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Preparation and characterization of human keratinocyte–fibroblast cell sheets constructed using PNIAM-co-AM grafted surfaces for burn wound healing

  • Tissue Engineering Constructs and Cell Substrates
  • Original Research
  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

Autologous skin grafting, the standard treatment for severe burns, is sometimes not possible due to the limited available skin surfaces for the procedure. With advances in tissue engineering, various cell-based skin substitutes have been developed to serve as skin replacements and to promote tissue regeneration and healing. In this work, we propose the use of cell sheet technology to fabricate keratinocyte–fibroblast tissue constructs from the temperature-responsive poly(N-isoproprylacrylamide-co-acrylamide) (PNIAM-co-AM) grafted surfaces for the treatment of burn wounds. The characteristics of the human keratinocyte and fibroblast cell sheets harvested using PNIAM-co-AM grafted surfaces were similar to those cell sheets detached from the commercially-available UpCellTM plates. Upon lowering the incubation temperature, confluent keratinocytes and fibroblasts could be detached as intact sheets, consisting of biologically active cells, as indicated by their high cell viability and their reattachment, migratory, and proliferative activities. A histological analysis of the stratified keratinocyte–fibroblast cell sheets revealed the evidence of cell migration and tissue reorganization to form two distinct epidermal and dermal layers, quite similar to the skin tissue’s structure. In addition, the keratinocyte–fibroblast sheets could synthesize and release significant amounts of essential cytokines and growth factors involved in regulating the wound healing process, including IL-1α, IL-6, TNF-α, VEGF, and bFGF, implying the therapeutic effect of these cell sheets, which could be beneficial to accelerate tissue repair and regeneration, leading to faster wound healing.

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Acknowledgements

This work was financially supported by Petchara Pra Jom Klao Doctoral Scholarship, King Mongkut’s University of Technology Thonburi (KMUTT), and Innovation Hub-Ageing Society: Translational Research (TR068), Council of University Presidents of Thailand.

Author contributions

KB carried out and analyzed all the experimental work in this study. She also wrote the paper with support from KVP, WS, and KC. PS and WS developed a method and fabricated the PNIAM-co-AM grafted surfaces. PM and KC provided human skin samples for this study and useful comments to help develop tissue-engineered skin equivalents. KVP supervised the work and approved the final draft.

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Authors and Affiliations

  1. Biological Engineering Program, Faculty of Engineering, King Mongkut’s University of Technology Thonburi, Bangkok, 10140, Thailand

    Kanokaon Benchaprathanphorn & Kwanchanok Viravaidya-Pasuwat

  2. School of Bio-Chemical Engineering and Technology, Sirindhorn International Institute of Technology, Thammasat University, Pathumthani, 12120, Thailand

    Phongphot Sakulaue & Wanwipa Siriwatwechakul

  3. Department of Surgery, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand

    Pornprom Muangman & Kusuma Chinaroonchai

  4. Department of Chemical Engineering, Faculty of Engineering, King Mongkut’s University of Technology Thonburi, Bangkok, 10140, Thailand

    Kwanchanok Viravaidya-Pasuwat

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  1. Kanokaon Benchaprathanphorn
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Correspondence to Kwanchanok Viravaidya-Pasuwat.

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Benchaprathanphorn, K., Sakulaue, P., Siriwatwechakul, W. et al. Preparation and characterization of human keratinocyte–fibroblast cell sheets constructed using PNIAM-co-AM grafted surfaces for burn wound healing. J Mater Sci: Mater Med 31, 126 (2020). https://doi.org/10.1007/s10856-020-06469-4

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