Abstract
Insulin-like growth factors (IGFs) plays various roles, including differentiation and mitogenesis, and IGFs are reported to regulate the bone growth and maintenance. This study was performed to analyze the enhancing effects of IGF-2 on osteogenic differentiation and the mineralization of stem cells cultured on deproteinized bovine bone mineral. Stem cell loaded bone graft material was cultured in the presence of the IGF-2 at final concentrations of 10 and 100 ng/mL and the morphology of the cells was observed on Days 1, 3, and 7. The commercially available, two-color assay based on plasma membrane integrity and esterase activity was also used for qualitative analyses on Days 1, 3, and 7. The level of alkaline phosphatase activity and anthraquinone dye assay were used to evaluate osteogenic differentiation on Days 7 and 14. Real-time polymerase chain reaction was applied in order to identify the mRNA expression of BGLAP, Runx2, and β-catenin. The stem cells were well-attached with fibroblast morphology and most of the stem cells produced a high intensity of green fluorescence, indicating that there were live cells on Day 1. The relative cellular viability assay values for IGF-2 groups at 0, 10, and 100 ng/mL on Day 1 were 0.419 ± 0.015, 0.427 ± 0.013, and 0.500 ± 0.030, respectively (p < 0.05). The absorbance values at 405 nm for alkaline phosphatase activity on Day 7 for IGF-2 at 0, 10, and 100 ng/mL were 2.112 ± 0.152, 1.897 ± 0.144, and 2.067 ± 0.128, respectively (p > 0.05). The mineralization assay results at Day 7 showed significantly higher values for IGF-2 groups at 10 and 100 ng/mL concentration when compared to the control (p < 0.05). The application of IGF-2 groups of 10 and 100 ng/mL produced a significant increase of BGLAP. Conclusively, this study indicates that the use of IGF-2 on stem cell loaded bone graft increased cellular viability, Alizarin red staining, and BGLAP expression of stem cells. This report suggests the combined approach of stem cells and IGF-2 with scaffold may have synergistic effects on osteogenesis.
Highlights
Bone loss is caused by various causes, such as infection, deterioration of function due to aging, trauma, cyst, and tumor in oral and maxillofacial areas [1]
This study indicates that the use of Insulin-like growth factors (IGFs)-2 on stem cell loaded bone graft increased cellular viability, Alizarin red staining, and bone gamma-carboxyglutamate protein (BGLAP) expression of stem cells
2020, 10, 5471real-time polymerase chain reaction (PCR) revealed that the mRNA levels of BGLAP
Summary
Bone loss is caused by various causes, such as infection, deterioration of function due to aging, trauma, cyst, and tumor in oral and maxillofacial areas [1]. Sci. 2020, 10, x FOR PEER REVIEW that is similar to bone, biocompatibility that does not cause immune or inflammatory reactions, can be appropriately replaced by regenerated tissues, and radiopacity if applicable [3]. Greater interest has been given to the bone graft material that can replace autogenous or allogenic bone to recover ornot reconstruct the defect area [4]. Many successful results is similar to bone, biocompatibility that does cause immune or inflammatory reactions, absorbability are being reported regarding the use of deproteinized bovine bone in oral and maxillofacial surgery so they can be appropriately replaced by regenerated tissues, and radiopacity if applicable [3].
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