• Title/Summary/Keyword: Extracellular matrix (ECM)

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Hydrothermal Synthesis of $TiO_2$ Nanowire Array for Osteoblast Adhesion

  • Yun, Young-Sik;Kang, Eun-Hye;Hong, Min-Eui;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.275-275
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    • 2013
  • Osteoblast is one of cells related with osseointegration and many research have conducted the adhesion of osteoblast onto the surface of implant. In the osseointegration, biocompatibility of the implant and cell adhesion to the surface are important factors. The researches related to cell adhesion have a direction from micro-scaled surface roughness to nano-scaled surface roughness with advancing nanotechnology. A cell reacts and sense to stimuli from extracellular matrix (ECM) and topography of the ECM [1]. Thus, for better osseointegration, we should provide an environment similar to ECM. In this study, we synthesize TiO2 nanowires using hydrothermal reaction because TiO2 provides inertness to titanium on its surface and enables it used as an implant material for the orthopedic treatment such as fixation of the bone fracture [2]. Ti substrate is immersed into NaOH aqueous solution. The solution are heated at $140{\sim}200^{\circ}C$ for various time (10~720 minutes). After heat treatment, we take out the sample and immerse it into HCl aqueous solution for 1 hour. The acid treated sample is heated again at $500^{\circ}C$ for 3 hours [3]. Then, we culture osteoblast on the TiO2 nanowires. For investigating cell adhesion onto nanostructured surface, we conduct several tests such as MTT assay, ALP (Alkaline phosphatase) activity assay, measuring calcium expression, and so on. These preliminary results of the cell culture on the nanowires are foundation for investigating cell-material interaction especially with nanostructure interaction.

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Functionalization of Electrospun Nano/Micro-fibrous Scaffolds Using Gamma-ray Irradiation (감마선 조사법을 이용한 전기방사 나노/마이크로 섬유 지지체의 표면 기능화)

  • Lim, Jong-Young;Shin, Young Min;Choi, Jong-Bae;Jeong, Jin-Oh;Gwon, Hui-Jeong;Jeong, Sung In;Park, Jong-Seok;Lim, Youn-Mook
    • Journal of Radiation Industry
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    • v.7 no.1
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    • pp.45-49
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    • 2013
  • In tissue engineering application, a fibrous structure of scaffolds has been issued as an alternative system to regulate cell survival and tissue regeneration, and electrospinning technique has been popularly used to generate fibrous meshes or sheets mimicking the structure of native extracellular matrix (ECM). However, recent strategy in the scaffold development is expanded to provide the structural property as well as a biological property of native ECM, a variety of surface modification techniques have been used to introduce biological property. In this study, we developed biomimetic poly(L-lactide-co-${\varepsilon}$-caprolactone) (PLCL) nano- and micro-fibrous scaffolds as a unique platform with structural and biological properties with native ECM using electrospinning method and gamma-ray irradiation. Surface morphology of the scaffolds was observed by scanning electron microscopy, and alteration of surface property was evaluated with toluidine blue O staining, water contact angle measurement and ATR-FTIR analysis.

Small Molecule-Based Strategy Promotes Nucleus Pulposus Specific Differentiation of Adipose-Derived Mesenchymal Stem Cells

  • Hua, Jianming;Shen, Ning;Wang, Jingkai;Tao, Yiqing;Li, Fangcai;Chen, Qixin;Zhou, Xiaopeng
    • Molecules and Cells
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    • v.42 no.9
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    • pp.661-671
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    • 2019
  • Adipose tissue-derived mesenchymal stem cells (ADSCs) are promising for regenerating degenerated intervertebral discs (IVDs), but the low efficiency of nucleus pulposus (NP)-specific differentiation limits their clinical applications. The Sonic hedgehog (Shh) signaling pathway is important in NP-specific differentiation of ADSCs, and Smoothened Agonist (SAG) is a highly specific and effective agonist of Shh signaling. In this study, we proposed a new differentiation strategy with the use of the small molecule SAG. The NP-specific differentiation and extracellular matrix (ECM) synthesis of ADSCs were measured in vitro, and the regenerative effects of SAG pretreated ADSCs in degenerated IVDs were verified in vivo. The results showed that the combination of SAG and transforming growth factor-${\beta}3$ ($TGF-{\beta}3$) is able to increase the ECM synthesis of ADSCs. In addition, the gene and protein expression levels of NP-specific markers were increased by treatment with SAG and $TGF-{\beta}3$. Furthermore, SAG pretreated ADSCs can also improve the disc height, water content, ECM content, and structure of degenerated IVDs in vivo. Our new differentiation scheme has high efficiency in inducing NP-specific differentiation of ADSCs and is promising for stem cell-based treatment of degenerated IVDs.

Syringaresinol derived from Panax ginseng berry attenuates oxidative stress-induced skin aging via autophagy

  • Choi, Wooram;Kim, Hyun Soo;Park, Sang Hee;Kim, Donghyun;Hong, Yong Deog;Kim, Ji Hye;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.46 no.4
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    • pp.536-542
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    • 2022
  • Background: In aged skin, reactive oxygen species (ROS) induces degradation of the extracellular matrix (ECM), leading to visible aging signs. Collagens in the ECM are cleaved by matrix metalloproteinases (MMPs). Syringaresinol (SYR), isolated from Panax ginseng berry, has various physiological activities, including anti-inflammatory action. However, the anti-aging effects of SYR via antioxidant and autophagy regulation have not been elucidated. Methods: The preventive effect of SYR on skin aging was investigated in human HaCaT keratinocytes in the presence of H2O2, and the keratinocyte cells were treated with SYR (0-200 ㎍/mL). mRNA and protein levels of MMP-2 and -9 were determined by real-time PCR and Western blotting, respectively. Radical scavenging activity was researched by 2,2 diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) assays. LC3B level was assessed by Western blotting and confocal microscopy. Results: SYR significantly reduced gene expression and protein levels of MMP-9 and -2 in both H2O2-treated and untreated HaCaT cells. SYR did not show cytotoxicity to HaCaT cells. SYR exhibited DPPH and ABTS radical scavenging activities with an EC50 value of 10.77 and 10.35 ㎍/mL, respectively. SYR elevated total levels of endogenous and exogenous LC3B in H2O2-stimulated HaCaT cells. 3-Methyladenine (3-MA), an autophagy inhibitor, counteracted the inhibitory effect of SYR on MMP-2 expression. Conclusion: SYR showed antioxidant activity and up-regulated autophagy activity in H2O2-stimulated HaCaT cells, lowering the expression of MMP-2 and MMP-9 associated with skin aging. Our results suggest that SYR has potential value as a cosmetic additive for prevention of skin aging.

Polymer brush: a promising grafting approach to scaffolds for tissue engineering

  • Kim, Woonjung;Jung, Jongjin
    • BMB Reports
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    • v.49 no.12
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    • pp.655-661
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    • 2016
  • Polymer brush is a soft material unit tethered covalently on the surface of scaffolds. It can induce functional and structural modification of a substrate's properties. Such surface coating approach has attracted special attentions in the fields of stem cell biology, tissue engineering, and regenerative medicine due to facile fabrication, usability of various polymers, extracellular matrix (ECM)-like structural features, and in vivo stability. Here, we summarized polymer brush-based grafting approaches comparing self-assembled monolayer (SAM)-based coating method, in addition to physico-chemical characterization techniques for surfaces such as wettability, stiffness/elasticity, roughness, and chemical composition that can affect cell adhesion, differentiation, and proliferation. We also reviewed recent advancements in cell biological applications of polymer brushes by focusing on stem cell differentiation and 3D supports/implants for tissue formation. Understanding cell behaviors on polymer brushes in the scale of nanometer length can contribute to systematic understandings of cellular responses at the interface of polymers and scaffolds and their simultaneous effects on cell behaviors for promising platform designs.

Expression and Preparation of Periostin FAS1 Domains for NMR Structure Determination

  • Yun, Hyosuk;Kim, Jae Il;Lee, Chul Won
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.1
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    • pp.17-21
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    • 2016
  • Periostin, a component of extracellular matrix (ECM) protein, is produced and secreted by the fibroblasts that are involved in chronic allergic inflammation diseases and various types of human cancers. Periostin protein is composed of multiple domains including four FAS1 domains which play important roles in cell adhesion and tumor metastasis by interacting with integrins. In spite of their important biological role, the structural information of periosin FAS1 domains was not revealed yet. Recently we systemically prepared various constructs of the FAS1 domains and tried to express them in E. coli. Of them, only single FAS1-II and -IV domains were highly soluble. Circular dichroism (CD) and nuclear magnetic resonance (NMR) studies revealed that the FAS1-IV domain might be suitable for three-dimensional structure determination using NMR spectroscopy.

