치조골 줄기세포 증식에 저강도 초음파 자극의 효과

이응태 ( Eung Tae Lee ),임기택 ( Ki Taek Lim ),김장호 ( Jang Ho Kim ),임애리 ( Ae Li Im ),손현목 ( Hyun Mok Son ),조종수 ( Chong Su Cho ),정필훈 ( Pill Hoon Choung ),정종훈 ( Jong Hoon Chung ) 한국조직공학·재생의학회 2008 조직공학과 재생의학 Vol.5 No.4

Low-intensity ultrasound stimulation produces significant multi-functional effects that are directly relevant to bone formation. It was previously found that low-intensity ultrasound stimulation enhanced bone regeneration although the exact cellular mechanism is not clear. The aim of the present study is to investigate the effect of low-intensity ultrasound stimulation on proliferation of alveolar bone marrow stem cells. Before low-intensity ultrasound stimulation, alveolar bone marrow stem cells were cultured for 24h to facilitate their attachment. The cells were cultured in medium with or without low-intensity ultrasound stimulation. The ultrasound frequency was 1 MHz. Cell cultures stimulated with ultrasound were conducted by three treatment groups - group 1: intensity(100, 200, 300, 400 and 500 mW/cm2), group 2: duty cycle(5, 10, 30 and 50%) and group 3: duration time(1, 3, 5, 10, 20 and 30 min). The effect of low-intensity ultrasound stimulation were evaluated by cell number and morphological changes. The proliferation rates of alveolar bone marrow stem cells for the particular stimulated groups were larger than those of control groups. After low-intensity ultrasound stimulation(intensity: 100 mW/cm2, duty cycle: 30% and duration time: 10 min), the alveolar bone marrow stem cell counts were significantly increased(p < 0.05). This study suggested that the cell growth could be enhanced by appropriate low-intensity ultrasound stimulation.

A pilot clinical study of low-intensity transcranial focused ultrasound in Alzheimer’s disease

정현석,Im Jooyeon Jamie,Park Jong-Sik,Na Seunghee,Lee Wonhye,Yoo Seung-Schik,송인욱,Chung Yong-An 대한초음파의학회 2021 ULTRASONOGRAPHY Vol.40 No.4

Purpose: Increasing attention has been paid to low-intensity transcranial focused ultrasound (tFUS) for its potential therapeutic effects in Alzheimer's disease (AD). While preclinical studies have shown promising therapeutic effects of low-intensity tFUS in AD models, its efficacy and safety remain unclear in humans. In this pilot study, we investigated the effects of low-intensity tFUS on blood-brain barrier opening, the regional cerebral metabolic rate of glucose (rCMRglu), and cognition in patients with AD. Methods: After receiving institutional review board approval, four patients with AD received tFUS to the hippocampus immediately after an intravenous injection of a microbubble ultrasound contrast agent. Sonication was delivered at low-intensity, at a pressure level below the threshold for blood-brain barrier opening. Patients underwent brain magnetic resonance imaging, 18F-fluoro-2-deoxyglucose positron emission tomography, and neuropsychological assessments before and after the tFUS procedure. A whole-brain voxel-wise paired t test was conducted to compare rCMRglu before and after tFUS. Results: The sonication, as anticipated, did not show evidence of active blood-brain barrier opening on T1 dynamic contrast-enhanced magnetic resonance imaging. rCMRglu in the superior frontal gyrus (P<0.001), middle cingulate gyrus (P<0.001), and fusiform gyrus increased after tFUS (P=0.001). Patients demonstrated mild improvement in measures of memory, executive, and global cognitive function following tFUS. No adverse events were reported. Conclusion: These results suggest that hippocampal sonication with low-intensity tFUS may have beneficial effects on cerebral glucose metabolism and cognitive function in patients with AD. Further larger studies are needed to confirm the therapeutic efficacy of tFUS in AD. Purpose: Increasing attention has been paid to low-intensity transcranial focused ultrasound (tFUS) for its potential therapeutic effects in Alzheimer's disease (AD). While preclinical studies have shown promising therapeutic effects of low-intensity tFUS in AD models, its efficacy and safety remain unclear in humans. In this pilot study, we investigated the effects of low-intensity tFUS on blood-brain barrier opening, the regional cerebral metabolic rate of glucose (rCMRglu), and cognition in patients with AD.Methods: After receiving institutional review board approval, four patients with AD received tFUS to the hippocampus immediately after an intravenous injection of a microbubble ultrasound contrast agent. Sonication was delivered at low-intensity, at a pressure level below the threshold for blood-brain barrier opening. Patients underwent brain magnetic resonance imaging, 18F-fluoro-2-deoxyglucose positron emission tomography, and neuropsychological assessments before and after the tFUS procedure. A whole-brain voxel-wise paired t test was conducted to compare rCMRglu before and after tFUS.Results: The sonication, as anticipated, did not show evidence of active blood-brain barrier opening on T1 dynamic contrast-enhanced magnetic resonance imaging. rCMRglu in the superior frontal gyrus (P<0.001), middle cingulate gyrus (P<0.001), and fusiform gyrus increased after tFUS (P=0.001). Patients demonstrated mild improvement in measures of memory, executive, and global cognitive function following tFUS. No adverse events were reported.Conclusion: These results suggest that hippocampal sonication with low-intensity tFUS may have beneficial effects on cerebral glucose metabolism and cognitive function in patients with AD. Further larger studies are needed to confirm the therapeutic efficacy of tFUS in AD.

