不同存储液对人离体牙牙周膜成纤维细胞的影响

Effects of Storage Medias on Human Periodontal Ligament Fibroblasts in vitro

目的:比较5种离体牙储存液,在不同温度、不同储存时间条件下,体外维持人牙周膜成纤维细胞(periodontal ligament fibroblasts,PDLF)活性的能力,为外伤后离体牙的保存提供实验依据。方法:选取5种离体牙存储液:牛奶、汉克斯液(Hank's balanced salt solution,HBSS)、HBSS1(HBSS+阿仑膦酸钠)、HBSS2(HBSS+还原型谷胱甘肽)、HBSS3(HBSS+碱性成纤维细胞生长因子),用cck-8试剂测定人PDLF不同温度下,在上述溶液中保存不同时间后的细胞活性。结果:4℃条件下,各实验组的PDLF细胞活性随时间延长均呈下降趋势。在不同时间点,牛奶组均表现出相对较高的细胞活性,尤其在6 h时与其他实验组相比,差异有统计学意义(P<0.05)。20℃条件下,各实验组的PDLF细胞活性随时间延长也呈下降趋势,其中HBSS3组活性下降相对缓慢。在不同时间点,HBSS组和HBSS3组均表现出相对较高的细胞活性,尤其在72 h时,HBSS3组与HBSS1组相比,差异有统计学意义(P<0.05)。总体上,HBSS组、HBSS1组和HBSS2组的存储效果不受温度影响,而牛奶组和HBSS3组的存储效果与温度有关。HBSS3组PDLF细胞活性20℃时高于4℃,其中48 h、72 h差异有统计学意义(P<0.05),而牛奶组PDLF细胞活性20℃时低于4℃,其中6 h、24 h差异有统计学意义(P<0.05)。结论:牛奶可作为一种方便有效的储存介质用于脱位牙的短期保存,且低温时更有优势。当脱位牙需要长期储存时,HBSS和HBSS3可作为合适的存储液。并且,HBSS在4℃和20℃时存储效果相当,而HBSS3在室温下优势更明显。

Objective： The aim of this study was to evaluate the capacity of temperature and storage media in maintaining the viability of human periodontal ligament fibroblasts（PDLF）. Methods： PDLF in vitro were kept in whole milk, Hank＇s balanced salt solution（HBSS）, 1 mmol/L alendronate in HBSS（HBSS-1）, 5 mmol/L glutathione in HBSS（HBSS-2）,10 ng/m L basic fibroblast growth factor in HBSS（HBSS-3）respectively. The temperature of solutions were at 4℃ and 20℃respectively. After 3, 6, 24, 48 and 72 h in storage media, cell viability of PDLF were examined by CCK-8 assay. Results：At 4℃, the PDLF viabilities in every storage solution showed a downward trend with time, The milk exhibited better than other storage media in maintaining cell viability at different storage time, especially at 6 h（P0.05）. At 20℃, the PDLF viabilities in every storage media were still decreasing over time. Among them, HBSS-3 had a relatively slow decline.HBSS and HBSS-3 were better than others in maintaining cell viability at different storage time. HBSS-3 was significantly better than HBSS-1 at 72 h（P0.05）. In general, the performance of HBSS-1, HBSS-2 and HBSS were not influenced by storage temperature. However, the higher temperature decreased the effectiveness of whole milk, especially at 6 and 24 h（P0.05）. But, favored the HBSS-3, especially at 48 and 72 h（P0.05）. Conclusion： Whole milk may be an effective and convenient storage medium for avulsed teeth in a short time period and at lower temperature condition. HBSS and HBSS-3 may be suitable for long term storage in normal temperature.