Basic Study
Copyright ©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Jul 26, 2023; 15(7): 713-733
Published online Jul 26, 2023. doi: 10.4252/wjsc.v15.i7.713
Transplantation of human induced pluripotent stem cell derived keratinocytes accelerates deep second-degree burn wound healing
Li-Jun Wu, Wei Lin, Jian-Jiang Liu, Wei-Xin Chen, Wen-Jun He, Yuan Shi, Xiao Liu, Ke Li
Li-Jun Wu, Department of Plastic and Aesthetic Surgery, The Second Affiliated Hospital of Soochow University, Suzhou 215004, Jiangsu Province, China
Wei Lin, Jian-Jiang Liu, Wei-Xin Chen, Wen-Jun He, Yuan Shi, Xiao Liu, Ke Li, Department of Burn and Plastic Surgery, The First Affiliated Hospital of Soochow University, Suzhou 215006, Jiangsu Province, China
Author contributions: Wu LJ contributed to methodology, investigation, data curation, original draft; Lin W contributed to review and editing; Liu JJ contribute to methodology, investigation, data curation, original draft; Chen WX contribute to writing, review & editing; He WJ contribute to investigation; Shi Y contribute to writing; Liu X contribute to review & editing; Li K contribute to idea, supervision, review & editing.
Supported by the Hospital Research Fund, No. SDFEYBS1805, No. SDFEYGJ2013 and No. XKTJ-HRC20210015; Suzhou Science and Technology Development Project, No. SYS2020105, No. SKJY2021078 and No. 2022SS43; the Special Project of “Technological Innovation” Project of CNNC Medical Industry Co. Ltd, No. ZHYLZD2021002; CNNC Elite Talent Program; 2022 State Key Laboratory of Radiological Medicine and Radiation Protection jointly built by Province and Ministry, No. GZK1202244.
Institutional review board statement: All extractions were reviewed and approved by the medical ethics committee of the First Affiliated Hospital of Soochow University.
Institutional animal care and use committee statement: Animal experiment procedures were performed in strict accordance with protocols approved by the Institutional Animal Care and Use Committee.
Informed consent statement: All study participants, or their legal guardian, provided informed written consent prior to study enrollment.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request at likefggf@163.com.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: https://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Ke Li, MM, Academic Research, Department of Burn and Plastic Surgery, The First Affiliated Hospital of Soochow University, No. 188 Shizi Street, Suzhou 215006, Jiangsu Province, China. likefggf@163.com
Received: March 15, 2023
Peer-review started: March 15, 2023
First decision: April 13, 2023
Revised: May 15, 2023
Accepted: June 26, 2023
Article in press: June 26, 2023
Published online: July 26, 2023
Processing time: 130 Days and 2.3 Hours
Abstract
BACKGROUND

Current evidence shows that human induced pluripotent stem cells (hiPSCs) can effectively differentiate into keratinocytes (KCs), but its effect on skin burn healing has not been reported.

AIM

To observe the effects of hiPSCs-derived KCs transplantation on skin burn healing in mice and to preliminarily reveal the underlying mechanisms.

METHODS

An analysis of differentially expressed genes in burn wounds based on GEO datasets GSE140926, and GSE27186 was established. A differentiation medium containing retinoic acid and bone morphogenetic protein 4 was applied to induce hiPSCs to differentiate into KCs. The expression of KCs marker proteins was detected using immunofluorescence staining. A model of a C57BL/6 mouse with deep cutaneous second-degree burn was created, and then phosphate buffered saline (PBS), hiPSCs-KCs, or hiPSCs-KCs with knockdown of COL7A1 were injected around the wound surface. The wound healing, re-epithelialization, engraftment of hiPSCs-KCs into wounds, proinflammatory factor level, and the NF-κB pathway proteins were assessed by hematoxylin-eosin staining, carboxifluorescein diacetate succinimidyl ester (CFSE) fluorescence staining, enzyme linked immunosorbent assay, and Western blotting on days 3, 7, and 14 after the injection, respectively. Moreover, the effects of COL7A1 knockdown on the proliferation and migration of hiPSCs-KCs were confirmed by immunohistochemistry, EdU, Transwell, and damage repair assays.

RESULTS

HiPSCs-KCs could express the hallmark proteins of KCs. COL7A1 was down-regulated in burn wound tissues and highly expressed in hiPSCs-KCs. Transplantation of hiPSCs-KCs into mice with burn wounds resulted in a significant decrease in wound area, an increase in wound re-epithelialization, a decrease in proinflammatory factors content, and an inhibition of NF-κB pathway activation compared to the PBS group. The in vitro assay showed that COL7A1 knockdown could rescue the inhibition of hiPSCs-KCs proliferation and migration, providing further evidence that COL7A1 speeds up burn wound healing by limiting cell proliferation and migration.

CONCLUSION

In deep, second-degree burn wounds, COL7A1 can promote KC proliferation and migration while also suppressing the inflammatory response.

Keywords: Induced pluripotent stem cell; Keratinocytes; Cell transplantation; Burn wound healing; COL7A1

Core Tip: Current evidence shows that human induced pluripotent stem cells (hiPSCs) can effectively differentiate into keratinocytes (KCs), but its effect on skin burn healing has not been reported. Therefore, this study was intended to observe the effects of hiPSCs-derived KCs transplantation on skin burn healing in mice and to preliminarily reveal the underlying mechanisms. Transplantation of hiPSCs-KCs into mice with burn wounds resulted in a significant decrease in wound area, an increase in wound re-epithelialization, a decrease in proinflammatory factors content, and an inhibition of NF-κB pathway activation, which rescued by COL7A1 knockdown.