scholarly journals A Sensitive Chemotaxis Assay Using a Novel Microfluidic Device

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Chen Zhang ◽  
Sunyoung Jang ◽  
Ovid C. Amadi ◽  
Koichi Shimizu ◽  
Richard T. Lee ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Microfluidic Device ◽  
Muscle Cells ◽  
Random Motion ◽  
Cellular Migration ◽  
Sensitive Assay ◽  
Chemotaxis Assay ◽  
Chemokine Gradient ◽  
Over Time

Existing chemotaxis assays do not generate stable chemotactic gradients and thus—over time—functionally measure only nonspecific random motion (chemokinesis). In comparison, microfluidic technology has the capacity to generate a tightly controlled microenvironment that can be stably maintained for extended periods of time and is, therefore, amenable to adaptation for assaying chemotaxis. We describe here a novel microfluidic device for sensitive assay of cellular migration and show its application for evaluating the chemotaxis of smooth muscle cells in a chemokine gradient.

Cell-based microfluidic device for screening anti-proliferative activity of drugs in vascular smooth muscle cells

2012 ◽  
Vol 14 (6) ◽  
pp. 1129-1140 ◽  
Author(s):  
R. Rodriguez-Rodriguez ◽  
X. Muñoz-Berbel ◽  
S. Demming ◽  
S. Büttgenbach ◽  
M. D. Herrera ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Microfluidic Device ◽  
Proliferative Activity ◽  
Muscle Cells

scholarly journals Influences of surface chemistry and swelling of salt-treated polyelectrolyte multilayers on migration of smooth muscle cells

2012 ◽  
Vol 9 (77) ◽  
pp. 3455-3468 ◽  
Author(s):  
Lulu Han ◽  
Zhengwei Mao ◽  
Jindan Wu ◽  
Yuying Zhang ◽  
Changyou Gao
Keyword(s):  
Smooth Muscle ◽  
Cell Migration ◽  
Surface Chemistry ◽  
Smooth Muscle Cells ◽  
Focal Adhesion ◽  
Chemical Properties ◽  
Muscle Cells ◽  
Cellular Migration ◽  
Gene Expressions ◽  
And Migration

The cell migration plays a crucial role in a variety of physiological and pathological processes and can be regulated by the cell–substrate interactions. We found previously that the poly(sodium 4-styrenesulphonate) (PSS)/poly(diallyldimethylammonium) chloride (PDADMAC) multilayers post-treated in 1–5 M NaCl solutions result in continuous changes of their physico-chemical properties such as thickness, chemical composition, surface charge, swelling ratio and wettability. In this study, the responses of human smooth muscle cells (SMCs) on these salt-treated multilayers, particularly the governing factors of cellular migration that offer principles for designing therapeutics and implants, were disclosed. The cell migration rate was slowest on the 3 M NaCl-treated multilayers, which was comparable with that on tissue culture plates, but it was highest on 5 M NaCl-treated multilayers. To elucidate the intrinsic mechanisms, cell adhesion, proliferation, adhesion and related gene expressions were further investigated. The SMCs preferred to attach, spread and proliferate on the PSS-dominated surfaces with well-organized focal adhesion and actin fibres, especially on the 3 M NaCl-treated multilayers, while were kept round and showed low viability on the PDADMAC-dominated surfaces. The relative mRNA expression levels of adhesion-related genes such as fibronectin, laminin and focal adhesion kinase, and migration-related genes such as myosin IIA and Cdc42 were compared to explain the different cellular behaviours. These results reveal that the surface chemistry and the swelling of the salt-treated multilayers govern the cell migration behaviours.

scholarly journals Galectin-3 is expressed in vascular smooth muscle cells and promotes pulmonary hypertension through changes in proliferation, apoptosis, and fibrosis

2019 ◽  
Vol 316 (5) ◽  
pp. L784-L797 ◽  
Author(s):  
Scott A. Barman ◽  
Xueyi Li ◽  
Stephen Haigh ◽  
Dmitry Kondrikov ◽  
Keyvan Mahboubi ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Molecular Mechanisms ◽  
Muscle Cells ◽  
Cellular Migration ◽  
Rat Models ◽  
Galectin 3 ◽  
Genetic Deletion

