Transforming Growth Factor-Beta Binds Reversibly in Vitro to Guglielmi Detachable Coils

1998 ◽  
Vol 4 (1) ◽  
pp. 21-26 ◽  
Author(s):  
D.F. Kallmes ◽  
G.R. Hankins ◽  
M.K. Borland ◽  
H.J. Cloft ◽  
M.E. Jensen ◽  
...  

We determined the propensity for and reversibility of transforming growth factor-ß (TGFß) binding to uncoated Guglielmi Detachable Coils (GDC) and to GDC coated with extracellular matrix (ECM) proteins. Three 1.0 centimetre samples each of uncoated GDC-18 and of GDC-18 coated with either poly-L-lysine, laminin, type I collagen, type IV collagen, fibronectin, or poly-L-lysine and laminin were prepared. These samples were immersed briefly in a solution containing I125-labelled TGFß at a concentration of 0.225 μg/ml with initial specific activity of 123.3 mCi/mg (DuPont-NEN, Billerica, MA), and were counted using a scintillation counter. Each sample was then placed in a vial containing saline, shaken for 60 seconds, and counted again. Selected samples were immersed for varying periods within the TGFß solution and counted before and after saline rinse. Samples were rinsed one week after initial rinsing and counted again. The amount of binding between coil types was compared using the Student t test. For all samples initial binding of TGFß was in the order of 60–120 pg/cm. For the pre-rinse data there were no statistically significant differences between the amount bound to any single coil coating type relative to other coatings. Compared to the initial accumulations, the amount remaining after rinsing ranged from 40% (poly-L-lysine) to 63% (poly-L-lysine with laminin), with a mean of 55% among the seven coil types. After rinsing there was more growth factor remaining on uncoated coils than on poly-L-lysine-coated coils (p=0.05), fibronectin-coated coils (p=0.01), and type IV collagen-coated coils (p=0.04). There was a trend toward greater residual growth factor on coils coated with poly-L-lysine and laminin compared to coils coated with poly-L-lysine alone (p=0.10). Delayed, second rinsing of the samples one week after initial testing demonstrated only minor incremental loss of TGFß from the coil surfaces. After five minutes of immersion, accumulation was approximately 200% greater than that noted with brief submersion, but immersions lasting over five minutes did not yield increasing levels of TGFß binding. TGFß binds to GDC coils. Binding is not improved with ECM protein-coated coils compared to uncoated coils. The absolute amount of TGFß bound to the coil will likely result in local concentrations of growth factor in the order of those required for biological activity in vivo.

2017 ◽  
Vol 131 (5) ◽  
pp. 411-423 ◽  
Author(s):  
Bo Wang ◽  
Kevin Yao ◽  
Andrea F. Wise ◽  
Ricky Lau ◽  
Hsin-Hui Shen ◽  
...  

The regulatory role of a novel miRNA, miR-378, was determined in the development of fibrosis through repression of the MAPK1 pathway, miR-378 and fibrotic gene expression was examined in streptozotocin (STZ)-induced diabetic mice at 18 weeks or in unilateral ureteral obstruction (UUO) mice at 7 days. miR-378 transfection of proximal tubular epithelial cells, NRK52E and mesangial cells was assessed with/without endogenous miR-378 knockdown using the locked nucleic acid (LNA) inhibitor. NRK52E cells were co-transfected with the mothers against decapentaplegic homolog 3 (SMAD3) CAGA reporter and miR-378 in the presence of transforming growth factor-β (TGF-β1) was assessed. Quantitative polymerase chain reaction (qPCR) showed a significant reduction in miR-378 (P<0.05) corresponding with up-regulated type I collagen, type IV collagen and α-smooth muscle actin (SMA) in kidneys of STZ or UUO mice, compared with controls. TGF-β1 significantly increased mRNA expression of type I collagen (P<0.05), type IV collagen (P<0.05) and α-SMA (P<0.05) in NRK52E cells, which was significantly reduced (P<0.05) following miR-378 transfection and reversed following addition of the LNA inhibitor of endogenous miR-378. Overexpression of miR-378 inhibited mesangial cell expansion and proliferation in response to TGF-β1, with LNA–miR-378 transfection reversing this protective effect, associated with cell morphological alterations. The protective function of MAPK1 on miR-378 was shown in kidney cells treated with the MAPK1 inhibitor, selumetinib, which inhibited mesangial cell hypertrophy in response to TGF-β1. Taken together, these results suggest that miR-378 acts via regulation of the MAPK1 pathway. These studies demonstrate the protective function of MAPK1, regulated by miR-378, in the induction of kidney cell fibrosis and mesangial hypertrophy.


2003 ◽  
Vol 51 (9) ◽  
pp. 1177-1189 ◽  
Author(s):  
Christine V. Whiting ◽  
John F. Tarlton ◽  
Michael Bailey ◽  
Clare L. Morgan ◽  
Paul W. Bland

Transforming growth factor-β (TGF-β) depresses mucosal inflammation and upregulates extracellular matrix (ECM) deposition. We analyzed TGF-β receptors RI and RII as well as ECM components using the CD4+ T-cell-transplanted SCID mouse model of colitis. The principal change in colitis was an increased proportion of TGF-β RII+ mucosal mesenchymal cells, predominantly α-smooth muscle actin (SMA)+ myofibroblasts, co-expressing vimentin and basement membrane proteins, but not type I collagen. TGF-β RII+ SMA− fibroblasts producing type I collagen were also increased, particularly in areas of infiltration and in ulcers. Type IV collagen and laminin were distributed throughout the gut lamina propria in disease but were restricted to the basement membrane in controls. In areas of severe epithelial damage, type IV collagen was lost and increased type I collagen was observed. To examine ECM production by these cells, mucosal mesenchymal cells were isolated. Cultured cells exhibited a similar phenotype and matrix profile to those of in vivo cells. The data suggested that there were at least two populations of mesenchymal cells responsible for ECM synthesis in the mucosa and that ligation of TGF-β receptors on these cells resulted in the disordered and increased ECM production observed in colitic mucosa.


