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Embryonic stem cell extracts improve wound healing in diabetic mice

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Abstract

Aims/hypothesis

Impaired wound healing significantly impacts morbidity and mortality in diabetic patients, necessitating the development of novel treatments to improve the wound healing process. We here investigated the topical use of acellular embryonic stem cell extracts (EXTs) in wound healing in diabetic db/db mice.

Methods

Wounds were induced in diabetic db/db mice, which were subsequently treated with EXTs, with 3T3 fibroblast cell line protein extracts (3T3XTs) or with saline as a control. Pathology and mechanistic assays were then performed.

Results

The in vivo topical administration of EXTs facilitates wound closure, contraction and re-epithelialization. Moreover, EXTs reduced the number of wound-infiltrating CD45+ inflammatory cells and increased the rate of repair and of angiogenesis as compared to controls. Interestingly, the EXT effect was partly enhanced by the use of a collagen-based biocompatible scaffold. In vivo, topical administration of EXTs increased the percentage of regulatory T cells in the wounded tissue, while in vitro EXT treatment reduced T cell-mediated IFN-γ production. Proteomic screening revealed 82 proteins differentially segregating in EXTs as compared to 3T3 extracts, with APEX1 identified as a key player for the observed immunomodulatory effect of EXTs.

Conclusions

EXTs are endowed with immunoregulatory and anti-inflammatory properties; their use improves wound healing in diabetic preclinical models.

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Availability of data and material

All data generated or analyzed during this study are available from the corresponding author on reasonable request.

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Acknowledgements

We are thankful to Prof. Nicholas Zavazava (Carver College of Medicine, University of Iowa) for providing embryonic cell extracts. We thank Mollie Jurewicz for editorial assistance. We thank the Fondazione Romeo and Enrica Invernizzi for the extraordinary support.

Funding

Paolo Fiorina is supported by an Italian Ministry of Health Grant RF-2016-02362512 and by Linea-2 2019 funding from Università di Milano. Francesca D’Addio is the recipient of a Società Italiana di Diabetologia (SID) Lombardia Grant and of the EFSD/JDRF/Lilly Programme on Type 1 Diabetes Research 2019. Vera Usuelli is supported by Fondazione Diabete Ricerca (FO.DI.RI) Società Italiana di Diabetologia (SID) fellowship.

Author information

Author notes

  1. Cristian Loretelli and Moufida Ben Nasr have contributed equally to this work.

Authors and Affiliations

  1. International Center for T1D, Pediatric Clinical Research Center Romeo ed Enrica Invernizzi, DIBIC L. Sacco, Università degli Studi di Milano, Milan, Italy

    Cristian Loretelli, Moufida Ben Nasr, Francesca D’Addio, Ahmed Abdelsalam, Elio Ippolito, Emma Assi, Vera Usuelli, Gian Vincenzo Zuccotti & Paolo Fiorina

  2. Division of Nephrology, Boston Children’s Hospital, Harvard Medical School, Enders Building 5th Floor, Rm EN530, 300 Longwood Avenue, Boston, MA, 02115, USA

    Moufida Ben Nasr, Roberto Bassi, Alessandro Valderrama-Vasquez & Paolo Fiorina

  3. Tissue Engineering and Wound Healing Laboratory, Division of Plastic Surgery, Department of Surgery, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Giorgio Giatsidis, Saja Sandra Scherer, Giorgio Pietramaggiori & Dennis Paul Orgill

  4. Department of Plastic Surgery, St. John’s Hospital, NHS Lothian, Edinburgh, Scotland, UK

    Luca Lancerotto

  5. Department of Engineering for Innovation, University of Salento, Lecce, Italy

    Luca Salvatore, Marta Madaghiele & Alessandro Sannino

  6. Department of Biochemistry and Biotechnology, Heliopolis University, Cairo, Egypt

    Ahmed Abdelsalam

  7. Transplantation Research Center, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, USA

    Basset El Essawy

  8. Medicine, Al-Azhar University, Cairo, Egypt

    Basset El Essawy

  9. Department of Pediatrics, Buzzi Children’s Hospital, Milan, Italy

    Gian Vincenzo Zuccotti

  10. Division of Endocrinology, ASST Fatebenefratelli-Sacco, Milan, Italy

    Paolo Fiorina

Authors
  1. Cristian Loretelli
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  2. Moufida Ben Nasr
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  3. Giorgio Giatsidis
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  4. Roberto Bassi
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  9. Luca Salvatore
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  13. Emma Assi
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  17. Giorgio Pietramaggiori
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Contributions

C.L. and M.B.N. designed the study, performed experiments, analyzed data, and wrote the paper; G.G., L.L. and SS.S. performed immunostaining and histological analysis of the wounds; R.B. performed proteomic analysis; F.D., A.VV., L.S., M.M., A.A., E.I., E.A. and V.U. performed experiments and analyzed data; B.EE., A.S., G.P. and GV.Z. coordinated and designed the research and edited the paper; DP.O. designed animal studies and reviewed the manuscript; P.F. conceived the idea, designed the study, and wrote and edited the paper. All authors were given full access to all data presented in this study and are responsible for the integrity of the data and accuracy of the data analysis. All authors have given their permission for submission of this manuscript.

Corresponding author

Correspondence to Paolo Fiorina.

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Conflict of interest

The authors declare no present or potential conflicts of interest. This study was performed without the support or involvement of any external funding source or study sponsor in any phase of the investigation, or in the writing or submission of the manuscript.

Human and animal rights disclosure

All procedures performed in the study were in accordance with the ethical standards as laid down in the 1964 Declaration of Helsinki and its later amendments or comparable ethical standards.

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Written informed consent was obtained from all individual participants included in the study.

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Loretelli, C., Ben Nasr, M., Giatsidis, G. et al. Embryonic stem cell extracts improve wound healing in diabetic mice. Acta Diabetol 57, 883–890 (2020). https://doi.org/10.1007/s00592-020-01500-0

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  • DOI: https://doi.org/10.1007/s00592-020-01500-0

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