Interaction between fibroblasts and keratinocytes in the wound edge skin tissue of a diabetic foot patient and the mechanism

Ruan Qiongfang; Zhang Siyu; Xi Maomao; Ruan Jingjing; Liu Shuhua; Li Binghui; Xie Weiguo

doi:10.3760/cma.j.cn501225-20240221-00067

Volume 40 Issue 8

Aug. 2024

Turn off MathJax

Article Contents

Abstract

References

Article Navigation > CHINESE JOURNAL OF BURNS AND WOUNDS > 2024 > 40(8): 762-771

Ruan QF,Zhang SY,Xi MM,et al.Interaction between fibroblasts and keratinocytes in the wound edge skin tissue of a diabetic foot patient and the mechanism[J].Chin J Burns Wounds,2024,40(8):762-771.DOI: 10.3760/cma.j.cn501225-20240221-00067.

Citation:

PDF( 12514 KB)

Interaction between fibroblasts and keratinocytes in the wound edge skin tissue of a diabetic foot patient and the mechanism

doi: 10.3760/cma.j.cn501225-20240221-00067

1.
Institute of Burns, Tongren Hospital of Wuhan UniversityWuhan Third Hospital, Wuhan 430060, China
2.
Department of Burns, Tongren Hospital of Wuhan UniversityWuhan Third Hospital, Wuhan 430060, China
3.
Department of Wound Repair, Liyuan Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430077, China

Funds:

Wuhan Science and Technology Bureau Knowledge Innovation Project 2023020201010192

General Program of Hubei Natural Science Foundation 2021CFB532

Scientific Research Program of Health Commission of Hubei Province of China WJ2021M260

Shanghai Wang Zhengguo Foundation for Traumatic Medicine Growth Factor Rejuvenation Plan SZYZ-TR-10

More Information

Corresponding author: Xie Weiguo, Email: wgxie@hotmail.com
Received Date: 2024-02-21

Abstract

Abstract

Objective To investigate the interaction between fibroblasts (Fb) and keratinocytes (KC) in the wound edge skin tissue of a diabetic foot patient and the mechanism. Methods This was an experimental research. The wound edge skin tissue from a diabetic foot patient (male and 33 years old) admitted to the Department of Wound Repair of Liyuan Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology in August 2021 and from an acute foot injury patient (male and 50 years old) admitted to the Department of Hand Surgery of the hospital in September 2021 was collected. The single-cell transcriptome sequencing was performed to analyze the interaction between chemokine ligands of Fb subgroup and chemokine receptors of KC subgroup. The supernatant was collected after human foreskin fibroblast (HFF) was cultured routinely and with high concentration of glucose for 7 days as normal conditioned medium (CM) and high glucose CM, respectively. HaCaT cells were collected and divided into normal CM group cultured with normal CM and high glucose CM group cultured with high glucose CM, the scratch test was performed to calculate the cell migration rates at 24 and 48 h after scratch (n=3). The content of cytokines in the two kinds of CM was detected by liquid suspension chip (n=5). HFF was collected and divided into normal group cultured routinely and high glucose group cultured with high concentration of glucose for 7 days, and the mRNA expressions of C-X-C motif chemokine ligand 1 (CXCL1), CXCL2, CXCL8, and CXCL12 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (n=6). HaCaT cells in normal CM group and high glucose CM group were collected to detect the protein expressions of C-X-C motif chemokine receptor 4 (CXCR4) in cells cultured for 48 h by Western blotting (n=3). HaCaT cells were collected and divided into normal CM group, high glucose CM group, normal CM+CXCL12 group, and high glucose CM+CXCL12 group. The first two groups of cells were treated as before, and the latter two groups of cells were cultured with normal CM and high glucose CM containing recombinant human CXCL12, respectively. Scratch test was performed, and cell migration rates were calculated at 24 and 48 h after scratch (n=3); the protein expression of CXCR4 in cells cultured for 48 h was detected by Western blotting (n=3). Results Compared with those in the wound edge skin tissue of acute foot injury, the interactions between chemokine ligands (CXCL1, CXCL2, CXCL3, CXCL8, and CXCL12) of Fb subgroup and chemokine receptors (CXCR2 and CXCR4) of KC subgroup were significantly weakened in the wound edge skin tissue of diabetic foot. At 24 and 48 h after scratch, the migration rates of HaCaT cells in high glucose CM group were significantly lower than those in normal CM group (with t values of 23.50 and 15.65, respectively, P<0.05). Compared with that in normal CM, the content of CXCL1 in high glucose CM was significantly increased (P<0.05), and the content of CXCL12 was significantly decreased (P<0.05). After 7 days of culture, compared with those in normal group, the mRNA expressions of CXCL1, CXCL2, and CXCL8 in HFF in high glucose group were significantly increased (with t values of 4.25, 4.98, and 10.04, respectively, P<0.05), while the mRNA expression of CXCL12 was significantly decreased (t=4.10, P<0.05). After 48 h of culture, the CXCR4 protein expression in HaCaT cells in high glucose CM group was significantly lower than that in normal CM group (t= 5.13, P<0.05). At 24 and 48 h after scratch, the migration rates of HaCaT cells in high glucose CM group were significantly lower than those in normal CM group and high glucose CM+CXCL12 group (with P values all <0.05); at 24 h after scratch, the migration rate of HaCaT cells in normal CM+CXCL12 group was significantly lower than that in normal CM group (P<0.05); at 48 h after scratch, the migration rate of HaCaT cells in normal CM+CXCL12 group was significantly higher than that in high glucose CM+CXCL12 group (P<0.05). At 48 h of culture, the CXCR4 protein expression of HaCaT cells in high glucose CM+CXCL12 group was 0.446±0.050, which was significantly higher than 0.247±0.010 in high glucose CM group (P<0.05) and similar to 0.522±0.082 in normal CM+CXCL12 group (P>0.05); the CXCR4 protein expression in HaCaT cells in normal CM group was 0.509±0.055, which was significantly higher than that in high glucose CM group (P<0.05). Conclusions The interactions between chemokine ligands of Fb subgroup and chemokine receptors of KC subgroup were significantly weakened in the wound edge skin tissue of diabetic foot. High glucose can inhibit CXCL12 secretion of HFF, and the stimulation of its cell culture supernatant can decrease HaCaT cell migration ability and CXCR4 expression. Exogenous CXCL12 protein can increase the CXCR4 protein expression in HaCaT cells and enhance the cell migration ability.
- Cell communication,
- Chemokines, CXC,
- Receptors, chemokine,
- Fibroblasts,
- Keratinocytes,
- Cell migration,
- Wound repair

