Email Alerts
RSS
Volume 37 Issue 11
Nov.  2021
Turn off MathJax
Article Contents
Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  > 2021  >  37(11): 1061-1069
Huang SB,Hu ZC,Zhang Y,et al.Effects and mechanisms of allogeneic epidermal stem cells on the survival of allogeneic full-thickness skin grafts in nude mice with full-thickness skin defect wounds[J].Chin J Burns,2021,37(11):1061-1069.DOI: 10.3760/cma.j.cn501120-20200704-00339.
Citation: Huang SB,Hu ZC,Zhang Y,et al.Effects and mechanisms of allogeneic epidermal stem cells on the survival of allogeneic full-thickness skin grafts in nude mice with full-thickness skin defect wounds[J].Chin J Burns,2021,37(11):1061-1069.DOI: 10.3760/cma.j.cn501120-20200704-00339.
shu

Effects and mechanisms of allogeneic epidermal stem cells on the survival of allogeneic full-thickness skin grafts in nude mice with full-thickness skin defect wounds

doi: 10.3760/cma.j.cn501120-20200704-00339
  • 1.

    Department of Burns, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China

  • 2.

    Department of Burns and Plastic Surgery, Affiliated Hospital of Nantong University, Nantong 226001, China

  • 3.

    Department of Burns and Plastic Surgery, the First Affiliated Hospital of Shenzhen University, Shenzhen 518037, China

Funds:

General Program of National Natural Science Foundation of China 81871565

Science and Technology Planning Project of Guangdong Province of China 2016B090916001

Foundation for Basic and Applied Basic Research of Guangdong Province of China 2019A1515012208