토끼모델에서 저강도 맥동초음파가 골절치유에 미치는 영향

김종만(Kim Jong man),이충휘(Yi Chung Hwi),조상현(Cho Sang Hyun),박정미(Park Jung mi),권혁철(Kwon Hyuk cheol),황태선(Hwang Tae sun) 한국전문물리치료학회 2002 한국전문물리치료학회지 Vol.9 No.1

The purpose of this research was to determine the effects on the healing of fibular fractures in rabbits of low-intensity pulsed ultrasound (50 mW/㎠ and 500 mW/㎠) applied for periods of 4, 14 and 24 days following fibular osteotomy. Thirty-six male Japanese white rabbits were randomly randomly divided into three groups of twelve for three treatment protocols: (1) ultrasound treatment at intensities of mW/㎠ and 500 mW/㎠ until the 4th following fibular osteotomy, (2) ultrasound treatment at intensities of 50 mW/㎠ and 500 mW/㎠ until the 14th day following fibular osteotomy, and (3) ultrasound treatment at intensities of 50mW/㎠ and 500mW/㎠ until the 24th day following fibular osteotomy. The low-intensity pulsed ultrasound was applied to only one fibular of each rabbit (these served as the experimental group). The other fibular of each rabbit served as the control group. The selection of which fibular was to be treated was made randomly. The animals were sacrificed on the 4th, 14th and 24th day after the start of ultrasound treatments. Percent of trabecular bone area and fibular radiography were carried out to compare the degree of fibular bone healing. A microscope was also used to determine any histologic changes. For statistical differences in radiological changes due to length of treatment period (4, 14 and 24 days respectively), the wilcoxon signed-ranks test was used to compare the experimental and control groups. For statistical differences in fracture healing due to differences in ultrasound intensity, radiological studies were compared using the Mann-Whitney Test. And, to compute percentage differences in areas of trabecular bone, Two-way analysis of variance (ultrasound intensity x each group) was used. Experiment results were as follows: 1. In animals sacrificed in the experimental and control groups (p>.05). However, experimental groups showed more rapid bone repair than control group. 2. Both radiographic and percent of trabecular bone area studies showed significant differences in rabbits sacrificed after 14 days. Fracture healing was significantly increased in the experimental group(p<.05) 3. In the animals sacrificed on the 24th day, histologic study showed rapid bone repair but fibular radiologic studies did not show statistical differences between the two groups(p>.05). 4. On the 14th day, bone union on radiograph was significantly more rapid in the treatment group with pulsed ultrasound of 50mW/㎠ than the group with 500mW/㎠ (p<.05). Histologic studies showed that both the 14 and 24 days groups had more rapid bone repair in animals treated with 50mW/㎠ ultrasound intensity than those treated with 500 mW/㎠ intensity. In conclusion, it has been shown that the low-intensity pulsed ultrasound has a positive effect on bone fracture healing in the early stage and the range of pulse ultrasound from 50mW/㎠ to 500mW/㎠ is effective for fracture healing. Further study is needed to investigate the influence of pulsed ultrasound on delayed union and non-union in bone fractures and also for the clinical use of low-intensity pulsed ultrasound for bone healing in humans.

