• 제목/요약/키워드: Fibronectin adhesion

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Fabrication of Micro Patterned Fibronectin for Studying Adhesion and Alignment Behavior of Human Dermal Fibroblasts

  • Lee, Seung-Jae;Son, Young-Sook;Kim, Chun-Ho;Choi, Man-Soo
    • Macromolecular Research
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    • 제15권4호
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    • pp.348-356
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    • 2007
  • The aim of this study was to fabricate a submicro-and micro-patterned fibronectin coated wafer for a cell culture, which allows the positions and dimensions of the attached cells to be controlled. A replica molding was made into silicon via a photomask in quartz, using E-beam lithography, and then fabricated a polydimethylsiloxane stamp using the designed silicon mold. Hexadecanethiol $[HS(CH_2){_{15}}CH_3]$, adsorbed on the raised plateau of the surface of polydimethylsiloxane stamp, was contact-printed to form self-assembled monolayers (SAMs) of hexadecanethiolate on the surface of an Au-coated glass wafer. In order to form another SAM for control of the surface wafer properties, a hydrophilic hexa (ethylene glycol) terminated alkanethiol $[HS(CH_2){_{11}}(OCH_2CH_2){_6}OH]$ was also synthesized. The structural changes were confirmed using UV and $^1H-NMR$ spectroscopies. A SAM terminated in the hexa(ethylene glycol) groups was subsequently formed on the bare gold remaining on the surface of the Aucoated glass wafer. In order to aid the attachment of cells, fibronectin was adsorbed onto the resulting wafer, with the pattern formed on the gold-coated wafer confirmed using immunofluorescence staining against fibronectin. Fibronectin was adsorbed only onto the SAMs terminated in the methyl groups of the substrate. The hexa (ethylene glycol)-terminated regions resisted the adsorption of protein. Human dermal fibroblasts (P=4), obtained from newborn foreskin, only attached to the fibronectin-coated, methyl-terminated hydrophobic regions of the patterned SAMs. N-HDFs were more actively adhered, and spread in a pattern spacing below $14{\mu}m$, rather than above $17{\mu}m$, could easily migrate on the substrate containing spacing of $10{\mu}m$ or less between the strip lines.

인간치은섬유아세포의 다양한 세포행동 관련 유전자발현에 마이크로그루브-파이브로넥틴 복합 티타늄표면이 미치는 영향에 대한 연구 (A study on the effect of microgroove-fibronectin complex titanium plate on the expression of various cell behavior-related genes in human gingival fibroblasts)

  • 황유정;이원중;이성복;이석원
    • 구강회복응용과학지
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    • 제38권3호
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    • pp.150-161
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    • 2022
  • 목적:인간 치은섬유아세포의 세포활동에 관련된 다양한 유전자들의 발현에 마이크로그루브-파이브로넥틴 복합 표면이 미치는 영향을 확인하고자 하였다. 연구 재료 및 방법: 평활한 티타늄시편(NE0), 산처리만 시행한 티타늄시편(E0), 마이크로그루브 및 산처리된 티타늄시편(E60/10), 파이브로넥틴을 고정시킨 평활한 티타늄시편(NE0FN), 산처리 및 파이브로넥틴을 고정시킨 티타늄시편(E0FN), 그리고 마이크로그루브 및 산처리 후 파이브로넥틴을 고정시킨 티타늄시편(E60/10FN) 실험군 제작 후 인간 치은섬유아세포의 세포행동 관련 44개 유전자에 대한 실시간 중합효소연쇄반응 실험을 진행하였다. 결과: 인간치은섬유아세포 부착과 증식 등에 관여하는 4종류 신호전달 경로가 활성화되었다. Focal adhesion 경로에 속하는 Integrin α5, Integrin β1, Integrin β3, Talin-2 유전자들, PI3K-AKT 신호 전달 경로에 속하는 AKT1, AKT2, NF-κB 유전자들, MAPK 신호전달 경로에 속하는 MEK2, ERK1, ERK2 유전자들, 세포주기 신호 전달 경로에 속하는 cyclin D1, CDK4, CDK6 유전자들이 마이크로그루브-파이브로넥틴 복합 티타늄표면(E60/10FN)에서 상향조절되었다. 결론: 마이크로그루브-파이브로넥틴 복합 티타늄표면이 세포행동에 관여하는 다양한 유전자들을 상향조절할 수 있다.

Sesquicillin, an Extracellular Matrix Adhesion Inhibitor, Inhibits the Invasion of B16 Melanoma Cells In vitro

  • Lee, Ho-Jae;Chun, Hyo-Kon;Chung, Myung-Chul;Lee, Choong-Hwan;Kho, Yung-Hee
    • Journal of Microbiology and Biotechnology
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    • 제9권1호
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    • pp.119-121
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    • 1999
  • Tumor cell interaction with the extracellular matrix is defined as the critical event of tumor invasion that signals the initiation of a metastatic cascade. Sesquicillin has been identified as an inhibitor of melanoma cell adhesion to the components of the extracellular matrix (ECM) in cultured broth of fungal strain F60063. Sesquicillin strongly inhibited the adhesion of B16 melanoma cells to laminin, fibronectin, and typeIV collagen. It also inhibited B16 melanoma cell invasion of reconstituted basement membrane Matrigel in vitro in a dose-dependent manner. These results suggest that sesquicillin is a new class of nonpeptidic ECM adhesion inhibitor having anti-invasive activity.

