Influence and mechanism of bone marrow mesenchymal stem cells overexpressing growth arrest specific 6 on full-thickness skin defect wounds in diabetic mice

Liu Pei; Wang Chao; Wei Qijian; Li Yuteng; Cui Lijun; Wang Changchuan; Zhang Fan; Ma Ling; Tian Xuan

doi:10.3760/cma.j.cn501225-20241024-00409

Volume 41 Issue 2

Feb. 2025

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Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2025 > 41(2): 145-154

Jiang WQ,Pan F,Chai M,et al.A prospective study on the development and application verification of the quantitative evaluation software for three-dimensional morphology of pathological scars based on photo modeling technology[J].Chin J Burns Wounds,2023,39(2):158-164.DOI: 10.3760/cma.j.cn501225-20220513-00184.

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Liu P,Wang C,Wei QJ,et al.Influence and mechanism of bone marrow mesenchymal stem cells overexpressing growth arrest specific 6 on full-thickness skin defect wounds in diabetic mice[J].Chin J Burns Wounds,2025,41(2):145-154.DOI: 10.3760/cma.j.cn501225-20241024-00409.

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Influence and mechanism of bone marrow mesenchymal stem cells overexpressing growth arrest specific 6 on full-thickness skin defect wounds in diabetic mice

doi: 10.3760/cma.j.cn501225-20241024-00409

1.
Department of Plastic Surgery, Qilu Hospital of Shandong University, Jinan 250012, China
2.
Department of Surgery, Mudan District Hospital of Traditional Chinese Medicine of Heze City, Heze 274000, China

Funds:

Youth Science Fund Program of National Natural Science Foundation of China 82400767

Youth Fund Program of Shandong Provincial Natural Science Foundation ZR2023QH243

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Corresponding author: Tian Xuan, Email: tianxuan0530@foxmail.com
Received Date: 2024-10-24

Abstract

Abstract

Objective To investigate the influence and mechanism of bone marrow mesenchymal stem cells (BMSCs) overexpressing growth arrest specific 6, i.e. GAS6/BMSCs on full-thickness skin defect wounds in diabetic mice. Methods This study was an experimental study. Twelve 8-week-old male C57BL/6J mice were divided into a control wound group with only full-thickness skin defects and a diabetic wound group with diabetic full-thickness skin defects according to the random number table method, with 6 mice in each group. The wound healing rates were calculated at 3, 7, 14, and 21 days after injury. At 21 days after injury, wound tissue specimens were collected for hematoxylin-eosin staining to observe the histopathological conditions; Masson staining was performed to detect collagen deposition; immunohistochemical staining was performed to detect the number of proliferating cell nuclear antigen (PCNA)-positive cells and CD31-positive cells, representing cell proliferation and capillary density, respectively; immunofluorescence staining was performed to detect the number of F4/80 and myeloperoxidase (MPO) double-positive cells, indicating efferocytosis. Two 4-week-old male C57BL/6J mice were used to extract BMSCs, and GAS6/BMSCs were constructed through adenovirus transfection and successfully identified. Eighteen 8-week-old male C57BL/6J mice were used to create diabetic full-thickness skin defect wound models and divided into phosphate buffered solution (PBS) group, BMSC group, and GAS6/BMSC group (with 6 mice in each group) according to the random number table method. Immediately after injury, PBS, BMSC single-cell suspension, and GAS6/BMSC single-cell suspension were injected locally into the wounds of the three groups of mice, respectively. The wound healing rates were calculated, and the cell proliferation, capillary density, and efferocytosis were detected at the same time points as the previous experiments. Results At 3, 7, 14, and 21 days after injury, the wound healing rates of mice in diabetic wound group were significantly lower than those in control wound group (with t values of 7.99, 8.62, 9.80, and 5.85, respectively, P<0.05). Compared with those in control wound group, the wound tissue of mice in diabetic wound group showed the infiltration of a large number of inflammatory cells and reduced collagen deposition at 21 days after injury. At 21 days after injury, the number of PCNA-positive cells and CD31-positive cells in the wound tissue of mice in diabetic wound group were significantly less than that in control wound group (with t values of 6.61 and 5.38, respectively, P<0.05). At 21 days after injury, the number of F4/80 and MPO double-positive cells in the wound tissue of mice in diabetic wound group was 3.3±0.8, which was significantly less than 12.7±1.8 in control wound group (t=11.00, P<0.05). At 14 and 21 days after injury, the wound healing rates of mice in BMSC group were significantly higher than those in PBS group (P<0.05); at 3, 7, 14, and 21 days after injury, the wound healing rates of mice in GAS6/BMSC group were significantly higher than those in BMSC group (P<0.05). At 21 days after injury, the number of PCNA-positive cells in the wound tissue of mice in BMSC group was significantly higher than that in PBS group (P<0.05), and the number of PCNA-positive cells and CD31-positive cells in the wound tissue of mice in GAS6/BMSC group were significantly higher than that in BMSC group (P<0.05). At 21 days after injury, the number of F4/80 and MPO double-positive cells in the wound tissue of mice in BMSC group was 4.2±1.2, which was similar to 3.5±1.1 in PBS group (P>0.05); the number of F4/80 and MPO double-positive cells in the wound tissue of mice in GAS6/BMSC group was 8.2±1.2, which was significantly more than that in BMSC group (P<0.05). Conclusions Dysfunctional efferocytosis of macrophage exists in the full-thickness skin defect wounds of diabetic mice, while GAS6/BMSC can promote wound healing by restoring the efferocytosis of macrophages.
- Diabetes mellitus, type 2,
- Ulcer,
- Mesenchymal stem cells,
- Macrophages,
- Efferocytosis,
- Growth arrest specific 6,
- Wound repair

