Ultrasound stimulation increases proliferation of MC3T3-E1 preosteoblast-like cells.
Ultrasound stimulation increases proliferation of MC3T3-E1 preosteoblast-like cells.
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DOI：
10.1186/2050-5736-2-1
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Sarkar K
中科院分区：
文献类型：
--
作者：
Katiyar A;Duncan RL;Sarkar K
骨的机械刺激至少部分地通过增加成骨细胞和骨祖细胞的增殖来增加骨量和骨折愈合。研究人员以前进行了超声诱导成骨细胞增殖的体外研究，但大多使用固定的超声设置，并报告了广泛变化和不确定的结果。在这里，我们严格调查的激励参数的低强度脉冲超声（LIPUS）刺激对增殖的MC 3 T3-E1前成骨细胞在单层培养的影响。我们使用定制设计的超声暴露系统来改变关键的超声参数-强度、频率和激发持续时间。将MC 3 T3-E1细胞接种在12孔细胞培养板中。除非另有说明，否则在细胞接种后，将三个一组的处理细胞激发两次，持续10分钟，间隔24小时。在最后一次LIPUS激发后24小时，使用BrdU和MTS测定确定这些细胞的增殖率。所有数据均表示为平均值±标准误差。使用Student双样本双尾t检验确定统计学显著性。使用离散的LIPUS强度范围从1到500 mW/cm 2（SATA，空间平均时间平均），我们发现，约75 mW/cm 2产生的成骨细胞增殖的最大增加。较高强度（约465 mW/cm 2）的超声暴露显著降低了MC 3 T3-E1细胞的增殖，表明高强度脉冲超声可能会增加这些细胞的凋亡或粘附丧失。LIPUS频率从0.5 MHz到5 MHz的变化表明成骨细胞增殖率不是频率依赖性的。我们发现，如果LIPUS应用30分钟/天或10分钟/天，增殖率的增加没有差异，表明习惯性反应。这项研究得出结论，具有最佳强度的短期刺激可以增强前成骨细胞样骨细胞的增殖，这在骨形成和加速骨折愈合中起着重要作用，也表明可能的治疗方法用于减少骨量。
Mechanical stimulation of bone increases bone mass and fracture healing, at least in part, through increases in proliferation of osteoblasts and osteoprogenitor cells. Researchers have previously performed in vitro studies of ultrasound-induced osteoblast proliferation but mostly used fixed ultrasound settings and have reported widely varying and inconclusive results. Here we critically investigated the effects of the excitation parameters of low-intensity pulsed ultrasound (LIPUS) stimulation on proliferation of MC3T3-E1 preosteoblastic cells in monolayer cultures. We used a custom-designed ultrasound exposure system to vary the key ultrasound parameters—intensity, frequency and excitation duration. MC3T3-E1 cells were seeded in 12-well cell culture plates. Unless otherwise specified, treated cells, in groups of three, were excited twice for 10 min with an interval of 24 h in between after cell seeding. Proliferation rates of these cells were determined using BrdU and MTS assays 24 h after the last LIPUS excitation. All data are presented as the mean ± standard error. The statistical significance was determined using Student's two-sample two-tailed t tests. Using discrete LIPUS intensities ranging from 1 to 500 mW/cm2 (SATA, spatial average-temporal average), we found that approximately 75 mW/cm2 produced the greatest increase in osteoblast proliferation. Ultrasound exposures at higher intensity (approximately 465 mW/cm2) significantly reduced proliferation in MC3T3-E1 cells, suggesting that high-intensity pulsed ultrasound may increase apoptosis or loss of adhesion in these cells. Variation in LIPUS frequency from 0.5 MHz to 5 MHz indicated that osteoblast proliferation rate was not frequency dependent. We found no difference in the increase in proliferation rate if LIPUS was applied for 30 min/day or 10 min/day, indicating a habituation response. This study concludes that a short-term stimulation with optimum intensity can enhance proliferation of preosteoblast-like bone cells that plays an important role in bone formation and accelerated fracture healing, also suggesting a possible therapeutic treatment for reduced bone mass.