Single-cell analysis reveals Cxcl14+ progenitor fibroblast accumulation in regenerating diabetic wound after carbon monoxide-releasing hydrogel therapy

Chronic and non-healing skin wound is one of grievous complications of diabetes. In this study, we demonstrated a gas (carbon monoxide)-releasing hyaluronic acid hydrogel (i.e. COHAG) in promoting diabetic wound healing. Our results show that the COHAG significantly accelerated the healing process in diabtic rat full-thickness skin defect model, as compared with hydrogel without gas-releasing molecule and untreated group. Single-cell analysis of regenerating skin samples revealed that Cxcl14-overexpressing (Cxcl14+) fibroblast with progenitor properties are abundantly accumulated at the wound site after COHAG therapy. Overall design: Rat skin tissue from diabetic SD rats at 8-10 weeks. Rat skin specimens(7 days of hydrogel treated wound)were cleaned with PBS and digested using the Epidermis Dissociation Kit (Miltenyi biotec, 130-095-928) for ~1 hours at 37? to obtain cell suspension. Single-cell RNA-seq libraries were made with the single cell 3 'Library and Gel Bead Kit V3.1(10x Genomics, 1000121) and sequenced using an Illumina NovaSeq6000 (150bp x2, 25~40k reads/cell). It is estimated that approximately 8000 cells will be recovered per sample.