Exosomes from adipose-derived stem cells overexpressing Nrf2 accelerate cutaneous wound healing by promoting vascularization in a diabetic foot ulcer rat model

Mishra²,

et al. 2018

IJMS

Adipose-derived stromal/stem cells (ASCs) seems to be a promising regenerative therapeutic agent due to the minimally invasive approach of their harvest and multi-lineage differentiation potential. The harvested adipose tissues are further digested to extract stromal vascular fraction (SVF), which is cultured, and the anchorage-dependent cells are isolated in order to characterize their stemness, surface markers, and multi-differentiation potential. The differentiation potential of ASCs is directed through manipulating culture medium composition with an introduction of growth factors to obtain the desired cell type. ASCs have been widely studied for its regenerative therapeutic solution to neurologic, skin, wound, muscle, bone, and other disorders. These therapeutic outcomes of ASCs are achieved possibly via autocrine and paracrine effects of their secretome comprising of cytokines, extracellular proteins and RNAs. Therefore, secretome-derivatives might offer huge advantages over cells through their synthesis and storage for long-term use. When considering the therapeutic significance and future prospects of ASCs, this review summarizes the recent developments made in harvesting, isolation, and characterization. Furthermore, this article also provides a deeper insight into secretome of ASCs mediating regenerative efficacy.

Section: Asc Secretome and Its Therapeutic Effectmentioning

confidence: 99%

Revisiting the Advances in Isolation, Characterization and Secretome of Adipose-Derived Stromal/Stem Cells

Mishra²,

et al. 2018

IJMS

“…Nrf2 (NF-E2-related factor) is an important transcription factor that maintains redox homeostasis in cells, participates in the development of various diseases, and plays an important role in the regulation of oxidative stress [16,42,45].At the same time, related reports indicate that Nrf2 is involved in the proliferation, differentiation, migration and apoptosis of various types of cells [5,46]. The exosomes of adipose-derived stem cells that highly express Nrf2 can promote the healing of diabetic foot ulcer skin wounds in rats [47,48].In vivo and in vitro experiments, we found that LPS-Exos can activate Nrf2 / HO-1 pathway-related proteins, while reducing in ammation-related proteins, promoting endothelial cell vascularization and collagen synthesis.Interestingly, after treatment with exosome-speci c inhibitors (GW4869), the symptoms improved by LPS-Exos were all reversed.…”

Section: Discussionmentioning

confidence: 99%

“…The cells pretreated with high glucose (HG, 30 mm / L) were used as experimental control group. At the same time, in order to better explore the role of LPS stimulated macrophage exosomes, the experimental group used the combination of exosomes inhibitor (GW4869) [5,[21][22][23] and different concentrations of exosomes to treat cells.…”

Section: Cell Culture and Sample Treatmentmentioning

confidence: 99%

“…Diabetes mellitus (DM) is a chronic metabolic disease with hyperglycemia as the main manifestation, in which the impaired wound healing is one of the main complications of diabetes [1,2].There are many reasons for the delayed healing of diabetic wounds, including neuropathy, peripheral vascular disease and immune system damage, which ultimately lead to the prolongation of in ammatory phase, proliferation and shortening of remodeling phase of skin wounds [3].At the same time, the production of a large number of reactive oxygen species (ROS) aggravates the in ammatory response of cells, promotes the apoptosis of injured tissues, and delays the healing of skin wounds [4].Therefore, reducing the production of reactive oxygen species and the continuous effect of in ammation in skin wound can be an important strategy to improve the wound healing of diabetes [5].…”

Section: Introductionmentioning

confidence: 99%

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LPS-stimulated Macrophage Exosomes Inhibit Inflammation by Activating the Nrf2 / HO-1 Defense Pathway and Promote Wound Healing in Diabetic Rats

