scholarly journals Omidenepag, a non-prostanoid EP2 receptor agonist, induces enlargement of the 3D organoid of 3T3-L1 cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yosuke Ida ◽  
Fumihito Hikage ◽  
Araya Umetsu ◽  
Haruka Ida ◽  
Hiroshi Ohguro
Keyword(s):  
Mrna Expression ◽  
Receptor Agonist ◽  
Collagen Type ◽  
Physical Property ◽  
Upper Eyelid ◽  
3D Cultures ◽  
Ep2 Receptor ◽  
Collagen Type 1 ◽  
2D And 3D

Abstract 2D and 3D cultures of 3T3-L1 cells were employed in a study of the effects of Omidenepag (OMD), interacting with a non-prostanoid EP2 receptor, on adipogenesis. Upon adipogenesis, the effects on lipid staining, the mRNA expression of adipogenesis-related genes (Pparγ, CEBPa, Ap2, and Glut4) and the extracellular matrix (ECM) including collagen type 1, 4 and 6, and fibronectin, and the size and physical property of 3D organoids were compared between groups that had been treated with EP2 agonists (butaprost and OMD) and PGF2α. Upon adipogenesis, these significantly suppressed lipid staining and the mRNA expression of related genes. EP2 agonists and PGF2α influenced the mRNA expression of ECM in different manners, and these effects were also different between 2 and 3D cultures. Examining the physical properties by a microsqueezer indicated that the solidity of the 3D organoids became significantly lowered upon adipogenesis and these effects were not affected by EP2 agonists. In contrast, 3D organoid stiffness was markedly enhanced by the presence of PGF2α. These observations indicate that EP2 agonists affect the adipogenesis of 3T3-L1 cells in different manners, as compared to PGF2α, suggesting that OMD may not induce PGF2α related orbital fat atrophy, called the deepening of the upper eyelid sulcus (DUES).

scholarly journals Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Stephan Payr ◽  
Elizabeth Rosado-Balmayor ◽  
Thomas Tiefenboeck ◽  
Tim Schuseil ◽  
Marina Unger ◽  
...  
Keyword(s):  
Gene Expression ◽  
Collagen Type ◽  
3D Culture ◽  
Osteogenic Potential ◽  
Donor Age ◽  
Collagen Type 1 ◽  
2D And 3D ◽  
Human Primary Osteoblasts ◽  
Gene Expression Levels

Abstract Background The aim of this study was the investigation of the osteogenic potential of human osteoblasts of advanced donor age in 2D and 3D culture. Methods Osteoblasts were induced to osteogenic differentiation and cultivated, using the same polystyrene material in 2D and 3D culture for 2 weeks. Samples were taken to evaluate alkaline phosphatase (ALP) activity, mineralization and gene expression. Results Osteoprotegerin (OPG) levels were significantly increased (8.2-fold) on day 7 in 3D compared to day 0 (p < 0.0001) and 11.6-fold higher in 3D than in 2D (p < 0.0001). Both culture systems showed reduced osteocalcin (OC) levels (2D 85% and 3D 50% of basic value). Collagen type 1 (Col1) expression was elevated in 3D on day 7 (1.4-fold; p = 0.009). Osteopontin (OP) expression showed 6.5-fold higher levels on day 7 (p = 0.002) in 3D than in 2D. Mineralization was significantly higher in 3D on day 14 (p = 0.0002). Conclusion Advanced donor age human primary osteoblasts reveal significantly higher gene expression levels of OPG, Col1 and OP in 3D than in monolayer. Therefore, it seems that a relatively high potential of bone formation in a natural 3D arrangement is presumably still present in osteoblasts of elderly people. Trial registration 5217/11 on the 22nd of Dec. 2011.

scholarly journals Vascular composition, apoptosis, and expression of angiogenic factors in the corpus luteum during prostaglandin F2α-induced regression in sheep

Reproduction ◽  
2006 ◽  
Vol 131 (6) ◽  
pp. 1115-1126 ◽  
Author(s):  
Kimberly A Vonnahme ◽  
Dale A Redmer ◽  
Ewa Borowczyk ◽  
Jerzy J Bilski ◽  
Justin S Luther ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Growth Factor ◽  
Mrna Expression ◽  
Collagen Type ◽  
Angiogenic Factors ◽  
Cell Marker ◽  
Luteal Regression ◽  
Endothelial Cell Marker ◽  
Collagen Type 1

