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低频脉冲超声波治疗绵羊模型中的软骨缺陷。

Treatment of cartilage defects by Low-intensity pulsed ultrasound in a sheep model.

机构信息

Department of Anatomical Sciences, Maragheh University of Medical Sciences, Maragheh, Iran.

Department of Basic Sciences, Maragheh University of Medical Sciences, Maragheh, Iran.

出版信息

Cell Tissue Bank. 2021 Sep;22(3):369-378. doi: 10.1007/s10561-020-09880-x. Epub 2020 Nov 11.

Abstract

Aim of this study was to evaluate effects of Low-intensity pulsed ultrasound on repair of articular cartilage defects. Low-intensity pulsed ultrasound (Lipus) can induce the differentiation and activation of chondrocytes. This study was designed to evaluate the effect of Lipus on articular cartilage defects in a sheep. Eight sheep were divided in to two groups. The animals received bilateraly, articular cartilage defects 4 mm in diameter and 2 mm in deep on the patellar groove and experimental groups were treated with intensity 200 mW/cm, 20 min/day with low-intensity pulsed ultrasound for 2 month. Then both knee joints underwent surgery for remove of formed tissue sample from defects.The samples were evaluated by Quantitative real-time polymerase chain reaction (qRT-PCR), Safranin-o staining, Immunofluorescence Staining and Morphological characterization. The best and worst sample per group according to Macroscopic and micriscopic scoring were icentified. The results showed that the operated groups with-Lipus-treatment and without-Lipus treatment had considered statistically significant. Gross photography revealed that the defects in experimental groups were filled with proliferative tissue, while in control groups, a thin layer of proliferative tissue was formed in defects. qRT-PCR results showed the expression of coll2, sox9, aggrecan and Osteocalcin in experimental groups. Intense safranin-O staining show the formation cartilage tissue in ultrasound treated group, while loose safranin-o-staining were observed at the control groups. Immunofluorescence staining showed the type 2 Collagen protein expression. We suggest that low-intensity pulsed ultrasound provide the mechanistic basis force for articular cartilage repair and effective treatment modality for improving of articular cartilage defects.

摘要

本研究旨在评估低强度脉冲超声对关节软骨缺损修复的影响。低强度脉冲超声(Lipus)可诱导软骨细胞的分化和激活。本研究旨在评估 Lipus 对绵羊关节软骨缺损的影响。将 8 只绵羊分为两组。动物双侧膝关节髁间沟处形成直径 4mm、深 2mm 的关节软骨缺损,实验组采用强度为 200mW/cm 的低强度脉冲超声治疗,每天 20 分钟,共 2 个月。然后,两组均行手术切除缺损处形成的组织样本。采用实时定量聚合酶链反应(qRT-PCR)、番红 O 染色、免疫荧光染色和形态学特征对样本进行评估。根据宏观和微观评分,确定每组中最佳和最差的样本。结果显示,有-Lipus 治疗和无-Lipus 治疗的实验组差异有统计学意义。大体摄影显示实验组缺损处有增生组织填充,而对照组缺损处形成了一层较薄的增生组织。qRT-PCR 结果显示实验组中 coll2、sox9、aggrecan 和 Osteocalcin 的表达。番红 O 染色显示实验组形成软骨组织,而对照组染色疏松。免疫荧光染色显示 2 型胶原蛋白的表达。我们认为,低强度脉冲超声为关节软骨修复提供了机制基础,是改善关节软骨缺损的有效治疗方法。

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