scholarly journals Increasing the Medium Osmolarity Reduces the Inflammatory Status of Human OA Chondrocytes and Increases Their Responsiveness to GDF-5

2020 ◽  
Vol 21 (2) ◽  
pp. 531
Author(s):  
Tanja Mang ◽  
Sven Lindemann ◽  
Anne Gigout
Keyword(s):  
Type I Collagen ◽  
3D Culture ◽  
Receptor Expression ◽  
Type I ◽  
Cytokine Release ◽  
Inflammatory Status ◽  
Medium Osmolarity ◽  
Oa Chondrocytes ◽  
Matrix Production ◽  
Protease Expression

The environment surrounding chondrocytes changes drastically in osteoarthritis (OA). For instance, the osmolarity in cartilage (ranging from 350 to 460 mOsm in healthy tissue) decreases during the progression of OA, reaching 270 mOsm. The objective of this study was to evaluate how osmolarity influences human OA chondrocytes. For this purpose, the osmolarity of the culture medium (340 mOsm) was increased to 380, 420 or 460 mOsm and its effect on the phenotype, matrix production, protease expression, cytokine release and growth and differentiation factor-5 (GDF-5) receptor expression in human OA chondrocytes was evaluated in a monolayer. Afterwards, the same parameters, as well as the responsiveness to GDF-5, were evaluated in 3D culture at 340 and 380 mOsm. Our results revealed that increasing the medium osmolarity increased matrix production but also reduced cytokine release, type I collagen and protease expression. It was also demonstrated that at 380 mOsm, the response to GDF-5 in 3D culture was more robust than at 340 mOsm. For the first time, it was established that a decreased osmolarity plays a role in sustaining inflammation and catabolic activities in OA chondrocytes and decreases their responsiveness to GDF-5. This indicates that osmolarity is a critical aspect of OA pathobiology.

Regulation of extracellular matrix production by chemically synthesized subfragments of type I collagen carboxy propeptide

Biochemistry ◽  
1991 ◽  
Vol 30 (29) ◽  
pp. 7097-7104 ◽  
Author(s):  
Kou Katayama ◽  
Jerome M. Seyer ◽  
Rajendra Raghow ◽  
Andrew H. Kang
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Type I ◽  
Extracellular Matrix Production ◽  
Matrix Production

Periodic high extracellular glucose enhances production of collagens III and IV by mesangial cells

1995 ◽  
Vol 268 (1) ◽  
pp. F13-F19 ◽  
Author(s):  
A. Takeuchi ◽  
D. C. Throckmorton ◽  
A. P. Brogden ◽  
N. Yoshizawa ◽  
H. Rasmussen ◽  
...  
Keyword(s):  
Glucose Concentration ◽  
Cellular Proliferation ◽  
Type I Collagen ◽  
Mesangial Cells ◽  
Blood Glucose Concentration ◽  
Type Iii Collagen ◽  
Type I ◽  
Extracellular Glucose Concentration ◽  
Extracellular Glucose ◽  
Matrix Production

We examined the effects of periodic changes in extracellular glucose concentration on matrix production and proliferation using three groups of cultured rat mesangial cells (MCs): 1) MCs in medium with continuous 5 mM glucose (CL), 2) MCs in medium alternating daily between 5 and 25 mM glucose (PH), and 3) MCs in medium with continuous 25 mM glucose (CH). MCs cultured in PH for 10 days produced 329 and 110% more type III collagen protein than MCs cultured in CL and CH, respectively. MCs cultured in PH induced 31 and 14% more type IV collagen than MCs cultured in CL and CH, respectively. Extracellular glucose concentration had no effect on the amount of type I collagen produced. MCs cultured in PH or CH for 5 days also expressed increased levels of type I, III, and IV collagen mRNA compared with MCs cultured in CL. MCs cultured in PH for 8-10 days also produced significantly more DNA than MCs in CL or CH. These data suggest that the temporal pattern of exposure to high extracellular glucose plays a role in regulating matrix formation and cellular proliferation by MCs. Furthermore, periodic elevations of extracellular glucose had a greater stimulatory effect on collagen production than a sustained elevation. These results suggest that decreasing the variability of blood glucose concentration may decrease the adverse effect of elevated glucose levels on MC matrix production and the progression of diabetic glomerulopathy.

scholarly journals Cyclic Tensile Strain Induces Tenogenic Differentiation of Tendon-Derived Stem Cells in Bioreactor Culture

2015 ◽  
Vol 2015 ◽  
pp. 1-13 ◽  
Author(s):  
Yuan Xu ◽  
Qiang Wang ◽  
Yudong Li ◽  
Yibo Gan ◽  
Pei Li ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Tensile Strain ◽  
Type I Collagen ◽  
3D Culture ◽  
Receptor Interaction ◽  
Type I ◽  
Bioreactor Culture ◽  
Cyclic Tensile Strain ◽  
Tenogenic Differentiation

