Chen Mei-Hsiu, Sun Jui-Sheng, Liao Shao-Yu, Tai Po-An, Li Tin-Chou, Chen Ming-Hong
Department of Internal Medicine, Far Eastern Memorial Hospital , Taipei , Taiwan .
Connect Tissue Res. 2015 Jun;56(3):219-27. doi: 10.3109/03008207.2015.1016609. Epub 2015 Mar 6.
There are limited strategies to restore the damaged annulus fibrosus (AF) of the intervertebral disc. Low-intensity pulsed ultrasound (LIPUS) has positive effects on the proliferation of several types of cells and the repair of damage tissue in vivo. However, scientific evidence of therapeutic effects of LIPUS on AF cells remains limited. The purpose of this study is to evaluate the feasibility of applying LIPUS to the repair of the AF.
We used an in vitro model of human AF cells subjected to LIPUS stimulation to examine its effects on cell proliferation and matrix metabolism. Cell viability, synthesis of collagen and glycosaminoglycan (GAG), expression of matrix metalloproteinases (MMPs) and transforming growth factor β1 and pathways involving mitogen-activated protein kinases (MAPKs) were investigated.
LIPUS significantly enhanced proliferation of AF cells after 5 days of treatment. LIPUS with an intensity of 0.5 W/cm(2) increased the collagen and GAG synthesis and decreased the expressions of MMP-1 and -3 of human AF cells. Real-time polymerase chain reactions and western blotting analysis revealed that LIPUS could increase transforming growth factor β1 (TGF-β1) and activate extracellular signal-regulated kinase (ERK) pathway. In addition, TGF-β receptor kinase inhibitor could suppress the ultrasound-induced alterations in cell viability and matrix metabolism.
The findings suggested that LIPUS could be useful as a physical stimulation of cell metabolism for the repair of the AF.
修复受损的椎间盘纤维环(AF)的策略有限。低强度脉冲超声(LIPUS)对多种细胞的增殖以及体内损伤组织的修复具有积极作用。然而,LIPUS对AF细胞治疗效果的科学证据仍然有限。本研究的目的是评估应用LIPUS修复AF的可行性。
我们使用体外人AF细胞模型,对其进行LIPUS刺激,以研究其对细胞增殖和基质代谢的影响。研究了细胞活力、胶原蛋白和糖胺聚糖(GAG)的合成、基质金属蛋白酶(MMPs)和转化生长因子β1的表达以及涉及丝裂原活化蛋白激酶(MAPKs)的信号通路。
治疗5天后,LIPUS显著增强了AF细胞的增殖。强度为0.5W/cm²的LIPUS增加了人AF细胞的胶原蛋白和GAG合成,并降低了MMP-1和-3的表达。实时聚合酶链反应和蛋白质印迹分析表明,LIPUS可增加转化生长因子β1(TGF-β1)并激活细胞外信号调节激酶(ERK)通路。此外,TGF-β受体激酶抑制剂可抑制超声诱导的细胞活力和基质代谢变化。
研究结果表明,LIPUS可作为一种物理刺激用于AF修复的细胞代谢调节。