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References

[1]	McDermottK,FangM,BoultonAJM,et al.Etiology, epidemiology, and disparities in the burden of diabetic foot ulcers[J].Diabetes Care,2023,46(1):209-221.DOI: 10.2337/dci22-0043.
[2]	阮琼芳,章思语,席毛毛,等.糖尿病足患者创缘皮肤组织中成纤维细胞与角质形成细胞的相互作用及其机制[J].中华烧伤与创面修复杂志,2024,40(8):762-771.DOI: 10.3760/cma.j.cn501225-20240221-00067.
[3]	ChenT,SongP,HeM,et al.Sphingosine-1-phosphate derived from PRP-Exos promotes angiogenesis in diabetic wound healing via the S1PR1/AKT/FN1 signalling pathway[J/OL].Burns Trauma,2023,11:tkad003[2024-10-24].https://pubmed.ncbi.nlm.nih.gov/37251708/.DOI: 10.1093/burnst/tkad003.
[4]	ShepherdHM,GauthierJM,TeradaY,et al.Updated views on neutrophil responses in ischemia-reperfusion injury[J].Transplantation,2022,106(12):2314-2324.DOI: 10.1097/TP.0000000000004221.
[5]	KaltenmeierC,YazdaniHO,HanduS,et al.The role of neutrophils as a driver in hepatic ischemia-reperfusion injury and cancer growth[J].Front Immunol,2022,13:887565.DOI: 10.3389/fimmu.2022.887565.
[6]	Boada-RomeroE,MartinezJ,HeckmannBL,et al.The clearance of dead cells by efferocytosis[J].Nat Rev Mol Cell Biol,2020,21(7):398-414.DOI: 10.1038/s41580-020-0232-1.
[7]	MehrotraP,RavichandranKS.Drugging the efferocytosis process: concepts and opportunities[J].Nat Rev Drug Discov,2022,21(8):601-620.DOI: 10.1038/s41573-022-00470-y.
[8]	WangJ,HossainM,ThanabalasuriarA,et al.Visualizing the function and fate of neutrophils in sterile injury and repair[J].Science,2017,358(6359):111-116.DOI: 10.1126/science.aam9690.
[9]	SakuragiT,NagataS.Regulation of phospholipid distribution in the lipid bilayer by flippases and scramblases[J].Nat Rev Mol Cell Biol,2023,24(8):576-596.DOI: 10.1038/s41580-023-00604-z.
[10]	Zdżalik-BieleckaD,PoświataA,KozikK,et al. The GAS6-AXL signaling pathway triggers actin remodeling that drives membrane ruffling, macropinocytosis, and cancer-cell invasion[J].Proc Natl Acad Sci U S A,2021,118(28):e2024596118.DOI: 10.1073/pnas.2024596118.
[11]	NepalS,TiruppathiC,TsukasakiY,et al.STAT6 induces expression of Gas6 in macrophages to clear apoptotic neutrophils and resolve inflammation[J].Proc Natl Acad Sci U S A,2019,116(33):16513-16518.DOI: 10.1073/pnas.1821601116.
[12]	TangQ,DongC,SunQ.Immune response associated with ischemia and reperfusion injury during organ transplantation[J].Inflamm Res,2022,71(12):1463-1476.DOI: 10.1007/s00011-022-01651-6.
[13]	YaoZ,QiW,ZhangH,et al.Down-regulated GAS6 impairs synovial macrophage efferocytosis and promotes obesity-associated osteoarthritis[J].Elife,2023,12:e83069.DOI: 10.7554/eLife.83069.
