Effects of recombinant human metallothionein-Ⅲ combined with wound dressing on wound healing of full-thickness skin defects in mice

Shen Xin; Sun Zuoyi; Zhang Rui; Xue Yuying

doi:10.3760/cma.j.cn501225-20231031-00164

Volume 40 Issue 5

May 2024

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > 40(5): 425-432

Shen X,Sun ZY,Zhang R,et al.Effects of recombinant human metallothionein-Ⅲ combined with wound dressing on wound healing of full-thickness skin defects in mice[J].Chin J Burns Wounds,2024,40(5):425-432.DOI: 10.3760/cma.j.cn501225-20231031-00164.

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Effects of recombinant human metallothionein-Ⅲ combined with wound dressing on wound healing of full-thickness skin defects in mice

doi: 10.3760/cma.j.cn501225-20231031-00164

Key Laboratory of Environmental Medicine and Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing 210009, China

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General Program of National Natural Science Foundation of China 82273674

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Corresponding author: Xue Yuying, Email: yyxue@seu.edu.cn
Received Date: 2023-10-31

Abstract

Abstract

Objective To investigate the effects of recombinant human metallothionein-Ⅲ (rh-MT-Ⅲ) combined with wound dressing on wound healing of full-thickness skin defects in mice. Methods This study was an experimental study. Twenty-four half male and half female 6 weeks old Institute of Cancer Research mice were taken, and two symmetrical round full-thickness skin defect wounds were prepared on the back of each mouse. The mice were stratified and randomly divided into normal saline group, dressing group, rh-MT-Ⅲgroup (applying the corresponding solution on the wounds), and combined treatment group (applying a mixture of rh-MT-Ⅲ and wound dressing on the wounds) according to sex and body weight, with 6 mice in each group. From 1 to 7 d after injury, all mice were observed daily for changes in activity, diet, and fur growth, their body weights and wound areas were recorded, and the relative wound area percentages were calculated. On 7 d after injury, the wound tissue of mice was taken for hematoxylin-eosin staining to observe the newborn granulation tissue, for Masson staining to observe collagen fiber formation, and for immunofluorescence staining to detect cell proliferation (denoted as Ki67 relative fluorescence intensity) and cell apoptosis (denoted as TdT-mediated dUTP nick end labeling (TUNEL) relative fluorescence intensity). The sample size in the above experiments was 6. Results There were no abnormalities in activity, diet, or fur growth in the 4 groups of mice within 7 d after injury. There were no statistically significant differences in the overall comparison of the body weights of mice in the 4 groups from 1 to 7 d after injury (P>0.05). The relative wound area percentages of mice in combined treatment group were significantly lower than those in normal saline group and rh-MT-Ⅲ group on 2, 3, 4, 5, 6, and 7 d after injury (P<0.05), and the relative wound area percentages of mice in combined treatment group were significantly lower than those in dressing group on 3, 4, 5, 6, and 7 d after injury (P<0.05). The relative wound area percentages of mice in dressing group on 6 and 7 d after injury and in rh-MT-Ⅲ group on 7 d after injury were significantly lower than those in normal saline group (P<0.05). On 7 d after injury, a large number of capillaries and fibroblasts could be seen in wound tissue of mice in combined treatment group, and the growth of new epithelial tissue at the upper edge of the wound was better than that of the other three groups; the collagen fibers in the wound tissue of mice in combined treatment group had higher degree of density and arrangement in a more orderly manner than those of the other three groups. On 7 d after injury, the Ki67 relative fluorescence intensity in the wound tissue of mice in dressing group, rh-MT-Ⅲ group, and combined treatment group was (289±35)%, (197±17)%, and (389±56)%, which was significantly higher than (100±15)% in normal saline group, respectively (P<0.05), and the Ki67 relative fluorescence intensity in the wound tissue of mice in combined treatment group was significantly higher than that in dressing group and rh-MT-Ⅲ group, respectively (with both P values <0.05). On 7 d after injury, the TUNEL relative fluorescence intensity in the wound tissue of mice in dressing group, rh-MT-Ⅲ group, and combined treatment group was (55.5±5.7)%, (66.7±8.0)%, and (20.0±2.2)%, which was significantly lower than (100.0±12.9)% in normal saline group, respectively (P<0.05), and the TUNEL relative fluorescence intensity in the wound tissue of mice in combined treatment group was significantly lower than that in dressing group and rh-MT-Ⅲ group, respectively (with both P values <0.05). Conclusions rh-MT-Ⅲ combined with wound dressing can promote the growth of granulation tissue around the wound as well as collagen deposition, increase the cell proliferation vitality, reduce cell apoptosis, and promote the re-epithelialization of skin at the edge of the wounds, thus accelerating the healing of full-thickness skin defect wounds in mice.
- Wounds and injuries,
- Metallothionein,
- Biological dressings,
- Cell proliferation,
- Apoptosis,
- Wound repair

