Purpose to observe if the pre-existing root cementum could have an effect on the gene expression of cementum proteins by human periodontal ligament cells (hPdlcs). Methods Human Pdlcs were isolated from the premolars extracted due to orthodontic therapy and cultured. root slices were made by splitting the periodontal ligament denuded premolars into two halves. in control group, named Pdlcs+d, the cementum was scaled completely from the root surface and the dentin was exposed; while in the test group, named Pdlcs+c, the cementum was preserved in situ.the second passage hPdlcs were inoculated on root slices. after a 7-day co-culture, the cells growing on the root slices of each group were harvested together , along with cells growing in the culture plate named as group Pdlcs . total rna of the cells in three groups was extracted by using ttrizol reagent respectively. real-time reverse transcriptase polymerase chain reaction (rt-Pcr) was performed forthe measurement of mrna expression of cementum attachment protein (caP) and cementum protein23 (cP-23).β-actin was used as an internal control. the amount of mrna was calculated for each sample from the standard curve using the instrument software. Results the relative mrna expression of both cP-23 and caP in the test group were significantly more than that in the control group (p<0.01). Conclusions the results show that the pre-existing root cementum can enhance the gene expression of cementum proteins by human periodontal ligament cells. it suggests that the pre-existent cementum maybe promote the hPdlcs in vitro to differentiate towards ce-mentoblasts.% 目的探讨根面保留牙骨质对体外培养的人牙周韧带细胞表达牙骨质蛋白的影响,进一步揭示牙骨质的功能.材料和方法选用因正畸拔除的牙齿,培养人牙周韧带细胞.将牙根纵劈为两半,去除牙髓组织.一半彻底刮除根面的牙骨质,暴露牙本质,作为对照组；另一半保留牙骨质,不暴露其下的牙本质,作为实验组.接种第二代牙周韧带细胞至根片表面,复合培养7天后,收集根片上细胞和同样条件下接种在培养皿里细胞,提取 rna, real-timert-Pcr 检测3组细胞中牙骨质附着蛋白(caP)和牙骨质蛋白23(cP-23)基因表达情况,用β- actin 作为内参照.软件分析获取数据并进行 t-检验.结果人牙周韧带细胞在不同处理组中表达不同量的 cP-23和caP,实验组均高于对照组,具有统计学意义(p<0.01).结论根面保留牙骨质促进人牙周韧带细胞牙骨质蛋白基因表达,提示根面保留牙骨质可能促进体外培养的人牙周韧带细胞向成牙骨质细胞方向分化.
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