Microenvironmental profiles of wound tissues with accelerated healing properties by HAMA hydrogel
10.3969/j.issn.1674-8115.2025.08.004
- VernacularTitle:HAMA水凝胶促进皮肤创面愈合的组织局部微环境特征
- Author:
Qianyu JIANG
1
;
Chengcheng YAO
1
;
Ping JI
1
;
Ying WANG
1
Author Information
1. 上海交通大学医学院上海市免疫学研究所,上海 200025
- Publication Type:Journal Article
- Keywords:
wound healing;
macrophages;
fibroblasts;
hyaluronic acid methacryloyl(HAMA);
single-cell RNA transcriptome
- From:
Journal of Shanghai Jiaotong University(Medical Science)
2025;45(8):969-980
- CountryChina
- Language:Chinese
-
Abstract:
Objective·To explore the roles of hyaluronic acid methacryloyl(HAMA)hydrogel in skin wound healing and to characterize the microenvironmental landscape at wound sites.Methods·A full-thickness skin excision model was established in mice,which were randomly divided into a control group(n=3)and a HAMA group(n=3).The wound in the HAMA and control groups were covered with 100 μL of HAMA hydrogel and 100 μL of phenyl-2,4,6-trimethylbenzoylphosphonic acid lithium(LAP),respectively.Both groups were then irradiated with a UV lamp for 20 s.The residual wound areas was measured on days 0,3,7,10,and 14.Wound healing effects of HAMA hydrogel were analyzed by measuring the residual wound area and through H-E staining.Single-cell RNA sequencing technology was utilized to analyze the cellular profile of local wound skin tissues at day 14 post-injury.Immunofluorescence assay was used to detect the levels of type I collagen,type Ⅲ collagen,F4/80,CD206,and CD86 in the wound sites.The mRNA expression levels of Arg1,Nos2,Itgam,and Itgb2 in the mouse macrophage cell line Raw264.7 co-cultured with HAMA hydrogel for 24 h were detected by RT-qPCR.The fibroblasts and macrophages in the local skin of the mouse wound on day 14 were analyzed using the Seurat package,and the communication between fibroblasts and macrophages was analyzed using the CellChat package.Results·Mice treated with HAMA hydrogel exhibited a significantly faster rate of wound healing process compared to the control group.At day 14,wounds in the HAMA-treated mice had already healed,while those in the control group remained unhealed.Single-cell RNA sequencing analysis revealed a remarkable increase in the proportion of fibroblasts in the skin tissues of HAMA-treated wounds.The proportion of the Col3a1-high-expressing fibroblast subset increased(90.2%)compared to the control group(79.8%),while the proportion of the Col1a1-high-expressing fibroblast subset decreased(5.7%vs 15.9%).Immunofluorescence analysis confirmed that the level of type Ⅲ collagen in the wound tissues of the HAMA group was significantly higher than that in the control group(P=0.035),while the level of type Ⅰ collagen was significantly lower(P=0.044).Although there was no significant difference in the proportions of macrophages in the wound tissues between the HAMA-treated and control groups,scRNA sequencing data and in vitro experiments using Raw264.7 cells showed that HAMA hydrogel could induce the expression of Arg1 and decrease the expression of Nos2 in the macrophages(P<0.001).Additionally,macrophages in the HAMA-treated wounds expressed higher levels of CD206 and lower levels of CD86(P=0.042,P=0.011).The results of the CellChat analysis showed that,compared to the control group,increased communication intensity was observed between macrophages and fibroblasts subsets at the wound sites in the mice of HAMA group.Conclusion·The microenvironment after HAMA hydrogel treatment is conducive to skin wound healing,characterized by a local aggregation of anti-inflammatory macrophages and fibroblasts that secrete type Ⅲ collagen.
