Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and <italic style="font-style: italic">ex vivo</italic> growth of mouse embryonic metatarsals

Mi LIU; Yangli XIE; Junlan HUANG; Ruobin ZHANG; Xianding SUN; Wanling JIANG; Hangang CHEN; Zhenhong NI; Bin ZHANG; Lin CHEN; Xiaolan DU

doi:10.16016/j.1000-5404.201901169

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Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsals

Basic Medicine | 更新时间：2026-01-09

- Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsals
- Journal of Third Military Medical University Vol. 41, Issue 10, Pages: 907-912(2019)
- 作者机构：
  
  400042 重庆, 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
- 作者简介：
- 基金信息：
  
  General Program of National Natural Science Foundation of China(81772306);Innovative Project of Military Medicine(16CXZ2016)
- DOI：10.16016/j.1000-5404.201901169
  CLC： R318.0; R454.4; R684.3
- Received：15 January 2019，
  
  Revised：2019-02-23，
  
  Published：30 May 2019
- 稿件说明：
移动端阅览
Mi LIU, Yangli XIE, Junlan HUANG, et al. Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsals[J]. Journal of Third Military Medical University, 2019, 41(10): 907-912.
DOI：

Mi LIU, Yangli XIE, Junlan HUANG, et al. Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsals[J]. Journal of Third Military Medical University, 2019, 41(10): 907-912. DOI： 10.16016/j.1000-5404.201901169.

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/cm

-20%；LIPUSⅡ，30 mW/cm

-40%；LIPUSⅢ，50 mW/cm

-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(

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(

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/cm

、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/cm

-20% (LIPUS Ⅰ)

30 mW/cm

-40% (LIPUS Ⅱ)

and 50 mW/cm

-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 cho

ndrocytes (

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 (

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/cm

and a duty ratio of 40% can be optimal for LIPUS treatment.

关键词

Keywords

references

COLOMBO V, CADOVÁ M, GALLO L M. Mechanical behavior of bovine nasal cartilage under static and dynamic loading. J Biomech, 2013, 46(13): 2137-2144. DOI: 10.1016/j.jbiomech.2013.07.001.

BIAN L M, ZHAI D Y, ZHANG E C, et al . Dynamic compressive loading enhances cartilage matrix synthesis and distribution and suppresses hypertrophy in hMSC-laden hyaluronic acid hydrogels. Tissue Eng Part A, 2012, 18(7/8): 715-724. DOI: 10.1089/ten.TEA.2011.0455.

HARRISON A, LIN S, POUNDER N, et al . Mode & mechanism of low intensity pulsed ultrasound (LIPUS) in fracture repair. Ultrasonics, 2016, 70: 45-52. DOI: 10.1016/j.ultras.2016.03.016.

YANG K H, PARVIZI J, WANG S J, et al . Exposure to low-intensity ultrasound increases aggrecan gene expression in a rat femur fracture model. J Orthop Res, 1996, 14(5): 802-809. DOI: 10.1002/jor.1100140518.

GUO G P, MA Y, GUO Y, et al . Enhanced porosity and permeability of three-dimensional alginate scaffolds via acoustic microstreaming induced by low-intensity pulsed ultrasound. Ultrason Sonochem, 2017, 37: 279-285. DOI: 10.1016/j.ultsonch.2017.01.016.

HOSSEINI M S, TAFAZZOLI-SHADPOUR M, HAGHIGHIPOUR N, et al . The synergistic effects of shear stress and cyclic hydrostatic pressure modulate chondrogenic induction of human mesenchymal stem cells. Int J Artif Organs, 2015, 38(10): 557-564. DOI: 10.5301/ijao.5000433.

GHARRAVI A M, ORAZIZADEH M, HASHEMITABAR M. Fluid-induced low shear stress improves cartilage like tissue fabrication by encapsulating chondrocytes. Cell Tissue Bank, 2016, 17(1): 117-122. DOI: 10.1007/s10561-015-9529-2.

XIE Y L, SU N, JIN M, et al . Intermittent PTH (1-34) injection rescues the retarded skeletaldevelopment and postnatal lethality of mice mimicking human achondroplasia and thanatophoric dysplasia. Hum Mol Genet, 2012, 21(18): 3941-3955. DOI: 10.1093/hmg/dds181.

GOLDRING M B, TSUCHIMOCHI K, IJIRI K. The control of chondrogenesis. J Cell Biochem, 2006, 97(1): 33-44. DOI: 10.1002/jcb.20652.

NOLTE P A, KLEIN-NULEND J, ALBERS G H R, et al . Low-intensity ultrasound stimulates endochondral ossification in vitro. J Orthop Res, 2001, 19(2): 301-307. DOI: 10.1016/s0736-0266(00)00027-9.

KORSTJENS C M, VAN DER RIJT R H, ALBERS G H, et al . Low-intensity pulsed ultrasound affects human articular chondrocytes in vitro . Med Biol Eng Comput, 2008, 46(12): 1263-1270. DOI: 10.1007/s11517-008-0409-9..

NAITO K, WATARI T, MUTA T, et al . Low-intensity pulsed ultrasound (LIPUS) increases the articular cartilage type Ⅱ collagen in a rat osteoarthritis model. J Orthop Res, 2010, 28(3): 361-369. DOI: 10.1002/jor.20995.

ZHANG Z J, HUCKLE J, FRANCOMANO C A, et al . The effects of pulsed low-intensity ultrasound on chondrocyte viability, proliferation, gene expression and matrix production. Ultrasound Med Biol, 2003, 29(11): 1645-1651. DOI: 10.1016/j.ultrasmedbio.2003.08.011.

ZAHOOR T, MITCHELL R, BHASIN P, et al . Effect of low-intensity pulsed ultrasound on joint injury and post-traumatic osteoarthritis: An animal study. Ultrasound Med Biol, 2018, 44(1): 234-242. DOI: 10.1016/j.ultrasmedbio.2017.09.014.

SUN H B. Mechanical loading, cartilage degradation, and arthritis. Ann N Y Acad Sci, 2010, 1211: 37-50. DOI: 10.1111/j.1749-6632.2010.05808.x.

TIEN Y C, LIN S D, CHEN C H, et al . Effects of pulsed low-intensity ultrasound on human child chondrocytes. Ultrasound Med Biol, 2008, 34(7): 1174-1181. DOI: 10.1016/j.ultrasmedbio.2007.12.019.

NISHIDA T, KUBOTA S, AOYAMA E, et al . Low-intensity pulsed ultrasound (LIPUS) treatment of cultured chondrocytes stimulates production of CCN family protein 2 (CCN 2 ), a protein involved in the regeneration of articular cartilage: mechanism underlying this stimulation. Osteoarthr Cartil, 2017, 25(5): 759-769. DOI: 10.1016/j.joca.2016.10.003..

WILTINK A, NIJWEIDE P J, OOSTERBAAN W A, et al . Effect of therapeutic ultrasound on endochondral ossification. Ultrasound Med Biol, 1995, 21(1): 121-127. DOI: 10.1016/0301-5629(94)00092-1.

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Effects of low-intensity pulsed ultrasound of different acoustic parameters on viability of cultured chondrocytes and ex vivo growth of mouse embryonic metatarsals

DOI：10.16016/j.1000-5404.201901169

摘要

Abstract

关键词

Keywords

references