The role and mechanism of human umbilical cord mesenchymal stem cell exosomes in wounds with scabbing and skin grafting in scalded rats

Wang Di; Dou Shuqian; Wu Kongjia; Zhang Gaofei; Lou Hanxiao; Zhang Chenying; Yang Guoxun; Jin Chengbo; Que Ting; Liu Wenjun

doi:10.3760/cma.j.cn501225-20231201-00223

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > > Accepted Manuscript

Wang Di, Dou Shuqian, Wu Kongjia, et al. The role and mechanism of human umbilical cord mesenchymal stem cell exosomes in wounds with scabbing and skin grafting in scalded rats[J]. Chin J Burns Wounds. Doi: 10.3760/cma.j.cn501225-20231201-00223

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The role and mechanism of human umbilical cord mesenchymal stem cell exosomes in wounds with scabbing and skin grafting in scalded rats

doi: 10.3760/cma.j.cn501225-20231201-00223

Department of Burns, the Second Affiliated Hospital of Kunming Medical University, Kunming650106, China

Funds:

Regional Science Foundation of National Natural Science Foundation of China 82060349

Yunnan Province "Ten Thousand Talents Program" Famous Doctor Project YNWR-MY-2019-013

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Corresponding author: Liu Wenjun, Email: 86550558@qq.com
Received Date: 2023-12-01
Available Online: 2024-10-31

Abstract

Abstract

Objective To investigate the role and mechanism of human umbilical cord mesenchymal stem cell exosomes (hUCMSC-ex) in wounds with scabbing and skin grafting in scalded rats. Methods The study was an experimental study. Twelve male Sprague-Dawley rats aged 6-8weeks were divided into combined treatment group, fixed+allogeneic skin group, autologous skin+allogeneic skin group, and allogeneic skin group by random number table method (the same rats and grouping methods were used hereafter), with 3 rats in each group. The four groups of rats were inflicted with scalded wounds on the back and with scabbing, and then the wounds of rats in combined treatment group were fixed with a metal ring and transplanted with autologous skin grafts and allogeneic skin grafts, and the other three groups were fixed and/or transplanted with skin grafts corresponding to the group name. On 14, 21, and 28 d after surgery, the wound healing area in four groups of rats was measured. Another 15 rats were divided into normal group with no treatment, high exosome group, low exosome group, supernatant group, and phosphate buffered solution (PBS) group, with 3 rats in each group. The last 4 groups of rats were treated as that in the above-mentioned combined treatment group, and then were injected around the wounds with 200 μL of PBS containing 100 μg of hUCMSC-ex, 200 μL of PBS containing 50 μg of hUCMSC-ex, supernatant with no hUCMSC-ex, and PBS at 0 (immediately), 7, 14, and 21 d after surgery, respectively. On 14, 21, and 28 d after surgery, the wound healing area in four groups of rats was measured. The wound neo-epithelial tissue of rats in high exosome group and PBS group on 28 d after surgery and the normal skin tissue of rats in normal group at the same time point were taken, and the differentially expressed proteins were screened by label-free quantitative proteomics technology; the protein expressions of IgG1 heavy chain constant region (IGHG1) and cystatin A (CSTA) which were the differentially expressed proteins with the most obvious differences between high exosome group and PBS group were detected by Western blotting. The number of samples in all experiments was 3. Results On 14, 21, and 28 d after surgery, the wound healing area in combined treatment group, autologous skin+allogeneic skin group and allogeneic skin group of rats was significantly larger than that in fixed+allogeneic skin group (P<0.05), the wound healing area in autologous skin+allogeneic skin group of rats on 21 d after surgery and that in allogeneic skin group of rats on 14 and 21 d after surgery was significantly larger than that in combined treatment group (P<0.05), and the wound healing area in allogeneic skin group of rats on 14 d after surgery was significantly larger than that in autologous skin+allogeneic skin group (P<0.05). The wound healing area of rats in high exosome group and low exosome group on 14, 21, and 28 d after surgery and in supernatant group on 14 and 28 d after surgery were significantly larger than those in PBS group (P<0.05); the wound healing area in high exosome group of rats on 14 and 28 d after surgery was significantly larger than that in supernatant group (P<0.05), and that on 14 d after surgery was significantly larger than low exosome group (P<0.05); the wound healing area in low exosome group of rats on 14 d after surgery was significantly larger than that in supernatant group (P<0.05). Compared with that in PBS group, there were 332 differentially expressed proteins in the wound neo-epithelial tissue in high exosome group on 28 d after surgery (P<0.05), among which IgG1 heavy chain constant region IGHG1 and CSTA proteins were significantly up-regulated (with foldchange of 12.60 and 2.27, respectively, P<0.05). Compared with those of normal skin tissue in normal group, the differentially expressed proteins in the wound neo-epithelial tissue in high exosome group and PBS group of rats on 28 d after surgery were 1 400 and 1 057, respectively. The protein expressions of IGHG1 and CSTA in the wound neo-epithelial tissue in high exosome group of rats on 28 d after surgery were significantly larger than those in PBS group (P<0.05) and those of normal skin tissue of rats in normal group (P<0.05). Conclusions hUCMSC-ex may accelerate the repair process of wounds with scabbing and skin grafting and improve the wound healing quality in scalded rats by regulating the protein expressions of IGHG1 and CSTA.
- Mesenchymal stem cells,
- Exosomes,
- Skin transplantation,
- Proteomics,
- Escharectomy,
- Wound repair

