Du Mi, Pan Wan, Yang Pishan, Ge Shaohua
Department of Periodontology, School of Stomatology, Shandong University; Shandong Provincial Key Laboratory of Oral Tissue Regeneration, Jinan 250012, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2016 Mar;51(3):160-5. doi: 10.3760/cma.j.issn.1002-0098.2016.03.007.
To determine the effect of aspirin on cell proliferation, alkaline phosphatase (ALP) activity, cell cycle and apoptosis in murine bone marrow stromal cells, so as to explore an appropriate dose range to improve bone regeneration in periodontal treatment.
ST2 cells were stimulated with aspirin (concentrations of 1, 10, 100 and 1 000 μmol/L) for 1, 2, 3, 5 and 7 d. Cell proliferation was measured by methyl thiazolyl tetrazolium (MTT) assay. After ST2 cells were treated for 1, 3 and 7 d, ALP activity was measured by ALP kit, cell cycle and apoptosis were measured by flow cytometry (FCM) after treated for 48 h.
MTT assays showed that various doses of aspirin have different effects on the cell growth. Briefly, lower concentrations (1, 10 μmol/L) of aspirin promoted the cell growth, the A value of 0, 1 and 10 μmol/L aspirin 7-day-treated cells were 0.313±0.012, 0.413±0.010 and 0.387±0.017 respectively (P <0.01 vs control), and so did the ALP level ([4.3±0.9], [6.0±0.3] and [7.7±0.4] μmol·min(-1)·g(-1), P <0.05 vs control), while higher concentrations, especially 1000 μmol/L of aspirin might inhibit the cell growth with time going, A value and ALP level were 0.267±0.016, (4.3±1.3) μmol·min(-1)·g(-1) respectively (P <0.05 vs control). Cell cycle analysis revealed no changes in comparison to control cells after treatment with 1 or 10 μmol/L aspirin, but it was observed that cell mitosis from S phase to G2/M phase proceeded at higher concentrations of 100 μmol/L aspirin, and the cell cycle in phase G0/G1 arrested at 1000 μmol/L. Parallel apoptosis/necrosis studies showed that the percentage of cells in apoptosis decreased dramatically at all doses of aspirin, the apoptosis rates of ST2 cells responded to 0, 1, 10, 100 and 1000 μmol/L aspirin were (11.50±0.90)%, (5.30±0.10)%, (5.50±0.10)%, (4.90±0.90)% and (7.95±0.25)% respectively (P<0.05 vs control).
This study demonstrated that lower dosage of aspirin can promote ST2 cells growth, osteogenic activity and inhibit its apoptosis. Aspirin maybe used for the bone reconstruction with a proper concentration.
确定阿司匹林对小鼠骨髓基质细胞增殖、碱性磷酸酶（ALP）活性、细胞周期及凋亡的影响，以探索促进牙周治疗中骨再生的合适剂量范围。
用阿司匹林（浓度为1、10、100和1000 μmol/L）刺激ST2细胞1、2、3、5和7天。采用甲基噻唑基四氮唑（MTT）法检测细胞增殖。ST2细胞处理1、3和7天后，用ALP试剂盒检测ALP活性，处理48小时后用流式细胞术（FCM）检测细胞周期和凋亡。
MTT分析表明，不同剂量的阿司匹林对细胞生长有不同影响。简而言之，较低浓度（1、10 μmol/L）的阿司匹林促进细胞生长，1、10 μmol/L阿司匹林处理7天的细胞A值分别为0.313±0.012、0.413±0.010和0.387±0.017（与对照组相比P<0.01），ALP水平也是如此（分别为[4.3±0.9]、[6.0±0.3]和[7.7±0.4] μmol·min⁻¹·g⁻¹，与对照组相比P<0.05），而较高浓度，尤其是1000 μmol/L的阿司匹林可能随时间抑制细胞生长，A值和ALP水平分别为0.267±0.016、(4.3±1.3) μmol·min⁻¹·g⁻¹（与对照组相比P<0.05）。细胞周期分析显示，用1或10 μmol/L阿司匹林处理后与对照细胞相比无变化，但观察到100 μmol/L较高浓度阿司匹林处理时细胞从S期到G2/M期的有丝分裂进行，1000 μmol/L时细胞周期在G0/G1期停滞。平行的凋亡/坏死研究表明，所有剂量的阿司匹林处理后凋亡细胞百分比均显著降低，ST2细胞对0、1、10、100和1000 μmol/L阿司匹林的凋亡率分别为(11.50±0.90)%、(5.30±0.10)%、(5.50±0.
