Objective: To study the effect of the inhibitory concentration minocycline on the proliferation, differentiation, and expression of Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and osteopontin (OPN) mRNA of osteoblasts.
Methods: Primary osteoblasts were cultured in osteogenic induction medium containing 0, 0.1, 0.5, 1, 10 μg·mL⁻¹ minocycline. Cell counting kit-8 was used to observe cell proliferation. ALP activity assay, alizarin red S staining, and real-time quantitative polymerase chain reaction (PCR) were used to determine cell differentiation and mineralization.
Results: The groups with 0.1, 0.5, 1 μg·mL⁻¹ minocycline promoted cell proliferation. The mRNA expression levels of ALP and Runx2 were up-regulated. Osteoblast-mediated mineralization was increased. The group with 1 μg·mL⁻¹ showed maximal promotion effect (P<0.05). When the concentration increased to 10 μg·mL⁻¹, the promoting effect began to decline, and the ALP activity and OPN expression were significantly inhibited (P<0.01).
Conclusions: Appropriate concentration of minocycline can promote osteoblasts proliferation, up-regulate the expression levels of Runx2, ALP and OPN, and increase the differentiation and mineralization of osteoblasts.
目的 研究抑菌浓度的米诺环素对成骨细胞增殖、分化和矿化及Runt相关转录因子2（Runx2）、碱性磷酸酶（ALP）、骨桥蛋白（OPN）mRNA表达的影响。方法 将米诺环素溶液（0、0.1、0.5、1、10 μg·mL⁻¹）与原代成骨细胞共培养，CCK-8检测增殖活性，通过ALP活性检测、茜素红染色、荧光定量聚合酶链反应检测探讨米诺环素对成骨细胞分化、矿化的影响。结果 0.1、0.5、1 μg·mL⁻¹的米诺环素可以促进细胞的增殖，上调ALP、Runx2 mRNA的表达水平，增加钙含量及钙化结节的形成，其中1 μg·mL⁻¹具有最大促进作用（P<0.05）；当浓度为10 μg·mL⁻¹时这种促进作用开始下降，并对ALP活性和OPN表达有显著抑制作用（P<0.01）。结论 适宜抑菌浓度的米诺环素能促进成骨细胞增殖，上调Runx2、ALP、OPN的表达水平，促进成骨细胞分化和矿化。.
Keywords: alkaline phosphatase; minocycline; osteoblast; proliferation.