scholarly journals A novel method for sensor-based quantification of single/multicellular force dynamics and stiffening in 3D matrices

2021 ◽  
Vol 7 (15) ◽  
pp. eabf2629
Author(s):  
Bashar Emon ◽  
Zhengwei Li ◽  
Md Saddam H. Joy ◽  
Umnia Doha ◽  
Farhad Kosari ◽  
...  
Keyword(s):  
Self Assembly ◽  
Single Cells ◽  
Human Colon ◽  
Matrix Remodeling ◽  
Tissue Stiffness ◽  
Force Dynamics ◽  
Cell Functions ◽  
The Matrix ◽  
Cancer Tumor

Cells in vivo generate mechanical traction on the surrounding 3D extracellular matrix (ECM) and neighboring cells. Such traction and biochemical cues may remodel the matrix, e.g., increase stiffness, which, in turn, influences cell functions and forces. This dynamic reciprocity mediates development and tumorigenesis. Currently, there is no method available to directly quantify single-cell forces and matrix remodeling in 3D. Here, we introduce a method to fulfill this long-standing need. We developed a high-resolution microfabricated sensor that hosts a 3D cell-ECM tissue formed by self-assembly. This sensor measures cell forces and tissue stiffness and can apply mechanical stimulation to the tissue. We measured single and multicellular force dynamics of fibroblasts (3T3), human colon (FET) and lung (A549) cancer cells, and cancer-associated fibroblasts (CAF05) with 1-nN resolution. Single cells show notable force fluctuations in 3D. FET/CAF coculture system, mimicking cancer tumor microenvironment, increased tissue stiffness by three times within 24 hours.

scholarly journals A novel method for sensor-based quantification of single/multi-cellular traction dynamics and remodeling in 3D matrices

2020 ◽  
Author(s):  
Bashar Emon ◽  
Zhengwei Li ◽  
Md Saddam Hossain Joy ◽  
Umnia Doha ◽  
Farhad Kosari ◽  
...  
Keyword(s):  
Self Assembly ◽  
Single Cells ◽  
Human Colon ◽  
Matrix Remodeling ◽  
Tissue Stiffness ◽  
Force Dynamics ◽  
Cell Functions ◽  
The Matrix ◽  
Cancer Tumor

AbstractCells in vivo generate mechanical forces (traction) on surrounding 3D extra cellular matrix (ECM) and cells. Such traction and biochemical cues may remodel the matrix, e.g. increase stiffness, which in turn influences cell functions and forces. This dynamic reciprocity mediates development and tumorigenesis. Currently, there is no method available to directly quantify single cell traction and matrix remodeling in 3D. Here, we introduce a method to fulfil this long-standing need. We developed a high-resolution microfabricated sensor which hosts a 3D cell-ECM tissue formed by self-assembly. It measures cell forces and tissue-stiffness and can apply mechanical stimulation to the tissue. We measured single and multicellular force dynamics of fibroblasts (3T3), human colon (FET) and lung (A549) cancer cells and cancer associated fibroblasts (CAF05) with 1 nN resolution. Single cells show significant force fluctuations in 3D. FET/CAF co-culture system, mimicking cancer tumor microenvironment, increased tissue stiffness by 3 times within 24 hours.

scholarly journals Functional angiogenesis requires microenvironmental cues balancing endothelial cell migration and proliferation

2019 ◽  
Author(s):  
William Y. Wang ◽  
Daphne Lin ◽  
Evan H. Jarman ◽  
William J. Polacheck ◽  
Brendon M. Baker
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cell ◽  
Single Cells ◽  
Endothelial Cell Migration ◽  
Migration Speed ◽  
Parent Vessel ◽  
Extracellular Milieu ◽  
Cell Functions

ABSTRACTAngiogenesis is a complex morphogenetic process that involves intimate interactions between multicellular endothelial structures and their extracellular milieu. In vitro models of angiogenesis can aid in reducing the complexity of the in vivo microenvironment and provide mechanistic insight into how soluble and physical extracellular matrix cues regulate this process. To investigate how microenvironmental cues regulate angiogenesis and the function of resulting microvasculature, we multiplexed an established angiogenesis-on-a-chip platform that affords higher throughput investigation of 3D endothelial cell sprouting emanating from a parent vessel through defined biochemical gradients and extracellular matrix. We found that two fundamental endothelial cell functions, migration and proliferation, dictate endothelial cell invasion as single cells vs. multicellular sprouts. Microenvironmental cues that elicit excessive migration speed incommensurate with proliferation resulted in microvasculature with poor barrier function and an inability to transport fluid across the microvascular bed. Restoring the balance between migration speed and proliferation rate rescued multicellular sprout invasion, providing a new framework for the design of pro-angiogenic biomaterials that guide functional microvasculature formation for regenerative therapies.