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References(34)

References

[1]	MasonSA,NathensAB,ByrneJP,et al.Increased rate of long-term mortality among burn survivors: a population-based matched cohort study[J].Ann Surg,2019,269(6):1192-1199.DOI: 10.1097/SLA.0000000000002722.
[2]	中华医学会烧伤外科学分会.磨痂术在烧伤创面中的临床应用全国专家共识(2021版)[J].中华烧伤杂志,2021,37(6):501-507.DOI: 10.3760/cma.j.cn501120-20210110-00013.
[3]	YuhuaS,LigenL,JiakeC,et al.Effect of Poloxamer 188 on deepening of deep second-degree burn wounds in the early stage[J].Burns,2012,38(1):95-101.DOI: 10.1016/j.burns.2010.06.002.
[4]	KohlhauserM,LuzeH,NischwitzSP,et al.Historical evolution of skin grafting-a journey through time[J].Medicina (Kaunas),2021,57(4).DOI: 10.3390/medicina57040348.
[5]	汤文彬,李孝建,张志,等.大鼠全层皮肤缺损创面自、异体小皮片混合移植模型的建立[J].广东医学,2011,32(24):3173-3176.DOI: 10.3969/j.issn.1001-9448.2011.24.008.
[6]	IsaacR,ReisF,YingW,et al.Exosomes as mediators of intercellular crosstalk in metabolism[J].Cell Metab,2021,33(9):1744-1762.DOI: 10.1016/j.cmet.2021.08.006.
[7]	HaDH,KimHK,LeeJ,et al.Mesenchymal stem/stromal cell-derived exosomes for immunomodulatory therapeutics and skin regeneration[J].Cells,2020,9(5):1157.DOI: 10.3390/cells9051157.
[8]	LiY,JiangQL,Van der MerweL,et al.Preclinical efficacy of stem cell therapy for skin flap: a systematic review and meta-analysis[J].Stem Cell Res Ther,2021,12(1):28.DOI: 10.1186/s13287-020-02103-w.
[9]	沈括,王许杰,刘开拓,等.人脂肪间充质干细胞外泌体对小鼠RAW264.7细胞的炎症反应和小鼠全层皮肤缺损创面愈合的影响[J].中华烧伤与创面修复杂志,2022,38(3):215-226.DOI: 10.3760/cma.j.cn501120-20201116-00477.
[10]	ShariatiA,NematiR,SadeghipourY,et al.Mesenchymal stromal cells (MSCs) for neurodegenerative disease: a promising frontier[J].Eur J Cell Biol,2020,99(6):151097.DOI: 10.1016/j.ejcb.2020.151097.
[11]	MeuliM,Hartmann-FritschF,HügingM,et al.A cultured autologous dermo-epidermal skin substitute for full-thickness skin defects: a phase i, open, prospective clinical trial in children[J].Plast Reconstr Surg,2019,144(1):188-198.DOI: 10.1097/PRS.0000000000005746.
[12]	RenH,ZhaoF,ZhangQ,et al.Autophagy and skin wound healing[J/OL].Burns Trauma,2022,10:tkac003[2023-12-01].https://pubmed.ncbi.nlm.nih.gov/35187180/.DOI: 10.1093/burnst/tkac003.
[13]	陈瑞婧,冯韬锦,程实,等.3D培养人脐带间充质干细胞来源的外泌体对成骨细胞分化的作用[J].解放军医学杂志,2023,48(4):411-419.DOI: 10.11855/j.issn.0577-7402.2023.04.0411.
[14]	LiuW,RongY,WangJ,et al.Exosome-shuttled miR-216a-5p from hypoxic preconditioned mesenchymal stem cells repair traumatic spinal cord injury by shifting microglial M1/M2 polarization[J].J Neuroinflammation,2020,17(1):47.DOI: 10.1186/s12974-020-1726-7.
[15]	Nourian DehkordiA,Mirahmadi BabaheydariF,ChehelgerdiM,et al.Skin tissue engineering: wound healing based on stem-cell-based therapeutic strategies[J].Stem Cell Res Ther,2019,10(1):111.DOI: 10.1186/s13287-019-1212-2.
[16]	BianD,WuY,SongG,et al.The application of mesenchymal stromal cells (MSCs) and their derivative exosome in skin wound healing: a comprehensive review[J].Stem Cell Res Ther,2022,13(1):24.DOI: 10.1186/s13287-021-02697-9.
[17]	ViswanathanS,ShiY,GalipeauJ,et al.Mesenchymal stem versus stromal cells: International Society for Cell & Gene Therapy (ISCT^®) Mesenchymal Stromal Cell committee position statement on nomenclature[J].Cytotherapy,2019,21(10):1019-1024.DOI: 10.1016/j.jcyt.2019.08.002.