FullText(HTML)

References(35)

References

[1]	McDermottK,FangM,BoultonA,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]	LanT,LiZ,ChenJ.FusionSegNet: fusing global foot features and local wound features to diagnose diabetic foot[J].Comput Biol Med,2023,152:106456.DOI: 10.1016/j.compbiomed.2022.106456.
[3]	PatelS,SrivastavaS,SinghMR,et al.Mechanistic insight into diabetic wounds: pathogenesis, molecular targets and treatment strategies to pace wound healing[J].Biomed Pharmacother,2019,112:108615.DOI: 10.1016/j.biopha.2019.108615.
[4]	曹涛,肖丹,计鹏,等.肝细胞生长因子修饰的人脂肪间充质干细胞外泌体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,2022,38(11):1004-1013.DOI: 10.3760/cma.j.cn501225-20220731-00330.
[5]	王宁,鞠上.糖尿病足溃疡难愈合机制研究进展[J].中华烧伤与创面修复杂志,2022,38(11):1085-1089.DOI: 10.3760/cma.j.cn501225-20220227-00038.
[6]	SorgH, SorgCGG. Skin wound healing: of players, patterns, and processes[J]. Eur Surg Res, 2023,64(2):141-157. DOI: 10.1159/000528271.
[7]	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.
[8]	OrmazabalV,Nova-LampetiE,RojasD,et al.Secretome from human mesenchymal stem cells-derived endothelial cells promotes wound healing in a type-2 diabetes mouse model[J].Int J Mol Sci,2022,23(2):941.DOI: 10.3390/ijms23020941.
[9]	SharifiaghdamM, ShaabaniE, Faridi-MajidiR, et al. Macrophages as a therapeutic target to promote diabetic wound healing[J]. Mol Ther,2022,30(9):2891-2908. DOI: 10.1016/j.ymthe.2022.07.016.
[10]	LuD,XuY,LiuQ,et al.Mesenchymal stem cell-macrophage crosstalk and maintenance of inflammatory microenvironment homeostasis[J].Front Cell Dev Biol,2021,9:681171.DOI: 10.3389/fcell.2021.681171.
[11]	DiX,ChenJ,LiY,et al.Crosstalk between fibroblasts and immunocytes in fibrosis: from molecular mechanisms to clinical trials[J].Clin Transl Med,2024,14(1):e1545.DOI: 10.1002/ctm2.1545.
[12]	ChabeliMS,WangX,YinghaoL,et al.Similarities between wound re-epithelialization and metastasis in ESCC and the crucial involvement of macrophages: a review[J].Cancer Treat Res Commun,2022,32:100621.DOI: 10.1016/j.ctarc.2022.100621.
[13]	MaZ,DingY,DingX,et al.PDK4 rescues high-glucose-induced senescent fibroblasts and promotes diabetic wound healing through enhancing glycolysis and regulating YAP and JNK pathway[J].Cell Death Discov,2023,9(1):424.DOI: 10.1038/s41420-023-01725-2.
[14]	ZhangS,MengN,LiuS,et al.Targeting senescent HDF with the USP7 inhibitor P5091 to enhance DFU wound healing through the p53 pathway[J].Biochem Biophys Res Commun,2024,722:150149.DOI: 10.1016/j.bbrc.2024.150149.
[15]	VozaFA,HuertaCT,LeN,et al.Fibroblasts in diabetic foot ulcers[J].Int J Mol Sci,2024,25(4):2172. DOI: 10.3390/ijms25042172.
[16]	LiuY,LiuY,HeW,et al.Fibroblasts: immunomodulatory factors in refractory diabetic wound healing[J].Front Immunol,2022,13:918223.DOI: 10.3389/fimmu.2022.918223.
[17]	何秀娟,林燕,刘青武,等.皮肤成纤维细胞在创面愈合中的研究进展[J/CD].中华损伤与修复杂志(电子版),2021,16(1):74-77.DOI: 10.3877/cma.j.issn.1673-9450.2021.01.015.
[18]	XiaoY,QianJ,DengX,et al.Macrophages regulate healing-associated fibroblasts in diabetic wound[J].Mol Biol Rep,2024,51(1):203.DOI: 10.1007/s11033-023-09100-1.