Corpora lutea and blood samples were collected from superovulated ewes 0, 4, 8, 12 and 24 h after prostaglandin F2α(PGF) analog injection on day 10 of the estrous cycle. Changes in vascular cell and fibroblast composition, apoptosis and mRNA expression for several angiogenic factors in the corpus luteum (CL) were determined. While peripheral progesterone concentration decreased at 24 h after PGF injection, CL weight did not change. The area of positive BS-1 lectin staining (endothelial cell marker), smooth muscle cell actin (SMCA; pericyte and SMC marker), collagen type 1 (fibroblast marker), and the rate of cell death changed in luteal tissues after PGF treatment. In association with these cellular changes, mRNA for several angiogenic factors including vascular endothelial growth factor (VEGF) and receptors (FltandKDR), basic fibroblast growth factor (FGF2) and receptor,angiopoietin (ANGPT) 1and receptorTie-2,endothelial nitric oxide synthase(NOS3), andangiotensin II receptor 1(AT1) were altered. Changes in endothelial cell marker expression were positively correlated with changes in VEGF and NO systems. In addition, changes in mRNA expression forVEGF,FltandKDRwere positively correlated with changes inANGPT2,Tie-2, andNOS3,indicating a functional relationship. This data demonstrates that after an initial increase, the endothelial component of the vascular bed decreases during PGF-induced luteal regression. However, SMCA expression remained high during luteal regression, potentially indicating a role of pericytes and vascular SMC in luteolysis, likely to regulate tissue remodeling and to maintain the integrity of larger blood vessels. Further, it appears that early regression may increase collagen type 1 production and/or expression by fibroblasts. Expression of angiogenic factors is influenced by PGF-induced luteolysis and may serve to maintain vascular structure in order to aid luteal regression.

MMP-2 Inhibition Attenuates IL-13 Induced Collagen Type-1 mRNA Expression in Human Airway Fibroblasts in Asthma

2011 ◽  
Vol 127 (2) ◽  
pp. AB127-AB127
Author(s):  
R. Firszt ◽  
T.D. Church ◽  
D. Francisco ◽  
D. Beaver ◽  
J.L. Ingram ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Collagen Type ◽  
Human Airway ◽  
Collagen Type 1

scholarly journals Evaluation of Collagen Type-1 Production and Anti-Inflammatory Activities of Human Placental Extracts in Human Gingival Fibroblasts

2016 ◽  
Vol 25 (3) ◽  
pp. 277-281 ◽  
Author(s):  
Homare Akagi ◽  
Yasuhiro Imamura ◽  
Yoshimasa Makita ◽  
Hiroe Nakamura ◽  
Naomi Hasegawa ◽  
...  
Keyword(s):  
Collagen Type ◽  
Human Gingival Fibroblasts ◽  
Gingival Fibroblasts ◽  
Anti Inflammatory ◽  
Collagen Type 1

scholarly journals Evaluation of Secretome Tenogenic Potential from Adipose Stem Cells (ACS) in Hypoxic Condition with Fresh Frozen Tendon Scaffold Using Scleraxis (Scx), Insulin-Like Growth Factor 1 (IGF-1) and Collagen Type 1

2021 ◽  
Vol 49 ◽  
pp. 111-118
Author(s):  
Ramadhan Ananditia Putra ◽  
Heri Suroto
Keyword(s):  
Cell Culture ◽  
Collagen Type ◽  
Hypoxic Condition ◽  
Adipose Stem Cells ◽  
Immunohistochemical Examination ◽  
Hypoxic Conditions ◽  
Fresh Frozen ◽  
Collagen Type 1

Various studies have been conducted to see the scaffold that supports the regeneration of tendon. This study aims to analyze thein vitrosecretome tenogenic potential produced by ASCs culture with fresh frozen tendon scaffold in hypoxic conditions. ELISA tests for Scx and IGF-1 levels in secretome were obtained from ASC culture with fresh frozen tendon scaffold under normoxic (21%) and hypoxia (2%) conditions. The immunohistochemical examination of COL-1 was also carried out on the 2ndand 6thdays of cell culture. The secretion of Scx and IGF-1 was increased in secretome from ASC cultures using a fresh frozen tendon scaffold compared with those which did not (p <0.05). In the normoxia condition, Scx and IGF-1 in secretome with fresh frozen tendons had better results than hypoxic conditions (p <0.05). The highest Scx levels were obtained in culture on the 6thday (p <0.05), while the highest IGF-1 levels were obtained in the culture on the 2ndday (p <0.05). There was an increase in the secretion of Scx and IGF-1 from ASC cultures with fresh frozen tendon scaffold under the hypoxic condition of 2%.

scholarly journals 1,25-Dihydroxyvitamin D3 Reduces TGF-β3-Induced Fibrosis-Related Gene Expression in Human Uterine Leiomyoma Cells

2011 ◽  
Vol 96 (4) ◽  
pp. E754-E762 ◽  
Author(s):  
Sunil K. Halder ◽  
J. Shawn Goodwin ◽  
Ayman Al-Hendy
Keyword(s):  
Protein Expression ◽  
Vitamin D3 ◽  
Uterine Leiomyoma ◽  
Collagen Type ◽  
Plasminogen Activator Inhibitor ◽  
Plasminogen Activator Inhibitor 1 ◽  
Dihydroxyvitamin D3 ◽  
Collagen Type 1 ◽  
Activator Inhibitor

