<로물루스 대제> 연구

송병욱 상명대학교 어문학연구소 1998 語文學硏究 Vol.8 No.-

Multiplexed targeting of microRNA in stem cell-derived extracellular vesicles for regenerative medicine

Byeong-Wook Song,Sekyung Oh,장우철 생화학분자생물학회 2022 BMB Reports Vol.55 No.2

Regenerative medicine is a research field that develops methodsto restore damaged cell or tissue function by regeneration, repairor replacement. Stem cells are the raw material of the bodythat is ultimately used from the point of view of regenerativemedicine, and stem cell therapy uses cells themselves or theirderivatives to promote responses to diseases and dysfunctions,the ultimate goal of regenerative medicine. Stem cell-derivedextracellular vesicles (EVs) are recognized as an attractive sourcebecause they can enrich exogenous microRNAs (miRNAs) bytargeting pathological recipient cells for disease therapy and canovercome the obstacles faced by current cell therapy agents. However, there are some limitations that need to be addressedbefore using miRNA-enriched EVs derived from stem cells formultiplexed therapeutic targeting in many diseases. Here, wereview various roles on miRNA-based stem cell EVs that caninduce effective and stable functional improvement of stemcell-derived EVs. In addition, we introduce and review the implicationsof several miRNA-enriched EV therapies improvedby multiplexed targeting in diseases involving the circulatorysystem and nervous system. This systemic review may offerpotential roles for stem cell-derived therapeutics with multiplexedtargeting.

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.

Multiplexed targeting of miRNA-210 in stem cell-derived extracellular vesicles promotes selective regeneration in ischemic hearts

Song Byeong-Wook,Lee Chang Youn,Kim Ran,Kim Won Jung,Lee Hee Won,Lee Min Young,Kim Jongmin,Jeong Jee-Yeong,장우철 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

Extracellular vesicles (EVs) are cell derivatives containing diverse cellular molecules, have various physiological properties and are also present in stem cells used for regenerative therapy. We selected a “multiplexed target” that demonstrates multiple effects on various cardiovascular cells, while functioning as a cargo of EVs. We screened various microRNAs (miRs) and identified miR-210 as a candidate target for survival and angiogenic function. We confirmed the cellular and biological functions of EV-210 (EVs derived from ASC miR-210 ) secreted from adipose-derived stem cells (ASCs) transfected with miR-210 (ASC miR-210 ). Under hypoxic conditions, we observed that ASC miR-210 inhibits apoptosis by modulating protein tyrosine phosphatase 1B (PTP1B) and death-associated protein kinase 1 (DAPK1). In hypoxic endothelial cells, EV-210 exerted its angiogenic capacity by inhibiting Ephrin A (EFNA3). Furthermore, EV-210 enhanced cell survival under the control of PTP1B and induced antiapoptotic effects in hypoxic H9c2 cells. In cardiac fibroblasts, the fibrotic ratio was reduced after exposure to EV-210, but EVs derived from ASC miR-210 did not communicate with fibroblasts. Finally, we observed the functional restoration of the ischemia/reperfusion-injured heart by maintaining the intercommunication of EVs and cardiovascular cells derived from ASC miR-210 . These results suggest that the multiplexed target with ASC miR-210 is a useful tool for cardiovascular regeneration.

Regulation of alternative macrophage activation by MSCs derived hypoxic conditioned medium, via the TGF-β1/Smad3 pathway

Byeong-Wook Song,Min-Ji Kim,Minji Kim,Hee Won Lee,Min Young Lee,Jongmin Kim,장우철 생화학분자생물학회 2020 BMB Reports Vol.53 No.11

Macrophages are re-educated and polarized in response to myocardial infarction (MI). The M2 anti-inflammatory phenotype is a known dominator of late stage MI. Mesenchymal stem cells (MSCs) represent a promising tool for cell therapy, particularly heart related diseases. In general, MSCs induce alteration of the macrophage subtype from M1 to M2, both in vitro and in vivo. We conjectured that hypoxic conditions can promote secretome productivity of MSCs. Hypoxia induces TGF-β1 expression, and TGF-β1 mediates M2 macrophage polarization for anti-inflammation and angiogenesis in infarcted areas. We hypothesized that macrophages undergo advanced M2 polarization after exposure to MSCs in hypoxia. Treatment of MSCs derived hypoxic conditioned medium (hypo-CM) promoted M2 phenotype and neovascularization through the TGF-β1/Smad3 pathway. In addition, hypo-CM derived from MSCs improved restoration of ischemic heart, such as attenuating cell apoptosis and fibrosis, and ameliorating microvessel density. Based on our results, we propose a new therapeutic method for effective MI treatment using regulation of macrophage polarization.

