• Title/Summary/Keyword: fibronectin

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ADSORPTION OF HUMAN RECOMBINANT FIBRONECTIN TO TITANIUM IN VITRO

  • Kim H.J.;Chang I.T.;Koak J.Y.;Heo S.J.;Yim S.A.;Jang J.H.
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.301-306
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    • 2004
  • Statement of problem. Fibronectin mediates its biological effects by binding to integrins on cell membranes through a consensus site including the Arg-Gly-Asp (RGD) sequence within tenth type III module. Purpose. The purpose of our study was to investigate the adsorption affinity of human recombinant fibronectin peptide (hFNIII 9-10) to titanium and to investigate the effect of the surrounding ionic composition on the adsorption process. Material and methods. As for evaluating the affinity of hFNIII 9-10 to Ti, titanium disks were incubated in 40, 80 and $120{\mu}g/ml$ hFNIII 9-10 solution at $37^{\circ}C$ overnight, repectively. As for evaluating the effect of surrounding ionic concentration, hFNIII 9-10 was dissolved in distilled water, phosphate buffered saline and RPMI 1640. Optical density (O.D.) was measured in ELISA reader. Results. The results were as follows; 1. The adsorption of hFNIII 9-10 showed significantly highest mean optical density (O.D.) value in $80{\mu}g/ml$. 2. The difference of ionic composition in DW, PBS and RPMI did not influence the adsorption amount of hFNIII 9-10.

Effects of the Chestnut Inner Shell Extract on the Expression of Adhesion Molecules, Fibronectin and Vitronectin, of Skin Fibroblasts in Culture

  • Chi, Yeon-Sook;Heo, Moon-Young;Chung, Ji-Hun;Jo, Byoung-Kee;Kim, Hyun-Pyo
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.469-474
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    • 2002
  • The inner shell of the chestnut (Castanea crenata S. et Z., Fagaceae) has been used as an anti-wrinkle/skin firming agent in East Asia, and preliminary experiments have found that a 70% ethanol extract from this plant material can prevent cell detachment of skin fibroblasts from culture plates. In order to examine the molecular mechanisms underlying this phenomenon, its effects on the expression of adhesion molecules, such as fibronectin and vitronectin, were investigated using the mouse skin fibroblast cell line, NIH/3T3. Using fixed-cell ELISA, Western blotting and immunofluorescence cell staining, it was clearly demonstrated that the chestnut inner shell extract enhanced the expression of the cell-associated fibronectin and vitronectin. Scoparone (6,7-dimethoxycoumarin), isolated from the extract, also possessed similar properties. These findings suggest that the enhanced expression of the adhesion molecules may be one of the molecular mechanisms for how the chestnut inner shell extract preventing cell detachment and may be also responsible for its anti-wrinkle/skin firming effect.

Effect of Retinoic Acid on Membrane Fusion and Expression of Fibronectin in Chick Embryonic Myoblasts (Retinoic Acid가 배양게배 근원세포의 융합과 Fibronictin의 발현에 미치는 영향)

  • 김혜선;정필중;강만식;정진하;하두봉
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.483-489
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    • 1995
  • Retinoic acid was found to block membrane fusion of chick embryonic myoblasts in culture. This effed was dosedependent and could he reversed upon removal of the agent from the culture medium. Furthermore, the retinoic acid-mediated inhibition of membrane fusion was observed with the fusion competent cells but not with the cells that had already been committed for fusion, indicating that the effect of RA is differentiation stage-specific. However, retinoic acid showed little or no effect on the ability of the cells to form bipolar shape and to align along their axes. Neither the cell proliferation nor accumulation of muscle specific proteins, such as creatine kinase and tropomyosin, was impaired significantly. On the other hand, retinoic acid blocked the differentiation time~ependent loss of fibronectin, whose process is prerequisite for myoblast fusion. These results suggest that retinoic add acts as a specific inhibitor of membrane fusion by preventing the loss of fibronectin from the differentiating myoblasts.