Sun Yat-sen University Clinical Research 5010 Program 2013001, 2018003

Fundamental Scientific Research of Sun Yat-sen University 19ykpy66

More Information
  • Corresponding author: Zhu Jiayuan, Email: zhujiay@mail.sysu.edu.cn
  • Received Date: 2020-07-04
    Abstract
  •   Objective  To investigate the effects and mechanisms of allogeneic epidermal stem cells (ESCs) on the survival of allogeneic full-thickness skin grafts in nude mice with full-thickness skin defect wounds.  Methods  Experimental research methods were applied. Primary ESCs that appeared paving stone-like after being cultured for 7 d were obtained by enzymatic digestion method from one 4-week-old male BALB/c-NU nude mouse (the same strain, age, and sex below). The cells of third passage were identified by flow cytometry to positively express ESC marker CD44 and negatively express CD45, meanwhile, the positive expression of ESC markers of p63 and integrin 6α, and negative expression of CD71 were identified by immunofluorescence method. The ESCs of third passage in the logarithmic growth phase were used for the following experiments. Twenty-six nude mice were equally divided into phosphate buffered saline (PBS) group and ESCs group according to the random number table. A full-thickness skin defect wound was made on the back of each nude mouse, and then the wounds of the two groups were sprayed with equal volumes of PBS and ESCs, respectively. The wounds were transplanted with full-thickness skin grafts cut from the backs of 4 other nude mice. Each ten nude mice from the two groups were selected, the wound healing and skin survival on post surgery day (PSD) 0 (immediately), 3, 7, 14, and 21 were observed, and the survival ratio and shrinkage rate of skin grafts on PSD 3, 7, 14, and 21 were calculated (the number of sample was the number of surviving skin grafts at each time point); the blood perfusion in the skin grafts on PSD 3, 7, and 14 was detected by the laser speckle blood flow imager, and the blood flow ratio of nude mice skin grafts in ESCs group to PBS group at each time point was calculated (the number of sample was the pair number of surviving skin grafts in group pairing at each time point). The skin graft tissue of each 3 nude mice remained in the two groups were collected on PSD 7, and the mRNA expressions and protein expressions of tumor necrosis factor α (TNF-α), interleukin 8 (IL-8), IL-10, type Ⅰ collagen, type Ⅲ collagen, and matrix metalloproteinase 9 (MMP-9) in the tissue were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction and Western blotting, respectively. Data were statistically analyzed with Log-rank test, analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, and Bonferroni correction.  Results  Taking the condition on PSD 0 as a reference, the wounds of nude mice in the two groups healed gradually on PSD 3, 7, 14, and 21, and the shrinkage of skin grafts was gradually obvious. Among them, the shrinkage healing of wound of nude mice in PBS group was more significant than that in ESCs group. On PSD 3, the skin graft of 1 nude mouse failed in ESCs group, while the skin graft of 3 nude mice failed in PBS group. On PSD 7, the skin graft of another nude mouse failed in PBS group. The survival ratio of skin grafts of nude mice in the two groups was similar on PSD 3, 7, 14, and 21 (P>0.05). On PSD 3, 7, 14, and 21, the shrinkage rates of skin grafts of nude mice in ESCs group were (9.2±0.4)%, (19.7±1.2)%, (53.6±3.5)%, and (62.2±5.1)%, respectively, which was significantly lower than (11.0±0.9)%, (47.8±2.8)%, (86.1±7.1)%, and (89.7±9.0)% in PBS group (t=5.719, 26.650, 11.940, 7.617, P<0.01). On PSD 3, 7, and 14, blood perfusion signals were observed in the skin grafts of nude mice in the two groups. The average blood perfusion ratios of the skin grafts of nude mice in ESCs group to PBS group were greater than 1, and there was no statistically significant difference in the overall comparison of 3 time points (P>0.05). On PSD 7, compared with those of PBS group, the mRNA and protein expressions of TNF-α, IL-8, type Ⅰ collagen, and type Ⅲ collagen in the skin graft tissue of nude mice in ESCs group were significantly reduced, while the mRNA and protein expressions of IL-10 and MMP-9 in the skin graft tissue of nude mice in ESCs group were significantly increased (in mRNA comparison, t=2.823, 2.934, 2.845, 2.860, 3.877, 2.916, P<0.05).  Conclusions  Allogeneic ESCs can reduce the shrinkage of allogeneic full-thickness skin grafts transplanted on full-thickness skin defect wounds in nude mice, promote the formation of new blood vessels between the skin graft and the wound, reduce inflammation and collagen protein expression, and promote the expression of MMP-9, thus improving the survival quality of skin grafts.

     

