Hyunjin Lee1, Hyuna Lee1, Chae-Bin Na1, Jun-Beom Park2. 1. Researcher, Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea. 2. Associate Professor, Department of Periodontics, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Abstract
PURPOSE: This study evaluates the effects of simvastatin on the morphology, viability, secretion of vascular endothelial growth factor (VEGF) and expression of stemness markers and messenger RNA of cell spheroids cultured in growth media. MATERIALS AND METHODS: Three-dimensional cell spheroids with stem cells and osteoblast-like cells were fabricated using concave, silicon, elastomer-based microwells in the presence of simvastatin at concentrations of 1 and 10 μM. Qualitative cellular viability was determined with a confocal microscope, and quantitative cellular viability was evaluated using a Cell Counting Kit-8 assay. The expression of stem cell surface markers was tested, and a quantitative real-time polymerase chain reaction was performed to evaluate the expression of collagen I and Runx2. RESULTS: The cell spheroids were well formed in the microwells, but the addition of simvastatin produced significant changes in the morphology of spheroids. No significant changes in cellular viability were noted with the addition of simvastatin on day 1, but the addition of simvastatin significantly decreased cellular viability on day 5. The addition of simvastatin significantly increased the secretion of VEGF. The expression of the CD90 surface marker was seen regardless of whether simvastatin was added. The addition of simvastatin significantly decreased the expression of collagen I. CONCLUSIONS: Based on these findings, the application of simvastatin clearly decreased the cellular viability of the cell spheroids made with stem cells and osteoblast-like cells but increased the secretion of VEGF by the cell spheroids.
PURPOSE: This study evaluates the effects of simvastatin on the morphology, viability, secretion of vascular endothelial growth factor (VEGF) and expression of stemness markers and messenger RNA of cell spheroids cultured in growth media. MATERIALS AND METHODS: Three-dimensional cell spheroids with stem cells and osteoblast-like cells were fabricated using concave, silicon, elastomer-based microwells in the presence of simvastatin at concentrations of 1 and 10 μM. Qualitative cellular viability was determined with a confocal microscope, and quantitative cellular viability was evaluated using a Cell Counting Kit-8 assay. The expression of stem cell surface markers was tested, and a quantitative real-time polymerase chain reaction was performed to evaluate the expression of collagen I and Runx2. RESULTS: The cell spheroids were well formed in the microwells, but the addition of simvastatin produced significant changes in the morphology of spheroids. No significant changes in cellular viability were noted with the addition of simvastatin on day 1, but the addition of simvastatin significantly decreased cellular viability on day 5. The addition of simvastatin significantly increased the secretion of VEGF. The expression of the CD90 surface marker was seen regardless of whether simvastatin was added. The addition of simvastatin significantly decreased the expression of collagen I. CONCLUSIONS: Based on these findings, the application of simvastatin clearly decreased the cellular viability of the cell spheroids made with stem cells and osteoblast-like cells but increased the secretion of VEGF by the cell spheroids.