A defining characteristic of pulmonary hypertension (PH) is the extensive remodeling of pulmonary arteries (PAs), which results in progressive increases in vascular resistance and stiffness and eventual failure of the right ventricle. There is no cure for PH and identification of novel molecular mechanisms that underlie increased proliferation, reduced apoptosis, and excessive extracellular matrix production in pulmonary artery smooth muscle cells (PASMCs) is a vital objective. Galectin-3 (Gal-3) is a chimeric lectin and potent driver of many aspects of fibrosis, but its role in regulating PASMC behavior in PH remains poorly understood. Herein, we evaluated the importance of increased Gal-3 expression and signaling on PA vascular remodeling and cardiopulmonary function in experimental models of PH. Gal-3 expression was quantified by qRT-PCR, immunoblotting, and immunofluorescence imaging, and its functional role was assessed by specific Gal-3 inhibitors and CRISPR/Cas9-mediated knockout of Gal-3 in the rat. In rat models of PH, we observed increased Gal-3 expression in PASMCs, which stimulated migration and resistance to apoptosis, whereas silencing or genetic deletion reduced cellular migration and PA fibrosis and increased apoptosis. Gal-3 inhibitors attenuated and reversed PA remodeling and fibrosis, as well as hemodynamic indices in monocrotaline (MCT)-treated rats in vivo. These results were supported by genetic deletion of Gal-3 in both MCT and Sugen Hypoxia rat models. In conclusion, our results suggest that elevated Gal-3 levels contribute to inappropriate PA remodeling in PH by enhancing multiple profibrotic mechanisms. Therapeutic strategies targeting Gal-3 may be of benefit in the treatment of PH.

An immunohistochemical study of arterial lesions due to pulmonary hypertension in patients with congenital heart defects

1994 ◽  
Vol 4 (1) ◽  
pp. 37-43
Author(s):  
Vera Demarchi Aiello ◽  
Maria de Lourdes Higuchi ◽  
Edgard Augusto Lopes ◽  
Antonio Augusto Barbosa Lopes ◽  
Miguel Barbero-Marcial ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Heart Defects ◽  
Muscle Cells ◽  
Cellular Migration ◽  
Cellular Mechanisms ◽  
Formalin Fixed Paraffin ◽  
Arterial Lesions ◽  
Formalin Fixed Paraffin Embedded ◽  
Lung Biopsies

AbstractIn order to understand some of the cellular mechanisms of interaction in secondary pulmonary vaso-occlusive disease, we studied 21 lung biopsies from patients with different types of congenital cardiac defects. Their ages ranged from four to 248 months (mean 71.5 months; median 41 months). Changes in the cytoskeleton and extracellular matrix were assessed in the arterial wall. Immunostaining was applied to formalin-fixed, paraffin- embedded tissue, using antibodies to muscle-specific actin, vimentin and fibronectin in supra-optimal dilution. The staining for muscle-specific actin in the medial layer revealed a heterogenous pattern, with areas exhibiting low or absent labelling, reflecting a process of dedifferentiation of the smooth muscle cells in those segments. Within intimal proliferative lesions, the expression of muscle-specific actin was variable, being weak in some lesions and strong in those showing concentrically arranged intimal smooth muscle cells, suggesting a reversion of the migrated cells to the contractile phenotype. The endothelial cells of arteries from cases presenting severe qualitative lesions exhibited strong expression of vimentin, reflecting their heightened regenerative activity and/or their necessity to maintain their shape. The expression of fibronectin was greater in the predominantly cellular lesions of the intima when compared to the fibrotic lesions, indicating the role of that matrix glycoprotein in cellular migration and in replicative processes.

scholarly journals A Pivotal Role for AP-1-Mediated Osteopontin Expression in the Increased Migration of Vascular Smooth Muscle Cells Stimulated With HMGB1

2021 ◽  
Vol 12 ◽  
Author(s):  
Eun Yeong Jeon ◽  
Seung Eun Baek ◽  
Ji On Kim ◽  
Jong Min Choi ◽  
Eun Jeong Jang ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Vascular Remodeling ◽  
Molecular Mechanisms ◽  
Muscle Cells ◽  
Cellular Migration ◽  
Pivotal Role ◽  
In Cells

Migration of vascular smooth muscle cells (VSMCs) plays an essential role in the development of vascular remodeling in the injured vasculatures. Previous studies have identified high-mobility group box 1 (HMGB1) as a principal effector mediating vascular remodeling; however, the mechanisms involved have not been fully elucidated. Thus, this study investigated the role of HMGB1 on VSMC migration and the underlying molecular mechanisms involved. VSMCs were ex plant cultured using rat thoracic aorta, and the cellular migration was measured using wound-healing assay. Osteopontin (OPN) mRNA and protein were determined by reverse transcription polymerase chain reaction (RT-PCR) and Western blot, respectively. The OPN promoter was cloned into pGL3 basic to generate a pLuc-OPN-2284 construct. Migration of VSMCs stimulated with HMGB1 (100ng/ml) was markedly increased, which was significantly attenuated in cells pretreated with MPIIIB10 (100–300ng/ml), a neutralizing monoclonal antibody for OPN as well as in cells deficient of OPN. In VSMCs stimulated with HMGB1, OPN mRNA and protein levels were significantly increased in association with an increased promotor activity of OPN gene. Putative-binding sites for activator protein 1 (AP-1) and CCAAT/enhancer-binding protein beta (C/EBPβ) in the indicated promoter region were suggested by TF Search, and the HMGB1-induced expression of OPN was markedly attenuated in cells transfected with siRNA for AP-1. VSMC stimulated with HMGB1 also showed an increased expression of AP-1. Results of this study suggest a pivotal role for AP-1-induced OPN expression in VSMC migration induced by HMGB1. Thus, the AP-1-OPN signaling axis in VSMC might serve as a potential therapeutic target for vascular remodeling in the injured vasculatures.

