Department of Periodontics, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
J Surg Res. 2012 Dec;178(2):737-42. doi: 10.1016/j.jss.2012.03.072. Epub 2012 Apr 21.
The tetracycline (TC) family, including minocycline and doxycycline (DC), has long been used in the medical field owing to its well-characterized antimicrobial properties. Moreover, TCs have been reported to have effects on bone formation and bone metabolism. Results have shown that more new bone formation is achieved when TCs have been applied in combination with bone graft material.
In the present study, the effects of TCs (TC, minocycline, and DC) on osteoprecursor cells were evaluated. The cell viability was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Differentiation and mineralization were evaluated using an alkaline phosphatase activity test and alizarin red-S staining. In addition, the expression of proteins related to bone formation, such as estrogen receptor (ER)-α, bone morphogenetic protein receptor-IA, bone morphogenetic protein-2, and phospho-Smad1/5, were evaluated by using Western blot analysis.
The morphology of the cells seemed normal, and their viability was not affected in the treated groups compared with the control. Alkaline phosphatase activity significantly increased in cultures grown in the presence of 0.1 and 1.0 μM of DC. No statistically significant increase in the mineralization was seen the treated groups. The results of the Western blot analysis revealed that the addition of DC upregulated ER-α, bone morphogenetic protein-2, and phospho-Smad1/5 expression with a statistically significant difference in ER-α expression.
From our findings, it was concluded that a low dose of DC could produce positive effects on the differentiation of osteoprecursor cells at an early stage. Our results also suggested that DC has osteoinductive effects that were achieved mainly through the ER pathway by enhancing the expression of ER-α.
四环素（TC）家族，包括米诺环素和强力霉素（DC），由于其具有良好特征的抗菌特性，长期以来一直在医学领域使用。此外，已经报道 TC 对骨形成和骨代谢有影响。结果表明，当 TC 与骨移植物材料联合应用时，会有更多的新骨形成。
在本研究中，评估了 TC（TC、米诺环素和 DC）对成骨前体细胞的影响。使用 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑溴盐测定法测定细胞活力。通过碱性磷酸酶活性试验和茜素红 S 染色评估分化和矿化。此外，通过 Western blot 分析评估与骨形成相关的蛋白质的表达，如雌激素受体（ER）-α、骨形态发生蛋白受体-IA、骨形态发生蛋白-2 和磷酸化 Smad1/5。
与对照组相比，处理组细胞形态似乎正常，其活力不受影响。在存在 0.1 和 1.0 μM 的 DC 的培养物中，碱性磷酸酶活性显著增加。处理组未见矿化有统计学显著增加。Western blot 分析的结果表明，添加 DC 可上调 ER-α、骨形态发生蛋白-2 和磷酸化 Smad1/5 的表达，其中 ER-α 的表达有统计学显著差异。
根据我们的发现，可以得出结论，低剂量的 DC 可以在早期对成骨前体细胞的分化产生积极影响。我们的结果还表明，DC 具有成骨作用，主要通过增强 ER-α 的表达通过 ER 途径实现。