Tian

et al. 2020

Preprint

Background:Impaired wound healing is one of the important complications of diabetes. However, the specific pathogenesis is still unclear, and there is no effective treatment. Macrophages pretreated with chemical or biological factors may increase the biological activity of macrophage-derived exosomes, which is expected to become a new effective treatment method. The purpose of this study was to investigate whether the exosomes secreted by macrophages pretreated with lipopolysaccharide (LPS) have better anti-inflammatory and angiogenic abilities in the treatment of diabetic wound healing and their underlying molecular mechanisms.Methods:In this study, macroscopical, biochemical, histological, immunofluorescence and molecular biology methods were used to evaluate the potential protective mechanism and effect of lipopolysaccharide stimulated macrophage exosomes (LPS-Exos) on wound healing in streptozotocin induced hyperglycemia rats.In the in vivo experiment, the percentages of wound closure and contraction were compared and analyzed on the 7th, 14th, and 21st day after treatment by grouping treatments with different concentrations of LPS-Exos.At the same time, hematoxylin and eosin staining (HE), Masson staining, immunofluorescence staining and Western blotting (WB) were used for histological analysis of the wound tissue on the 14th day after injury to evaluate the impact of different treatment methods on wound healing.In in vitro experiments, the ability of endothelial cells related to proliferation, migration, tube formation and the expression of vascular endothelial growth factor (VEGF) were tested.At the same time, in vivo and in vitro experiments, the effect of Nrf2/HO-1 signaling pathway on LPS-Exos in inhibiting inflammation and promoting angiogenesis was evaluated by using exosome-specific inhibitors.Results:LPS-Exos reduced the content of ROS and MDA in the wound tissue of hyperglycemic rats, increased the activity of SOD and the production of GSH-Px, activated the Nrf2/HO-1 pathway, inhibited the expression of inflammation-related proteins, and promoted blood vessels generate. The group given exosome inhibitors reversed this phenomenon.Conclusions: LPS-Exos may activate the Nrf2/HO-1 defense pathway, improve endothelial cell function, inhibit oxidative damage and inflammation, and promote wound healing in diabetic rats, thereby having the potential to treat diabetic skin defects.

“…After co-incubation for 24 h with PKH26-labeled ASC-Exos, red fluorescence was localized within the cytosol of HDFs, thus confirming the Exo internalization. Li et al [81] suggested that the pro-healing action of ASC-Exos could be mediated by overexpression of nuclear factor-E2-related factor 2 (Nrf2), a transcription factor with a protective role against oxidative stress [82]. The authors isolated Exos from the CM of ASCs cultured in EGM-2MV medium deprived of FBS and supplemented with serum replacement solution for an additional 24 h. CM was centrifuged at 300× g for 10 min and 2000× g for 10 min to remove cell debris.…”

Section: In Vitro Studiesmentioning

confidence: 99%

Secretome of Adipose Tissue-Derived Stem Cells (ASCs) as a Novel Trend in Chronic Non-Healing Wounds: An Overview of Experimental In Vitro and In Vivo Studies and Methodological Variables

Lombardi

Palumbo

Augello

et al. 2019

IJMS

Wound healing is a complex process with a linear development that involves many actors in a multistep timeline commonly divided into four stages: Hemostasis, inflammation, proliferation, and remodeling. Chronic non-healing wounds fail to progress beyond the inflammatory phase, thus precluding the next steps and, ultimately, wound repair. Many intrinsic or extrinsic factors may contribute to such an occurrence, including patient health conditions, age-related diseases, metabolic deficiencies, advanced age, mechanical pressure, and infections. Great interest is being focused on the adipose tissue-derived stem cell’s (ASC) paracrine activity for its potential therapeutic impact on chronic non-healing wounds. In this review, we summarize the results of in vitro and in vivo experimental studies on the pro-wound healing effects of ASC-secretome and/or extracellular vesicles (EVs). To define an overall picture of the available literature data, experimental conditions and applied methodologies are described as well as the in vitro and in vivo models chosen in the reported studies. Even if a comparative analysis of the results obtained by the different groups is challenging due to the large variability of experimental conditions, the available findings are undoubtedly encouraging and fully support the use of cell-free therapies for the treatment of chronic non-healing wounds.