2022, Vol. 26 ›› Issue (20): 3164-3172

Effects of concentrated growth factor combined with epidermal growth factor on the proliferation and aging of oral mucosa equivalents

Li Tengyan1, 2, Nie Minhai1, 2, Liu Xuqian1, 2   

Abstract: BACKGROUND: Concentrated growth factor (CGF) and epidermal growth factor (EGF) are biological materials that have been extensively studied in recent years. Studies have shown that they can affect the biological behavior of cells, thereby promoting tissue repair and injury healing.
OBJECTIVE: To explore the effects of CGF combined with EGF on the proliferation and aging of human gingival fibroblasts, epithelial cells, and human normal melanocytes.
METHODS: CGF/EGF was co-cultured with human gingival fibroblasts, epithelial cells and human normal melanocytes in vitro. The MTT assay was used to screen the optimal concentrations of CGF, EGF and their combination. Experiments were divided into: human gingival fibroblast control group, 50% CGF group, 10 μg/L EGF group, 50% CGF+10 μg/L EGF group; epithelial cell control group, 30% CGF group, 10 μg/L EGF, 30% CGF + 10 μg/L EGF group; human skin melanocyte control group, 30% CGF group, 20 μg/L EGF group, 30% CGF + 10 μg/L EGF group, 30% CGF + 20 μg/L EGF group. Cell counting kit-8 assay was used to detect the proliferation of cells, Transwell technology used to detect the migration of cells, and Annexin V/PI double staining method used to detect the apoptosis of cells. An ethical approval was obtained from the Biomedical Science Research Ethics Committee of the Affiliated Stomatological Hospital of Southwest Medical University (approval No. 20181008001) and written informed consents were obtained from patients and volunteers.
RESULTS AND CONCLUSION: The results of MTT assay suggested that the optimal concentrations of CGF on the cell proliferation were 50% for human gingival fibroblasts, 30% for epithelial cells and 30% for human normal melanocytes, while the optimal concentrations of EGF on the cell proliferation were 10 μg/L for human gingival fibroblasts, 10 μg/L for epithelial cells, and 20 μg/L for human normal melanocytes. Cell counting kit-8 assay, Transwel assay and Annexin V/PI double staining method showed that CGF combined with EGF could promote the proliferation and migration of human gingival fibroblasts, epithelial cells and human normal melanocytes and inhibit cell apoptosis. Their effects were ranked as follows: EGF > CGF + EGF > CGF. To conclude, CGF combined with EGF can promote cell proliferation and migration and inhibit cell apoptosis, and its effect is stronger than that of CGF alone but weaker than that of EGF alone.
Key words: concentrated growth factor, epidermal growth factor, proliferation, migration, apoptosis, aging, oral mucosa