Abstract
Background and Objectives: Vascular endothelial growth factor (VEGF) is a protein which stimulates the formation of new blood vessels, playing a crucial role in processes such as wound healing and tumor growth. Methods: This study investigated the effects of VEGF on cell viability and osteogenic differentiation in mesenchymal stem cell (MSC) spheroids. Stem cell spheroids were fabricated using concave microwells and cultured with VEGF at concentrations of 0, 0.01, 0.1, 1, and 10 ng/mL. Morphological assessments were conducted on days 1, 3, 5, and 7, while cell viability was evaluated using the LIVE/DEAD assay and Cell Counting Kit-8. Alkaline phosphatase activity (ALP) and calcium deposition were measured to assess osteogenic differentiation, and qPCR was used to analyze osteogenic marker expression. Results: The spheroids maintained their shape across all VEGF concentrations, with the largest diameter being at 0.01 ng/mL on day 1, which decreased over time. Cell viability was highest at 0.01 ng/mL VEGF, while calcium deposition peaked at 0.1 ng/mL. Osteogenic markers, including RUNX2, osteocalcin, and COL1A1, showed significant upregulation at 1 ng/mL VEGF. Conclusions: These results suggest that VEGF enhances early osteogenic differentiation in MSC spheroids, indicating its potential for bone repair and tissue regeneration. VEGF could be applied in clinical settings for bone healing, fracture repair, and regenerative dentistry treatments.