scholarly journals The role of extracellular matrix biomolecules on endometrial epithelial cell attachment and cytokeratin 18 expression on gelatin hydrogels

2021 ◽  
Author(s):  
Samantha G Zambuto ◽  
Ishita Jain ◽  
Kathryn Clancy ◽  
Gregory Underhill ◽  
Brendan Harley
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Menstrual Cycle ◽  
Epithelial Cell ◽  
Basement Membrane ◽  
Collagen I ◽  
Cytokeratin 18 ◽  
Tenascin C ◽  
Endometrial Epithelial Cell ◽  
Collagen Iii

The endometrium undergoes profound changes in tissue architecture and composition, both during the menstrual cycle as well as in the context of pregnancy. Dynamic remodeling processes of the endometrial extracellular matrix (ECM) are a major element of endometrial homeostasis, including changes across the menstrual cycle. A critical element of this tissue microenvironment is the endometrial basement membrane, a specialized layer of proteins that separates the endometrial epithelium from the underlying endometrial ECM. Bioengineering models of the endometrial microenvironment that present an appropriate endometrial ECM and basement membrane may provide an improved environment to study endometrial epithelial cell (EEC) function. Here, we exploit a tiered approach using two-dimensional high throughput microarrays and three-dimensional gelatin hydrogels to define patterns of EEC attachment and cytokeratin 18 (CK18) expression in response to combinations of endometrial basement membrane proteins. We identify combinations (collagen IV + tenascin C; collagen I + collagen III; hyaluronic acid + tenascin C; collagen V; collagen V + hyaluronic acid; collagen III; collagen I) that facilitate increased EEC attachment, increased CK18 intensity, or both. We also identify significant EEC mediated remodeling of the GelMA matrix environment via analysis of nascent protein deposition. Together, we report efforts to tailor the localization of basement membrane-associated proteins and proteoglycans in order to investigate tissue engineered models of the endometrial microenvironment.

scholarly journals Immunohistochemical evaluation of fibrillar components of the extracellular matrix of transversalis fascia and anterior abdominal rectus sheath in men with inguinal hernia

2014 ◽  
Vol 41 (1) ◽  
pp. 23-29 ◽  
Author(s):  
Rogério De Oliveira Gonçalves ◽  
Evandro De Moraes e Silva ◽  
Gaspar De Jesus Lopes Filho
Keyword(s):  
Extracellular Matrix ◽  
Inguinal Hernia ◽  
Rectus Sheath ◽  
Staining Technique ◽  
Collagen I ◽  
Elastic Fibers ◽  
Collagen Iii ◽  
Extracellular Matrix Components ◽  
Matrix Components ◽  
Age Range

OBJECTIVE: to evaluate the role of fibrillar extracellular matrix components in the pathogenesis of inguinal hernias. METHODS: samples of the transverse fascia and of the anterior sheath of the rectus abdominis muscle were collected from 40 men aged between 20 and 60 years with type II and IIIA Nyhus inguinal hernia and from 10 fresh male cadavers (controls) without hernia in the same age range. The staining technique was immunohistochemistry for collagen I, collagen III and elastic fibers; quantification of fibrillar components was performed with an image analysis processing software. RESULTS: no statistically significant differences were found in the amount of elastic fibers, collagen I and collagen III, and the ratio of collagen I / III among patients with inguinal hernia when compared with subjects without hernia. CONCLUSION: the amount of fibrillar extracellular matrix components did not change in patients with and without inguinal hernia.

scholarly journals Physiological regulation of extracellular matrix collagen and elastin in the arterial wall of rats by noradrenergic tone and angiotensin II

2011 ◽  
Vol 13 (1) ◽  
pp. 19-28 ◽  
Author(s):  
Houcine Dab ◽  
Kamel Kacem ◽  
Rafik Hachani ◽  
Nadra Dhaouadi ◽  
Wassim Hodroj ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Angiotensin Ii ◽  
Vascular Diseases ◽  
Vascular Wall ◽  
Collagen I ◽  
Ang Ii ◽  
Physiological Regulation ◽  
Collagen Iii ◽  
Wistar Kyoto

The interactions between the effects of the sympathetic nervous system (SNS) and angiotensin II (ANG II) on vascular extracellular matrix (ECM) synthesis were determined in rats. The mRNA and protein content of collagen I, collagen III and elastin in the abdominal aorta (AA) and femoral artery (FA) was investigated in Wistar–Kyoto rats treated for 5 weeks with guanethidine, a sympathoplegic, losartan, an ANG II AT1 receptor (AT1R) blocker, or both. The effects of noradrenaline (NE) and ANG II on collagen III and elastin mRNA, and the receptor involved, were tested in cultured vascular smooth muscle cells (VSMCs) in vitro. Guanethidine increased collagen types I and III and decreased elastin, while losartan had an opposite effect, although without effect on collagen III. The combination of treatments abrogated changes induced by simple treatment with collagen I and elastin, but increased collagen III mRNA in AA and not in FA. NE stimulated collagen III mRNA via β receptors and elastin via α1 and α2 receptors. ANG II stimulated collagen III but inhibited elastin mRNA via AT1R. Overall, SNS and ANG II exert opposite and antagonistic effects on major components of ECM in the vascular wall. This may be of relevance for the choice of a therapeutic strategy in vascular diseases.

scholarly journals Tripeptidyl peptidase I promotes human endometrial epithelial cell adhesive capacity implying a role in receptivity

2020 ◽  
Vol 18 (1) ◽  
Author(s):  
Leilani L. Santos ◽  
Cheuk Kwan Ling ◽  
Evdokia Dimitriadis
Keyword(s):  
Menstrual Cycle ◽  
Epithelial Cell ◽  
Stromal Cells ◽  
Embryo Implantation ◽  
Unexplained Infertility ◽  
Implantation Failure ◽  
Secretory Phase ◽  
Endometrial Epithelial Cell ◽  
Tripeptidyl Peptidase ◽  
Adhesive Capacity

AbstractThe endometrium undergoes cyclic remodelling throughout the menstrual cycle in preparation for embryo implantation which occurs in a short window during the mid-secretory phase. It is during this short ‘receptive window’ that the endometrial luminal epithelium acquires adhesive capacity permitting blastocysts firm adhesion to the endometrium to establish pregnancy. Dysregulation in any of these steps can compromise embryo implantation resulting in implantation failure and infertility. Many factors contribute to these processes including TGF-β, LIF, IL-11 and proteases. Tripeptidyl peptidase 1 (TPP1) is a is a lysosomal serine-type protease however the contribution of the TPP1 to receptivity is unknown. We aimed to investigate the role of TPP1 in receptivity in humans.In the current study, TPP1 was expressed in both epithelial and stromal compartments of the endometrium across the menstrual cycle. Expression was confined to the cytoplasm of luminal and glandular epithelial cells and stromal cells. Staining of mid-secretory endometrial tissues of women with normal fertility and primary unexplained infertility showed reduced immunostaining intensity of TPP1 in luminal epithelial cells of infertile tissues compared to fertile tissues. By contrast, TPP1 levels in glandular epithelial and stromal cells were comparable in both groups in the mid-secretory phase. Inhibition of TPP1 using siRNA compromised HTR8/SVneo (trophoblast cell line) spheroid adhesion on siRNA-transfected Ishikawa cells (endometrial epithelial cell line) in vitro. This impairment was associated with decreased sirtuin 1 (SIRT1), BCL2 and p53 mRNA and unaltered, CD44, CDH1, CDH2, ITGB3, VEGF A, OSTEOPONTIN, MDM2, CASP4, MCL1, MMP2, ARF6, SGK1, HOXA-10, LIF, and LIF receptor gene expression between treatment groups. siRNA knockdown of TPP1 in primary human endometrial stromal cells did not affect decidualization nor the expression of decidualization markers prolactin (PRL) and insulin-like growth factor-binding protein 1 (IGFBP1). Taken together, our data strongly suggests a role for TPP1 in endometrial receptivity via its effects on epithelial cell adhesion and suggests reduced levels associated with unexplained infertility may contribute to implantation failure.

scholarly journals Human foreskin-derived dermal stem/progenitor cell-conditioned medium combined with hyaluronic acid promotes extracellular matrix regeneration in diabetic wounds

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yu Xin ◽  
Peng Xu ◽  
Xiangsheng Wang ◽  
Yunsheng Chen ◽  
Zheng Zhang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Matrix Metalloproteinases ◽  
Conditioned Medium ◽  
Progenitor Cell ◽  
Bioactive Molecules ◽  
Published Data ◽  
Collagen Iii ◽  
Diabetic Wounds

