OBJECTIVE: To explore the relationship between the expression of EIIIA+ fibronectin in incised wound of rat's skin and injury time. METHODS: The wounding model was established by cutting the dorsal skin of 48 adult SD rats. The rats were sacrificed at the pre-set injury time as immediately, 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, and 8 h. The skin samples were taken at the margin of wound. The expression of the EIIIA? fibronectin was detected by immunohistochemistry and Western blotting and the relationship be- tween its expression and injury time was observed. Results The expression of EIIIA+ fibronectin was not observed immediately. The basal cell of skin began to show positive expression 0.5 h after injury. With the extension of injury time, positive staining became stronger. The value of relative optical density was gradually increased with prolonged injury time by the Western blotting analysis. CONCLUSION The expression of EIIIA+ fibronectin could be used for estimation of injury time in the early stage of skin injury.
Animals ; Fibronectins/metabolism* ; Immunohistochemistry ; Proteins/metabolism* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Staining and Labeling ; Time Factors

Cell adhesion is a basic and very important tissue in the field of tissue engineering. Fibronectin and integrins are the most important elements to cell adhesion. Some surface receptors of fibroblast can also conjugate with type I collagen in extracellular matrix (ECM) directly. Laminin receptors on the surface of fibroblast bound to laminin also play a role in cell adhesion. In this paper are reviewed a number of related articles. The structures and function of fibronectin and integrins are discussed in detail; the tendon cell's adhesion structures are also discussed. Yet, there was scarcely any paper on the effects which the preservation of tissue engineered products may have on cells' adhesion fo ECM. Therefore, researching on cell adhesion and finding a way of preservation that has no or very little adverse effect on cell adhesion is an important topic. Results from expected advanced researches on cell adhesion may probably find promising applications in the field of tissue engineering.
Cell Adhesion ; Extracellular Matrix ; metabolism ; Fibroblasts ; cytology ; Fibronectins ; metabolism ; Humans ; Integrins ; metabolism ; Laminin ; metabolism ; Tendons ; metabolism ; Tissue Engineering

OBJECTIVETo investigate the effect of different dynamic tensional and compressive stress on the mRNA expression of collagen type I and fibronectin in cultured human periodontal ligament fibroblasts (hPDLF), and explore the regularity of functional change in hPDLF.

METHODSA new cyclic strain loading apparatus was used for mechanically loading. Cells cultured in vitro were loaded with three levels (1000 microstrain, 2000 microstrain, 4000 microstrain) of tensional and compressive forces and collected at different time (0 h, 0.5 h, 1 h, 4 h, 8 h,12 h) course after strain loading. The quantity of collagen type I and fibronectin mRNA was analyzed by means of quantitative real-time PCR with special primers of up- and down-regulated genes. Data were analyzed using SPSS version 10.0 software.

RESULTSDifferent magnitude and different kinds of mechanical forces as well as the force application time significantly changed the expression of collagen type I and fibronectin mRNA in hPDLF.

CONCLUSIONSDynamic mechanical forces could regulate the expression of collagen type I and fibronectin mRNA in hPDLF. Collagen type I and fibronectin participated in the mechanical signal transduction in human periodontal ligament fibroblasts.

Cells, Cultured ; Collagen Type I ; metabolism ; Fibroblasts ; metabolism ; Fibronectins ; metabolism ; Humans ; Periodontal Ligament ; cytology ; metabolism ; RNA, Messenger ; genetics ; Stress, Mechanical

OBJECTIVETo observe the expression of laminin and fibronectin in alkali-burned corneas in rats.

METHODSA total of 18 normal Wistar rats were randomly divided into 6 groups (n = 3 in each group). For each rat, one eye was injured by alkali burn, the other one was taken as the normal control. Then all the corneas were surgically removed and the expression of laminin and fibronectin was observed with immunohistochemistry respectively at 7 hours, 1 day, 3 days, 7 days, 14 days and 28 days after alkali burn.

RESULTSCompared with that of the normal controls, the expression of laminin and fibronectin of the burned eyes was dramatically higher at 7 hours, reached peak at 14 days and decreased to the normal level at 28 days after alkali burn.

CONCLUSIONSIn the process of wound healing after alkali burn, the expression of laminin and fibronectin increases dramatically, which suggests that laminin and fibronectin may participate in the process of corneal wound healing.