[14]	KalluriR,LeBleuVS.The biology, function, and biomedical applications of exosomes[J].Science,2020,367(6478):eaau6977.DOI: 10.1126/science.aau6977.
[15]	PiL,YangL,FangBR,et al.Exosomal microRNA-125a-3p from human adipose-derived mesenchymal stem cells promotes angiogenesis of wound healing through inhibiting PTEN[J].Mol Cell Biochem,2022,477(1):115-127.DOI: 10.1007/s11010-021-04251-w.
[16]	ChristB,BrücknerS,WinklerS.The therapeutic promise of mesenchymal stem cells for liver restoration[J].Trends Mol Med,2015,21(11):673-686.DOI: 10.1016/j.molmed.2015.09.004.
[17]	MeloniM,IzzoV,GiuratoL,et al.Prevalence, clinical aspects and outcomes in a large cohort of persons with diabetic foot disease: comparison between neuropathic and ischemic ulcers[J].J Clin Med,2020,9(6):1780.DOI: 10.3390/jcm9061780.
[18]	DoranAC,YurdagulAJr,TabasI.Efferocytosis in health and disease[J].Nat Rev Immunol,2020,20(4):254-267.DOI: 10.1038/s41577-019-0240-6.
[19]	WangY,WangJ,ZhangJ,et al.Stiffness sensing via Piezo1 enhances macrophage efferocytosis and promotes the resolution of liver fibrosis[J].Sci Adv,2024,10(23):eadj3289.DOI: 10.1126/sciadv.adj3289.
[20]	RaziS,Yaghmoorian KhojiniJ,KargarijamF,et al. Macrophage efferocytosis in health and disease[J].Cell Biochem Funct,2023,41(2):152-165.DOI: 10.1002/cbf.3780.
[21]	SchilperoortM,NgaiD,SukkaSR,et al.The role of efferocytosis-fueled macrophage metabolism in the resolution of inflammation[J].Immunol Rev,2023,319(1):65-80.DOI: 10.1111/imr.13214.
[22]	LiW,TeradaY,TyurinaYY,et al.Necroptosis triggers spatially restricted neutrophil-mediated vascular damage during lung ischemia reperfusion injury[J].Proc Natl Acad Sci U S A,2022,119(10):e2111537119.DOI: 10.1073/pnas.2111537119.
[23]	PoonIKH,RavichandranKS.Targeting efferocytosis in inflammaging[J].Annu Rev Pharmacol Toxicol,2024,64:339-357.DOI: 10.1146/annurev-pharmtox-032723-110507.
[24]	SchilperoortM,NgaiD,KaterelosM,et al.PFKFB2-mediated glycolysis promotes lactate-driven continual efferocytosis by macrophages[J].Nat Metab,2023,5(3):431-444.DOI: 10.1038/s42255-023-00736-8.
[25]	JunJI,KimKH,LauLF.The matricellular protein CCN1 mediates neutrophil efferocytosis in cutaneous wound healing[J].Nat Commun,2015,6:7386.DOI: 10.1038/ncomms8386.
[26]	GrégoireM,UhelF,LesouhaitierM,et al.Impaired efferocytosis and neutrophil extracellular trap clearance by macrophages in ARDS[J].Eur Respir J,2018,52(2):1702590.DOI: 10.1183/13993003.02590-2017.
[27]	邢楠,霍然,王海涛,等.脂肪干细胞基质胶促进创面愈合的研究进展[J].中华烧伤与创面修复杂志,2023,39(1):81-84.DOI: 10.3760/cma.j.cn501120-20211204-00404.