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BAY11-7082에 의한 U937 세포의 CD29-매개성 세포간 유착과정 조절 효과 (Modulatory Effect of BAY11-7082 on CD29-mediated Cell-cell Adhesion in Monocytic U937 Cells)

  • 김병훈;조재열
    • 약학회지
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    • 제52권5호
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    • pp.412-417
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    • 2008
  • BAY11-7082 was initially found to be an anti-inflammatory drug with NF-${\kappa}B$ inhibitory property. In this study, we evaluated modulatory function of BAY11-7082 on U937 cell-cell adhesion induced by CD29 (${\beta}1$-integrins). BAY11-7082 strongly blocked functional activation of CD29 (${\beta}1$-integrins), as assessed by cell-cell adhesion assay. However, this compound did not block a simple activation of CD29, as assessed by cell-fibronectin adhesion assay. In particular, to understand molecular mechanism of BAY11-7082-mediated inhibition, the regulatory roles of CD29-induced actin cytoskeleton rearrangement under cell-cell adhesion and surface level of CD29 were examined using confocal and flow cytometic analysis. Interestingly, this compound strongly suppressed the molecular association of actin cytoskeleton with CD29 at cell-cell adhesion site. Moreover, BAY11-7082 also diminished surface levels of CD29 as well as its-associated adhesion molecule CD147, but not other adhesion molecules such as CD18 and CD43. Therefore, our data suggest that BAY11-7082 may be involved in regulating immune responses managed by CD29-mediated cell-cell adhesion.

Early bone healing onto implant surface treated by fibronectin/oxysterol for cell adhesion/osteogenic differentiation: in vivo experimental study in dogs

  • Lee, Jung-Seok;Yang, Jin-Hyuk;Hong, Ji-Youn;Jung, Ui-Won;Yang, Hyeong-Cheol;Lee, In-Seop;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • 제44권5호
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    • pp.242-250
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    • 2014
  • Purpose: This study aimed to evaluate the effects of fibronectin and oxysterol immobilized on machined-surface dental implants for the enhancement of cell attachment and osteogenic differentiation, on peri-implant bone healing in the early healing phase using an experimental model in dogs. Methods: Five types of dental implants were installed at a healed alveolar ridge in five dogs: a machined-surface implant (MI), apatite-coated MI (AMI), fibronectin-loaded AMI (FAMI), oxysterol-loaded AMI (OAMI), and sand-blasted, large-grit, acid-etched surface implant (SLAI). A randomly selected unilateral ridge was observed for 2 weeks, and the contralateral ridge for a 4-week period. Histologic and histometric analyses were performed for the bone-to-implant contact proportion (BIC) and bone density around the dental implant surface. Results: Different bone healing patterns were observed according to the type of implant surface 2 weeks after installation; newly formed bone continuously lined the entire surfaces in specimens of the FAMI and SLAI groups, whereas bony trabecula from adjacent bone tissue appeared with minimal new bone lining onto the surface in the MI, AMI, and OAMI groups. Histometric results revealed a significant reduction in the BIC in MI, AMI, and OAMI compared to SLAI, but FAMI demonstrated a comparable BIC with SLAI. Although both the BIC and bone density increased from a 2- to 4-week healing period, bone density showed no significant difference among any of the experimental and control groups. Conclusions: A fibronectin-coated implant surface designed for cell adhesion could increase contact osteogenesis in the early bone healing phase, but an oxysterol-coated implant surface designed for osteoinductivity could not modify early bone healing around implants in normal bone physiology.

Glycoportein IIb/IIIa와 칼슘동원의 조절을 통한 Steppogenin의 혈소판활성 억제효과 (Inhibitory Actions of Steppogenin on Platelet Activity Through Regulation of Glycoprotein IIb/IIIa and Ca2+ Mobilization)

  • 신정해;하주연;권혁우
    • 생약학회지
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    • 제51권2호
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    • pp.100-106
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    • 2020
  • The extract of Cudrania tricuspidata is used in ethnomedicine throughout Eastern Asia in China, Korea and Japan. In Korean traditional medicine, Cudrania tricuspidata has been used to treat eczema, mumps, tuberculosis, contusions, insomnia and acute arthritis. In addition, it has been reported that root extract of Cudrania tricuspidata has anti-platelet effects. Therefore, we investigated which compound in Cudrania tricuspidata has inhibitory effect on platelet aggregation. In this study, we tried to explain the inhibitory mechanism of steppogenin from Cudrania tricuspidata on human platelet aggregation. Collagen-induced human platelet aggregation and [Ca2+]i mobilization were dose-dependently inhibited by steppogenin and we determined the inhibition by steppogenin is due to the down regulation of extracellular-signal-regulated kinase(ERK) and inositol-1,4,5-triphosphate receptor type I(IP3RI) phosphorylation. In addition, steppogenin inhibited collagen-induced fibronectin adhesion to αIIb/β3 and thromboxane A2 generation. Thus, in the present study, steppogenin showed an inhibitory effect on human platelet aggregation, suggesting its potential use for preventing platelet-induced cardiovascular disease.

