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An Important Role of Macrophages for Wound Margin Regeneration in a Murine Flap Model

  • Original Article
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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Macrophages have been known to have diverse roles either after tissue damage or during the wound healing process; however, their roles in flap wound healing are poorly understood. In this study, we aimed to evaluate how macrophages contribute to the flap wound regeneration.

Methods:

A murine model of a pedicled flap was generated, and the time-course of the wound healing process was determined. Especially, the interface between the flap and the residual tissue was histopathologically evaluated. Using clodronate liposome, a macrophage-depleting agent, the functional role of macrophages in flap wound healing was investigated. Coculture of human keratinocyte cell line HaCaT and monocytic cell line THP-1 was performed to unveil relationship between the two cell types.

Results:

Macrophage depletion significantly impaired flap wound healing process showing increased necrotic area after clodronate liposome administration. Interestingly, microscopic evaluation revealed that epithelial remodeling between the flap tissue and residual normal tissue did not occurred under the lack of macrophage infiltration. Coculture and scratch wound healing assays indicated that macrophages significantly affected the migration of keratinocytes.

Conclusion:

Macrophages play a critical role in the flap wound regeneration. Especially, epithelial remodeling at the flap margin is dependent on proper macrophage infiltration. These results implicate to support the cellular mechanisms of impaired flap wound healing.

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References

  1. Hanasono MM, Matros E, Disa JJ. Important aspects of head and neck reconstruction. Plast Reconstr Surg. 2014;134:968e–80.

    Article  CAS  Google Scholar 

  2. Cannady SB, Rosenthal EL, Knott PD, Fritz M, Wax MK. Free tissue transfer for head and neck reconstruction: a contemporary review. JAMA Facial Plast Surg. 2014;16:367–73.

    Article  Google Scholar 

  3. Chang EI, Zhang H, Liu J, Yu P, Skoracki RJ, Hanasono MM. Analysis of risk factors for flap loss and salvage in free flap head and neck reconstruction. Head Neck. 2016;38:E771–5.

    Article  Google Scholar 

  4. Pattani KM, Byrne P, Boahene K, Richmon J. What makes a good flap go bad? A critical analysis of the literature of intraoperative factors related to free flap failure. Laryngoscope. 2010;120:717–23.

    Article  Google Scholar 

  5. Rosado P, Cheng HT, Wu CM, Wei FC. Influence of diabetes mellitus on postoperative complications and failure in head and neck free flap reconstruction: a systematic review and meta-analysis. Head Neck. 2015;37:615–8.

    Article  Google Scholar 

  6. Herle P, Shukla L, Morrison WA, Shayan R. Preoperative radiation and free flap outcomes for head and neck reconstruction: a systematic review and meta-analysis. ANZ J Surg. 2015;85:121–7.

    Article  Google Scholar 

  7. Paderno A, Piazza C, Bresciani L, Vella R, Nicolai P. Microvascular head and neck reconstruction after (chemo)radiation: facts and prejudices. Curr Opin Otolaryngol Head Neck Surg. 2016;24:83–90.

    Article  Google Scholar 

  8. Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature. 2008;453:314–21.

    Article  CAS  Google Scholar 

  9. Singer AJ, Clark RA. Cutaneous wound healing. N Engl J Med. 1999;341:738–46.

    Article  CAS  Google Scholar 

  10. Martin P, Leibovich SJ. Inflammatory cells during wound repair: the good, the bad and the ugly. Trends Cell Biol. 2005;15:599–607.

    Article  CAS  Google Scholar 

  11. Wynn TA, Vannella KM. Macrophages in tissue repair, regeneration, and fibrosis. Immunity. 2016;44:450–62.

    Article  CAS  Google Scholar 

  12. Minutti CM, Knipper JA, Allen JE, Zaiss DM. Tissue-specific contribution of macrophages to wound healing. Semin Cell Dev Biol. 2017;61:3–11.

    Article  CAS  Google Scholar 

  13. Kim KE, Koh YJ, Jeon BH, Jang C, Han J, Kataru RP, et al. Role of CD11b+ macrophages in intraperitoneal lipopolysaccharide-induced aberrant lymphangiogenesis and lymphatic function in the diaphragm. Am J Pathol. 2009;175:1733–45.

    Article  CAS  Google Scholar 

  14. Li S, Li B, Jiang H, Wang Y, Qu M, Duan H, et al. Macrophage depletion impairs corneal wound healing after autologous transplantation in mice. PLoS One. 2013;8:e61799.