Combination Therapy Comprising a Static Magnetic Field with Contractility Improves Skin Wounds

Song, Byeong-Wook,Hong, Hyunki,Jung, Yu Jin,Lee, Ju Hyung,Kim, Bong Soo,Lee, Hoon-Bum Mary Ann Liebert 2018 Tissue engineering. Part A Vol.24 No.17

<P>Cutaneous wounds can present significant clinical problems because of abnormal healing after deep dermal damage. Despite technical advances in wound care, there are still unmet needs that result from inefficient treatment. In this study, we aimed to improve skin wound healing using a contractibility band with static magnetic field (SMF), termed a magnetic band (Mb). To examine the effect of the Mb on wound healing, full-thickness 15x35mm excision wounds were surgically created on the dorsum of rats. An elastic and contractile band (nontreatment), or one neodymium magnet (Nd-1) or two magnets with an elastic and contractile band (Nd-2) were topically applied to the wound daily and the wound size was measured from day 1 to 7 after surgery. Nd-2 showed a significant (95%) reduction in the wound size on day 3. Histological analysis showed that proinflammatory cytokine levels were diminished by Nd-2, and granulation tissue and microvessels were increased compared with those in the sham group. During Mb-induced wound healing, apoptosis was significantly reduced and matrix remodeling-related factors were initially regulated. The results suggest that combination therapy comprising an SMF and an elastic and contractile band could be a promising tool to heal cutaneous wounds rapidly.</P>

Integrin-Linked Kinase Is Required in Hypoxic Mesenchymal Stem Cells for Strengthening Cell Adhesion to Ischemic Myocardium

Song, Suk-Won,Chang, Woochul,Song, Byeong-Wook,Song, Heesang,Lim, Soyeon,Kim, Hye-Jung,Cha, Min-Ji,Choi, Eunju,Im, Sin-Hyeog,Chang, Byung-Chul,Chung, Namsik,Jang, Yangsoo,Hwang, Ki-Chul Wiley (John WileySons) 2009 Stem Cells Vol.27 No.6

<P>Mesenchymal stem cells (MSCs) therapy has limitations due to the poor viability of MSCs after cell transplantation. Integrin-mediated adhesion is a prerequisite for cell survival. As a novel anti-death strategy to improve cell survival in the infarcted heart, MSCs were genetically modified to overexpress integrin-linked kinase (ILK). The survival rate of ILK-transfected MSCs (ILK-MSCs) was augmented by about 1.5-fold and the phosphorylation of ERK1/2 and Akt in ILK-MSCs were increased by about three and twofold, respectively. ILK-MSCs demonstrated an increase of twofold in the ratio of Bcl-2/Bax and inhibited caspase-3 activation, compared with hypoxic MSCs. The adhesion rate of ILK-MSCs also had a 32.2% increase on the cardiac fibroblast-derived three-dimensional matrix and ILK-MSCs showed higher retention by about fourfold compared to unmodified MSCs. Six animals per group were used for the in vivo experiments analyzed at 1 week after occlusion of the left coronary artery. ILK-MSC transplanted rats had a 12.0% +/- 3.1% smaller infarct size than MSC-treated rats after ligation of left anterior descending coronary artery. Transplantation of ILK-MSCs not only led to a 16.0% +/- 0.4% decrease in the fibrotic heart area, but also significantly reduced the apoptotic positive index by two-thirds when compared with ligation only. The mean microvessel count per field in the infarcted myocardium of ILK-MSCs group was increased relative to the sham group and MSCs group. In conclusion, the ILK gene transduction of MSCs further assisted cell survival and adhesion, and improved myocardial damage when compared with MSC only after transplantation.</P>

Abies koreana 유래 정유의 항주름 및 미백 효과

송병욱(Byeong-Wook Song),송민정(Min-Jeong Song),박미진(Mi-Jin Park),최돈하(Don-Ha Choi),이성숙(Sung-Suk Lee),김명길(Myungkil Kim),황기철(Ki-Chul Hwang),김일권(Il-Kwon Kim) 한국생명과학회 2018 생명과학회지 Vol.28 No.5