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An Immunohistological and Immunogold Study on the Fibronectin Reacions in Rat Lung Differentiation (흰쥐 폐의 분화과정에서 폐포막내 Fibronectin발현에 대한 면역조직학적 연구)

  • 문광덕;지행옥;정호삼
    • Journal of Chest Surgery
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    • v.32 no.12
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    • pp.1078-1086
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    • 1999
  • 배경 :폐포내 fibronectin(FN)의 분포와 역할은 많은 연구자에 의하여 연구되어왔다. 흰주에서 폐의 분화시 FN은 태자에서 폐포의 기저막에 주로 분포되고 간엽조직에서도 관찰되면 분화가 진행되면 폐포막의 간질조직에 FN의 함량이 높아진다. 또 FN은 일반적으로 폐포대식세포(alveolar macrophage)에서 분비되고 폐에 질병이 발생하였을 때 다량의 FN이 폐포대식세포에서 분비된다고 보고되어 있다(Schoenberger 등 1984: Ozaki 등 1990; Rom 등 1987 ; Cordier 등 1990) 저자는 흰쥐의 폐포발생이 진행중인 폐포기 후반에서의 폐포막내 정상적인 FN이니 분포의 변화와폐포를 구성하는 큰 폐포세포(type II pneumocyte)에서의 FN의 분비여부를 면역조직염색법과 전자현미경을 이용하여 추적하고자하였다 실험대상 및 방법: 청정동물실에서 사육한 SPF 흰쥐(Sprague-Dawley 계)를 임신시켜 질도말법을 이용하여 태령을 정한 뒤 태아제 17일 및 20일 출생 제 1일, 2일, 3일, 5일, 및 7일의 신생흰쥐를 실험동물로 사용하였으며 대조군의 흰쥐는 체중 200ㅎ의 건강한 수컷을 사용하였다 흰쥐의 폐조직은 면역조직염색을 위해 rabbit anti rat fibronectin polyclonal antibody를 일차항체로 biotinylated goat anti rabbit IgG를 이차항체로 사용하여 폐실질세포내 FN의 분포를 LM으로 관찰하였고 한편 폐포막을 구성하는 세포 중 큰폐포세포가 FN을 분비하는 세포인지를 추적하기 위해 금과립을 첨가한 항체를 사용하여 큰 폐포세포내 FN의 분포를 EM을 이용해서 추적한 결과 다음과 같은 결과를 얻었다 결과 : 제 17일 및 20일 태아시기의 폐에서의 혈관주위에 강한 FN반응이 관찰되었다 출생후 폐포막의 FN의 활성은 출생후 5일 및 7일에 최고주에 달했다. 출생직후 1-2일경에 혈관의 조직내 FN의 활성이 양성을 나타내지만 3일이후 활성이감소되었다. 폐포대식세포내 FN의 활성은 출생후 증가되었다. 폐조직내 소기관지의 FN의 활성은 출생후 완만하게 상승되었다. 큰 폐포세포는 출생 1-3일에 일정량의 FN 반응이 세포질과 미세융모내에 관찰되었다. 결론 : 이상과 같은 결과로 흰쥐의 폐포의 분화과정이 계속되는 출생후 폐에서 FN의 분비는 7일이내에 성숙흰쥐의 폐포내 반응과 비슷한 반응으르 보이며 이때 폐의 실질조직은 분화가 거의 완료되었을 것으로 사료되었고 큰 폐포세포에서도 FN이 분비되는 것으로 결론지울수 있다.

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Diagnostic and Prognostic Relevance of Bone Marrow Microenvironment Components in Non Hodgkin's Lymphoma Cases Before and After Therapy

  • Soliman, Amira H
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5273-5280
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    • 2016
  • Objective: To evaluate stromal cells of the bone marrow microenvironment (BMM) in bone marrow trephine biopsy (BMTB) specimens, with a focus on fibronectin, tumor necrosis factor- alpha (TNF-${\alpha}$) and L-selectin in Non-Hodgkin's lymphoma (NHL) patients, before and after therapy. Materials and Methods: A total of 80 de novo NHL patients, 64 with B-cell lymphomas 80%, (follicular cell lymphoma (FCL) in 32, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) in 12, and diffuse large cell lymphoma in 20) and 16 with T-cell lymphomas (20%) all diagnosed as T-Lymphoblastic lymphomas, were evaluated before and after therapy. For comparison, 25 age and sex matched BM donors, were included as a control group. BMTB material and BM aspirates were taken for morphological assessment of stromal cells, the plasma of these samples being examined for $TNF{\alpha}$ and L-selectin by ELISA, and fibronectin by radial immunodiffusion (RID). Results: BM stromal cells comprising reticular macrophages and fibroblasts were elevated in 53.3% of NHL cases at diagnosis, while BM fibronectin levels were decreased and BM $TNF{\alpha}$ and L-selectin were higher than in controls (p<0.05). In NHL cases, elevated values of BM $TNF{\alpha}$ and BM L-selectin were associated with signs of aggressive disease, including >1 extra nodal sites, detectable B symptoms, high grade, BM and CNS invasion, and a high International prognostic index (IPI) (p<0.05). Conclusion: BMM components, $TNF{\alpha}$, L-selectin and fibronectin, in NHL can be useful in evaluating disease activity, extent and response to treatment and as prognostic markers according to the IPI.

