A Novel Bioreactor for Stimulating Skeletal Muscle In Vitro
A Novel Bioreactor for Stimulating Skeletal Muscle In Vitro
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DOI：
10.1089/ten.tec.2009.0125
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发表时间：
2010-08-01
影响因子：
3
通讯作者：
Baar, Keith
中科院分区：
文献类型：
--
作者：
Donnelly, Kenneth;Khodabukus, Alastair;Baar, Keith
300多年来，科学家们一直明白，以电脉冲形式的刺激是正常肌肉功能所必需的。最近，电脉冲的特定参数(即脉冲幅度、脉冲宽度和作息比)的作用已得到更好的认识。然而，大多数现有的生物反应器系统不允许对这些参数进行足够的控制。因此，本研究的目的是设计一种廉价的肌肉电刺激生物反应器，将生理相关的电刺激模式应用于培养中的组织工程肌肉和单层。使用低功率微控制器和DC-DC转换器为脉冲电路供电，该电路将4.5V输入转换为高达50V的输出，脉冲宽度从0.05到4ms，频率高达100赫兹(具有一定的操作限制)。当二维培养细胞受到高频(100 Hz)刺激时，蛋白质合成速率增加(12h时，对照组[CTL]=5.0+/-0.16；10 Hz5.0+/-0.07；100 Hz5.5+/-0.13fmoL/min/mg)，表明这是一个合成代谢信号。当刺激时间为0.1ms，刺激1~2倍时，刺激可提高肌肉的作用力(CTL=0.07±/-0.009；1.25V/mm=0.10±0.011；2.5V/mm=0.14146+/-0.012；5V/mm=0.03756+/-0.008kN/mm(2))和兴奋性(CTL=0.53+/-0.022；1.25V/mm=0.44+/-0.025；2.5V/mm=0.41+/-0.012；5V/mm=0.60+/-0.021V/mm)，提示成熟度增强。总之，这些数据表明，使用生物反应器可以在体外改善肌肉的生理和功能，这种生物反应器允许控制脉冲幅度、脉冲宽度、脉冲频率和作息比。
For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5 V input to outputs of up to 50 V, with pulse widths from 0.05 to 4 ms, and frequencies up to 100 Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100 Hz), this resulted in an increase in the rate of protein synthesis (at 12 h, control [CTL] = 5.0 +/- 0.16; 10 Hz 5.0 +/- 0.07; and 100 Hz 5.5 +/- 0.13 fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1 ms and one or two times rheobase, stimulation improved force production (CTL = 0.07 +/- 0.009; 1.25V/mm = 0.10 +/- 0.011; 2.5V/mm = 0.14146 +/- 0.012; and 5V/mm = 0.03756 +/- 0.008kN/mm(2)) and excitability (CTL = 0.53 +/- 0.022; 1.25V/mm = 0.44 +/- 0.025; 2.5V/mm = 0.41 +/- 0.012; and 5V/mm = 0.60 +/- 0.021V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio.