• 대한의용생체공학회:의공학회지
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• 제40권1호
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• pp.1-6
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• 2019

Human mesenchymal stem cells (hMSC) are multipotent stromal cells that have great potential to differentiate into a variety of cell types such as osteocytes, chondrocytes, and myocytes. Although there have been many studies on their clinical availability, little is known about how intracellular signals can be modulated by topographic features of the extracellular matrix (ECM). In this study, we investigated whether and how microwavy-patterned extracellular matrix (ECM) could affect the signaling activity of focal adhesion kinase (FAK), a key cellular adhesion protein. The fluorescence resonance energy transfer (FRET)-based FAK biosensor-transfected cells are incubated on microwavy-patterned surfaces and then platelet derived growth factor (PDGF) are treated to trigger FAK signals, followed by monitoring through live-cell FRET imaging in real time. As a result, we report that PDGF-induced FAK was highly activated in cells cultured on microwavy-patterned surface with L or M type, while inhibited by H type-patterned surface. In further studies, PDGF-induced FAK signals are regulated by functional support of actin filaments, microtubules, myosin-related proteins, suggesting that PDGF-induced FAK signals in hMSC upon microwavy surfaces are dependent on cytoskeleton (CSK)-actomyosin networks. Thus, our findings not only provide new insight on molecular mechanisms on how FAK signals can be regulated by distinct topographical cues of the ECM, but also may offer advantages in potential applications for regenerative medicine and tissue engineering.

• 대한치과보철학회지
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• 제44권5호
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• pp.574-583
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• 2006

Statement of problem : The use of permanent magnetics is increasing in implant dentistry. Purpose : This study is to know the effect of permanent magnetics on bone matrix formation of osteoblasts. Materials and methods : The konus abutment-shaped permanent magnetics were connected to the implant fixture, and placed on the culture plate. The osteoblast-like cell Mc3T3E1 were used for cell culture. As the control group, the implants were connected to titanium healing caps, and cultured in the same conditions of experimental group. After 3. 7, 14 days, cells were cultured, and we measured and compared the amount of collagen type I, osteocalcin, which is bone matrix protein by Western immunoblotting analysis. Results: As a result of Western immunoblotting analysis for estimating the amount of bone extracellular matrix, there was no difference between osteoblast of the experimental group and the control group during 3 and 7day-osteoblast culturing. However when cells were cultured for 14days, the amount of bone extracellular matrix was increased, on the experimental group. Conclusion: From these results, magnetic field of permanent magnetics might have effect on bone formation of osteoblast, especially at initial stage of implant placement. Therefore, their clinical application for implant or bone graft could be possible.

• BMB Reports
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• 제29권5호
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• pp.405-410
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• 1996

Previous studies have shown that transforming growth factor beta ($TGF-{\beta}$) is a potent regulator of cell growth and differentiation. To study the effects of $TGF-{\beta}$ on cell morphology and cytoskeleton reorganization, we conducted a survey using Mv1Lu mink lung epithelial cells with antibodies to cytoskeletal proteins and an extracellular matrix protein. While the untreated cells showed a cuboidal shape of typical epithelia, the Mv1Lu cells displayed a drastic shape change in the presence of $TGF-{\beta}$. This alteration was most prominent when near-confluent cells were treated with $TGF-{\beta}$. Since the morphology alteration is known to be accompanied by the reorganization of cytoskeletal proteins in other cell types, we investigated the intracellular distribution of the three major cytoskeletal structures: microfilaments, microtubules, and intermediate filaments. In the microfilament system, $TGF-{\beta}$ induced new stress fiber formation, which was caused primarily by the polymerization of cytoplasmic G-actin. However, $TGF-{\beta}$ appeared not to induce any significant changes in microtubular structures and vimentin filaments as determined by indirect fluorescence microscopy. Finally we confirmed the rapid accumulation of fibronectin by immunoblot analysis and chased the protein locations by immunofluorescence microscopy.