저강도 초음파치료의 골절치유 효과

황태선,김종만,이충휘 한국전문물리치료학회 1998 한국전문물리치료학회지 Vol.5 No.3

AbstractNoninvasive low intensity ultrasound has been shown to be an effective means of accelerating bone fracture repair in both animal and clinical studies. The effects of ultrasound stimulation on bone repair after fibular osteotomy were assessed in a rabbit fibular fracture model. Bilateral closed fibular fractures were made in skeletally mature male White Japanese rabbits. In this study, 24 subjects were randomly divided into 2 groups: experimental group 1 (n=12), and experimental group 2 (n=12). Experimental group 1 received 0.875MHz continuous ultrasound and Experimental group 2 was treated with 3 MHz continuous ultrasound. The ultrasound intensity was 50 mW/ciif and treatment time was 10 minutes for every session in both groups. In each rabbit, one fibula served as a control and the other was subjected to ultrasound treatment 5 times per week for 3 weeks. After 3 weeks, rabbits were sacrificed and the ratios of the area between the trabeculae and bone marrow of the fibulae were calculated. At the end of the experimental period, 14 of the 24 rabbits were excluded due to complications from surgery or inadequate fracture status for this study. There was no statistically significant difference in the trabeculae area between experimental leg and control leg in experimental group 1 and experimental group 2 (p>0.05). And there was also no statistic- statistically significant difference between experimental group 1 and experimental group 2 according to ultrasound treatment frequencies, 0.875MHz and 3 MHz (p>0.05). These data suggest that in Japanese white rabbits, low intensity ultrasound stimulation does not facilitate fracture repair nor is there any difference in fracture repair results between ultrasound frequencies, 0.875 MHz and 3 MHz.

저강도 초음파치료의 골절치유 효과

이충휘,김종만,황태선 연세대학교 보건과학연구소 1998 보건과학논집 Vol.8 No.-

Low-Intensity Ultrasound (LIUS) Attenuates Myogenic and Adipogenic Differentiations of Rat Bone Marrow-Derived Mesenchymal Stem Cells (MSCs)

( Kil Hwan Kim ),( Mi Ae Lee ),( Byung Hyune Choi ),( So Ra Park ) 한국조직공학·재생의학회 2011 조직공학과 재생의학 Vol.8 No.1

Mesenchymal stem cells (MSCs) are multipotent stem cells that can differentiate into a variety of cell types, including chondrocytes, osteocytes, adipocytes, myocytes, and neurons. However, more efficient methods for specific lineage differentiation from MSCs are still required for their clinical application. In the present work we examined the effects of low intensity ultrasound (LIUS) on myogenic and adipogenic differentiation of MSCs isolated from the bone marrow of rats. We found that the LIUS stimulation has inhibitory effects on both differentiation toward myocyte and adipocyte from rat MSCs (rMSCs). bFGF enhanced the myogenic and adipogenic differentiation when MSCs were cultured in each differentiation medium, inducing the mRNA expression of myogenic and adipogenic markers. The LIUS stimulation showed the decrease in the expression of myogenic markers, desmin and troponin C, in the condition of myogenic differentiation containing bFGF. Similarly, the LIUS stimulation also showed the decrease in the expression of PPAR and the Oil Red O staining in adipogenic differentiation. These results reveal that mechanical stimulation by LIUS attenuates both myogenic and adipogenic differentiation of rMSCs in vitro.

저강도 파동형 초음파를 이용한 골 재생 촉진

강경리,정종혁 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.13

Ultrasound has been studied for stimulation of bone fracture healing for a long time. After many researches of high and low intensity ultrasound, researchers became focused on low-intensity pulsed ultrasound(LIPUS) recently. One of the effects of LIPUS is acceleration of bone regeneration without unfavorable results such as bone necrosis and fibrous or delayed healing, but the exact mechanism is not elucidated yet. The purpose of this feature article is to review researches of LIPUS up to now and to know the trend and future research focuses.

A Combinational Therapy of Articular Cartilage Defects: Rapid and Effective Regeneration by Using Low-Intensity Focused Ultrasound After Adipose Tissue-Derived Stem Cell Transplantation

Song Byeong-Wook,Park Jun-Hee,Kim Bomi,Lee Seahyoung,Lim Soyeon,Kim Sang Woo,Choi Jung-Won,Lee Jiyun,Kang Misun,Hwang Ki-Chul,Chae Dong-Sik,Kim Il-Kwon 한국조직공학과 재생의학회 2020 조직공학과 재생의학 Vol.17 No.3

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.

