不同离体时间和保存液对犬离体牙牙周膜细胞活力影响的实验研究

Effects of different extra-oral storage time and storage medium on the viability of periodontal ligament cells from avulsed dog teeth

目的:探讨完全脱位牙不同离体时间和保存液对牙周膜细胞活力的影响。方法:麻醉拔除犬牙35个,首先将20个牙随机分为5组,分别为室温干燥放置0、30、60、120、240 min组,另15个牙室温干燥放置30 min后,随机分为3组,分别放入牛奶、HBSS液,100 g/L蜂胶液中浸泡2 h。各组处理完成后,采用全牙消化法获得牙周膜细胞,并通过4 g/L台盼蓝染色法检测各组牙周膜活细胞数和存活率。结果:室温干燥放置30、60、120、240 min后,牙周膜细胞存活率依次为33.6%、23.6%、18.5%、0.8%,而0 min的牙周膜细胞存活率可达95.5%。拔后30 min,经牛奶、HBSS液和100 g/L蜂胶液中保存2 h后,牙周膜细胞均有活力,其细胞存活率大小依次为100 g/L蜂胶液、HBSS液和牛奶,其中100 g/L蜂胶液与HBSS液相比无统计学差异(P>0.05),但与牛奶组相比,均有统计学差异(P<0.05)。结论:随着离体时间延长,完全脱位牙根面牙周膜细胞活力明显下降。100 g/L蜂胶液和HBSS液保存犬牙牙周膜细胞活力优于牛奶液。

AIM：To evaluate the effects of different extra-oral storage time and storage medium on the viability of periodontal ligament cells from avulsed dog teeth.METHODS：Thirty-five dog teeth were extracted under local anesthesia and were randomly assigned to different treatment groups.Twenty teeth were stored at room temperature for 0,30,60,120,240 min,with 4 teeth in each time point group.The PDL cells were subsequently isolated by direct enzymatic digestion method.The viabilities of the cells were examined by 0.4% Trypan Blue Stains.The other 15 teeth were stored at room temperature for 30 min.Then they were divided into HBSS,milk and 10% Propolis group（with 5 teeth in each group） and stored in the respective medium for 2 hours.Cell viability were assessed in the same way.RESULTS： The number of viable PDL cells decreased with the increase of storage time.At 0,30,60,120 and 240 minutes,the survival rate of PDL cells was 95.5%,33.6%,23.6%,18.5% and 0.8% respectively.The viability of PDL cells from teeth stored in 10% Propolis showed higher viability than those in HBSS and milk.There was no significant difference between 10% Propolis and HBSS,however,significant difference was observed between milk and other groups.CONCLUSION：The number of viable PDL cells remarkably decreases with the increase of dry storage time.10% Propolis and HBSS are better in maintaining PDL cell viability than milk.