Effects of nerve cells and adhesion molecules on nerve conduit for peripheral nerve regeneration

  • Chung, Joo-Ryun;Choi, Jong-Won;Fiorellini, Joseph P.;Hwang, Kyung-Gyun;Park, Chang-Joo
    • Journal of Dental Anesthesia and Pain Medicine
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    • 제17권3호
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    • pp.191-198
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    • 2017
  • Background: For peripheral nerve regeneration, recent attentions have been paid to the nerve conduits made by tissue-engineering technique. Three major elements of tissue-engineering are cells, molecules, and scaffolds. Method: In this study, the attachments of nerve cells, including Schwann cells, on the nerve conduit and the effects of both growth factor and adhesion molecule on these attachments were investigated. Results: The attachment of rapidly-proliferating cells, C6 cells and HS683 cells, on nerve conduit was better than that of slowly-proliferating cells, PC12 cells and Schwann cells, however, the treatment of nerve growth factor improved the attachment of slowly-proliferating cells. In addition, the attachment of Schwann cells on nerve conduit coated with fibronectin was as good as that of Schwann cells treated with glial cell line-derived neurotrophic factor (GDNF). Conclusion: Growth factor changes nerve cell morphology and affects cell cycle time. And nerve growth factor or fibronectin treatment is indispensable for Schwann cell to be used for implantation in artificial nerve conduits.

Antiplatelet Effect of Cudraxanthone L Isolated from Cudrania tricuspidata via Inhibition of Phosphoproteins

  • Shin, Jung-Hae;Rhee, Man Hee;Kwon, Hyuk-Woo
    • Natural Product Sciences
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    • 제26권4호
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    • pp.295-302
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    • 2020
  • Cudrania tricuspidata (C. tricuspidata) is a deciduous tree found in Japan, China and Korea. The root, stems, bark and fruit of C. tricuspidata has been used as traditional herbal remedies such as eczema, mumps, acute arthritis and tuberculosis. In this study, we investigated the potential efficacies of this natural compound by focusing on the inhibitory effect of cudraxanthone L (CXL) isolated from the roots of C. tricuspidata on human platelet aggregation. Our study focused on the action of CXL on collagen-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding, intracellular calcium mobilization, fibronectin adhesion, dense granule secretion, and thromboxane A2 secretion. In addition, we investigated the inhibitory effect of CXL on thrombin-induced clot retraction. Our results showed that CXL inhibited collagen-induced human platelet aggregation, intracellular calcium mobilization, fibrinogen binding, fibronectin adhesion and clot retraction without cytotoxicity. Therefore, we confirmed that CXL has inhibitory effects on human platelet activities and has potential value as a natural substance for preventing thrombosis.

다양한 세포외기질이 배양 골아세포의 이동에 미치는 영향 (The Effects of Various Extracellular Matrices on Motility of Cultured MC3T3-E1 Cell)

  • 박병윤;서상우;이원재;류창우;나동균;손현주;박종철
    • Archives of Plastic Surgery
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    • 제32권2호
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    • pp.143-148
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    • 2005
  • Chemotactic migration of bone forming cell, osteoblast, is an important event during bone formation, bone remodeling, and fracture healing. Migration of cells is mediated by adhesion receptors, such as integrins, that link the cell to extracellular matrix ligands, type I collagen, fibronectin, laminin and depend on interaction between integrin and extracellular ligand. Our study was designed to investigate the effect of extracellular matrix like fibronectin, laminin, type I collagen on migration of osteoblast. Migration distance and speed of MC3T3-E1 cell on extracellular matrix-coated glass were measured for 24 hours using 0.01% type I collagen, 0.01% fibronectin, 100 microliter/ml laminin. The migration distance and speed of MC3T3-E1 cell was compared using a video-microscopy system. To determine migration speed, cells were viewed with a 4 phase- contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. The migration distance on 0.01% type I collagen or 0.01% fibronectin was longer than that on $100{\mu}l/ml$ laminin-coated glass. The migration speed on fibronectin-coated glass was 68 micrometer/hour which was fastest. The migration speed on type I collagen-coated glass was similar with that on fibronectin-coated glass. The latter two migration speeds were faster than that on no-coated glass. On the other hand, the average migration speed on laminin-coated glass was 37micrometer/hour and not different from that of control group. In conclusion, the extracelluar matrix ligands such as type I collagen and fibronectin seem to play an important role in cell migration. The type I collagen or fibronectin coated scaffold is more effective for migration of osteoblast in tissue engineering process.