scholarly journals Measuring dynamic cell–material interactions and remodeling during 3D human mesenchymal stem cell migration in hydrogels

2015 ◽  
Vol 112 (29) ◽  
pp. E3757-E3764 ◽  
Author(s):  
Kelly M. Schultz ◽  
Kyle A. Kyburz ◽  
Kristi S. Anseth
Keyword(s):  
Stem Cell ◽  
3D Culture ◽  
Human Mesenchymal Stem Cell ◽  
Matrix Remodeling ◽  
Tissue Engineering Scaffolds ◽  
Encapsulated Cells ◽  
3D Microenvironment ◽  
The Matrix ◽  
Biomaterial Scaffolds

Biomaterials that mimic aspects of the extracellular matrix by presenting a 3D microenvironment that cells can locally degrade and remodel are finding increased applications as wound-healing matrices, tissue engineering scaffolds, and even substrates for stem cell expansion. In vivo, cells do not simply reside in a static microenvironment, but instead, they dynamically reengineer their surroundings. For example, cells secrete proteases that degrade extracellular components, attach to the matrix through adhesive sites, and can exert traction forces on the local matrix, causing its spatial reorganization. Although biomaterials scaffolds provide initially well-defined microenvironments for 3D culture of cells, less is known about the changes that occur over time, especially local matrix remodeling that can play an integral role in directing cell behavior. Here, we use microrheology as a quantitative tool to characterize dynamic cellular remodeling of peptide-functionalized poly(ethylene glycol) (PEG) hydrogels that degrade in response to cell-secreted matrix metalloproteinases (MMPs). This technique allows measurement of spatial changes in material properties during migration of encapsulated cells and has a sensitivity that identifies regions where cells simply adhere to the matrix, as well as the extent of local cell remodeling of the material through MMP-mediated degradation. Collectively, these microrheological measurements provide insight into microscopic, cellular manipulation of the pericellular region that gives rise to macroscopic tracks created in scaffolds by migrating cells. This quantitative and predictable information should benefit the design of improved biomaterial scaffolds for medically relevant applications.

scholarly journals The R-enantiomer of ketorolac reduces ovarian cancer tumor burden in vivo

BMC Cancer ◽  
2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Martha M. Grimes ◽  
S. Ray Kenney ◽  
Dayna R. Dominguez ◽  
Kathryn J. Brayer ◽  
Yuna Guo ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Tumor Cells ◽  
High Throughput Screening ◽  
Tumor Burden ◽  
Migration And Invasion ◽  
Rna Seq ◽  
Downstream Signaling ◽  
Cell Functions ◽  
Cancer Tumor

Abstract Background Rho-family GTPases, including Ras-related C3 botulinum toxin substrate 1 (Rac1) and cell division control protein 42 (Cdc42), are important modulators of cancer-relevant cell functions and are viewed as promising therapeutic targets. Based on high-throughput screening and cheminformatics we identified the R-enantiomer of an FDA-approved drug (ketorolac) as an inhibitor of Rac1 and Cdc42. The corresponding S-enantiomer is a non-steroidal anti-inflammatory drug (NSAID) with selective activity against cyclooxygenases. We reported previously that R-ketorolac, but not the S-enantiomer, inhibited Rac1 and Cdc42-dependent downstream signaling, growth factor stimulated actin cytoskeleton rearrangements, cell adhesion, migration and invasion in ovarian cancer cell lines and patient-derived tumor cells. Methods In this study we treated mice with R-ketorolac and measured engraftment of tumor cells to the omentum, tumor burden, and target GTPase activity. In order to gain insights into the actions of R-ketorolac, we also performed global RNA-sequencing (RNA-seq) analysis on tumor samples. Results Treatment of mice with R-ketorolac decreased omental engraftment of ovarian tumor cells at 18 h post tumor cell injection and tumor burden after 2 weeks of tumor growth. R-ketorolac treatment inhibited tumor Rac1 and Cdc42 activity with little impact on mRNA or protein expression of these GTPase targets. RNA-seq analysis revealed that R-ketorolac decreased expression of genes in the HIF-1 signaling pathway. R-ketorolac treatment also reduced expression of additional genes associated with poor prognosis in ovarian cancer. Conclusion These findings suggest that R-ketorolac may represent a novel therapeutic approach for ovarian cancer based on its pharmacologic activity as a Rac1 and Cdc42 inhibitor. R-ketorolac modulates relevant pathways and genes associated with disease progression and worse outcome.