Inhibitory Effect of Rhein on Renal Fibrosis in Diabetic Nephropathy Rats (대황산(大黃酸)의 당뇨병쥐 신장조직섬유화 억제 효과에 관한 실험연구)

  • Zhao, Rongjie;Zhao, Zhenglin;Zhang, Jie;Liu, Hongfeng;Cui, Rongjun;Kim, Sang Chan;Kim, Sun-Hyung
    • Herbal Formula Science
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    • v.21 no.1
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    • pp.154-160
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    • 2013
  • Objectives : To investigate the therapeutic effect and underlying mechanisms of rhein on renal fibrosis in diabetic rats. Methods : Diabetic nephropathy (DN) was induced in adult Wistar rats via introperitoneal injection of streptozotocin (STZ) (20 mg/kg/d) for three consecutive days. Two days after the last dose of STZ, rhein was administered to the diabetic rats at a dose of 25 mg/kg or 50 mg/kg, twice a day by gavage, respectively. Following 28 days treatment with rhein, the plasma glucose and creatinine levels were measured, the renal levels of TGF-${\beta}1$ protein and mRNA were examined, and the fibronectin mRNA levels were also determined. Results : Rhein significantly inhibited the increased plasma glucose and creatinine levels of diabetic rats in a dose- and a time-dependent way. Immunohistochemical analysis showed both doses of rhein markedly attenuated elevated induction of renal TGF-${\beta}1$ protein expressions in diabetic rats. Additionally, the high dose of rhein improved both TGF-${\beta}1$ and fibronectin mRNA expressions, while the low dose of rhein only alleviated fibronectin mRNA expressions. Conclusions : Rhein can improve renal fibrosis in diabetic nephropathy rats, and which may be mediated through inhibition of the renal mRNA expressions of TGF-${\beta}1$ and fibronectin.

Renal mRNA Expression of Renin, $AT_1$ Receptor, TGF-${\beta}1$ and Fibronectin in Obstructive Nephropathy

  • Yang, Eun-Kyoung;Kim, In-Kyeom
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.55-63
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    • 1997
  • The present study was designed to quantify the alterations of renal renin, angiotensin type I receptor ($AT_1$), $TGF-{\beta}1$, and fibronectin gene expression in rats with unilateral ureteral obstruction (UUO). We also investigated the change of $AT_1$ density during UUO. Reverse transcription-polymerase chain reaction (RT-PCR) technique and receptor binding assay were used to detect mRNA expression and receptor density, respectively. At one day after UUO, renin mRNA level of the obstructed kidneys was decreased transiently and then subsequently increased to the level of sham kidneys. In the contralateral kidneys of the same rats, on the contrary, renin mRNA level was gradually decreased. Then, at 9 days after UUO, it was significantly lower than that of sham kidneys. The expressions of both $AT_1$ subtypes, called $AT_{1A}$ and $AT_{1B}$, mRNAs did not change at any time. UUO led to a significant decrease in $AT_1$ density in the obstructed kidneys compared with the sham kidneys at 1 and 3 days $(66\;{\pm}\;11.6%\;(p<0.005)\;and\;73\;{\pm}\;4.0%$ (p<0.01), respectively). Thereafter, $AT_1$ density was gradually increased and at 9 days it showed a marked elevation in the obstructed kidneys compared to the sham kidneys. In contrast, in the contralateral kidneys $AT_1$ density was significantly reduced from 3 to 9 days after UUO. The $TGF-{\beta}$1 mRNA level of the obstructed kidneys was unexpectedly decreased at 6 days after UUO. Then, at 9 days it was followed by a significant increase in the obstructed kidneys, whereas it showed an obvious decrease in the contralateral kidneys. In addition, fibronectin mRNA level was also significantly increased in the obstructed kidneys after UUO compared to the sham or the contralateral kidneys of the same rats. These results suggest a differential regulation of renal renin, $AT_1$ receptor, $TGF-{\beta}$1 and fibronectin mRNA levels at different stages of UUO.

