USING LOW-INTENSITY PULSED ULTRASOUND TO IMPROVE MUSCLE HEALING AFTER LACERATION INJURY: AN IN VITRO AND IN VIVO STUDY
USING LOW-INTENSITY PULSED ULTRASOUND TO IMPROVE MUSCLE HEALING AFTER LACERATION INJURY: AN IN VITRO AND IN VIVO STUDY
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DOI：
10.1016/j.ultrasmedbio.2010.02.010
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发表时间：
2010-05-01
影响因子：
2.9
通讯作者：
Ueng, Steve Wen-Neng
中科院分区：
文献类型：
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作者：
Chan, Yi-Sheng;Hsu, Kuo-Yao;Ueng, Steve Wen-Neng
本研究的目的是确定低强度脉冲超声（LIPUS）是否可以增强肌纤维的再生并缩短受伤肌肉的愈合时间。 NIH C2C12 细胞是一种众所周知的成肌细胞系，是源自正常成年 C3H 小鼠腿部肌肉建立的小鼠成肌细胞系的亚克隆。细胞迅速分化并产生广泛收缩的肌管，表达特征性的肌肉蛋白。我们使用 EXOGEN 2000+ 系统超声设备（Exogen Inc.，Piscataway，新泽西州，美国）将 C2C12 细胞暴露于 LIPUS 治疗，每 24 小时总治疗 20 分钟。每隔2、4、6和8天，通过细胞数量的增加以及肌细胞生成素和肌动蛋白的蛋白质印迹分析来测量细胞生长。将四十只小鼠 (C57BL10J+/+) 分为五组，每组八只动物，并用于已发表的撕裂伤模型。所有动物的左腿腓肠肌均被撕裂。对照组（假超声）不接受 LIPUS 治疗。超声 7 天、14 天、21 天和 28 天组（仅改变超声应用于受伤肌肉的天数）分别用 LIPUS（20 分钟/天）治疗连续 7、14、21 和 28 天。受伤后4周处死所有动物。评估方法包括肌肉再生和肌肉收缩特性。 LIPUS 治疗在第 6 天和第 8 天产生显着更高的增殖率和细胞数量 (p < 0.05)。光密度评估显示，在 4、6 和 8 天组中，用 LIPUS 处理的细胞中肌细胞生成素和肌动蛋白有所增加。与对照肌肉相比，经 LIPUS 处理的肌肉（21 天和 28 天）的肌纤维、快肌纤维和破伤风的再生明显更大。正常未受伤的肌肉和接受 LIPUS 治疗 28 天的组之间没有明显的力量差异。总之，这是第一项实验研究表明，LIPUS 疗法能够增强撕裂后受伤小鼠肌肉的肌纤维再生，并具有更好的生理性能，尤其是在术后第 4 周之前。这项研究的结果为未来的临床试验提供了科学依据，并为 LIPUS 增强撕裂伤后肌肉愈合的适应症奠定了基础。 (E-mail: chan512@adm.cgmh.org.tw) (c) 2010 世界超声医学与生物学联合会。
The purpose of this study was to determine whether low-intensity pulsed ultrasound (LIPUS) could enhance the regeneration of myofibers and shorten the healing time in injured muscle. NIH C2C12 cells, a well-known myoblastic cell line, are subclones derived from the mouse myoblast cell line established from normal adult C3H mouse leg muscle. The cells differentiate rapidly and produce extensive contracting myotubes expressing characteristic muscle proteins. We exposed C2C12 cells to LIPUS therapy using the EXOGEN 2000+ system ultrasound apparatus (Exogen Inc., Piscataway, NJ, USA) with a total treatment of 20 min every 24 h. At intervals of 2, 4, 6 and 8 days, cell growth was measured by the increase in cell number and western blot analysis of myogenin and actin. Forty mice (C57BL10J+/+) were divided into five groups of eight animals each and used in the published laceration injury model. The gastrocnemius muscle of the left leg was lacerated in all the animals. The control group (sham ultrasound) did not undergo LIPUS therapy. The ultrasound 7-, 14-, 21- and 28-day groups (only changing the number of days during which the ultrasound was applied to the injured muscle) were treated with LIPUS (20 min/day) for 7, 14, 21 and 28 consecutive days, respectively. All animals were sacrificed at 4 weeks after the injury. Evaluation methods included muscle regeneration and muscle contractile properties. LIPUS therapy produced a significantly higher proliferative rate and cell number at days 6 and 8 (p < 0.05). Densitometric evaluation revealed an increase in myogenin and actin proteins in cells treated with LIPUS in the 4-, 6- and 8-day groups. The regeneration of myofibers, fast-twitch and tetanus of LIPUS-treated muscles (21 and 28 days) was significantly greater relative to control muscles. There was no major strength difference between the normal non-injured muscle and the group treated with LIPUS for 28 days. In conclusion, this was the first experimental study to show that LIPUS therapy is able to enhance the regeneration of myofibers with better physiologic performance in injured mice muscles after laceration, especially prior to postoperative week 4. Findings of this study demonstrate a scientific basis for future clinical trials and establish an indication for LIPUS in enhancing muscle healing after laceration injury. (E-mail: chan512@adm.cgmh.org.tw) (c) 2010 World Federation for Ultrasound in Medicine & Biology.