    • FullText(HTML)
  • loading
    • References(33)
  • [1]
    AkitaS.Wound repair and regeneration: mechanisms, signaling[J].Int J Mol Sci,2019,20(24):6328.DOI: 10.3390/ijms20246328.
    [2]
    WerdinF,TenenhausM,RennekampffHO.Chronic wound care[J].Lancet,2008,372(9653):1860-1862.DOI: 10.1016/S0140-6736(08)61793-6.
    [3]
    SorgH,TilkornDJ,HagerS,et al.Skin wound healing: an update on the current knowledge and concepts[J].Eur Surg Res,2017,58(1/2):81-94.DOI: 10.1159/000454919.
    [4]
    KaplaniK,KoutsiS,ArmenisV,et al.Wound healing related agents: ongoing research and perspectives[J].Adv Drug Deliv Rev,2018,129:242-253.DOI: 10.1016/j.addr.2018.02.007.
    [5]
    AndreassiA,BilenchiR,BiagioliM,et al.Classification and pathophysiology of skin grafts[J].Clin Dermatol,2005,23(4):332-337.DOI: 10.1016/j.clindermatol.2004.07.024.
    [6]
    ConverseJM,SmahelJ,BallantyneDLJr,et al.Inosculation of vessels of skin graft and host bed: a fortuitous encounter[J].Br J Plast Surg,1975,28(4):274-282.DOI: 10.1016/0007-1226(75)90031-4.
    [7]
    GkotsouliasE.Split thickness skin graft of the foot and ankle bolstered with negative pressure wound therapy in a diabetic population: the results of a retrospective review and review of the literature[J].Foot Ankle Spec,2020,13(5):383-391.DOI: 10.1177/1938640019863267.
    [8]
    BoyceST,SimpsonPS,RiemanMT,et al.Randomized, paired-site comparison of autologous engineered skin substitutes and split-thickness skin graft for closure of extensive, full-thickness burns[J].J Burn Care Res,2017,38(2):61-70.DOI: 10.1097/BCR.0000000000000401.
    [9]
    TakabayashiY,IshiharaM,KuwabaraM,et al.Improved survival of full-thickness skin graft with low-molecular weight heparin-protamine micro/nanoparticles including platelet-rich plasma[J].Ann Plast Surg,2017,78(5):562-568.DOI: 10.1097/SAP.0000000000001051.
    [10]
    NicaO,PopaDG,GrecuAF,et al.Histological aspects of full-thickness skin grafts augmented with platelet-rich fibrin in rat model[J].Rom J Morphol Embryol,2019,60(2):581-588.
    [11]
    BehrB,KoSH,WongVW,et al.Stem cells[J].Plast Reconstr Surg,2010,126(4):1163-1171.DOI: 10.1097/PRS.0b013e3181ea42bb.
    [12]
    KiranaS,StratmannB,PranteC,et al.Autologous stem cell therapy in the treatment of limb ischaemia induced chronic tissue ulcers of diabetic foot patients[J].Int J Clin Pract,2012,66(4):384-393.DOI: 10.1111/j.1742-1241.2011.02886.x.
    [13]
    MarandaEL,Rodriguez-MenocalL,BadiavasEV.Role of mesenchymal stem cells in dermal repair in burns and diabetic wounds[J].Curr Stem Cell Res Ther,2017,12(1):61-70.DOI: 10.2174/1574888x11666160714115926.
    [14]
    KølleSF,Fischer-NielsenA,MathiasenAB,et al.Enrichment of autologous fat grafts with ex-vivo expanded adipose tissue-derived stem cells for graft survival: a randomised placebo-controlled trial[J].Lancet,2013,382(9898):1113-1120.DOI: 10.1016/S0140-6736(13)61410-5.
    [15]
    TengM,HuangYS,ZhangHS.Application of stems cells in wound healing--an update[J].Wound Repair Regen,2014,22(2):151-160.DOI: 10.1111/wrr.12152.
    [16]
    ChenCY,RaoSS,RenL,et al.Exosomal DMBT1 from human urine-derived stem cells facilitates diabetic wound repair by promoting angiogenesis[J].Theranostics,2018,8(6):1607-1623.DOI: 10.7150/thno.22958.
    [17]
    LiZ,MaitzP.Cell therapy for severe burn wound healing[J/OL].Burns Trauma,2018,6:13[2020-07-04].http://burnstrauma.biomedcentral.com/track/pdf/ 10.1186/s41038-018-0117-0.DOI: 10.1186/s41038-018-0117-0.
    [18]
    ShawkyLM,El BanaEA,MorsiAA.Stem cells and metformin synergistically promote healing in experimentally induced cutaneous wound injury in diabetic rats[J].Folia Histochem Cytobiol,2019,57(3):127-138.DOI: 10.5603/FHC.a2019.0014.
