSCs) and then loading them to the chitosan/silk hydrogel sponge to evaluate the effects of this novel non-invasive method on skin defects in diabetic rats.Methods: GMSCs were isolated from human gingival connective tissue and characterized by surface antigen analysis and in vitro multipotent differentiation. The cell supernatant was collected to isolate the exosomes. The exosomes were characterized by transmission electron microscopy, Western blot and size distribution analysis. The chitosan/silk-based hydrogel sponge was prepared using the freeze-drying method and then structural and physical properties were characterized. Then, the exosomes were added to the hydrogel and tested in a diabetic rat skin defect model. The effects were evaluated by wound area measurement, histological, immunohistochemical and immunofluorescence analysis.Results: We have successfully isolated GMSCs and exosomes with a mean diameter of 127 nm. The chitosan/silk hydrogel had the appropriate properties of swelling and moisture retention capacity. The in vivo studies showed that the incorporating of GMSC-derived exosomes to hydrogel could effectively promote healing of diabetic skin defects. The histological analysis revealed more neo-epithelium and collagen in the hydrogel-exosome group. In addition, the hydrogel-exosome group had the highest microvessel density and nerve density.Conclusions: The combination of GMSC-derived exosomes and hydrogel could effectively promote skin wound healing in diabetic rats by promoting the re-epithelialization, deposition and remodeling of collagen and by enhancing angiogenesis and neuronal ingrowth. These findings not only provide new information on the role of the GMSC-derived exosomes in wound healing but also provide a novel non-invasive application method of exosomes with practical value for skin repair.

Heparin‐poloxamer hydrogel‐encapsulated rhFGF21 enhances wound healing in diabetic mice

“…However, growth factors rapidly degrade in vivo, and therefore, we needed a suitable delivery vehicle to maintain the biologic activity and bioavailability of growth factors, transport them safely to the wound, and control their release. A previous report revealed that exosomes combined with a chitosan or silk hydrogel can accelerate wound healing in diabetic rats (39), and our previous studies confirmed that growth factor and hydrogel combinations, such as heparin-FGF2@PEAD coacervate (40) and HP-FGF2 (24), can greatly promote skin wound healing in normal mice. HP is a thermosensitive hydrogel micelle material composed of poloxamer 407 and heparin combined via a polycondensation reaction.…”

Dental Stem Cell-Derived Secretome/Conditioned Medium: The Future for Regenerative Therapeutic Applications

“…(Table 1). Various investigations suggested that gingival MSC-derived EXs, EVs, or CM could represent novel therapeutic interventions in managing peripheral nerve injury [168,169], motor neuron injury [170], and skin [171] and bone defects [172]. The results were comparable with effects conferred by direct transplantation of gingival MSCs [168,169].…”

“…The implementation of gingival MSC-derived EXs in skin repair proved to be of practical value. Isolated EXs with an average diameter of 127 nm derived from gingival MSCs loaded on chitosan/silk hydrogel sponge effectively promoted healing of skin defects in diabetic rats detected by the formation of neoepithelium and collagen as well as a rise in the microvessels' number detected by CD34 in the wound 17 Stem Cells International bed and neuronal ingrowth detected by neurofilament heavy chain (NEFH), two weeks postsurgery [171]. (Table 4).…”

Oral Mesenchymal Stem/Progenitor Cells: The Immunomodulatory Masters

“…In an acute lung injury mouse model, a single intravenous administration of SHED-secreted factors alleviated lung injury, weight loss, and inflammatory response, accompanied by an increased number of anti-inflammatory M2-like lung macrophages [100]. GMSC-derived exosomes were capable of accelerating wound healing in diabetic mice by promoting reepithelialization, angiogenesis and neuronal ingrowth, and collagen remodeling [109]. Particularly, exosomes derived from healthy BM-MSCs alleviated radiation-induced bone loss by recovering the adipogenic and osteogenic differentiation potential of radiation-injured BM-MSCs via Wnt/β-catenin activation [110].…”