[19]	HeS,LiZ,WangL,et al.A nanoenzyme-modified hydrogel targets macrophage reprogramming-angiogenesis crosstalk to boost diabetic wound repair[J].Bioact Mater,2024,35:17-30.DOI: 10.1016/j.bioactmat.2024.01.005.
[20]	HollJ,KowalewskiC,ZimekZ,et al.Chronic diabetic wounds and their treatment with skin substitutes[J].Cells,2021,10(3):655.DOI: 10.3390/cells10030655.
[21]	WilkinsonHN,HardmanMJ.Wound healing: cellular mechanisms and pathological outcomes[J].Open Biol,2020,10(9):200223.DOI: 10.1098/rsob.200223.
[22]	GrzelakEM,ElshanNGRD,ShaoS,et al.Pharmacological YAP activation promotes regenerative repair of cutaneous wounds[J].Proc Natl Acad Sci U S A,2023,120(28):e2305085120.DOI: 10.1073/pnas.2305085120.
[23]	JohnJV,SharmaNS,TangG,et al.Nanofiber aerogels with precision macrochannels and LL-37-mimic peptides synergistically promote diabetic wound healing[J].Adv Funct Mater,2023,33(1):2206936. DOI: 10.1002/adfm.202206936.
[24]	张喻平,张琼,邓芳,等.高糖微环境下P62对人表皮细胞株HaCaT迁移和运动性的影响及其机制[J].中华烧伤与创面修复杂志,2022,38(11):1014-1022.DOI: 10.3760/cma.j.cn501225-20220630-00272.
[25]	TheocharidisG,BaltzisD,RoustitM,et al.Integrated skin transcriptomics and serum multiplex assays reveal novel mechanisms of wound healing in diabetic foot ulcers[J].Diabetes,2020,69(10):2157-2169.DOI: 10.2337/db20-0188.
[26]	DinhHQ,PanF,WangG,et al.Integrated single-cell transcriptome analysis reveals heterogeneity of esophageal squamous cell carcinoma microenvironment[J].Nat Commun,2021,12(1):7335.DOI: 10.1038/s41467-021-27599-5.
[27]	曾帅丹,杨磊.各种组学分析在体表慢性难愈合创面中的研究进展[J].中华烧伤与创面修复杂志,2023,39(1):75-80.DOI: 10.3760/cma.j.cn501225-20220216-00030.
[28]	WangZ,WeiD,LiS,et al.Healing mechanism of diabetic foot ulcers using single-cell RNA-sequencing[J].Ann Transl Med,2023,11(5):210.DOI: 10.21037/atm-23-240.
[29]	WilkinsonHN,ClowesC,BanyardKL,et al.Elevated local senescence in diabetic wound healing is linked to pathological repair via CXCR2[J].J Invest Dermatol,2019,139(5):1171-1181.e6.DOI: 10.1016/j.jid.2019.01.005.
[30]	XuJ,HuJ,Idlett-AliS,et al.Discovery of small molecule activators of chemokine receptor CXCR4 that improve diabetic wound healing[J].Int J Mol Sci,2022,23(4):2196.DOI: 10.3390/ijms23042196.
[31]	WilkinsonHN,HardmanMJ.Senescence in wound repair: emerging strategies to target chronic healing wounds[J].Front Cell Dev Biol,2020,8:773.DOI: 10.3389/fcell.2020.00773.
[32]	PeddibhotlaS,CaplesK,MehtaA,et al.Triazolothiadiazine derivative positively modulates CXCR4 signaling and improves diabetic wound healing[J].Biochem Pharmacol,2023,216:115764.DOI: 10.1016/j.bcp.2023.115764.
[33]	RestivoTE,MaceKA,HarkenAH,et al.Application of the chemokine CXCL12 expression plasmid restores wound healing to near normal in a diabetic mouse model[J].J Trauma,2010,69(2):392-398.DOI: 10.1097/TA.0b013e3181e772b0.
[34]	YeboahA,CohenRI,FaulknorR,et al.The development and characterization of SDF1α-elastin-like-peptide nanoparticles for wound healing[J].J Control Release,2016,232:238-247.DOI: 10.1016/j.jconrel.2016.04.020.
[35]	ShafiqM,YuanZ,RafiqueM,et al.Combined effect of SDF-1 peptide and angiogenic cues in co-axial PLGA/gelatin fibers for cutaneous wound healing in diabetic rats[J].Colloids Surf B Biointerfaces,2023,223:113140.DOI: 10.1016/j.colsurfb.2023.113140.

