Abstract
The differentiation, migration, and proliferation of skin fibroblasts are identified as key factors in cutaneous wound healing. Adipose-derived mesenchymal stem cells (ADMSCs) and their exosomes (ADMSC-Exos) have been considered as potential therapeutic tools for tissue regeneration; however, the underlying mechanisms on cutaneous wound healing are still not well understood. In this study, we successfully obtained ADMSC-Exos and found ADMSC-Exos significantly promoted the migration and proliferation of fibroblasts in a dose-dependent manner in vitro. The expression levels of COL-I and COL-III in fibroblasts treated with ADMSC-Exos were significantly increased, while the expression level of α-SMA was decreased. In addition, the enhanced protein expression of WNT2b and β-catenin confirmed the activation of the WNT/β-catenin signaling pathway and the WNT/β-catenin inhibitor (XAV939) reversed the promoting effect of ADMSC-Exos on wound healing and the β-catenin expression. Taken together, our study partially elucidates the mechanism of ADMSC-Exos in wound healing, illustrating the potential of ADMSC-Exos as a new therapeutic approach to promote skin wound healing.
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Introduction
Cutaneous wound healing is a complex and continuous process that includes haemostatic, inflammatory, proliferative, and remodelling stages. Various factors, such as high wound tension, infection, radiation damage, and metabolic disease, lead to prolonged wound healing time, causing physical or mental pain and imposing a severe treatment burden on patients [1, 39]; collagen deposition early in wound healing contributes to wound healing, but later collagen deposition leads to the formation of unsightly scars and organ dysfunction [40]. As for our results, the high concentration of ADMSC-Exos reduced collagen formation and increased the migration and proliferation of fibroblasts, which may be one of the reasons why ADMSC-Exos accelerated wound healing and weakened scar formation. Unfortunately, the specific mechanism is still unclear and needs further study. We hypothesized that these effects were controlled by the complex contents of exosomes, which was shown by our concentration gradient assay.
For another, α-SMA is the marker of fibroblast differentiation into myofibroblast [41], which is closely related to the mechanical tension of wound surface, and excessive α-SMA expression is also the cause of promoting scar formation. Regarding the decreased expression of α-SMA in our research, we predicted that the application of ADMSC-Exos to the wound increased the migration rate of fibroblasts from the periphery to the centre of the wound surface, thus reducing the surface tension. Meanwhile, we speculated that ADMSC-Exos may have been able to inhibit the conversion of fibroblasts to myofibroblasts, thereby inhibiting excessive scar formation.
The WNT/β-catenin signaling pathway serves a vital role in proliferation, differentiation, movement, and morphology of cells, and it is also involved in mediating stem cell pluripotency [20]. β‐Catenin, a subunit of the cadherin protein complex, is an integral component of the classic WNT signaling pathway [42]. Accumulating evidence has confirmed that activation of the WNT/β‐catenin signaling pathway plays an essential role in the proliferative phase of wound healing [42, 43]. Our previous studies showed that WNT-responsive oral mucosal stem cells are activated in response to oral mucosal injury, thereby accelerating wound healing [21]. Thus, according to the conclusion that ADMSC-Exos activated WNT/β-catenin signaling pathway in this study, we have reason to deduce that whether ADMSC-Exos can also promote skin stem cell differentiation through the WNT signaling pathway during wound healing remains to be further studied. Moreover, our previous studies also showed that WNT-responsive stem cells can regulate periodontal membrane fibrosis and alveolar bone density to adapt to changes in mechanical stress [22]. Therefore, it remains to be further studied whether the effect of ADMSC-Exos on skin wound healing can also regulate the effect of mechanical tension by regulating WNT-responsive stem cells, which would be significant. At present, exosomes are an emerging platform and a critical factor in facilitating WNT secretion and transport. Zhang et al. found that exosomes from human umbilical cord MSCs can deliver WNT4 to target cells and activate β-catenin nuclear translocation, which has a positive effect on wound healing [44]. Ma et al. found that ADMSC-Exos could increase the protein expression of WNT and β-catenin in HaCaT cells treated with hydrogen peroxide to simulate skin damage[35]. However, their study did not research the effect of exosomes on fibroblasts during wound healing, and did not investigate the expression of ECM. Our study just complemented this and further refined the mechanism of exosomes through WNT signaling pathway. In this study, the elevated expression of β‐catenin in vitro and in vivo under treatment with ADMSC-Exos suggested that WNT/β‐catenin signaling may be related to the underlying mechanism of ADMSC‐Exos in wound healing. However, because of the complexity of wound tissue healing, more studies should be carried out on the association between the WNT/β-catenin signaling pathway and cutaneous wound healing.
Conclusion
In summary, ADMSC-Exos implies a pivotally stimulative role for wound healing process, in which this study revealed that WNT/β-catenin signaling pathway was involved. Furthermore, ADMSC-Exos had a preventive effect on cicatrization by boosting the proliferation and migration of fibroblasts, downregulating the expression of fibroblast α-SMA, and promoting the rearrangement of collagen fibres concurrently, which showed great possibilities for wound applications, providing novel treatment strategies for wound healing.
Data availability
The data underlying this article will be shared on reasonable request to the corresponding author.
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Acknowledgements
The authors wish to thank all the staff of the Burn and Plastic Surgery Department of the Affiliated Hospital of Qingdao University for their support of this study.
Funding
This paper was supported by the Shandong Provincial Natural Science Foundation (Grant/Award Number: ZR2017MH083, ZR2020MH183).
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Cong Li, Yu An: Conception and design, experiments, manuscript writing, data analysis and interpretation; Yu Sun, Fan Yang: Experiments, Collection of data; Zhiguo Wang, Quanchen Xu: Conception and design, manuscript writing, final approval of manuscript.
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The Ethics Committee of the Affiliated Hospital of Qingdao University approved the study (QYFY WZLL 26708), and it was conducted in accordance with the Declaration of Helsinki.
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The fat tissues and foreskin tissue were collected after obtaining informed consent from the donors.
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Cong Li and Yu An contributed equally to this work.
Cong Li and Yu An are co-first authors.
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Li, C., An, Y., Sun, Y. et al. Adipose Mesenchymal Stem Cell-Derived Exosomes Promote Wound Healing Through the WNT/β-catenin Signaling Pathway in Dermal Fibroblasts. Stem Cell Rev and Rep 18, 2059–2073 (2022). https://doi.org/10.1007/s12015-022-10378-0
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DOI: https://doi.org/10.1007/s12015-022-10378-0