    [19]
    WangP,HuZC,CaoXL,et al.Fibronectin precoating wound bed enhances the therapeutic effects of autologous epidermal basal cell suspension for full-thickness wounds by improving epidermal stem cells' utilization[J].Stem Cell Res Ther,2019,10(1):154.DOI: 10.1186/s13287-019-1236-7.
    [20]
    HuangSB,HuZC,WangP,et al.Rat epidermal stem cells promote the angiogenesis of full-thickness wounds[J].Stem Cell Res Ther,2020,11(1):344.DOI: 10.1186/s13287-020-01844-y.
    [21]
    HarrisJE,DhupaS.Treatment of degloving injuries with autogenous full thickness mesh scrotal free grafts[J].Vet Comp Orthop Traumatol,2008,21(4):378-381.DOI: 10.3415/vcot-07-04-0029.
    [22]
    ZografouA,TsigrisC,PapadopoulosO,et al.Improvement of skin-graft survival after autologous transplantation of adipose-derived stem cells in rats[J].J Plast Reconstr Aesthet Surg,2011,64(12):1647-1656.DOI: 10.1016/j.bjps.2011.07.009.
    [23]
    XiaoSE,LiuZY,YaoYZ,et al.Diabetic human adipose-derived stem cells accelerate pressure ulcer healing by inducing angiogenesis and neurogenesis[J].Stem Cells Dev,2019,28(5):319-328.DOI: 10.1089/scd.2018.0245.
    [24]
    PhinneyDG,PittengerMF.Concise review: MSC-derived exosomes for cell-free therapy[J].Stem Cells,2017,35(4):851-858.DOI: 10.1002/stem.2575.
    [25]
    LeeDE,AyoubN,AgrawalDK.Mesenchymal stem cells and cutaneous wound healing: novel methods to increase cell delivery and therapeutic efficacy[J].Stem Cell Res Ther,2016,7:37.DOI: 10.1186/s13287-016-0303-6.
    [26]
    RustadKC,WongVW,SorkinM,et al.Enhancement of mesenchymal stem cell angiogenic capacity and stemness by a biomimetic hydrogel scaffold[J].Biomaterials,2012,33(1):80-90.DOI: 10.1016/j.biomaterials.2011.09.041.
    [27]
    BadiavasEV,FalangaV.Treatment of chronic wounds with bone marrow-derived cells[J].Arch Dermatol,2003,139(4):510-516.DOI: 10.1001/archderm.139.4.510.
    [28]
    ZhugeY,RegueiroMM,TianRX,et al.The effect of estrogen on diabetic wound healing is mediated through increasing the function of various bone marrow-derived progenitor cells[J].J Vasc Surg,2018,68(6S): S127-135.DOI: 10.1016/j.jvs.2018.04.069.
    [29]
    MesaKR,KawaguchiK,CockburnK,et al.Homeostatic epidermal stem cell self-renewal is driven by local differentiation[J].Cell Stem Cell,2018,23(5):677-686.e4.DOI: 10.1016/j.stem.2018.09.005.
    [30]
    D'ArcangeloD,TinaburriL,DellambraE.The role of p16INK4a pathway in human epidermal stem cell self-renewal, aging and cancer[J].Int J Mol Sci,2017,18(7):1591.DOI: 10.3390/ijms18071591.
    [31]
    JacksonCJ,TønsethKA,UtheimTP.Cultured epidermal stem cells in regenerative medicine[J].Stem Cell Res Ther,2017,8(1):155.DOI: 10.1186/s13287-017-0587-1.
    [32]
    HuZ,GuoD,LiuP,et al.Randomized clinical trial of autologous skin cell suspension for accelerating re-epithelialization of split-thickness donor sites[J].Br J Surg,2017,104(7):836-842.DOI: 10.1002/bjs.10508.
    [33]
    HuZC,ChenD,GuoD,et al.Randomized clinical trial of autologous skin cell suspension combined with skin grafting for chronic wounds[J].Br J Surg,2015,102(2):e117-123.DOI: 10.1002/bjs.9688.
    • Relative Articles
    • Supplements(0)
    • Cited By
  • 加载中

Catalog

    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Figures(7)  / Tables(3)

    Get Citation
    PDF
    XML

    Article Metrics

    Article views (280) PDF downloads(20) Cited by()
    Proportional views
    Related

    Copyright © Chinese Journal of Burns京ICP备07035254号-14

    E-mail：shaoshangzazhi@163.com

    Website：https://zhsszz.xml-journal.net/

    Supported by: Beijing Renhe Information Technology Co. Ltd    

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return