Abstract Background Diabetic wounds remain a challenging clinical problem, which requires further treatment development. Published data showed that dermis-derived stem/progenitor cells (DSPCs) display superior wound healing in vitro. The beneficial effects of DSPCs are mediated through paracrine secretion, which can be obtained from conditioned medium (CM). Hyaluronic acid (HA) is especially suitable for skin regeneration and delivering bioactive molecules in CM. This study investigated the effect of human foreskin-derived dermal stem/progenitor cell (hFDSPC)-CM combined with HA on a diabetic mouse model and relevant mechanism in vitro. Methods hFDSPCs and human adipose-derived stem cells (hADSCs) were identified, and the respective CM was prepared. PBS, HA, hFDSPC-CM combined with HA, or hADSC-CM combined with HA was topically applied to mice. HE, CD31, CD68, CD86, and CD206 staining was performed to evaluate gross wound condition, angiogenesis, and inflammation, respectively. Masson and Picrosirius red staining was performed to evaluate collagen deposition and maturation. The effects of hFDSPC-CM and hADSC-CM on human keratinocyte cells (HaCaT) and fibroblasts were evaluated in vitro using CCK-8 and EdU assays to determine cell viability and proliferation, respectively. The scratch assay was performed to evaluate cell migration. Tube formation assay was performed on human umbilical vein endothelial cells (HUVECs) to confirm angiogenesis. Extracellular matrix (ECM) metabolic balance-related genes and proteins, such as collagen I (COL 1), collagen III (COL 3), fibronectin (FN), α-SMA, matrix metalloproteinases 1 (MMP-1), matrix metalloproteinases 3 (MMP-3), and transforming growth factor-beta 1 (TGF-β1), were analysed. Results hFDSPC-CM combined with HA showed superior wound closure rate over hADSC-CM. Histologically, the hFDSPC-CM combined with HA group showed significantly improved re-epithelialisation, angiogenesis, anti-inflammation, collagen regeneration, and maturation compared to hADSC-CM combined with HA group. In vitro assays revealed that hFDSPC-CM displayed significant advantages on cell proliferation, migration, and ECM regeneration through a TGF-β/Smad signalling pathway compared with hADSC-CM. Conclusions hFDSPC-CM combined with HA was superior for treating diabetic wounds. The underlying mechanism may promote proliferation and migration of epidermal cells with fibroblasts, thus leading to ECM deposition and remodelling. Reduced inflammation may be due to the above-mentioned mechanism.

scholarly journals Matrilin-2 interacts with itself and with other extracellular matrix proteins

2002 ◽  
Vol 367 (3) ◽  
pp. 715-721 ◽  
Author(s):  
Dorothea PIECHA ◽  
Charlotte WIBERG ◽  
Matthias MÖRGELIN ◽  
Dieter P. REINHARDT ◽  
Ferenc DEÁK ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Basement Membrane ◽  
Extracellular Matrix Proteins ◽  
Collagen I ◽  
Matrix Proteins ◽  
Hek 293 ◽  
Protein Ligands ◽  
293 Cells ◽  
Physiological Relevance ◽  
Kd Values

Matrilin-2 is a component of extracellular filamentous networks. To study the interactions by which it can be integrated into such assemblies, full-length and truncated forms of matrilin-2 were recombinantly expressed in HEK-293 cells and purified from conditioned medium. The recombinant proteins, when used in interaction assays, showed affinity to matrilin-2 itself, but also to other collagenous and non-collagenous extracellular matrix proteins. The interaction between matrilin-2 and collagen I was studied in greater detail and could be shown to occur at distinct sites on the collagen I molecule and to have a KD of about 3×10-8M. Interactions with some non-collagenous protein ligands were even stronger, with matrilin-2 binding to fibrillin-2, fibronectin and laminin-1—nidogen-1 complexes, with KD values in the range of 10-8—10-11M. Co-localization of matrilin-2 with these ligands in the dermal-epidermal basement membrane, in the microfibrils extending from the basement membrane into the dermis, and in the dermal extracellular matrix, indicates a physiological relevance of the interactions in the assembly of supramolecular extracellular matrix structures.

scholarly journals Extracellular Matrix Components HAPLN1, Lumican, and Collagen I Cause Hyaluronic Acid-Dependent Folding of the Developing Human Neocortex

Neuron ◽  
2018 ◽  
Vol 99 (4) ◽  
pp. 702-719.e6 ◽  
Author(s):  
Katherine R. Long ◽  
Ben Newland ◽  
Marta Florio ◽  
Nereo Kalebic ◽  
Barbara Langen ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Hyaluronic Acid ◽  
Collagen I ◽  
Human Neocortex ◽  
Extracellular Matrix Components ◽  
Matrix Components

Extracellular matrix and morphogenesis in cnidarians: a tightly knit relationship

2019 ◽  
Vol 63 (3) ◽  
pp. 407-416 ◽  
Author(s):  
Bruno Gideon Bergheim ◽  
Suat Özbek
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cell ◽  
Basement Membrane ◽  
Genetic Manipulation ◽  
Body Plan ◽  
Body Axis ◽  
Basic Composition ◽  
Cell Layers ◽  
Simple Body ◽  
Matrix Components