    Article  CAS  Google Scholar 

  15. Kawanishi N, Mizokami T, Niihara H, Yada K, Suzuki K. Macrophage depletion by clodronate liposome attenuates muscle injury and inflammation following exhaustive exercise. Biochem Biophys Rep. 2016;5:146–51.

    PubMed  Google Scholar 

  16. Wang TY, Serletti JM, Kolasinski S, Low DW, Kovach SJ, Wu LC. A review of 32 free flaps in patients with collagen vascular disorders. Plast Reconstr Surg. 2012;129:421e–7.

    Article  Google Scholar 

  17. Leoni G, Neumann PA, Sumagin R, Denning TL, Nusrat A. Wound repair: role of immune–epithelial interactions. Mucosal Immunol. 2015;8:959–68.

    Article  CAS  Google Scholar 

  18. Rousselle P, Braye F, Dayan G. Re-epithelialization of adult skin wounds: cellular mechanisms and therapeutic strategies. Adv Drug Deliv Rev. 2018. https://doi.org/10.1016/j.addr.2018.06.019.

    Article  PubMed  Google Scholar 

  19. Murray PJ, Wynn TA. Protective and pathogenic functions of macrophage subsets. Nat Rev Immunol. 2011;11:723–37.

    Article  CAS  Google Scholar 

  20. Genin M, Clement F, Fattaccioli A, Raes M, Michiels C. M1 and M2 macrophages derived from THP-1 cells differentially modulate the response of cancer cells to etoposide. BMC Cancer. 2015;15:577.

    Article  Google Scholar 

  21. Matsumoto NM, Aoki M, Nakao J, Peng WX, Takami Y, Umezawa H, et al. Experimental rat skin flap model that distinguishes between venous congestion and arterial ischemia: the reverse u-shaped bipedicled superficial inferior epigastric artery and venous system flap. Plast Reconstr Surg. 2017;139:79e–84.

    Article  CAS  Google Scholar 

  22. Casal D, Pais D, Iria I, Mota-Silva E, Almeida MA, Alves S, et al. A model of free tissue transfer: the rat epigastric free flap. J Vis Exp. 2017. https://doi.org/10.3791/55281.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Lee MS, Ahmad T, Lee J, Awada HK, Wang Y, Kim K, et al. Dual delivery of growth factors with coacervate-coated poly(lactic-co-glycolic acid) nanofiber improves neovascularization in a mouse skin flap model. Biomaterials. 2017;124:65–77.

    Article  CAS  Google Scholar 

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Acknowledgements

This study was supported by research Grants from Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2018R1D1A1A02043691, NRF-2016R1D1A1B03932867).

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Author notes

  1. Jeon Yeob Jang and Yoo Seob Shin have contributed equally to this work.

Authors and Affiliations

  1. Department of Otorhinolaryngology - Head and Neck Surgery, Chungnam National University Hospital, 282 Munhwa-ro, Jung-gu, Daejeon, 35015, Republic of Korea

    Ho-Ryun Won

  2. Department of Otolaryngology, Ajou University School of Medicine, 164 Worldcup Street, Yeongtong-gu, Suwon, 16499, Republic of Korea

    Chorong Seo, Hye-Young Lee, Chul-Ho Kim, Jeon Yeob Jang & Yoo Seob Shin

  3. Department of Pathology, Ajou University School of Medicine, 164 Worldcup Street, Yeongtong-gu, Suwon, 16499, Republic of Korea

    Jin Roh

  4. Department of Molecular Science and Technology, Ajou University, 164 Worldcup Street, Yeongtong-gu, Suwon, 16499, Republic of Korea

    Chul-Ho Kim & Yoo Seob Shin

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  1. Ho-Ryun Won
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  2. Chorong Seo
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Corresponding authors

Correspondence to Jeon Yeob Jang or Yoo Seob Shin.

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The authors declare that they have no conflict of interest.

Ethical statement

The animal studies were performed after receiving approval of the Institutional Animal Care and Use Committee (IACUC) in Ajou University (IACUC approval No. 2017-0025).

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Won, HR., Seo, C., Lee, HY. et al. An Important Role of Macrophages for Wound Margin Regeneration in a Murine Flap Model. Tissue Eng Regen Med 16, 667–674 (2019). https://doi.org/10.1007/s13770-019-00214-x

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  • DOI: https://doi.org/10.1007/s13770-019-00214-x

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