Abstract Background: Uterine leiomyomas (fibroids) are the most common benign estrogen-dependent tumors of premenopausal women. TGF-β3 up-regulates the synthesis of many of extracellular matrix proteins that are associated with tissue fibrosis. Objective: To examine the effect of 1,25-dihydroxyvitamin D3 (vitamin D3) on TGF-β3-induced fibrosis-related protein expression in immortalized human uterine leiomyoma (HuLM) cells. Methods: HuLM cells were treated with TGF-β3 with or without vitamin D3. Western blot analyses were employed to test the effect of vitamin D3 on TGF-β3-induced protein expression of collagen type 1, fibronectin, and plasminogen activator inhibitor-1 proteins. Western blots as well as immunofluorescence analyses were used to verify the effect of vitamin D3 on TGF-β3-induced Smad activation involved in extracellular matrix protein synthesis and deposition, which ultimately lead to tissue fibrosis. Results: We observed that TGF-β3 induced fibronectin and collagen type 1 protein expression in HuLM cells, and that effect was suppressed by vitamin D3. TGF-β3 also induced protein expression of plasminogen activator inhibitor-1, an important TGF-β target, in HuLM cells, which was also inhibited by vitamin D3. Additionally, TGF-β3 induced phosphorylation of Smad2 as well as nuclear translocation of Smad2 and Smad3 in HuLM cells, whereas vitamin D significantly reduced all these TGF-β3-mediated effects. Therefore, our results suggest that vitamin D3 has consistently reduced TGF-β3 effects that are involved in the process of fibrosis in human leiomyoma cells. Conclusion: Vitamin D3 is an antifibrotic factor that might be potentially useful as a novel therapeutic for nonsurgical treatment of benign uterine fibroids.

scholarly journals β1-Integrin binding to collagen type 1 transmits breast cancer cells into chemoresistance by activating ABC efflux transporters

2020 ◽  
Vol 1867 (5) ◽  
pp. 118663 ◽  
Author(s):  
Fabian Baltes ◽  
Vladlena Pfeifer ◽  
Katja Silbermann ◽  
Julia Caspers ◽  
Kathleen Wantoch von Rekowski ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Collagen Type ◽  
Efflux Transporters ◽  
Β1 Integrin ◽  
Collagen Type 1

Cystathionine-β-synthase gene transfer and 3-deazaadenosine ameliorate inflammatory response in endothelial cells

2007 ◽  
Vol 293 (6) ◽  
pp. C1779-C1787 ◽  
Author(s):  
Utpal Sen ◽  
Neetu Tyagi ◽  
Munish Kumar ◽  
Karni S. Moshal ◽  
Walter E. Rodriguez ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Inflammatory Response ◽  
Protein Expression ◽  
Adhesion Molecules ◽  
Adhesion Molecule ◽  
Vascular Remodeling ◽  
Collagen Type ◽  
Gene Transfection ◽  
Collagen Type 1

Although elevated levels of homocysteine (Hcy) known as hyperhomocysteinemia (HHcy) are associated with increased inflammation and vascular remodeling, the mechanism of Hcy-mediated inflammation and vascular remodeling is unclear. The matrix metalloproteinases (MMPs) and adhesion molecules play an important role in vascular remodeling. We hypothesized that HHcy induces inflammation by increasing adhesion molecules and matrix protein expression. Endothelial cells were supplemented with high methionine, and Hcy accumulation was measured by HPLC. Nitric oxide (NO) bioavailability was detected by a NO probe. The protein expression was measured by Western blot analysis. MMP-9 activity was detected by gelatin-gel zymography. We demonstrated that methionine supplement promoted upregulation of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) through increased Hcy accumulation. In addition, increased synthesis of collagen type-1 was also observed. MMP-9 gene expression and protein activity were increased in methionine supplement groups. 3-Deazaadenosine (DZA), an adenosine analogue, prevented high methionine-induced ICAM-1 and VCAM-1 expression and collagen type-1 synthesis. Transfection of endothelial cells with cystathionine-β-synthase (CBS) gene construct, which converts Hcy to cystathionine, reduced Hcy accumulation in high methionine-fed cells. CBS gene transfection reduced the inflammatory response, as evident by attenuated ICAM-1 and VCAM-1 expression. Furthermore, collagen type-1 expression and MMP-9 activity were dramatically attenuated with CBS gene transfection. These results suggested that methionine supplement increased Hcy accumulation, which was associated with inflammatory response and matrix remodeling such as collagen type-1 synthesis and MMP-9 activity. However, in vitro DZA and CBS gene therapy successfully treated the HHcy-induced inflammatory reaction in the methionine metabolism pathway.

Sign in / Sign up

Export Citation Format

Share Document