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miR-21 Expressed by Dermal Fibroblasts Enhances Skin Wound Healing Through the Regulation of Inflammatory Cytokine Expression

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Abstract

The management of skin wound healing is still a challenge. MicroRNA-21 (miR-21) has been reported to play important roles in wound repair; however, the underlying mechanism needs to be further clarified. The present study aimed to study the direct role of miR-21 in skin wound healing in miR-21 KO mice and to investigate the role of miR-21 in controlling the migration and proliferation of primary human skin cells and its underlying mechanism(s). miR-21 KO and wild-type (WT) mice were used for in vivo wound healing assays, while mouse and human primary skin cells were used for in vitro assays. miR-21 inhibitors or mimics or negative control small RNAs were transfected to either inhibit or enhance miR-21 expression in the human primary dermal fibroblasts or epidermal cells. RNA sequencing analysis was performed to identify the potential molecular pathways involved. We found that the loss of miR-21 resulted in slower wound healing in miR-21 KO mouse skin and especially delayed the healing of dermal tissue. In vitro assays demonstrated that the reduced expression of miR-21 caused by its inhibitor inhibited the migration of human primary dermal fibroblasts, which could be enhanced by increased miR-21 expression caused by miR-21 mimics. RNA-sequence analysis revealed that the inhibition of miR-21 expression downregulated the inflammatory response pathways associated with the decreased expression of inflammatory cytokines, and the addition of IL-1β into the culture medium enhanced the migration and proliferation of dermal fibroblasts in vitro. In conclusion, miR-21 in dermal fibroblasts can promote the migration and growth of epidermal and dermal cells to enhance skin wound healing through controlling the expression of inflammatory cytokines.

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Acknowledgements

We gratefully acknowledge Xunwei Wu’s lab members for helpful discussions and technical assistance.

Funding

This work was supported by the General Program of the National Natural Science Foundation of China (82273554, 82073470) and the Shandong Provincial Key R&D Program (ZR2019ZD36) to X.W., and the Key R&D Program of Shandong Province (2018GSF118240) and “Fan 3315 plan” Medical and Health Innovative talents of Ningbo City (2020B-30-G) to J.G.

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Authors and Affiliations

  1. Engineering Laboratory for Biomaterials and Tissue Regeneration, Ningbo Stomatology Hospital, Savaid Stomatology School, Hangzhou Medical College, Ningbo, China

    Chang Liu, Dexuan Zhuang, Huiting Deng, Yuxin Shi, Jianfeng Sun, Jing Guo & Xunwei Wu

  2. Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, Jinan, China

    Chang Liu, Qun Zhang, Zhenan Liu, Dexuan Zhuang, Shuangshuang Wang & Xunwei Wu

  3. Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, No. 44-1 Wenhua Road West, Jinan, Shandong, China

    Jing Guo & Fulan Wei

  4. Suzhou Research Institute, Shandong University, No. 388 Ruoshui Road, Suzhou, Jiangsu, China

    Xunwei Wu

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  1. Chang Liu
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Contributions

Data curation, C.L., Q.Z., and Z.L.; formal analysis, C.L. and X.W.; funding acquisition, J.G. and X.W.; investigation, C.L., Q.Z., Z.L., D.Z., S.W., H.D., J.S., and Y.S.; methodology, Q.Z., Z.L., and X.W.; supervision, J.G., F.W., and X.W.; validation, F.W. and X.W.; writing original draft, C.L.; writing review and editing, X.W. All authors have read and agreed to the published version of the manuscript.

Corresponding authors

Correspondence to Fulan Wei or Xunwei Wu.

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Ethics Approval and Consent to Participate

The studies involving human participants were reviewed and approved for the use of human tissues collected from discarded hospital specimens without any personal identity information and the protocol (protocol no. GR201711, date February 27, 2017) was approved by the Ethics Committee of the Hospital of Stomatology, Shandong University. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements. The animal protocol carried out in this study was reviewed and approved by the Ethics Committee of the Hospital of Stomatology, Shandong University (protocol no. GR201720, date February 27, 2017).

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Liu, C., Zhang, Q., Liu, Z. et al. miR-21 Expressed by Dermal Fibroblasts Enhances Skin Wound Healing Through the Regulation of Inflammatory Cytokine Expression. Inflammation 47, 572–590 (2024). https://doi.org/10.1007/s10753-023-01930-2

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