[18]	付文,王向臣,王延桂,等.脂肪源性间充质干细胞外泌体在大鼠全层皮肤缺损创面愈合中的机制研究[J].组织工程与重建外科杂志,2023,19(4):342-351,379.DOI: 10.3969/j.issn.1673-0364.2023.04.003.
[19]	KoikeY,YozakiM,UtaniA,et al.Fibroblast growth factor 2 accelerates the epithelial-mesenchymal transition in keratinocytes during wound healing process[J].Sci Rep,2020,10(1):18545.DOI: 10.1038/s41598-020-75584-7.
[20]	BashirovaAA,ZhengW,AkdagM,et al.Population-specific diversity of the immunoglobulin constant heavy G chain (IGHG) genes[J].Genes Immun,2021,22(7-8):327-334.DOI: 10.1038/s41435-021-00156-2.
[21]	LiY,WangP,YeD,et al.IGHG1 induces EMT in gastric cancer cells by regulating TGF-β/SMAD3 signaling pathway[J].J Cancer,2021,12(12):3458-3467.DOI: 10.7150/jca.56056.
[22]	LiY,YunX,LiJ,et al.CSTF2T up-regulates IGHG1 by binding to ZEB1 to promote melanoma cell proliferation, migration, and invasion[J].Tissue Cell,2023,81:102029.DOI: 10.1016/j.tice.2023.102029.
[23]	LiX,ChenW,YangC,et al.IGHG1 upregulation promoted gastric cancer malignancy via AKT/GSK-3β/β-Catenin pathway[J].Cancer Cell Int,2021,21(1):397.DOI: 10.1186/s12935-021-02098-1.
[24]	TianY,HanW,FuL,et al.Silencing of IGHG1 reverses the resistance of pancreatic cancer to multidrug chemotherapy by modulating autophagy[J].Environ Toxicol,2023,38(8):1835-1845.DOI: 10.1002/tox.23810.
[25]	MarconiGD,FonticoliL,RajanTS,et al.Epithelial-mesenchymal transition (EMT): the Type-2 EMT in wound healing, tissue regeneration and organ fibrosis[J].Cells,2021,10(7):1587. DOI: 10.3390/cells10071587.
[26]	TakahashiH,KomatsuN,IbeM,et al.Cystatin A suppresses ultraviolet B-induced apoptosis of keratinocytes[J].J Dermatol Sci,2007,46(3):179-187.DOI: 10.1016/j.jdermsci.2007.02.003.
[27]	GuptaA,NitoiuD,Brennan-CrispiD,et al.Cell cycle- and cancer-associated gene networks activated by DSG2: evidence of cystatin A deregulation and a potential role in cell-cell adhesion[J].PLoS One,2015,10(3):e0120091.DOI: 10.1371/journal.pone.0120091.
[28]	MaY,ChenY,LiY,et al.Cystatin A suppresses tumor cell growth through inhibiting epithelial to mesenchymal transition in human lung cancer[J].Oncotarget,2018,9(18):14084-14098.DOI: 10.18632/oncotarget.23505.
[29]	WangL,HuL,ZhouX,et al.Author Correction: Exosomes secreted by human adipose mesenchymal stem cells promote scarless cutaneous repair by regulating extracellular matrix remodelling[J].Sci Rep,2021,11(1):3245.DOI: 10.1038/s41598-021-82225-0.
[30]	吴涛,徐圣洁,李晓,等.外泌体在恶病质炎症反应中的作用及机制研究进展[J].解放军医学杂志,2023,48(5):602-608.DOI: 10.11855/j.issn.0577-7402.2023.05.0602.
[31]	MonyMP,HarmonKA,HessR,et al.An updated review of hypertrophic scarring[J].Cells,2023,12(5):678.DOI: 10.3390/cells12050678.
[32]	WangY,HuangB,JinT,et al.Intestinal fibrosis in inflammatory bowel disease and the prospects of mesenchymal stem cell therapy[J].Front Immunol,2022,13:835005.DOI: 10.3389/fimmu.2022.835005.
[33]	WuP,ZhangB,ShiH,et al.MSC-exosome: a novel cell-free therapy for cutaneous regeneration[J].Cytotherapy,2018,20(3):291-301.DOI: 10.1016/j.jcyt.2017.11.002.
[34]	YaoY,ChenR,WangG,et al.Exosomes derived from mesenchymal stem cells reverse EMT via TGF-β1/Smad pathway and promote repair of damaged endometrium[J].Stem Cell Res Ther,2019,10(1):225.DOI: 10.1186/s13287-019-1332-8.