Animals ; Burns, Chemical ; metabolism ; Corneal Injuries ; Eye Burns ; metabolism ; Fibronectins ; metabolism ; Immunohistochemistry ; Laminin ; metabolism ; Rats ; Rats, Wistar ; Wound Healing ; physiology

BACKGROUND: Recent clinial observations have suggested that colchicine, which is in frequent use in gout can affect the conneciive tissue metabolism in skin and other ti.ues. OBJECTIVE: This study suggest that further development of colchiine might provide a novel means of modulating collagen gen expression in patients with fibrotic disease. METHOD: We examed the effect of colchicine on the expres, on of collagen, fibronectin and collagenase gene by skin filroblast culture hy Nort.hern and dot-blot iybridization. Result : The rate of transcription of genomic DNA corresponding o type I collagen and libvonectin was reduced in colchicine-treated cultures but collagenase was not. reduced. Canclusion : The microti.ikile disruptive agent, colchicine, reduced the expression of type I collagen and fibronectin in a dose-(lependent manner. This st.udy suppor that colchicine is one of the promising antifibrogenic drugs curvently being tested as a treatment, of hun an fibrotic disease.
Colchicine* ; Collagen Type I ; Collagen* ; Collagenases ; DNA ; Fibroblasts ; Fibronectins ; Gene Expression* ; Gout ; Humans ; Metabolism ; Skin

OBJECTIVETo investigate the functions of human periodontal myofibroblast (MFB) in vitro.

METHODSHuman periodontal fibroblast (hPDLFs) was cultured and induced to MFB by transforming growth factor-β1 (TGF-β1). MFB was denoted as the experimental group, whereas the hPDLFs was the control group. The groups were continuously cultured and harvested at 0, 12, 24, 48, and 72 h. The MFB marker α-smooth muscle actin (α-SMA) was examined by immunocytochemistry. The expression of fibronectin (FN) between MFB was examined by immunocytochemistry to detect the MFB contact relationship. The mRNA expression levels of α-SMA, collagen (Col) I, and Col III were measured by reverse transcription-polymerase chain reaction (RT-PCT) to analyze extracellular matrix secretion. The protein expression levels of α-SMA and Col I were also assessed by Western blot.

RESULTSThe experimental group had significantly higher α-SMA expression than the control group at 0 h (P < 0.001). A positive expression of FN was found between MFB. The experimental group had significantly higher expression levels of Col I and Col III than the control group at 24 h (P < 0.001).

CONCLUSIONHuman periodontal MFB presents a continuous, high expression of α-SMA. MFB could interact through FN. MFB is significantly capable of extracellular matrix secretion.

Actins ; Epithelial Cells ; Extracellular Matrix ; Fibroblasts ; Fibronectins ; Humans ; Jaw ; metabolism ; Myofibroblasts ; Transforming Growth Factor beta1

BACKGROUNDThe efficacies of current treatments for invasive aspergillus (IA) are unsatisfactory and new therapeutic targets or regimens to treat IA are urgently needed. Previous studies have indicated that the ability of conidia to invade host cells is critical in IA development and fibronectin has a hand in the conidia adherence process. In the clinical setting, many patients who receive glucocorticoid for extended periods are susceptible to Aspergillus fumigatus (A. fumigatus) infection, for this reason we investigated the effect of glucocorticoid on conidia invasiveness by comparing the invasiveness of A. fumigatus conidia in the type II human alveolar cell line (A549) cultured with different concentrations of dexamethasone. We also explored the relationships between dexamethasone and fibronectin expression.

METHODSFollowing culture with anti-fibronectin antibodies and/or dexamethasone, type II human alveolar A549 cells were infected with conidia of A. fumigatus. After 4 hours, the extracellular free conidia were washed away and the remaining immobilized conidia were released using Triton-X 100 and quantified by counting the colony-forming units. The invasiveness of conidia was measured by calculating the invasion rate (%). The transcription of the fibronectin gene in cells cultured with different concentrations of dexamethasone for 24 hours was tested by fluorogenic quantitative RT-PCR while the expression of fibronectinin cells cultured for 48 hours was tested by Western blotting and immunocytochemistry.