Different loading regimens of cyclic tensile strain impose different effects on cell proliferation and tenogenic differentiation of TDSCs in three-dimensional (3D) culture in vitro, which has been little reported in previous literatures. In this study we assessed the efficacy of TDSCs in a poly(L-lactide-co-ε-caprolactone)/collagen (P(LLA-CL)/Col) scaffold under mechanical stimulation in the custom-designed 3D tensile bioreactor, which revealed that cyclic tensile strain with different frequencies (0.3 Hz, 0.5 Hz, and 1.0 Hz) and amplitudes (2%, 4%, and 8%) had no influence on TDSC viability, while it had different effects on the proliferation and the expression of type I collagen, tenascin-C, tenomodulin, and scleraxis of TDSCs, which was most obvious at 0.5 Hz frequency with the same amplitude and at 4% amplitude with the same frequency. Moreover, signaling pathway from microarray analysis revealed that reduced extracellular matrix (ECM) receptor interaction signaling initiated the tendon genius switch. Cyclic tensile strain highly upregulated genes encoding regulators of NPM1 and COPS5 transcriptional activities as well as MYC related transcriptional factors, which contributed to cell proliferation and differentiation. In particular, the transcriptome analysis provided certain new insights on the molecular and signaling networks for TDSCs loaded in these conditions.

scholarly journals Matrix metalloproteinase-2 governs lymphatic vessel formation as an interstitial collagenase

Blood ◽  
2012 ◽  
Vol 119 (21) ◽  
pp. 5048-5056 ◽  
Author(s):  
Benoit Detry ◽  
Charlotte Erpicum ◽  
Jenny Paupert ◽  
Silvia Blacher ◽  
Catherine Maillard ◽  
...  
Keyword(s):  
Matrix Metalloproteinase ◽  
Lymphatic Vessel ◽  
Type I Collagen ◽  
3D Culture ◽  
Matrix Composition ◽  
Matrix Metalloproteinase 2 ◽  
Type I ◽  
Vessel Formation

Abstract Lymphatic dysfunctions are associated with several human diseases, including lymphedema and metastatic spread of cancer. Although it is well recognized that lymphatic capillaries attach directly to interstitial matrix mainly composed of fibrillar type I collagen, the interactions occurring between lymphatics and their surrounding matrix have been overlooked. In this study, we demonstrate how matrix metalloproteinase (MMP)–2 drives lymphatic morphogenesis through Mmp2-gene ablation in mice, mmp2 knockdown in zebrafish and in 3D-culture systems, and through MMP2 inhibition. In all models used in vivo (3 murine models and thoracic duct development in zebrafish) and in vitro (lymphatic ring and spheroid assays), MMP2 blockage or down-regulation leads to reduced lymphangiogenesis or altered vessel branching. Our data show that lymphatic endothelial cell (LEC) migration through collagen fibers is affected by physical matrix constraints (matrix composition, density, and cross-linking). Transmission electron microscopy and confocal reflection microscopy using DQ-collagen highlight the contribution of MMP2 to mesenchymal-like migration of LECs associated with collagen fiber remodeling. Our findings provide new mechanistic insight into how LECs negotiate an interstitial type I collagen barrier and reveal an unexpected MMP2-driven collagenolytic pathway for lymphatic vessel formation and morphogenesis.

scholarly journals Crosstalk between invadopodia and the extracellular matrix

2020 ◽  
Author(s):  
Shinji Iizuka ◽  
Ronald P. Leon ◽  
Kyle P. Gribbin ◽  
Ying Zhang ◽  
Jose Navarro ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Complex Structure ◽  
Three Dimensional ◽  
3D Culture ◽  
Model Systems ◽  
Type I ◽  
Extracellular Matrix Components

ABSTRACTThe scaffold protein Tks5α is required for invadopodia-mediated cancer invasion both in vitro and in vivo. We have previously also revealed a role for Tks5 in tumor cell growth using three-dimensional (3D) culture model systems and mouse transplantation experiments. Here we use both 3D and high-density fibrillar collagen (HDFC) culture to demonstrate that native type I collagen, but not a form lacking the telopeptides, stimulated Tks5-dependent growth, which was dependent on the DDR collagen receptors. We used microenvironmental microarray (MEMA) technology to determine that laminin, collagen I, fibronectin and tropoelastin also stimulated invadopodia formation. A Tks5α-specific monoclonal antibody revealed its expression both on microtubules and at invadopodia. High- and super-resolution microscopy of cells in and on collagen was then used to place Tks5α at the base of invadopodia, separated from much of the actin and cortactin, but coincident with both matrix metalloprotease and cathepsin proteolytic activity. Inhibition of the Src family kinases, cathepsins or metalloproteases all reduced invadopodia length but each had distinct effects on Tks5α localization. These studies highlight the crosstalk between invadopodia and extracellular matrix components, and reveal the invadopodium to be a spatially complex structure.

scholarly journals 3D cell culture using a clinostat reproduces microgravity-induced skin changes

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Dong Hyun Choi ◽  
Byoungjun Jeon ◽  
Min Hyuk Lim ◽  
Dong Hun Lee ◽  
Sang-Kyu Ye ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Skin Irritation ◽  
3D Culture ◽  
3D Cell Culture ◽  
Tissue Growth ◽  
Microgravity Environment ◽  
Type I ◽  
Physiological Changes ◽  
3D Culture System