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References

[1]	王布雷白及多糖水凝胶促皮肤伤口愈合作用研究西安陕西师范大学2022DOI: 10.27292/d.cnki.gsxfu.2021.001160 王布雷. 白及多糖水凝胶促皮肤伤口愈合作用研究 [D]. 西安:陕西师范大学, 2022. DOI: 10.27292/d.cnki.gsxfu.2021.001160.
[2]	GuerraA,BelinhaJ,JorgeRN.Modelling skin wound healing angiogenesis: a review[J].J Theor Biol,2018,459:1-17.DOI: 10.1016/j.jtbi.2018.09.020.
[3]	KagiJH,ValeeBL.Metallothionein: a cadmium- and zinc-containing protein from equine renal cortex[J].J Biol Chem,1960,235:3460-3465.
[4]	LiuAL,ZhangZM,ZhuBF,et al.Metallothionein protects bone marrow stromal cells against hydrogen peroxide-induced inhibition of osteoblastic differentiation[J].Cell Biol Int,2004,28(12):905-911.DOI: 10.1016/j.cellbi.2004.09.004.
[5]	CherianMG,JayasuryaA,BayBH.Metallothioneins in human tumors and potential roles in carcinogenesis[J].Mutat Res,2003,533(1/2):201-209.DOI: 10.1016/j.mrfmmm.2003.07.013.
[6]	CoyleP,PhilcoxJC,CareyLC,et al.Metallothionein: the multipurpose protein[J].Cell Mol Life Sci,2002,59(4):627-647.DOI: 10.1007/s00018-002-8454-2.
[7]	何婧雯,熊海容,王靖,等.金属硫蛋白的研究现状及进展[J].农产品加工(上半月),2019(9):72-75. DOI: 10.16693/j.cnki.1671-9646(X).2019.09.018.
[8]	YouHJ,LeeKJ,JeongHG.Overexpression of human metallothionein-III prevents hydrogen peroxide-induced oxidative stress in human fibroblasts[J].FEBS Lett,2002,521(1/2/3):175-179.DOI: 10.1016/s0014-5793(02)02870-3.
[9]	燕艳,季志会,杜伟,等.金属硫蛋白应用研究进展[J].东北农业大学学报,2010,41(7):150-154.DOI: 10.3969/j.issn.1005-9369.2010.07.030.
[10]	ZhangW,XieY,LiuW,et al.Role of metallothionein in post-burn inflammation[J].Inflammation,2016,39(2):768-774.DOI: 10.1007/s10753-016-0305-7.
[11]	LansdownAB.Metallothioneins: potential therapeutic aids for wound healing in the skin[J].Wound Repair Regen,2002,10(3):130-132.DOI: 10.1046/j.1524-475x.2002.20101.x.
[12]	薛燕.金属硫蛋白在皮肤疾病中的研究进展[J].中国美容医学,2013,22(17):1823-1826.DOI: 10.3969/j.issn.1008-6455.2013.17.031.
[13]	徐连春,尚剑,孙晔,等.银纳米颗粒及载银抗菌涂层的研究与进展[J].中国组织工程研究,2016,20(25):3793-3800.DOI: 10.3969/j.issn.2095-4344.2016.25.022.
[14]	AlemdaroğluC,DeğimZ,CelebiN,et al.An investigation on burn wound healing in rats with chitosan gel formulation containing epidermal growth factor[J].Burns,2006,32(3):319-327.DOI: 10.1016/j.burns.2005.10.015.
[15]	王宏宇, 巴特, 周彪, 等. 不同途径应用人脐带间充质干细胞外泌体治疗小鼠全层皮肤缺损创面的效果[J]. 中华烧伤与创面修复杂志, 2024, 40(4): 314-322. DOI: 10.3760/cma.j.cn501225-20231123-00203.
[16]	姚梦云,张宁,张庆,等.白细胞介素4修饰的金纳米酶对糖尿病小鼠全层皮肤缺损的作用[J].中华烧伤与创面修复杂志,2023,39(1):15-24.DOI: 10.3760/cma.j.cn501225-20220630-00275.
[17]	谢军,毛玉洁,王思宇,等.紫草素对大鼠慢性皮肤溃疡创面愈合及新生血管形成的促进作用及其机制[J].解放军医学杂志,2022,47(1):39-45.DOI: 10.11855/j.issn.0577-7402.2022.01.0039.
[18]	PenkowaM,CarrascoJ,GiraltM,et al.CNS wound healing is severely depressed in metallothionein I- and II-deficient mice[J].J Neurosci,1999,19(7):2535-2545.DOI: 10.1523/JNEUROSCI.19-07-02535.1999.
[19]	MorelliniNM,GilesNL,ReaS,et al.Exogenous metallothionein-IIA promotes accelerated healing after a burn wound[J].Wound Repair Regen,2008,16(5):682-690.DOI: 10.1111/j.1524-475X.2008.00418.x.
