Department of Medical Sciences, College of Medicine, Catholic Kwandong University, 24, Beomil-ro 579beon-gil, Gangneung-si, Gangwon-do, 25601, Republic of Korea.
International St. Mary's Hospital, Catholic Kwandong University, 25, Simgok-ro 100beon-gil, Seo-gu, Incheon, 22711, Republic of Korea.
Tissue Eng Regen Med. 2020 Jun;17(3):313-322. doi: 10.1007/s13770-020-00256-6. Epub 2020 Apr 9.
Although low-intensity pulsed ultrasound has been reported to be potential cartilage regeneration, there still unresolved treatment due to cartilage fibrosis and degeneration by a lack of rapid and high-efficiency treatment. The purpose of this study was to investigate the effect of a combination therapy of focused acoustic force and stem cells at site for fast and efficient healing on cartilage regeneration.
Using a rat articular cartilage defects model, one million adipose tissue-derived stem cells (ASCs) were injected into the defect site, and low-intensity focused ultrasound (LOFUS) in the range of 100-600 mV was used for 20 min/day for 2 weeks. All experimental groups were sacrificed after 4 weeks in total. The gross appearance score and hematoxylin and eosin (H&E), Alcian blue, and Safranin O staining were used for measuring the chondrogenic potential. The cartilage characteristics were observed, and type II collagen, Sox 9, aggrecan, and type X collagen were stained with immunofluorescence. The results of the comprehensive analysis were calculated using the Mankin scoring method.
The gross appearance scores of regenerated cartilage and chondrocyte-like cells in H&E images were higher in LOFUS-treated groups compared to those in negative control or ASC-treated groups. Safranin O and Alcian blue staining demonstrated that the 100 and 300 mV LOFUS groups showed greater synthesis of glycosaminoglycan and proteoglycan. The ASC + LOFUS 300 mV group showed positive regulation of type II collagen, Sox 9 and aggrecan and negative regulation of type X collagen, which indicated the occurrence of cartilage regeneration based on the Mankin score result.
The combination therapy, which involved treatment with ASC and 300 mV LOFUS, quickly and effectively reduced articular cartilage defects.
虽然低强度脉冲超声已被报道具有潜在的软骨再生能力，但由于软骨纤维化和退化，缺乏快速高效的治疗方法，仍存在未解决的治疗问题。本研究旨在探讨聚焦声力学和干细胞联合治疗在软骨再生中的作用，以快速高效地促进软骨愈合。
采用大鼠关节软骨缺损模型，将 100 万个脂肪组织来源的干细胞（ASCs）注入缺损部位，使用 100-600 mV 的低强度聚焦超声（LOFUS）每天治疗 20 分钟，共治疗 2 周。所有实验组在 4 周后全部处死。采用大体观察评分和苏木精和伊红（H&E）、阿尔辛蓝、番红 O 染色评估软骨的成软骨能力。观察软骨特征，并通过免疫荧光染色检测 II 型胶原、Sox9、聚集蛋白聚糖和 X 型胶原。采用 Mankin 评分法对综合分析结果进行计算。
LOFUS 治疗组的再生软骨和软骨细胞样细胞的大体外观评分均高于阴性对照组或 ASC 治疗组。H&E 图像中的番红 O 和阿尔辛蓝染色表明，100 和 300 mV 的 LOFUS 组表现出更大的糖胺聚糖和蛋白聚糖合成。ASC+LOFUS 300 mV 组表现出 II 型胶原、Sox9 和聚集蛋白聚糖的阳性调节以及 X 型胶原的阴性调节，根据 Mankin 评分结果表明发生了软骨再生。
ASCs 和 300 mV LOFUS 联合治疗可快速有效地减少关节软骨缺损。