Abstract Cnidarians, members of an early-branching metazoan phylum, possess an extracellular matrix (ECM) between their two epithelial cell layers, called the mesoglea. The cnidarian ECM, which is best studied in Hydra, contains matrix components reflective of both interstitial matrix and basement membrane. The identification of core matrisome components in cnidarian genomes has led to the notion that the basic composition of vertebrate ECM is of highly conserved nature and can be traced back to pre-bilaterians. While in vertebrate classes ECM factors have often diverged and acquired specialized functions in the context of organ development, cnidarians with their simple body plan retained direct links between ECM and morphogenesis. Recent advances in genetic manipulation techniques have provided tools for systematically studying cnidarian ECM function in body axis patterning and regeneration.

scholarly journals Differential Expression of Extracellular Matrix Components in Nasal Polyp Endotypes

2019 ◽  
Vol 33 (6) ◽  
pp. 665-670 ◽  
Author(s):  
Xin Feng ◽  
Spencer C. Payne ◽  
Larry Borish ◽  
John W. Steinke
Keyword(s):  
Extracellular Matrix ◽  
Nasal Polyp ◽  
Collagen Iv ◽  
Disease State ◽  
Collagen I ◽  
Cellular Infiltrate ◽  
Matrix Deposition ◽  
Total Collagen ◽  
Collagen Iii ◽  
Extracellular Matrix Components

Background Chronic rhinosinusitis is a difficult-to-treat disease that is often characterized by recurrent nasal polyp (NP) growth following surgical removal. The disease has been separated into distinct phenotypes based on cellular infiltrate or underlying physiological mechanisms. NPs are composed in part of an inflammatory cellular infiltrate, blood vessels, and a large amount of extracellular matrix (ECM). Despite the recognition of prominent ECM deposition, few studies have examined the components in detail and how they might differ with disease state. Objective The purpose of this study was to quantitate the expression of ECM components in NPs. Methods NPs were stained with pico-sirius red to determine total collagen content, and immunofluorescence was used to detect collagen I, collagen III, collagen IV, fibronectin, and laminin. Expression of each was quantitated and analyzed in relation to rhinosinusitis phenotype and separately as a function of polyp eosinophil number. Results When analyzed by phenotype, collagen I, collagen III, and fibronectin were expressed at the highest levels in noneosinophilic sinus disease. Collagen IV was not different among any groups, and its location was found predominately around vessels. When analyzed as a function of polyp eosinophil number, total collagen and collagen III showed a significant inverse correlation. Conclusions NP ECM composition differs with disease state with higher expression in cases where eosinophil levels are low. This suggests that in eosinophilic polyps there is a loss of matrix deposition either through break down or a failure to produce the essential components. Understanding these differences may identify new therapeutic targets.

Denervation does not change the ratio of collagen I and collagen III mRNA in the extracellular matrix of muscle

2007 ◽  
Vol 292 (2) ◽  
pp. R983-R987 ◽  
Author(s):  
Ellen M. Arruda ◽  
Kevin Mundy ◽  
Sarah Calve ◽  
Keith Baar
Keyword(s):  
Extracellular Matrix ◽  
Type I Collagen ◽  
Lysyl Oxidase ◽  
Muscle Fibers ◽  
Fold Increase ◽  
Collagen I ◽  
Type I ◽  
Collagen Concentration ◽  
Collagen Iii ◽  
Picrosirius Red

Denervation or inactivity is known to decrease the mass and alter the phenotype of muscle and the mechanics of tendon. It has been proposed that a shift in the collagen of the extracellular matrix (ECM) of the muscle, increasing type III and decreasing type I collagen, may be partially responsible for the observed changes. We directly investigated this hypothesis using quantitative real-time PCR on muscles and tendons that had been denervated for 5 wk. Five weeks of denervation resulted in a 2.91-fold increase in collagen concentration but no change in the content of collagen in the muscle, whereas in the tendon there was no change in either the concentration or content of collagen. The expression of collagen I, collagen III, and lysyl oxidase mRNA in the ECM of muscle decreased (76 ± 1.6%, 73 ± 2.3%, and 83 ± 3.2%, respectively) after 5 wk of denervation. Staining with picrosirius red confirmed the earlier observation of a change in staining color from red to green. Taken with the observed equivalent decreases in collagen I and III mRNA, this suggests that there was a change in orientation of the ECM of muscle becoming more aligned with the axis of the muscle fibers and no change in collagen type. The change in collagen orientation may serve to protect the smaller muscle fibers from damage by increasing the stiffness of the ECM and may partly explain why the region of the tendon closest to the muscle becomes stiffer after inactivity.

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