[20]	AignerGP,PeerV,FiechtnerB,et al.Wound healing and Cadmium detoxification in the earthworm Lumbricus terrestris - a potential case for coelomocytes?[J].Front Immunol,2023,14:1272191.DOI: 10.3389/fimmu.2023.1272191.
[21]	SunZ,QinJ,YuanH,et al.Recombinant human metallothionein-III alleviates oxidative damage induced by copper and cadmium in Caenorhabditis elegans[J].J Appl Toxicol,2023,43(8):1242-1252.DOI: 10.1002/jat.4460.
[22]	Cortese-KrottMM,MünchowM,PirevE,et al.Silver ions induce oxidative stress and intracellular zinc release in human skin fibroblasts[J].Free Radic Biol Med,2009,47(11):1570-1577.DOI: 10.1016/j.freeradbiomed.2009.08.023.
[23]	PavlíkV,SobotkaL,PejchalJ,et al.Silver distribution in chronic wounds and the healing dynamics of chronic wounds treated with dressings containing silver and octenidine[J].FASEB J,2021,35(5):e21580.DOI: 10.1096/fj.202100065R.
[24]	LinX,JagadapillaiR,CaiJ,et al.Metallothionein induction attenuates the progression of lung injury in mice exposed to long-term intermittent hypoxia[J].Inflamm Res,2020,69(1):15-26.DOI: 10.1007/s00011-019-01287-z.
[25]	GurtnerGC, WernerS, BarrandonY, et al. Wound repair and regeneration[J]. Nature, 2008,453(7193):314-321. DOI: 10.1038/nature07039.
[26]	WangZ,ZhaoF,XuC,et al.Metabolic reprogramming in skin wound healing[J/OL].Burns Trauma,2024,12:tkad047[2023-10-31].https://pubmed.ncbi.nlm.nih.gov/38179472/.DOI: 10.1093/burnst/tkad047.
[27]	ManchandaM,TorresM,InuossaF,et al.Metabolic reprogramming and reliance in human skin wound healing[J].J Invest Dermatol,2023,143(10):2039-2051.e10.DOI: 10.1016/j.jid.2023.02.039.
[28]	EmingSA,MurrayPJ,PearceEJ.Metabolic orchestration of the wound healing response[J].Cell Metab,2021,33(9):1726-1743.DOI: 10.1016/j.cmet.2021.07.017.
[29]	韩莹,时玉峥,魏训东,等.Ki67法检测间充质干细胞对淋巴细胞增殖抑制能力[J].药物评价研究,2022,45(4):673-679.DOI: 10.7501/j.issn.1674-6376.2022.04.009.
[30]	李丽,牛钰清,陈菲,等.介绍一种细胞凋亡TUNEL检测与Ki-67 DAPI三色荧光染色操作流程[J].临床与实验病理学杂志,2019,35(8):980-981.DOI: 10.13315/j.cnki.cjcep.2019.08.026.
[31]	ChowdhuryD,AlrefaiH,Landero FigueroaJA,et al.Metallothionein 3 controls the phenotype and metabolic programming of alternatively activated macrophages[J].Cell Rep,2019,27(13):3873-3886.e7.DOI: 10.1016/j.celrep.2019.05.093.
[32]	ÅgrenMS,ChafranskaL,EriksenJO,et al.Spatial expression of metallothionein, matrix metalloproteinase-1 and Ki-67 in human epidermal wounds treated with zinc and determined by quantitative immunohistochemistry: a randomised double-blind trial[J].Eur J Cell Biol,2021,100(3):151147.DOI: 10.1016/j.ejcb.2020.151147.

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Effects of recombinant human metallothionein-Ⅲ combined with wound dressing on wound healing of full-thickness skin defects in mice

doi: 10.3760/cma.j.cn501225-20231031-00164

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Effects of recombinant human metallothionein-Ⅲ combined with wound dressing on wound healing of full-thickness skin defects in mice

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