Objective: To investigate the effects of storage media and temperature on the activity and proliferation of human periodontal ligament fibroblasts(hPDLFs) by simulating the storage model of an avulsed tooth in vitro cell culture. Methods: Three readily available storage media(milk, saliva, and purified water) were selected and applied to hPDLFs for 1 h at room temperature and 4℃. Then, 5-ethynyl-2'-deoxyuridine(EdU) and cell counting kit-8(CCK-8) were used to detect the viability and proliferative capacity of hPDLFs after treatment. Results: The EdU assay showed that after treating the cells with milk at 4℃ for 1 h, most cells were EdU-positive(green), accounting for about 50.9% of the total cells. The CCK-8 assay showed that hPDLFs viabilities in milk, saliva, and purified water were 62.8%, 17.8%, and 3.3% respectively, at 4℃, and 56.2%, 15.2%, and 0.7%, respectively, at room temperature. The growth curve of cells stored in milk at 4℃ was the closest to the normal control after 24, 48, and 72 h of culture. The proliferation rate of cells stored in milk was statistically significantly different compared with that of cells stored in saliva and purified water. Conclusion: hPDLFs showed the best cell viability and proliferative ability in milk at 4℃. Therefore, low-temperature milk is the preferred preservation solution for avulsed teeth. |
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