구상나무로부터 추출된 정유가 개발되어 있지만, 피부와 연관된 연구 관점에서 효능이 아직까지 밝혀지지 않았다. 본 연구의 목적은 B16F10 흑색종 세포(B16F10)와 인간 피부아세포주(HDF)에서 멜라닌과 주름 형성 상의 구상나무 추출물 효과를 알아보는데 있다. 정유는 수첨 증류법으로 추출하였고 무수 황산나트륨으로 정제 하였다. 10<SUP>-5</SUP>배의 정유농도에서, 두 세포의 생존능은 세포독성 평가에 의해 확인 되었다. B18F10의 항멜라닌 효과는 티로시나아제 억제 분석과 티로시나아제, 티로시나아제 연관 단백질 -1과 -2(TRP-1, TRP-2)의 유전자 발현 검증을 통해 이루어졌다. 정유는 알파-멜라닌세포 자극 호르몬 유도 그룹과 비교하여 티로시나아제 억제 활성이 5배, 아르부틴 유도 그룹과 비교하였을 때 30% 정도 감소되었다. 3가지 멜라닌 유래 인자들의 mRNA 수준도 각각 증가하였다. 항주름 효과를 분석하기 위해, 프로콜라겐 타입 I c 펩티드 합성(PIP) 분석과 웨스턴 블록을 수행하였다. 정유를 처리한 그룹에서 PIP가 증가되었고 콜라겐 타입 1과 기질 금속 단백질 분해 효소가 개선 되었다. 또한 정유는 60 mJ/㎠의 자외선 조건에서, 자외선-억제 콜라겐 타입 1의 증가와 기질 금속 단백질 분해 효소 생성 억제를 통해 항주름 효과를 보여줬다. 따라서 구상나무 추출물은 미백과 항주름을 위한 안전하고 효과적인 피부 제재로서 가능성을 가지고 있음을 암시한다. The essential oil from Abies koreana E.H. Wilson had been developed, however, its efficacy has not yet been studied especially in terms of skin care research. The aim of this study is to investigate the effects of Abies koreana extracts (AKE) on melanogenesis and wrinkle formation in B16F10 melanoma cells (B16F10) and human dermal fibroblast cell line (HDF). The essential oil was extracted by hydrodistillation method and purified by anhydrous sodium sulfate. At a concentration of 10<SUP>-5</SUP>-fold, viability in these cells had been defined by cytotoxicity assays. Anti-melanogenic effects on B16F10 were evaluated using tyrosinase inhibition assay, and real-time PCR for verifying gene expression of tyrosinase, tyrosinase related protein-1 and -2 (TRP-1 and -2). AKEs reduced about 5-fold of tyrosinase inhibitory activity compared to α-melanocyte-stimulating hormone (α-MSH)-induced group and about 30% reduction compared to Arbutin induced group. The mRNA levels of three melanin-related factors were increased, separately. To investigate the effects of anti-wrinkle, procollagen type I c peptide synthesis assay (PIP) and Western blot were performed. At AKE-treated group, PIP was up-regulated and the expression of collagen type 1 and matrix metalloproteinase (MMP)-1 were improved. Furthermore, AKE presented anti-wrinkle effects by increasing UVB-inhibited collagen type 1 expression, and reducing UVB-induced MMP-1 production at 60 mJ/cm² of UVB radiation. Therefore, Abies koreana extracts has potentials as a safe and an effective skin ingredient for whitening and anti-wrinkle.

In Vivo Assessment of Stem Cells for Treating Neurodegenerative Disease: Current Approaches and Future Prospects

Hindawi Publishing Corporation 2017 Stem cells international Vol.2017 No.-

<P>In recent years, stem cell-related therapies have been widely applied for treating neurodegenerative disease. Despite their potential, stem cell tracking and imaging techniques for the evaluation of in vivo proof-of-concept (PoC) therapies have not been sufficiently represented in the research area. This review summarizes the recent approaches that have been used for tracking and imaging engrafted stem cells in vivo. Furthermore, we introduce tissue clearing technology that can be applied to develop three-dimensional in vivo experiments. Monitoring stem cell survival and migration and graft-host relationships is a useful strategy to evaluate the therapeutic efficacy of regenerative medicine approaches in neurodegenerative disease.</P>

Cardiomyocytes from phorbol myristate acetate-activated mesenchymal stem cells restore electromechanical function in infarcted rat hearts.

Song, Heesang,Hwang, Hye Jin,Chang, Woochul,Song, Byeong-Wook,Cha, Min-Ji,Kim, Il-Kwon,Lim, Soyeon,Choi, Eun Ju,Ham, Onju,Lee, Chang Youn,Park, Jun-Hee,Lee, Se-Yeon,Choi, Eunmi,Lee, Chungkeun,Lee, Myo National Academy of Sciences 2011 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF Vol.108 No.1

<P>Despite the safety and feasibility of mesenchymal stem cell (MSC) therapy, an optimal cell type has not yet emerged in terms of electromechanical integration in infarcted myocardium. We found that poor to moderate survival benefits of MSC-implanted rats were caused by incomplete electromechanical integration induced by tissue heterogeneity between myocytes and engrafted MSCs in the infarcted myocardium. Here, we report the development of cardiogenic cells from rat MSCs activated by phorbol myristate acetate, a PKC activator, that exhibited high expressions of cardiac-specific markers and Ca(2+) homeostasis-related proteins and showed adrenergic receptor signaling by norepinephrine. Histological analysis showed high connexin 43 coupling, few inflammatory cells, and low fibrotic markers in myocardium implanted with these phorbol myristate acetate-activated MSCs. Infarct hearts implanted with these cells exhibited restoration of conduction velocity through decreased tissue heterogeneity and improved myocardial contractility. These findings have major implications for the development of better cell types for electromechanical integration of cell-based treatment for infarcted myocardium.</P>