The importance of both fibroblasts and keratinocytes in a bilayered living cellular construct used in wound healing

TLDR

Cross‑talk between fibroblasts and keratinocytes that maintains skin homeostasis is disrupted in chronic wounds, and a bilayered living cellular construct (BLCC) containing both cell types has proven safe and effective for healing venous leg ulcers and diabetic foot ulcers. The study aimed to demonstrate the importance of both fibroblasts and keratinocytes in BLCC. To do so, the authors generated constructs containing only fibroblasts or only keratinocytes and compared them directly to BLCC using histology, mechanical testing, gene/protein analysis, and angiogenesis assays. BLCC exhibited a fully differentiated epithelium, greater tensile strength, intact basement membrane and stratum corneum, modulated expression of wound‑healing genes, cytokines, and growth factors, and enhanced endothelial network formation, confirming that both cell types together improve epithelial stratification, mechanical integrity, and angiogenic properties compared with single‑cell constructs.

Abstract

Abstract Cross talk between fibroblasts and keratinocytes, which maintains skin homeostasis, is disrupted in chronic wounds. For venous leg ulcers and diabetic foot ulcers, a bilayered living cellular construct ( BLCC ), containing both fibroblasts and keratinocytes that participate in cross talk, is a safe and effective product in healing chronic wounds. To show the importance of both cell types in BLCC , constructs were generated containing only fibroblasts or only keratinocytes and compared directly to BLCC via histology, mechanical testing, gene/protein analysis, and angiogenesis assays. BLCC contained a fully differentiated epithelium and showed greater tensile strength compared with one‐cell‐type constructs, most likely due to formation of intact basement membrane and well‐established stratum corneum in BLCC . Furthermore, expression of important wound healing genes, cytokines, and growth factors was modulated by the cells in BLCC compared with constructs containing only one cell type. Finally, conditioned medium from BLCC promoted greater endothelial network formation compared with media from one‐cell‐type constructs. Overall, this study characterized a commercially available wound healing product and showed that the presence of both fibroblasts and keratinocytes in BLCC contributed to epithelial stratification, greater tensile strength, modulation of cytokine and growth factor expression, and increased angiogenic properties compared with constructs containing fibroblasts or keratinocytes alone.