[1]刘汨,谢杨丽,黄俊兰,等.不同声学参数的低强度脉冲超声对软骨细胞活性及培养骨胚生长的影响[J].第三军医大学学报,2019,41(10):907-912.
 LIU Mi,XIE Yangli,HUANG Junlan,et al.Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsal[J].J Third Mil Med Univ,2019,41(10):907-912.
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不同声学参数的低强度脉冲超声对软骨细胞活性及培养骨胚生长的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第10期
页码:
907-912
栏目:
基础医学
出版日期:
2019-05-30

文章信息/Info

Title:
Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsal
作者:
刘汨谢杨丽黄俊兰张若彬孙先定蒋宛凌陈涵纲倪振洪张斌陈林杜晓兰
陆军军医大学(第三军医大学)大坪医院野战外科研究所战创伤康复医学研究室,创伤实验室, 骨代谢与修复中心, 创伤、烧伤与复合伤国家重点实验室 ,全军军事训练伤防治与康复实验室
Author(s):
LIU Mi XIE Yangli HUANG Junlan ZHANG Ruobin SUN Xianding JIANG Wanling CHEN Hangang NI Zhenhong ZHANG Bin CHEN Lin DU Xiaolan

Laboratory for Rehabilitation of Traumatic Injuries, Laboratory of Trauma, Center of Bone Metabolism and Repair ,State Key Laboratory of Trauma, Burns and Combined Injury, Laboratory for Prevention and Rehabilitation of Military Training Related Injuries,  Institute of Surgery Research, Daping Hospital, Army Medical University(Third Military Medical University), Chongqing 400042, China

关键词:
低强度脉冲超声强度占空比软骨细胞骨培养小鼠
Keywords:
low-intensity pulsed ultrasound intensity duty ratio chondrocytes bone culture mice  
分类号:
R318.0;R454.4;R684.3
文献标志码:
A
摘要:

目的 探讨低强度脉冲超声(low-intensity pulsed ultrasound, LIPUS)对体外培养软骨细胞活性、增殖、分化和凋亡以及骨组织生长的影响。方法 分离新生3~5 d C57BL/6J小鼠的软骨细胞和胚胎期18.5 d小鼠跖骨,分为对照组(未处理组)及3种LIPUS处理组,其声强度和占空比的参数为:LIPUSⅠ,30 mW/cm2-20%;LIPUSⅡ,30 mW/cm2-40%;LIPUSⅢ,50 mW/cm2-20%;其余参数为:中心频率1.5 MHz,脉冲重复频率1 KHz,处理时间20 min/d;分别用CCK-8、EdU掺入及TUNEL检测软骨细胞的活性变化、增殖、凋亡;定量RT-PCR检测蛋白聚糖(Aggrecan,Acan)、Ⅱ型胶原(Collagen Ⅱ,ColⅡ)、Ⅹ型胶原(Collagen Ⅹ,ColⅩ)等软骨基质相关基因表达;采集图像后测量跖骨长度,计算跖骨全长增长率、软骨增长率。结果 3种参数的LIPUS处理能促进软骨细胞的活性和增殖,抑制软骨细胞凋亡,其中LIPUSⅡ组效果最明显;定量RT-PCR结果显示:各处理组软骨细胞的Acan、ColⅡ的mRNA表达均较对照组明显增高,ColⅩ表达降低(P<0.05);跖骨长度计算结果显示:LIPUSⅡ、Ⅲ组软骨和全长增长率均显著高于对照组(P<0.01),而LIPUSⅠ组与对照组无明显差异。结论  LIPUS可能通过促进软骨细胞活性、增殖以及细胞外基质合成,抑制细胞凋亡,加快骨组织生长,且效果与超声参数有关;其中声强度30 mW/cm2、占空比40%的LIPUS作用最明显。

Abstract:

Objective To investigate the effects of low-intensity pulsed ultrasound (LIPUS) on the viability, proliferation, differentiation and apoptosis of cultured chondrocytes and ex vivo development of embryonic mouse metatarsals. Methods Chondrocytes isolated from neonatal mice (3-5 days old) and metatarsals from mouse embryos (18 days of gestation) were both divided into control group and 3 LIPUS treatment groups for LIPUS exposures with acoustic intensity and duty ratio combinations of 30 mW/cm2-20% (LIPUS Ⅰ), 30 mW/cm2-40% (LIPUS Ⅱ), and 50 mW/cm2-20% (LIPUS Ⅲ); all the other ultrasound parameters were identical (central frequency of 1.5 MHz, repetition frequency of 1 kHz and exposure time of 20 min per day). After LIPUS treatments, the chondrocytes were examined for changes in cell viability, proliferation and apoptosis using cell counting kit-8, EdU assay and TUNEL assay, respectively. The mRNA expression levels of cartilage matrix-related genes were examined by qRT-PCR. The changes in the total length and cartilage length of the metatarsals were measured after the treatments. Results Exposure to LIPUS significantly enhanced the viability and proliferation and reduced the apoptotic rate of the chondrocytes, and the effect was the most obvious in LIPUS Ⅱ group. LIPUS treatment significantly up-regulated the mRNA expressions of Col Ⅱ and proteoglycan and reduced the mRNA expression of Col X in the chondrocytes (P<0.05). As for the metatarsals, the growth rates of the total length and cartilage length were significantly greater in LIPUS Ⅱ and LIPUS Ⅲ groups than in the control group (P<0.001), but no significant difference was found between LIPUS Ⅰ group and control group. Conclusion LIPUS treatment can promote bone growth by stimulating chondrocyte proliferation and extracellular matrix synthesis while inhibiting cell apoptosis. These effects are related with the acoustic parameters of LIPUS, and the parameter setting with an acoustic intensity of 30 mW/cm2 and a duty ratio of 40% can be optimal for LIPUS treatment.

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更新日期/Last Update: 2019-05-24