Relative Articles

[1]	Zhao Shihan, Jin Linbo, Zhang Jianghe, Zhang Yiming, Fan Dongli. Effects of tumor necrosis factor-alpha/extracellular signal-regulated kinase pathway on migration ability of HaCaT cells and full-thickness skin defects in mice[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2023, 39(2): 122-131. doi: 10.3760/cma.j.cn501225-20221019-00460
[2]	Sun Lixiang, Wu Shuai, Zhang Xiaowei, Liu Wenjie, Zhang Lingjuan. Investigation on the growth factor regulatory network of dermal fibroblasts in mouse full-thickness skin defect wounds based on single-cell RNA sequencing[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2022, 38(7): 629-639. doi: 10.3760/cma.j.cn501225-20220215-00029
[3]	Leng Min, Peng Ying, Wang Hong. Research advances on the biomechanical micro- environment facilitated wound repair through the regulation of cell migration[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2022, 38(1): 90-94. doi: 10.3760/cma.j.cn501120-20200921-00419
[4]	Jiang Yaonan, Wang Yuxiang, Zheng Yongjun, Hu Xiaoyan, He Fei, Shi Wenjun, Wu Qiong, Xia Zhaofan, Xiao Shichu. Clinical study of cell sheets containing allogeneic keratinocytes and fibroblasts for the treatment of partial-thickness burn wounds[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2020, 36(3): 171-178. doi: 10.3760/cma.j.cn501120-20191113-00426
[5]	Zhang Zhi, Kuang Fang, Liu Changling, Chen Bin, Tang Wenbin, Li Xiaojian. Effects of silencing Smad ubiquitination regulatory factor 2 on the function of human hypertrophic scar-derived fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2017, 33(3): 145-151. doi: 10.3760/cma.j.issn.1009-2587.2017.03.004
[6]	Liu Jie, Ren Xi, Guo Xiaowei, Sun Huanbo, Tang Yong, Luo Zhenghui, Zhang Qiong, Zhang Dongxia, Huang Yuesheng, Zhang Jiaping. Effects of direct current electric field on directional migration and arrangement of dermal fibroblasts in neonatal BALB/c mice and the mechanisms[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2016, 32(4): 224-231. doi: 10.3760/cma.j.issn.1009-2587.2016.04.007
[7]	Zhang Lu, Li Haisheng, Yao Zhihui, Yang Sisi, He Weifeng, Wu Jun, Luo Gaoxing. Interaction between P311 and transforming growth factor beta 1 and its effect on the function of murine fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2016, 32(4): 208-215. doi: 10.3760/cma.j.issn.1009-2587.2016.04.005
[8]	Wang Yang, Zhang Liangping, Lei Rui, Shen Yichen, Shen Hui, Wu Zhinan, Xu Jinghong. Effects of blocking two sites of transforming growth factor-β/Smads signaling on the formation of scar-related proteins in human skin fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2015, 31(5): 372-377. doi: 10.3760/cma.j.issn.1009-2587.2015.05.013
[9]	Jia Wenbin, Hu Dahai, Wang Hongtao, Chen Dongdong, Bai Xiaozhi, Li Na, Han Fu, Fang Xiaobing, Yang Longlong. Effects of mouse adipose-derived stem cell conditioned medium on the apoptosis of keratinocytes in- duced by thermal injury in vitro[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2014, 30(2): 102-108. doi: 10.3760/cma.j.issn.1009-2587.2014.02.003
[11]	DONG Zhi-wei, PENG Yi-zhi, ZHANG Shuai, CHEN Yu, DONG Feng-juan. Influence of infection of murine chemokine receptor-7 recombinant lentivirus on the immunogenicity and migration of DC 2.4 cells[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2013, 29(1): 41-45. doi: 10.3760/cma.j.issn.1009-2587.2013.01.015