1999 ◽  
Vol 111 (1) ◽  
pp. 171-177
Author(s):  
Toshihiko Hayashi ◽  
Kazunori Mizuno ◽  
Motohiro Hirose ◽  
Koichi Nakazato ◽  
Eijiro Adachi ◽  
...  

1985 ◽  
Vol 100 (6) ◽  
pp. 1941-1947 ◽  
Author(s):  
M K Skinner ◽  
P S Tung ◽  
I B Fritz

We examined the synthesis and deposition of extracellular matrix (ECM) components in cultures of Sertoli cells and testicular peritubular cells maintained alone or in contact with each other. Levels of soluble ECM components produced by populations of isolated Sertoli cells and testicular peritubular cells were determined quantitatively by competitive enzyme-linked immunoabsorbent assays, using antibodies shown to react specifically with Type I collagen, Type IV collagen, laminin, or fibronectin. Peritubular cells in monoculture released into the medium fibronectin (432 to 560 ng/microgram cell DNA per 48 h), Type I collagen (223 to 276 ng/microgram cell DNA per 48 h), and Type IV collagen (350 to 436 ng/microgram cell DNA per 48 h) during the initial six days of culture in serum-free medium. In contrast, Sertoli cells in monoculture released into the medium Type IV collagen (322 to 419 ng/microgram cell DNA per 48 h) but did not form detectable amounts of Type I collagen or fibronectin during the initial six days of culture. Neither cell type produced detectable quantities of soluble laminin. Immunocytochemical localization investigations demonstrated that peritubular cells in monoculture were positive for fibronectin, Type I collagen, and Type IV collagen but negative for laminin. In all monocultures most of the ECM components were intracellular, with scant deposition as extracellular fibrils. Sertoli cells were positive immunocytochemically for Type IV collagen and laminin but negative for fibronectin and Type I collagen. Co-cultures of peritubular cells and Sertoli cells resulted in interactions that quantitatively altered levels of soluble ECM components present in the medium. This was correlated with an increased deposition of ECM components in extracellular fibrils. The data correlated with an increased deposition of ECM components in extracellular fibrils. The data presented here we interpret to indicate that the two cell types in co-culture act cooperatively in the formation and deposition of ECM components. Results are discussed with respect to the nature of interactions between mesenchymal peritubular cell precursors and adjacent epithelial Sertoli cell precursors in the formation of the basal lamina of the seminiferous tubule.


2021 ◽  
pp. 109-115
Author(s):  
I. I. Yakovtsova ◽  
S. V. Danilyuk ◽  
P. M. Zamyatin ◽  
R. M. Mikhailusov ◽  
V. V. Negoduyko ◽  
...  

Summary. The aim of the study was to analyze the data of immunohistochemical studies of the lungs after a gunshot wound penetrating at different times. Materials and methods. Lung tissue removed from the wounded with an encapsulated foreign body or without a foreign body of fire origin in the period from 1 day to 10 months after injury was studied. Results and discussion. At 1 day after injury, pathological type IV collagen and transforming growth factor begin to accumulate in the damaged areas. On day 2 of the wound, there is a slight increase in the expression of TGF-β1 and Collagen IV, VEGF is practically not produced, the number of CD68 + macrophages increases. Day 10 was characterized by a progressive increase in the content of transforming growth factor in leukocytes, lymphocytes, fibroblasts, macrophages and collagen type IV in the surrounding wounds. Ten months after injury and surgery, the wound canal was a large area of dense connective tissue with single venous vessels, often without a muscular layer. Transforming growth factor showed pronounced expression in macrophages and vascular endothelium, while VEGF was manifested as a weak positive response in vascular endothelial cells and single macrophages. Type IV collagen remained between the fibers of dense fibrous tissue, which replaced the wound canal, and was also found in the walls of blood vessels, interalveolar membranes, fibroblasts of the adjacent lung parenchyma. Conclusions. 1. Traumatic disorders in the lung parenchyma in FPI in the first hours after injury are accompanied by the accumulation of CD68 + macrophages, which persist for 10 months after FPI and become the main producers of TGF-β1 and VEGF. 2. Transforming growth factor TGF-β1 and vascular-endothelial growth factor VEGF are found in large quantities in macrophages, leukocytes, lymphocytes, fibroblasts and vascular endothelium from the first day of trauma to the lung parenchyma; up to 10 days the expression of TGF-β1 becomes significant, while VEGF weakly marks capillary endotheliocytes, arterioles, single macrophages. This immunohistochemical picture explains the long healing period of FPI. 3. The content of pathological collagen type IV Collagen IV on the first day of gunshot wound is minimal, increases up to 10 days and is detected in the fields of fibrosis and dystelectasis of the lung parenchyma after 10 months. 4. Determination of IHC expression of CD68, TGF-β1, VEGF and Collagen IV in the first 10 days after FPI and in the subsequent stages of treatment of the wounded will help to predict the course of combat trauma and adjust the methods of treatment.


ChemInform ◽  
2010 ◽  
Vol 30 (30) ◽  
pp. no-no
Author(s):  
Toshihiko Hayashi ◽  
Kazunori Mizuno ◽  
Motohiro Hirose ◽  
Koichi Nakazato ◽  
Eijiro Adachi ◽  
...  

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