[28]	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.
[29]	SunD,CaoH,YangL,et al.MiR-200b in heme oxygenase-1-modified bone marrow mesenchymal stem cell-derived exosomes alleviates inflammatory injury of intestinal epithelial cells by targeting high mobility group box 3[J].Cell Death Dis,2020,11(6):480.DOI: 10.1038/s41419-020-2685-8.
[30]	丁健,宋亮,阮柏,等.间充质干细胞及其修饰体在肝纤维化治疗中的作用研究进展[J].解放军医学杂志,2023,48(5):609-614.DOI: 10.11855/j.issn.0577-7402.2023.05.0609.
[31]	纪鸥洋,方均燕,宋阿会,等.间充质干细胞外泌体对腹膜间皮细胞高糖损伤的作用研究[J].组织工程与重建外科杂志,2022,18(4):294-299.DOI: 10.3969/j.issn.1673-0364.2022.04.002.
[32]	WuL,CaoH,TianX,et al.Bone marrow mesenchymal stem cells modified with heme oxygenase-1 alleviate rejection of donation after circulatory death liver transplantation by inhibiting dendritic cell maturation in rats[J].Int Immunopharmacol,2022,107:108643.DOI: 10.1016/j.intimp.2022.108643.
[33]	YuanYG,WangJL,ZhangYX,et al. Biogenesis, composition and potential therapeutic applications of mesenchymal stem cells derived exosomes in various diseases[J].Int J Nanomedicine,2023,18:3177-3210.DOI: 10.2147/IJN.S407029.
[34]	FanH,AiR,MuS,et al.MiR-19a suppresses ferroptosis of colorectal cancer cells by targeting IREB2[J].Bioengineered,2022,13(5):12021-12029.DOI: 10.1080/21655979.2022.2054194.
[35]	DongJ,WuB,TianW.Human adipose tissue-derived small extracellular vesicles promote soft tissue repair through modulating M1-to-M2 polarization of macrophages[J].Stem Cell Res Ther,2023,14(1):67.DOI: 10.1186/s13287-023-03306-7.
[36]	胡佳,张海霞,苏婉露,等.间充质干细胞对2型糖尿病小鼠糖尿病肾病进展的影响及其机制[J].解放军医学杂志,2023,48(4):383-393.DOI: 10.11855/j.issn.0577-7402.2023.04.0383.
[37]	TianX,YanX,ZangN,et al.Injectable thermosensitive selenium-containing hydrogel as mesenchymal stem cell carrier to improve treatment efficiency in limb ischemia[J].Mater Today Bio,2024,25:100967.DOI: 10.1016/j.mtbio.2024.100967.
[38]	TianX,WuL,LiX,et al.Exosomes derived from bone marrow mesenchymal stem cells alleviate biliary ischemia reperfusion injury in fatty liver transplantation by inhibiting ferroptosis[J].Mol Cell Biochem,2024,479(4):881-894.DOI: 10.1007/s11010-023-04770-8.
[39]	孙皓,蒲胤瑄,刘佳林,等.过表达miR-378a修饰骨髓间充质干细胞复合胶原蛋白海绵支架对大鼠股骨缺损的修复作用[J].解放军医学杂志,2023,48(2):175-182.DOI: 10.11855/j.issn.0577-7402.2023.02.0175.
[40]	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.
[41]	WuL,TianX,ZuoH,et al.miR-124-3p delivered by exosomes from heme oxygenase-1 modified bone marrow mesenchymal stem cells inhibits ferroptosis to attenuate ischemia-reperfusion injury in steatotic grafts[J].J Nanobiotechnology,2022,20(1):196.DOI: 10.1186/s12951-022-01407-8.