Akademik Veri Yönetim Sistemi
Biomedical Research and Therapy, cilt.12, sa.10, ss.7865-7875, 2025 (ESCI, Scopus)
Introduction: This study examines the effects of varying L-ascorbic acid (L-AA) concentrations and initial seeding densities on cell-sheet formation by human dermal fibroblasts (HDFs) and ker-atinocytes (HaCaT). Both cell types were seeded at either 50,000 or 100,000 cells cm−2 and ex-posed to 0, 25, 50, or 100 µg mL−1 L-AA. Cell-sheet formation was assessed morphologically; cell viability was quantified by an MTT assay; and gene expression was measured by qRT-PCR using newly designed primers. Methods: In HDF cultures, the 50 µg mL−1 group at a seeding density of 50,000 cells cm−2 produced the most cohesive and viable sheets, accompanied by a marked up-regulation of COL1A1, reflecting enhanced extracellular-matrix synthesis. At 25 µg mL−1, HDF morphology and viability were similar to those of the untreated control, with no cohesive sheet formation, whereas 100 µg mL−1 induced pronounced cytotoxicity. Results: In HaCaT cultures, no cohesive sheets were observed at any concentration. Nonetheless, at 50 µg mL−1 and 100,000 cells cm−2, confluent HaCaT monolayers displayed increased expression of keratin 14 (KRT14), in-volucrin (IVL), and Ki-67, indicating enhanced proliferation and early differentiation despite absent sheet integrity. Conclusion: Collectively, these findings demonstrate that L-AA promotes sheet formation in fi-broblasts but not in keratinocytes under two-dimensional monolayer conditions. The limited Ha-CaT response suggests that co-culture or three-dimensional systems may be required for epithelial stratification. This study underscores the importance of tailoring L-AA concentrations and cell densities to each cell type in skin tissue engineering.