scholarly journals The Role of Thyroid Hormone 3,3 ‘, 5 - Triiodothyronine (T3) in the Expression of Osteocalcin and Lipocalin 2 (LCN2) in Osteoblasts

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A238-A238
Author(s):  
Amanda Fantini de Camargo Andrade ◽  
Thaís Silva Pinto ◽  
Geórgia Silva Feltran ◽  
Renato Ferretti ◽  
Willian Fernando Zambuzzi
Keyword(s):  
Thyroid Hormone ◽  
Marker Genes ◽  
Matrix Remodeling ◽  
Bone Homeostasis ◽  
Lipocalin 2 ◽  
Physiological Level ◽  
The Matrix ◽  
Perform Gene Expression

Abstract Introduction: The thyroid hormone 3,3 ‘, 5 - triiodothyronine (T3) has an important role in bone physiology and metabolism, stimulating osteodifferentiation and bone homeostasis. Although there is this evidence, little is known about the synergistic events of T3 together with the endocrine role of other players that interfere with osteoblast metabolism, such as Lipocalin 2 (LCN2) and its hypothalamic-located MCR4 receptor. It is also known that LCN2 interferes at a physiological level with the parameters of food intake. Objective: To evaluate the role of the thyroid hormone 3,3 ‘, 5 - Triiodothyronine (T3) in the control of the expression of both Osteocalcin (OCN) and Lipocalin 2 (LCN2) in osteoblasts. Materials and Methods:. In order to understand the molecular and physiological mechanisms about that probable synergism, the experiments were as follows: Mouse pre-osteoblasts were challenged with T3 hormone treatments in three levels (1nM; 1.5nM; 2.0nM) and different times (0h; 24h; 72h). The samples were collected to perform gene expression of the main osteodifferentiation markers (RUNX2, OTX, BSP), matrix remodeling (MMPs, TIMPs, BMPII) and zymography assay for analysis of metalloproteinase activities (MMPs). Results and Discussion: Preliminarily, it was founded a synergism between the hormone T3 and LCN2 expression in osteoblasts, occurring the modulation of the marker genes of differentiation, extracellular matrix and hormonal synthesis. The T3 hormone acts in the modulation of RUNX2, OTX, BMP2, favoring osteodifferentiation and the remodeling of the matrix, probably activating the TIMP1-MMP9 and TIMP2-MMP2 complexes, mainly in hyperthyroid conditions. In addition, we note a direct influence of T3 on the both expression of both Osteocalcin (OCN) and Lipocalin 2 (LCN2). Final Considerations: Our preliminary data indicates that the hormone T3 acts on the metabolism of osteoblasts through the modulation of LCN2 and BMPII. These molecular findings need to be confronted with further analyzes in vivo for more conclusive physiological conclusions.

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

Ultrastructural modification of cellular interactions in cancer tumor

1978 ◽  
Vol 36 (2) ◽  
pp. 318-319
Author(s):  
N. P. Dmitrieva
Keyword(s):  
Cancer Cells ◽  
Human Thyroid ◽  
Adjacent Cell ◽  
Main Role ◽  
Cellular Interactions ◽  
Electron Microscopic ◽  
Cancer Tumor ◽  
Normal Human ◽  
Characteristic Features

One of the most characteristic features of cancer cells is their ability to metastasia. It is suggested that the modifications of the structure and properties of cancer cells surfaces play the main role in this process. The present work was aimed at finding out what ultrastructural features apear in tumor in vivo which removal of individual cancer cells from the cell population can provide. For this purpose the cellular interactions in the normal human thyroid and cancer tumor of this gland electron microscopic were studied. The tissues were fixed in osmium tetroxide and were embedded in Araldite-Epon.In normal human thyroid the most common type of intercellular contacts was represented by simple junction formed by the parallelalignment of adjacent cell membranees leaving in between an intermembranes space 15-20 nm filled with electronlucid material (Fig. 1a). Sometimes in the basal part of cells dilatations of the intercellular space 40-50 nm wide were found (Fig. 1a). Here the cell surfaces may form single short microvilli.