Effects of Low-Intensity Ultrasound on Gramicidin D-Induced Erythrocyte Edema

Lim, Mi Hyun,Seo, A. Rum,Kim, Jiyoung,Min, Byoung-Hyun,Baik, Eun Joo,Park, So Ra,Choi, Byung Hyune American Institute of Ultrasound in Medicine 2014 Journal of ultrasound in medicine Vol.33 No.6

<P><B>Objectives—</B></P><P>To determine whether low-intensity ultrasound (US) can reduce red blood cell (RBC) edema and, if so, whether the US activity is associated with aquaporin 1 (AQP-1), a water channel in the cell membrane.</P><P><B>Methods—</B></P><P>Red blood cell edema was induced by gramicidin D treatment at 40 ng/mL for 20 minutes and evaluated by a hematocrit assay. Low-intensity continuous wave US at 1 MHz was applied to RBCs for the last 10 minutes of gramicidin D treatment. To determine whether US activity was associated with AQP-1, RBCs were treated with 40 μM mercuric chloride (HgCl<SUB>2</SUB>), an AQP-1 inhibitor, for 20 minutes at the time of gramicidin D treatment. Posttreatment morphologic changes in RBCs were observed by actin staining with phalloidin.</P><P><B>Results—</B></P><P>Red blood cell edema increased significantly with gramicidin D at 20 (1.8%), 40 (6.7%), 60 (16.7%), and 80 (11.3%) ng/mL, reaching a peak at 60 ng/mL, compared to the control group (20 ng/mL, <I>P</I> = .019; 40, 60, and 80 ng/mL, <I>P</I> < .001). No significant RBC hemolysis was observed in any group. Edema induced by gramicidin D at 40 ng/mL was significantly reduced by US at 30 (3.4%; <I>P</I> = .003), 70 (4.4%; <I>P</I> = .001), and 100 (2.9%; <I>P</I> = .001) mW/cm<SUP>2</SUP>. Subsequent experiments showed that edema reduction by US ranged from 7% to 10%. Cotreatment with HgCl<SUB>2</SUB> partially reversed the US effect and showed a significantly different level of edema compared to gramicidin D-alone and US-cotreated groups (<I>P</I> = .001). These results were confirmed by microscopic observation of RBC morphologic changes.</P><P><B>Conclusions—</B></P><P>Low-intensity US could reduce gramicidin D–induced RBC edema, and its effect appeared to at least partly involve regulation of AQP-1 activity. These results suggest that low-intensity US can be used as an alternative treatment to control edema and related disorders.</P>

Preconditioning of Rabbit Mesenchymal Stem Cells in Polyglycolic Acid (PGA) Scaffold using Low-Intensity Ultrasound Improved Regeneration of Cartilage in Rabbit Articular Cartilage Defect Model

( Ji Hao Cui ),( So Ra Park ),( Byung Hyune Choi ),( Byoung Hyun Min ) 한국조직공학·재생의학회 2010 조직공학과 재생의학 Vol.7 No.1

This study investigated the effect of low intensity ultrasound (LIUS) stimulation of mesenchymal stem cells (MSCs) in vitro on the repair of cartilage defect after implantation of the construct in vivo. Rabbit MSCs were cultured in the polyglycolic acid (PGA) scaffold and preconditioned with (MSCs/US+) or without (MSCs/US-) LIUS stimulation during the chondrogenic differentiation for 1 week in vitro. The LIUS stimulation was carried out at the intensity of 200 mW/Cm2 every day for 20 min over a week. The constructs were implanted into the cartilage defects created in the rabbit femoral trochlea. The defect only was used as a negative control and rabbit chondrocytes seeded in PGA was used as a positive control, respectively. The repair of cartilage defect was examined at 2, 4, 8 and 12 weeks after implantation, respectively. The gross observation showed that the articular cartilage defects were filled with the repaired tissue in all groups. Histological and immunohistochemical analyses revealed, however, more intensive and widespread expression of proteoglycans and type II collagen in the MSCs/US+ group than in the MSCs/US- group. Fibrous tissues were observed mainly in the defect only group. The chondrocytes groups showed efficient repair of the defect by hyaline cartilage. In conclusion, this study suggested that LIUS preconditioning of MSCs in vitro could be an effective method to promote chondrogenesis of MSCs and repair of cartilage defect in vivo.