AbstractExposure to microgravity affects human physiology in various ways, and astronauts frequently report skin-related problems. Skin rash and irritation are frequent complaints during space missions, and skin thinning has also been reported after returning to Earth. However, spaceflight missions for studying the physiological changes in microgravity are impractical. Thus, we used a previously developed 3D clinostat to simulate a microgravity environment and investigate whether physiological changes of the skin can be reproduced in a 3D in vitro setting. Our results showed that under time-averaged simulated microgravity (taSMG), the thickness of the endothelial cell arrangement increased by up to 59.75%, indicating skin irritation due to vasodilation, and that the diameter of keratinocytes and fibroblast co-cultured spheroids decreased by 6.66%, representing skin thinning. The α1 chain of type I collagen was upregulated, while the connective tissue growth factor was downregulated under taSMG. Cytokeratin-10 expression was significantly increased in the taSMG environment. The clinostat-based 3D culture system can reproduce physiological changes in the skin similar to those under microgravity, providing insight for understanding the effects of microgravity on human health before space exploration.

Temporal expression of PDGF receptors and PDGF regulatory effects on osteoblastic cells in mineralizing cultures

1997 ◽  
Vol 272 (5) ◽  
pp. C1709-C1716 ◽  
Author(s):  
X. Yu ◽  
S. C. Hsieh ◽  
W. Bao ◽  
D. T. Graves
Keyword(s):  
Osteoblast Differentiation ◽  
Type I Collagen ◽  
Receptor Expression ◽  
Osteoblastic Cells ◽  
Nodule Formation ◽  
Specific Cell ◽  
Type I ◽  
Apparent Contradiction ◽  
Tyrosyl Phosphorylation

Platelet-derived growth factor (PDGF) is mitogenic and chemotactic for osteoblastic cells in vitro. It is expressed during osseous wound healing and stimulates formation of new bone in vivo. PDGF stimulates cells by binding to specific cell surface receptors. The purpose of this study was to examine the effects of PDGF on osteoblastic proliferation and differentiation in long-term mineralizing cultures. Utilizing Northern blot analysis, we found that continuous PDGF treatment increased histone expression, indicative of enhanced proliferation, but suppressed osteoblast differentiation, demonstrated by inhibition of alkaline phosphatase, type I collagen, and osteocalcin expression. The inhibitory effect of PDGF on the differentiated function of osteoblasts was further established by findings that PDGF significantly inhibited nodule formation. The expression of PDGF receptors varied at different stages of culture. PDGF receptor mRNA expression increased when the cells had achieved a mature phenotype, during the stage of matrix maturation, and then decreased. However, as demonstrated by thymidine incorporation assays, the capacity of PDGF to stimulate DNA synthesis actually decreased during osteoblast maturation, as receptor expression increased. To investigate this apparent contradiction, tyrosyl phosphorylation and immunoblot assays were performed to assess changes in PDGF activation of their cognate receptors. The pattern of PDGF-induced tyrosyl phosphorylation remained relatively constant. This suggests that the diminished mitogenic activity of PDGF that occurs after osteoblast differentiation is regulated at a postreceptor level.

Microfluidics Bioreactor: A Platform for Studying Capillary Morphogenesis in Response to Biochemical and Biophysical Cues

2007 ◽  
Author(s):  
Vernella V. Vickerman Kelley ◽  
Roger D. Kamm
Keyword(s):  
Endothelial Cells ◽  
Type I Collagen ◽  
3D Culture ◽  
Control Flow ◽  
Type I ◽  
Interstitial Flow ◽  
Surface Shear ◽  
Capillary Morphogenesis

The in vivo microvasculature is a dynamic structure which is influenced by both biochemical (e.g. cytokines, growth factors) and biophysical factors (e.g. shear stress, interstitial flow). Important regulators of this structure are the endothelial cells which are normally quiescent but under certain conditions are able to form new vascular sprouts. Investigations into the mechanism of capillary morphogenesis of human endothelial cells warrant an in vitro model that closely mimics the physiological in vivo microenvironment. To this end, we have developed a novel microfabricated system which permits 2D and 3D culture of endothelial cells in biologically derived (e.g. type I collagen) or synthetic (self assembling peptides) scaffolds and delivers control flow rates and pressures. This system offers tremendous flexibility with regard to scaffold physical and chemical properties, physiologically relevant mechanical stress induced by surface shear and interstitial flow as well as chemotactic gradients. In addition we are able to directly monitor the progression of vascular networks in response to these critical factors.

scholarly journals Leptin stimulates type I collagen production in db/db mesangial cells: Glucose uptake and TGF-β type II receptor expression

2001 ◽  
Vol 59 (4) ◽  
pp. 1315-1323 ◽  
Author(s):  
Dong Cheol Han ◽  
Motohide Isono ◽  
Sheldon Chen ◽  
Alberto Casaretto ◽  
Soon Won Hong ◽  
...  
Keyword(s):  
Glucose Uptake ◽  
Type I Collagen ◽  
Mesangial Cells ◽  
Receptor Expression ◽  
Type I ◽  
Type Ii ◽  
Collagen Production
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