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Article Navigation >  CHINESE JOURNAL OF BURNS AND WOUNDS  >  > Accepted Manuscript
Jia Yanhui,Yuan Yixuan,Hu Dahai,et al.Mechanism of Wnt9a regulating fibroblast function to promote chronic wound healing[J].Chin J Burns Wounds,2026,42(3):1-9.DOI: 10.3760/cma.j.cn501225-20241122-00457.
Citation: Jia Yanhui,Yuan Yixuan,Hu Dahai,et al.Mechanism of Wnt9a regulating fibroblast function to promote chronic wound healing[J].Chin J Burns Wounds,2026,42(3):1-9.DOI: 10.3760/cma.j.cn501225-20241122-00457.
shu

Mechanism of Wnt9a regulating fibroblast function to promote chronic wound healing

doi: 10.3760/cma.j.cn501225-20241122-00457

  • Department of Burns and Cutaneous Surgery, the First Affiliated Hospital of Air Force Medical University, Xi'an 710032, China

Funds:

General Program of National Natural Science Foundation of China 81772071

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  • Corresponding author: Guan Hao, Email: guanhao@fmmu.edu.cn
  • Received Date: 2024-11-22
    Available Online: 2026-03-09
    Abstract
  •   Objective  To investigate the expression changes, role, and its possible mechanism of Wnt9a in chronic wound healing.  Methods  This study was a group-designed experimental research.The chronic wound tissue and adjacent normal skin tissue were collected from 8 patients with diabetic foot ulcers, including 5 males and 3 females, aged 45-72 years. The expression of Wnt9a was detected by enzyme-linked immunosorbent assay (ELISA) method and immunofluorescence method. Eight male C57BL/6 mice aged 6-8 weeks were used to establish a full-thickness skin defect wound on the back and were divided into control group with no additional treatment and chronic wound group subcutaneously injected with M1 macrophage derived exosomes at the wound edge to construct a chronic wound model using a random number table method (the same grouping method below), with 4 mice in each group. At 7 days after modeling, the Wnt9a expressions in the wound tissue of mice in two groups was detected by ELISA method. Additional 16 male C57BL/6 mice aged 6-8 weeks were used to establish the chronic wound model as before and were divided into blank control group and Wnt9a overexpression group, with 8 mice in each group, which were injected subcutaneously at the wound edge with enhanced green fluorescent protein empty adenovirus (AV-eGFP) and Wnt9a gene recombinant adenovirus expressing enhanced green fluorescent protein (AV-Wnt9a-eGFP), respectively. At 3, 7, and 14 days after modeling, the percentages of residual wound area were calculated; at 14 days after modeling, the expressions of type Ⅰ and type Ⅲ collagen protein were detected by Western blotting. The collected human normal skin tissue was used to extract fibroblasts (Fbs), which were divided into empty control group infected with AV-eGFP and Wnt9a overexpression group infected with AV-Wnt9a-eGFP. The protein expression of Wnt9a at 72 h after infection was detected by Western blotting, and the cell migration rate at 48 h after scratching was detected by scratch test. Additional human normal skin Fbs were collected and divided into Wnt9a specific small interfering RNA (siRNA-Wnt9a) group and negative control small interfering RNA (siRNA-NC) group, which were transfected with corresponding small interfering RNA (siRNA). Scratch test was conducted to detect the cell migration rate at 48 h after scratching. Additional human normal skin Fbs were taken and divided into empty control group and Wnt9a overexpression group treated as before. At 72 h after infection, transcriptome sequencing was performed to screen for differentially expressed genes (DEGs); the gene ontology (GO) and Kyoto encyclopedia of genes and genomes enrichment analysis were performed on DEGs. The sample number in all cell experiments was 3.  Results  The results of ELISA method and immunofluorescence method showed that the expression level of Wnt9a in human chronic wound tissue was significantly lower than that in normal skin tissue (with t values of 7.68 and 10.25, respectively, P<0.05). At 7 days after modeling, the expression level of Wnt9a in the wound tissue of mice in chronic wound group was significantly lower than that in control group (t=5.12, P<0.05). The percentages of residual wound area of mice in Wnt9a overexpression group were significantly lower than those in empty control group at 3, 7, and 14 days after modeling (with t values of 3.90, 6.62, and 5.73, respectively, P<0.05). At 14 days after modeling, the protein expression levels of type Ⅰ and type Ⅲ collagen in the wound tissue of mice in Wnt9a overexpression group were significantly lower than those in empty control group (with t values of 6.25 and 5.48, respectively, P<0.05). The Wnt9a protein expression level in Fbs in Wnt9a overexpression group was significantly higher than that in empty control group (t=6.96, P<0.05). At 48 h after scratching, the cell migration rate in Wnt9a overexpression group was (71.6±6.4)%, which were significantly higher than (38.5±2.4)% in empty control group (t=8.31, P<0.05). The cell migration rate in siRNA-Wnt9a group was (15±3)%, which were significantly lower than (32±4)% in siRNA-NC group (t=5.93, P<0.05). At 72 h after infection, compared with that in empty control group, the significantly downregulated DEGs in Fbs in Wnt9a overexpression group included multiple collagen family genes, and DEGs in Fbs in Wnt9a overexpression group were significantly enriched in the non-classical Wnt signaling pathway.  Conclusions  Wnt9a expression is downregulated in chronic wound tissue of human and mice, and the overexpressed Wnt9a may promote migration of Fbs and collagen remodeling through non-classical Wnt signaling pathway, thereby accelerating chronic wound healing.

     

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