Chang Chen-Jung, Hsu Shan-Hui, Lin Fang-Tsun, Chang Han, Chang Chao-Sheng
Department of Radiological Technology, Chung-Tai Institute of Health Science and Technology, Taichung, Taiwan, Republic of China.
J Biomed Mater Res B Appl Biomater. 2005 Oct;75(1):99-107. doi: 10.1002/jbm.b.30269.
This study investigated the effects of low intensity ultrasound on seeded Schwann cells within poly(DL-lactic acid-co-glycolic acid) (PLGA) conduits by in vitro and in vivo trials for peripheral nerve regeneration. The possible differences in the ultrasonic effects when using biodegradable and non-biodegradable materials as the conduits were also studied, using silicone rubber tubes as comparisons. In the in vitro study, seeded Schwann cells were cultured in serum deprivation culture medium that simulated the environment of mechanical trauma on injury nerve site. After 12, 24, and 48 h, only the PLGA conduit groups exposed to 0.05 W/cm(2), 3 min/treatment of ultrasound exhibited decreased LDH release and increased MTT values compared to the sham groups. Based on the results of the in vitro experiment in LDH and MTT testing, the silicone conduits with seeded Schwann cells group was ignored in the in vivo study. The PLGA nerve conduits seeded with Schwann cells (9 x 10(3) cells) were implanted to 15-mm right sciatic nerve defects in rats. Each conduit received 12 ultrasonic treatment sessions over 2 weeks after 1 day of rest. Ultrasound was applied as follows: frequency, 1MHz; intensity, 0.3 W/cm(2) (SATP); treatment, 5 min/day. Implanted graft specimens were harvested for histological analysis at 8 weeks following surgery. PLGA groups (with and without Schwann cells) treated with pulsed ultrasonic stimulation were found to have significantly greater number and area of regenerated axons at the mid-conduit of implanted grafts, as compared to the sham groups. Ultrasonic stimulation on silicone groups was found to induce a mass of fibrous tissues that covered the nerve conduits and retarded axon regeneration.
本研究通过体外和体内试验,探究了低强度超声对聚(DL-乳酸-乙醇酸共聚物)(PLGA)导管内接种的雪旺细胞促进周围神经再生的作用。还以硅橡胶管作为对照,研究了使用可生物降解和不可生物降解材料作为导管时超声效应的可能差异。在体外研究中,将接种的雪旺细胞培养在模拟损伤神经部位机械创伤环境的血清剥夺培养基中。12、24和48小时后,与假手术组相比,仅暴露于0.05W/cm²、每次治疗3分钟超声的PLGA导管组乳酸脱氢酶(LDH)释放减少,噻唑蓝(MTT)值增加。基于LDH和MTT测试的体外实验结果,体内研究忽略了接种雪旺细胞的硅橡胶导管组。将接种雪旺细胞(9×10³个细胞)的PLGA神经导管植入大鼠15毫米的右侧坐骨神经缺损处。在休息1天后的2周内,每个导管接受12次超声治疗。超声应用如下:频率1MHz;强度0.3W/cm²(空间平均时间平均声功率);治疗时间5分钟/天。术后8周采集植入移植物标本进行组织学分析。与假手术组相比,发现接受脉冲超声刺激的PLGA组(有和没有雪旺细胞)在植入移植物的导管中部有明显更多数量和更大面积的再生轴突。发现对硅橡胶组的超声刺激诱导了大量覆盖神经导管并阻碍轴突再生的纤维组织。