Recent Progress in Study and Development of Polymeric Scaffolds for Tissue Regeneration (조직재생을 위한 고분자 지지체의 최근 연구개발 동향)

  • Joung, Yoon-Ki;Park, Ki-Dong;Park, Kwi-Deok;Han, Dong-Keun
    • Journal of Biomedical Engineering Research
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    • v.29 no.4
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    • pp.255-266
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    • 2008
  • In tissue engineering, scaffolds play an important role in the growth of cells to 3-D organs or tissues. For the success of tissue engineering, they should be mimicked to meet the requirements of natural extracellular matrix (ECM) in the body, such as mechanical properties, adhesiveness, porosity, biodegradability, and growth factor release, etc. Contrary to other materials, polymeric materials are adequate to engineer scaffolds for tissue engineering because controlling the structure and the ratio of components and designing various shapes and size are possible. In this review, the importance, major characteristics, processes, and recent examples of polymeric scaffolds for tissue engineering applications are discussed.

TGF-$\beta$1 Releasing Chitosan Microgranules for Bone Regeneration

  • Lee, Jue-Yeon;Lee, Sun-Yoon;Han, Sang-Mun;Lee, Yong-Moo;Rhyu, In-Chul;Chung, Chong-Pyoung
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.303.2-304
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    • 2003
  • For the purpose of obtaining high bone forming efficacy, chitosan microgranules were developed as bone substitutes. Chitosan has been applied to conduct the extracellular matrix (ECM) formation in tissue regenerative therapy. Microgranules designed in this study confer drug releasingcapacity in bone defect over long period to enhance bone regeneration. (omitted)

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Myocardial tissue engineering using electrospun nanofiber composites

  • Kim, Pyung-Hwan;Cho, Je-Yoel
    • BMB Reports
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    • v.49 no.1
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    • pp.26-36
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    • 2016
  • Emerging trends for cardiac tissue engineering are focused on increasing the biocompatibility and tissue regeneration ability of artificial heart tissue by incorporating various cell sources and bioactive molecules. Although primary cardiomyocytes can be successfully implanted, clinical applications are restricted due to their low survival rates and poor proliferation. To develop successful cardiovascular tissue regeneration systems, new technologies must be introduced to improve myocardial regeneration. Electrospinning is a simple, versatile technique for fabricating nanofibers. Here, we discuss various biodegradable polymers (natural, synthetic, and combinatorial polymers) that can be used for fiber fabrication. We also describe a series of fiber modification methods that can increase cell survival, proliferation, and migration and provide supporting mechanical properties by mimicking micro-environment structures, such as the extracellular matrix (ECM). In addition, the applications and types of nanofiber-based scaffolds for myocardial regeneration are described. Finally, fusion research methods combined with stem cells and scaffolds to improve biocompatibility are discussed. [BMB Reports 2016; 49(1): 26-36]

A cost-effective and simple culture method for primary hepatocytes

  • Adaya, Sezin;Hasircib, Nesrin;Gurhana, Ismet Deliloglu
    • Animal cells and systems
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    • v.15 no.1
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    • pp.19-27
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    • 2011
  • Hepatocytes, the major epithelial cells of the liver, maintain their morphology in culture dishes coated with extracellular matrix (ECM) components such as collagen and fibronectin or biodegradable polymers (e.g. chitosan, gelatin). In these coated dishes, survival of cells and maintaining of liver-specific functions may increase. The aim of this study was to determine a suitable, cost-effective and simple system for hepatocyte isolation and culture which may be useful for various applications such as in vitro toxicology studies, hepatocyte transplantation and bioartificial liver (BAL) systems. In order to obtain primary cultures, hepatocytes were isolated from liver by an enzymatic method and cultured on plates coated with collagen, chitosan or gelatin. Collagen, gelatin-sandwich and gelatin-cell mixture methods were also evaluated. Morphology and attachment of the cells were observed by inverted microscope and scanning electron microscope (SEM). An MTT assay was used to determine cell viability and mitochondrial activity.