RESULTSA significant reduction in the invasiveness of conidia was seen in the cells cultured with anti-fibronectin antibody ((14.42 ± 1.68)% vs. (19.17 ± 2.53)%, P < 0.05), but no significant difference was observed in cells cultured with a combination of anti-fibronectin antibody and dexamethasone (6.37 ± 10(-5) mol/L). There was no correlation between the dexamethasone concentration and the invasiveness of conidia after dexamethasone pretreatment of cells for 4 hours. In contrast, after pretreated for 24 hours, the invasiveness of conidia in the presence of 6.37×10(-5) mol/L dexamethasone ((24.66 ± 2.41)%) was higher than for the control ((19.17 ± 2.53)%) and the 0.25×10(-5) mol/L group ((19.93 ± 3.06)%), and the invasiveness in the 1.27×10(-5) mol/L group ((22.47 ± 2.46)%) was also higher than in the control, P < 0.05. The relative transcripts of the fibronectin gene after exposure to 6.37×10(-5) mol/L dexamethasone (9.19×10(-3)±1.2×10(-3)) was higher than for the control (4.61×10(-3)± 1.54×10(-3)) and the 0.25×10(-5) mol/L group (6.20×10(-3)± 1.93×10(-3)), and expression in the 1.27×10(-5) mol/L group (7.94×10(-3)± 2.24×10(-3)) was also higher than for the control, P < 0.05. High concentrations of dexamethasone promoted fibronectin production after culture for 48 hours.

CONCLUSIONSDexamethasone can increase invasiveness of A. fumigatus conidia by promoting fibronectin expression. This may partially explain why patients who are given large doses of glucocorticoids for extended periods are more susceptible to A. fumigatus infection.

Aspergillus fumigatus ; pathogenicity ; Cell Line, Tumor ; Dexamethasone ; pharmacology ; Fibronectins ; genetics ; metabolism ; Gene Expression ; drug effects ; Humans

OBJECTIVE: To study the value of Fibronectin(Fn) immunohistochemical staining for diagnosing slight viral myocarditis. METHODS: The heart samples of human with myocarditis were studied by using LSAB immunohistochemical staining with anti-fibronectin antibody. RESULTS: Dense deposition was found in the myocardium of human with myocarditis. Some Fn-positive cardiomyocytes were observed. CONCLUSION Slight degeneration of cardiomyocytes could be identified by Fn-LSAB immunohistochemical staining and Fn-deposition is one of the reliable marks for inflammation in the myocardium.
Autopsy ; Death, Sudden/pathology* ; Diagnosis, Differential ; Fibronectins/metabolism* ; Humans ; Immunohistochemistry ; Myocarditis/virology* ; Myocardium/pathology* ; Staining and Labeling

OBJECTIVETo study the expression and significance of structural proteins, VEGF and Ang-1 in cavernous venous malformations of the body surface.

METHODSTissue samples came from 25 cases of cavernous venous malformations, 12 cases of normal moderate veins and 12 cases of normal small veins. Envision immunohistochemical stain was used to investigate the expression of IV collagen, fibronectin, laminin, VEGF and Ang-1. The results were analyzed semi-quantitatively.

RESULTSThe distribution of structural proteins in cavernous venous malformations is similar to moderate and small veins, but the expression in venous malformations is less obviously. VEGF expression in cavernous venous malformations and small veins is stronger obviously than moderate veins. Ang-1 expression in small veins is stronger remarkably than cavernous venous malformations and moderate veins.

CONCLUSIONThe abnormal expression of structural proteins may be an important factor in etiopathology and progress of cavernous venous malformations. There is disturbance of blood vessel remodelling in the sinusoid of cavernous venous malformations, with which the less expression of Ang-1 may be related.

Angiopoietin-1 ; metabolism ; Collagen Type IV ; metabolism ; Fibronectins ; analysis ; Hemangioma, Cavernous ; metabolism ; Humans ; Laminin ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Veins ; abnormalities ; metabolism

We examined the fibronectin (FN) secretion of cultured trabecular meshwork (TM) cells in a normal human eye by indirect immunofluorescent technique using mouse anti-human FN monoclonal antibody and FITC-conjugated goat anti-mouse IgG. To localize FN on frozen sections of normal TM, which were obtained from 7 enucleated eyes owing to traumatic eyeball rupture, the same indirect immunofluorescent method was used. Immunoelectron microscopy was applied to demonstrate the distribution pattern of FN in the normal TM of 2 human eyes using an avidin-biotin-peroxidase complex method. In the tissue culture of TM, the TM cell walls and extracellular matrices showed an intense staining with antibody to FN. Indirect immunofluorescent staining of FN on frozen sections of TM showed strong positive reactions in the subendothelial region. There was no reaction in the central core of the trabecular beam. Immunoelectron microscopy revealed the reaction products to FN in the areas lining the trabecular endothelial cells.
Adolescent ; Adult ; Antibodies, Monoclonal ; Cells, Cultured ; Fibronectins/biosynthesis/*metabolism ; Fluorescent Antibody Technique ; Humans ; Male ; Microscopy, Immunoelectron ; Trabecular Meshwork/*metabolism/ultrastructure