Byeong-Wook Song1, Jun-Hee Park2, Bomi Kim2, Seahyoung Lee2,3, Soyeon Lim2,3, Sang Woo Kim2,3, Jung-Won Choi3, Jiyun Lee3, Misun Kang3, Ki-Chul Hwang2,3, Dong-Sik Chae4, Il-Kwon Kim5,6. 1. Department of Medical Sciences, College of Medicine, Catholic Kwandong University, 24, Beomil-ro 579beon-gil, Gangneung-si, Gangwon-do, 25601, Republic of Korea. 2. International St. Mary's Hospital, Catholic Kwandong University, 25, Simgok-ro 100beon-gil, Seo-gu, Incheon, 22711, Republic of Korea. 3. Institute for Biomedical Convergence, College of Medicine, Catholic Kwandong University, 24, Beomil-ro 579beon-gil, Gangneung-si, Gangwon-do, 25601, Republic of Korea. 4. Department of Orthopedic Surgery, International St. Mary's Hospital, College of Medicine, Catholic Kwandong University, 25, Simgok-ro 100beon-gil, Seo-gu, Incheon, 22711, Republic of Korea. drchae@ish.ac.kr. 5. International St. Mary's Hospital, Catholic Kwandong University, 25, Simgok-ro 100beon-gil, Seo-gu, Incheon, 22711, Republic of Korea. ilkwonkim@empas.com. 6. Institute for Biomedical Convergence, College of Medicine, Catholic Kwandong University, 24, Beomil-ro 579beon-gil, Gangneung-si, Gangwon-do, 25601, Republic of Korea. ilkwonkim@empas.com.
Abstract
BACKGROUND: 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. METHODS: 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. RESULTS: 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. CONCLUSION: The combination therapy, which involved treatment with ASC and 300 mV LOFUS, quickly and effectively reduced articular cartilage defects.
BACKGROUND: 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. METHODS: Using a ratarticular 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. RESULTS: 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. CONCLUSION: The combination therapy, which involved treatment with ASC and 300 mV LOFUS, quickly and effectively reduced articular cartilage defects.
Authors: S Farshid Moussavi-Harami; Douglas R Pedersen; James A Martin; Stephen L Hillis; Thomas D Brown Journal: J Orthop Res Date: 2009-04 Impact factor: 3.494