[12]	YANG Chong-zhi, ZHANG Hui-tang, WANG Gong-sheng, ZHOU Hai-quan, MA Chi, HU Da-hai. Mechanism underlying the inhibitory effects of peroxisome proliferator-activated receptor γ agonists on transforming growth factor β1 in adult skin fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2010, 26(6): 448-451. doi: 10.3760/cma.j.issn.1009-2587.2010.06.015
[13]	BAI Xiao-zhi, HU Da-hai, ZHANG Wan-fu, ZHANG Zhan-feng, SHI Ji-hong, CAI Wei-xia, ZHU Hua-yu, ZHU Xiong-xiang, TANG Chao-wu. Effect of heat injured keratinocytes supernatant on biological behavior of fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2010, 26(2): 133-137. doi: 10．3760／cma．j．issn．1009-2587．2010．02．015
[14]	LIU Jia-qi, HU Da-hai, ZHANG Zhan-feng, GUAN Hao, SHE Tao, ZHANG Jun, BAI Xiao-zhi. Effects of interferon-gamma on the transforming growth factor beta/Smad pathway in keloid-derived fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2009, 25(6): 454-459. doi: 10.3760/cma.j.issn.1009-2587.2009.06.019
[15]	LI Zhe, LI Shi-rong, LIU Jian-yi, DAI Xia, TAO Ling. To transdifferentiate human hypertrophic scar fibroblasts induced by connective tissue growth factor mediated transforming growth factor-β1 in vitro[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2009, 25(1): 49-52. doi: 10.3760/cma.j.issn.1009-2587.2009.01.019
[16]	LI Jin-xi, LIU Xu-sheng, TANG Hui, ZHOU Xin, HUANG Yue-sheng. Influence of some topical antibiotics and FGF2, EGF and rhGH on the biological characteristics of fibroblasts in vitro[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2006, 22(1): 33-37.
[17]	CHENG Biao, LIU Hong-wei, FU Xiao-bing, SHENG Zhi-yong, ZHANG Ping, MENG Ji-hong. The role of in fibroblast growth factor-2 on expression of S100 protein in apoptosis of fibroblasts after heat stress[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2006, 22(2): 109-112.
[18]	ZHANG Zhi, LI Xiao-jian, LIANG Da-rong, LI Ye-yang, XU Wei-shi. The antagonistic effect of recombinant human decorin on TGF-β1 stimulation of fibroblasts in collagen lattices of scar[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2006, 22(3): 207-210.
[19]	LlU Jian-yi, LI Shi-rong, JI Shu-xing. Effects of antisense oligonucleotides on the expression of connective tissue growth factor gene and on the collagen synthesis in the cultured human keloid fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2004, 20(2): 72-75.
[20]	HUANG Yue-sheng, YANG Zhong-cheng. Effects of thrombin peptides and thrombin on the release of TGF-β and VEGF from human fibroblasts[J]. CHINESE JOURNAL OF BURNS AND WOUNDS, 2003, 19(Z1): 5-7. doi: 10.3760/cma.j.issn.1009-2587.2003.Z1.102

Supplements(0)

Cited By

Proportional views

Proportional views

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

Figures(7) / Tables(3)

Get Citation

PDF

XML

Article Metrics

Article views (280) PDF downloads(30)

Interaction between fibroblasts and keratinocytes in the wound edge skin tissue of a diabetic foot patient and the mechanism

doi: 10.3760/cma.j.cn501225-20240221-00067

Abstract

References

Relative Articles

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

Interaction between fibroblasts and keratinocytes in the wound edge skin tissue of a diabetic foot patient and the mechanism

doi: 10.3760/cma.j.cn501225-20240221-00067

Abstract

References

Relative Articles

Proportional views

Catalog

通讯作者: 陈斌, bchen63@163.com

Article Metrics

Proportional views

Related

Export File

Citation

Format

Content