• Asian-Australasian Journal of Animal Sciences
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• 제29권10호
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• pp.1398-1406
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• 2016

In general, the seminiferous tubule basement membrane (STBM), comprising laminin, collagen IV, perlecan, and entactin, plays an important role in self-renewal and spermatogenesis of spermatogonial stem cells (SSCs) in the testis. However, among the diverse extracellular matrix (ECM) proteins constituting the STBM, the mechanism by which each regulates SSC fate has yet to be revealed. Accordingly, we investigated the effects of various ECM proteins on the maintenance of the undifferentiated state of SSCs in pigs. First, an extracellular signaling-free culture system was optimized, and alkaline phosphatase (AP) activity and transcriptional regulation of SSC-specific genes were analyzed in porcine SSCs (pSSCs) cultured for 1, 3, and 5 days on non-, laminin- and collagen IV-coated Petri dishes in the optimized culture system. The microenvironment consisting of glial cell-derived neurotrophic factor (GDNF)-supplemented mouse embryonic stem cell culture medium (mESCCM) (GDNF-mESCCM) demonstrated the highest efficiency in the maintenance of AP activity. Moreover, under the established extracellular signaling-free microenvironment, effective maintenance of AP activity and SSC-specific gene expression was detected in pSSCs experiencing laminin-derived signaling. From these results, we believe that laminin can serve as an extracellular niche factor required for the in vitro maintenance of undifferentiated pSSCs in the establishment of the pSSC culture system.

• Molecules and Cells
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• 제42권3호
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• pp.218-227
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• 2019

This study was designed to determine the effects of the long non-coding RNA (lncRNA) plasmacytoma variant translocation 1 (PVT1) on vascular smooth muscle cell (VSMC) apoptosis and extracellular matrix (ECM) disruption in a murine abdominal aortic aneurysm (AAA) model. After injection of PVT1-silencing lentiviruses, AAA was induced in Apolipoprotein E-deficient ($ApoE^{-/-}$) male mice by angiotensin II (Ang II) infusion for four weeks. After Ang II infusion, mouse serum levels of pro-inflammatory cytokines were analysed, and aortic tissues were isolated for histological, RNA, and protein analysis. Our results also showed that PVT1 expression was significantly upregulated in abdominal aortic tissues from AAA patients compared with that in controls. Additionally, Ang II treatment significantly increased PVT1 expression, both in cultured mouse VSMCs and in AAA murine abdominal aortic tissues. Of note, the effects of Ang II in facilitating cell apoptosis, increasing matrix metalloproteinase (MMP)-2 and MMP-9, reducing tissue inhibitor of MMP (TIMP)-1, and promoting switching from the contractile to synthetic phenotype in cultured VSMCs were enhanced by overexpression of PVT1 but attenuated by knockdown of PVT1. Furthermore, knockdown of PVT1 reversed Ang II-induced AAA-associated alterations in mice, as evidenced by attenuation of aortic diameter dilation, marked adventitial thickening, loss of elastin in the aorta, enhanced aortic cell apoptosis, elevated MMP-2 and MMP-9, reduced TIMP-1, and increased pro-inflammatory cytokines. In conclusion, our findings demonstrate that knockdown of lncRNA PVT1 suppresses VSMC apoptosis, ECM disruption, and serum pro-inflammatory cytokines in a murine Ang II-induced AAA model.

• The Plant Pathology Journal
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• 제17권6호
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• pp.376.1-376
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• 2001

• Molecules and Cells
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• 제46권9호
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• pp.558-572
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• 2023

Chronic obstructive pulmonary disease (COPD) will be the third leading cause of death worldwide by 2030. One of its components, emphysema, has been defined as a lung disease that irreversibly damages the lungs' alveoli. Treatment is currently unavailable for emphysema symptoms and complete cure of the disease. Hyaluronan (HA) and proteoglycan link protein 1 (HAPLN1), an HA-binding protein linking HA in the extracellular matrix to stabilize the proteoglycan structure, forms a bulky hydrogel-like aggregate. Studies on the biological role of the full-length HAPLN1, a simple structure-stabilizing protein, are limited. Here, we demonstrated for the first time that treating human alveolar epithelial type 2 cells with recombinant human HAPLN1 (rhHAPLN1) increased TGF-β receptor 1 (TGF-β RI) protein levels, but not TGF-β RII, in a CD44-dependent manner with concurrent enhancement of the phosphorylated Smad3 (p-Smad3), but not p-Smad2, upon TGF-β1 stimulation. Furthermore, rhHAPLN1 significantly increased sirtuins levels (i.e., SIRT1/2/6) without TGF-β1 and inhibited acetylated p300 levels that were increased by TGF-β1. rhHAPLN1 is crucial in regulating cellular senescence, including p53, p21, and p16, and inflammation markers such as p-NF-κB and Nrf2. Both senile emphysema mouse model induced via intraperitoneal rhHAPLN1 injections and porcine pancreatic elastase (PPE)-induced COPD mouse model generated via rhHAPLN1-containing aerosols inhalations showed a significantly potent efficacy in reducing alveolar spaces enlargement. Preclinical trials are underway to investigate the effects of inhaled rhHAPLN1-containing aerosols on several COPD animal models.