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Plasminogen Activator Inhibitor-1 Antisense Oligodeoxynucleotides Abrogate Mesangial Fibronectin Accumulation

  • Park, Je-Hyun;Seo, Ji-Yeon;Ha, Hun-Joo
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.6
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    • pp.385-390
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    • 2010
  • Excessive extracellular matrix (ECM) accumulation is the main feature of chronic renal disease including diabetic nephropathy. Plasminogen activator inhibitor (PAI)-1 is known to play an important role in renal ECM accumulation in part through suppression of plasmin generation and matrix metalloproteinase (MMP) activation. The present study examined the effect of PAI-1 antisense oligodeoxynucleotide (ODN) on fibronectin upregulation and plasmin/MMP suppression in primary mesangial cells cultured under high glucose (HG) or transforming growth factor (TGF)-${\beta}1$, major mediators of diabetic renal ECM accumulation. Growth arrested and synchronized rat primary mesangial cells were transfected with $1\;{\mu}M$ phosphorothioate-modified antisense or control mis-match ODN for 24 hours with cationic liposome and then stimulated with 30 mM D-glucose or 2 ng/ml TGF-${\beta}1$. PAl-1 or fibronectin protein was measured by Western blot analysis. Plasmin activity was determined using a synthetic fluorometric plasmin substrate and MMP-2 activity analyzed using zymography. HG and TGF-${\beta}1$ significantly increased PAI-1 and fibronectin protein expression as well as decreased plasmin and MMP-2 activity. Transient transfection of mesangial cells with PAI-1 antisense ODN, but not mis-match ODN, effectively reversed basal as well as HG- and TGF-${\beta}1$-induced suppression of plasmin and MMP-2 activity. Both basal and upregulated fibronectin secretion were also inhibited by PAI-1 antisense ODN. These data confirm that PAI-1 plays an important role in ECM accumulation in diabetic mesangium through suppression of protease activity and suggest that PAI-1 antisense ODN would be an effective therapeutic strategy for prevention of renal fibrosis including diabetic nephropathy.

인공피부 개발을 위한 생채 적합성 지지체에 관한 연구

  • Kim, Chang-Hwan;Kim, Cheon-Ho;Park, Hyeon-Suk;Gang, Hyeon-Ju;Han, Eun-Suk;Kim, Yun-Yeong;Choe, Yeong-Ju;Lee, Su-Hyeon;Choe, Tae-Bu;Son, Yeong-Suk
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.429-432
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    • 2000
  • Chitosan scaffold is widely applied to drug delivery and tissue engineering. We have developed chitosan scaffolds, with various pore size, by differing freezing temperature and duration of ultraviolet (UV) irradiation, for reconstructing skin equivalent. Chitosan scaffold was coated with type I collagen, fibronectin and basic fibroblast growth factor (bFGF) in various combinations and concentrations, to evaluate the effect of extracellular matrix (ECM) and bFGF on cell adhesion, growth and differentiation of dermal fibroblasts. Human dermal fibroblasts, isolated from newborn foreskin and passaged between 3 and 5, were seeded on the top of scaffolds and cultivated for 2 weeks. We examined the morphology and the secretion of ECM of fibroblasts using scanning electron microsopy (SEM) and histochemistry. A stellate morphology of fibroblasts were seen in all groups. The scaffold coated with either type I collagen and bFGF or type I collagen and fibronectin, however, showed the best condtion of dermal fibroblasts, in that the highest cell number and ECM secretion were seen. On the contrary, scaffolds coated with all three factors, type I collagen, bFGF and fibronectin, showed lower number of cells and ECM secretion than scaffolds with two factors. There was a tendency of dose-dependence in all three factors for fibroblast growth and ECM secretion. In conclusion, we may suggest that chitosan scaffold coated with either type I collagen/bFGF or type I collagen/fibronectin could provide more favorable environment for the growth and differentiation of dermal fibroblasts.

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Agmatine Reduces Hydrogen Peroxide in Mesangial Cells under High Glucose Conditions

  • Lee, Geun-Taek;Ha, Hun-Joo;Lee, Hyun-Chul;Cho, Young-Dong
    • BMB Reports
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    • v.36 no.3
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    • pp.251-257
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    • 2003
  • Agmatine, an amine and organic cation, reduced $H_2O_2$ that was generated by hyperglycemia, and transcription factors such as NF-${\kappa}B$ and AP-1 activity in the mesangial cells that were exposed to high glucose. However, spermine which shares a strong nucleophilic structure with agmatine decreased the $H_2O_2$ levels and AP-1, but not the NF-${\kappa}B$ activity. Possible roles for agmatine and spermine in decreasing fibronectin are discussed, and the signaling pathway for agmatine-reduced fibronectin accumulation is presented.