Single-Cell Dissection of Fibroblast Heterogeneity in Diabetic Ulcers: Platelet-Rich Plasma (PRP) Therapy Activates Core Regenerative Programs via PLAGL1/RUNX2/ZKSCAN7 Networks.

OBJECTIVE

This integrated study aimed to characterize fibroblast heterogeneity in diabetic ulcers and evaluate the efficacy of platelet-rich plasma (PRP) using multi-omics approaches.

METHODS

We analyzed single-cell RNA sequencing (scRNA-seq) data (GSE165816) from healed (n = 9) and non-healed (n = 5) patients with diabetic foot ulcers (DFU) to characterize fibroblast dynamics, utilizing cell-cell communication analysis, transcription factor profiling, and pseudotime trajectory reconstruction. A streptozotocin-induced diabetic ulcer rat model was established to validate the therapeutic effects of PRP.

RESULTS

scRNA-seq identified 13 cell types, with fibroblasts showing the most significant proportional increase in healed DFU (32% versus 25% in non-healed tissue). Fibroblast-centric communication networks revealed synergistic interactions with endothelial and keratinocyte lineages. Three key transcription factors (PLAGL1, RUNX2, and ZKSCAN7) were upregulated in healed fibroblasts, regulating pathways related to extracellular matrix (ECM) synthesis, angiogenesis, and cell migration. Pseudotemporal analysis confirmed the differentiation of fibroblasts toward ECM-producing states, with enrichment of platelet-derived growth factor (PDGF) signaling pathways. In the rat model, PRP treatment resulted in epidermal/dermal thickening, reduced inflammatory infiltration, and transcriptomic reprogramming that converged with non-diabetic profiles. Venn analysis identified a 26-core gene signature (e.g., COL1A1, FN1) associated with fibroblast-mediated ECM reorganization.

CONCLUSION

Fibroblasts drive diabetic ulcer healing via transcription factor-regulated functional networks. PRP accelerates tissue repair by modulating fibroblast ECM-related gene expression, with the 26-gene signature providing a promising foundation for novel diagnostic and therapeutic targets.