• 생명과학회지
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• 제20권11호
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• pp.1603-1610
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• 2010

Fucoidan은 갈조류의 세포벽에 존재하는 황산화 다당류이다. 본 연구에서는 자외선 B를 인체각질형성세포에 조사하여 matrix metalloproteinase-1 (MMP-1)을 발현 시킨 후 Fucus evanescens fucoidan이 MMP-1 promoter, mRNA, 단백 발현과 mitogen-activated protein kinases (MAPKs)의 인산화에 미치는 영향을 확인하고자 하였다. 자외선 B에 의해 생성된 MMP-1의 promoter activity와 mRNA, 단백 발현은 fucoidan $10\;{\mu}g/ml$와 $100\;{\mu}g/ml$를 투여하였을 때 fucoidan을 투여하지 않고 자외선만 조사한 군에 비하여 유의하게 억제되었다. 그리고 F. evanescens fucoidan은 extracellular signal regulated kinase (ERK)의 활성은 현저히 억제시켰으나 c-JUN N-terminal kinase (JNK)와 p38의 활성에 미치는 영향은 약하였다. 따라서 이 연구결과들은 F. evanescens fucoidan이 피부 광노화의 예방과 치료에 도움이 될 가능성을 확인할 수 있었다.

• 대한화장품학회지
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• 제48권2호
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• pp.157-167
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• 2022

본 연구에서는 7 개의 아미노산으로 이루어진 헵타펩타이드의 섬유아세포 활성 증가 및 광노화 조건에서의 세포 손상 억제 효과를 확인하였다. 실험 결과 헵타펩타이드 처리 시 섬유아세포 증식 및 세포외기질(extracellular matrix, ECM) 구성 인자의 발현이 증가되었다. 그리고 자외선 A (ultraviolet A, UVA) 조사에 의해 유도된 광노화조건에서 감소된 세포 생존율이 헵타펩타이드에 의해 증가되었고, UVA 조사에 의해 유도된 세포 사멸, 기질금속단백질분해효소-1(matrix metalloproteinases-1, MMP-1) 발현 및 세포 내 활성산소종(reactive oxygen species, ROS) 수준이 헵타펩타이드에 의해 감소되었다. UVA 조사 시 나타나는 transforming growth factor-β (TGF-β)/smad 기전 억제와 그에 따른 ECM 구성 인자 발현 감소 또한 헵타펩타이드에 의해 회복되었다. 또 다른 광노화 유도 조건으로 heat shock을 주었고 헵타펩타이드를 전 처리 하였을 때 heat shock에 의한 mitogen-activated protein kinase (MAPK) 인산화 및 MMP-1 발현이 억제됨을 확인할 수 있었다. 이 결과를 종합해 볼 때, 본 연구의 헵타펩타이드는 섬유아세포의 활성을 촉진하며, 광노화 유도 모델로 사용된 UVA 조사 및 heat shock 조건에서도 세포 내 ROS 억제 효과를 보여 세포 손상에 대한 회복 및 보호 효과를 나타내는 것으로 보인다. 이러한 진피 보호 효과를 갖는 헵타펩타이드는 향 후 신규 화장품 소재로 응용될 수 있을 것으로 기대된다.

• 대한화장품학회지
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• 제28권1호
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• pp.31-41
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• 2002

The production of matrix matalloproteinases(MMPs) by the UV irradiated skin fibroblast and the degradation of extracellular matrix(ECM) by these enzymes is known as one of the main reasons of photoaging. Recently, Fisher group showed that the MMP expression is mainly regulated by the mitogen-activated protein(MAP) kinas family, such as extracellular signal-regulated kinase(ERK), c-Jun amino-terminal kinase(JNK) and p38, each of which forms a signaling pathway. In this work we first examined the effect of nitric oxide (NO) on the production of MMP-1 and MMP-2 by the human dermal fibroblasts (HDFs). NO is a multifunctional messenger molecule generated from L-arginine and involved in many kinds of signaling pathway. We found that the treatment of HDF with NO donor, sodium nitroprusside (SNP) enhanced the expression of MMPs with or without UV irradiation and the treatment with nitric oxide synthase (NOS) inhibitors resulted in the significant decrease of MMPs production. From these results, we concluded that the production of MMPs by the UV irradiated HDF is regulated through the signaling pathway involving NO and cyclic GMP.