Ultrastructural Study of a differentiating human colon carcinoma cell line

1990 ◽  
Vol 48 (3) ◽  
pp. 208-209
Author(s):  
Sylvie Polak-Charcon ◽  
Mehrdad Hekmati ◽  
Yehuda Ben Shaul
Keyword(s):  
Cell Line ◽  
Morphological Changes ◽  
Secretory Granules ◽  
Human Colon ◽  
Cell Types ◽  
Culture Conditions ◽  
Morphological Evidence ◽  
Human Colon Carcinoma Cell ◽  
Colon Cell

The epithelium of normal human colon mucosa “in vivo” exhibits a gradual pattern of differentiation as undifferentiated stem cells from the base of the crypt of “lieberkuhn” rapidly divide, differentiate and migrate toward the free surface. The major differentiated cell type of the intestine observed are: absorptive cells displaying brush border, goblet cells containing mucous granules, Paneth and endocrine cells containing dense secretory granules. These different cell types are also found in the intestine of the 13-14 week old embryo.We present here morphological evidence showing that HT29, an adenocarcinoma of the human colon cell line, can differentiate into various cell types by changing the growth and culture conditions and mimic morphological changes found during development of the intestine in the human embryo.HT29 cells grown in tissue-culture dishes in DMEM and 10% FCS form at late confluence a multilayer of morphologically undifferentiated cell culture covered with irregular microvilli, and devoid of tight junctions (Figs 1-3).

In Vitro and in Vivo Comparison of Binding of 99m-Tc-Iabeled Anti-CEA MAb F33-104 with 99m-Tc-labeled Anti-CEA MAb BW431/26

1999 ◽  
Vol 38 (04) ◽  
pp. 115-119
Author(s):  
N. Oriuchi ◽  
S. Sugiyama ◽  
M. Kuroki ◽  
Y. Matsuoka ◽  
S. Tanada ◽  
...  
Keyword(s):  
Nude Mice ◽  
Binding Capacity ◽  
Colon Adenocarcinoma ◽  
Human Colon ◽  
Cell Binding ◽  
Vitro Binding ◽  
Labeling Method ◽  
Human Colon Adenocarcinoma

Summary Aim: The purpose of this study was to assess the potential for radioimmunodetection (RAID) of murine anti-carcinoembryonic antigen (CEA) monoclonal antibody (MAb) F33-104 labeled with technetium-99m (99m-Tc) by a reduction-mediated labeling method. Methods: The binding capacity of 99m-Tc-labeled anti-CEA MAb F33-104 with CEA by means of in vitro procedures such as immunoradiometric assay and cell binding assay and the biodistribution of 99m-Tc-labeled anti-CEA MAb F33-104 in normal nude mice and nude mice bearing human colon adenocarcinoma LS180 tumor were investigated and compared with 99m-Tc-labeled anti-CEA MAb BW431/26. Results: The in vitro binding rate of 99m-Tc-labeled anti-CEA MAb F33-104 with CEA in solution and attached to the cell membrane was significantly higher than 99m-Tclabeled anti-CEA MAb BW431/261 (31.4 ± 0.95% vs. 11.9 ± 0.55% at 100 ng/mL of soluble CEA, 83.5 ± 2.84% vs. 54.0 ± 2.54% at 107 of LS 180 cells). In vivo, accumulation of 99m-Tc-labeled anti-CEA MAb F33-104 was higher at 18 h postinjection than 99m-Tc-labeled anti-CEA MAb BW431/26 (20.1 ± 3.50% ID/g vs. 14.4 ± 3.30% ID/g). 99m-Tcactivity in the kidneys of nude mice bearing tumor was higher at 18 h postinjection than at 3 h (12.8 ± 2.10% ID/g vs. 8.01 ± 2.40% ID/g of 99m-Tc-labeled anti-CEA MAb F33-104, 10.7 ± 1.70% ID/g vs. 8.10 ± 1.75% ID/g of 99m-Tc-labeled anti-CEA MAb BW431/26). Conclusion: 99m-Tc-labeled anti-CEA MAb F33-104 is a potential novel agent for RAID of recurrent colorectal cancer.

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