In Vitro Biocompatibility of Coaxial Electrospun Scaffolds for Cardiovascular Tissue Engineering

2013 ◽  
Author(s):  
Valerie M. Merkle ◽  
Kaitlyn R. Ammann ◽  
Katrina J. DeCook ◽  
Phat L. Tran ◽  
Marvin J. Slepian ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Endothelial Cells ◽  
Smooth Muscle ◽  
Polyvinyl Alcohol ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Cellular Adhesion ◽  
Cellular Migration ◽  
Cardiovascular Tissue ◽  
Cardiovascular Tissue Engineering

Coaxial electrospinning is a novel technique that allows the fabrication of composite nanofibers in a core-shell structure. This technique can be used to optimize the biological properties of a natural polymer (i.e. gelatin) and the mechanical properties of a synthetic polymer (i.e. polyvinyl alcohol). In this study, we fabricated coaxial nanofibers of gelatin and polyvinyl alcohol (PVA) for use in cardiovascular tissue engineering. Cellular adhesion and proliferation of human umbilical endothelial cells (HUVEC) and smooth muscle cells (SMC) is determined on coaxial nanofibers fabricated from a 1:1 and 3:1 (gelatin:PVA volumetric flow rate ratios), as well as nanofibers composed solely of gelatin or PVA. In addition, cellular migration on the coaxial nanofibers, gelatin nanofibers, and PVA nanofibers is determined for both endothelial cells and smooth muscle cells.

scholarly journals Resistin and High Glucose Concentrations-Activation of Human Smooth Muscle Cells Induces Enhanced Monocyte Chemotaxis

2012 ◽  
Vol 19 (1) ◽  
pp. 17-24
Author(s):  
Viorel Simion ◽  
Ana-Maria Gan ◽  
Daniela Stan ◽  
Monica Pirvulescu ◽  
Manuela Calin ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Conditioned Medium ◽  
High Glucose ◽  
Muscle Cells ◽  
Chemotaxis Assay ◽  
Inverted Microscope ◽  
Monocyte Chemotaxis ◽  
Receptor Cx3cr1

Resistin and High Glucose Concentrations-Activation of Human Smooth Muscle Cells Induces Enhanced Monocyte ChemotaxisObjectives. Recent data indicate that upon activation by resistin and high glucose concentrations (HG) vascular smooth muscle cells (SMC) acquire pro-inflammatory properties. We questioned whether resistin and HG-activated SMC generate an enhanced monocytes chemotaxis and if the chemokine fractalkine (Fk) is involved in the process. Material and Methods: SMC were incubated with resistin or/and HG and the conditioned medium was used for monocytes chemotaxis assays. The role of Fk was assessed by blocking the Fk receptor, CX3CR1, on monocytes (U937 cell lines) prior to the chemotaxis assay. The quantification of migrated monocytes was assayed under an inverted microscope and statistically analyzed. Results: (i) conditioned medium (CM) collected from SMC incubated with resistin in the presence or absence of HG triggered a significant increase (25 - 100 %) of monocytes chemotaxis as compared to controls; (ii) blocking the CX3CR1 receptor significantly decreased the monocyte chemotaxis towards resistin-treated SMC. Conclusions: Resistin±HG increases the expression of chemotaxis inductors in human SMC and the ensuing monocytes chemotaxis by a mechanism in which Fk plays a major role.

Immunogold localization of HMB-45 antigen in pulmonary lymphangiomyomatosis

1995 ◽  
Vol 53 ◽  
pp. 998-999
Author(s):  
J.M. Minda ◽  
E. Dessy ◽  
G. G. Pietra
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Room Temperature ◽  
Osmium Tetroxide ◽  
Muscle Cells ◽  
Ultrastructural Localization ◽  
Reproductive Age ◽  
Thin Sections ◽  
Smooth Muscle Proliferation ◽  
Hmb 45

Pulmonary lymphangiomyomatosis (PLAM) is a rare disease occurring exclusively in women of reproductive age. It involves the lungs, lymph nodes and lymphatic ducts. In the lungs, it is characterized by the proliferation of smooth muscle cells around lymphatics in the bronchovascular bundles, lobular septa and pleura The nature of smooth muscle proliferation in PLAM is still unclear. Recently, reactivity of the smooth muscle cells for HMB-45, a melanoma-related antigen has been reported by immunohistochemistry. The purpose of this study was the ultrastructural localization of HMB-45 immunoreactivity in these cells using gold-labeled antibodies.Lung tissue from three cases of PLAM, referred to our Institution for lung transplantation, was embedded in either Poly/Bed 812 post-fixed in 1% osmium tetroxide, or in LR White, without osmication. For the immunogold technique, thin sections were placed on Nickel grids and incubated with affinity purified, monoclonal anti-melanoma antibody HMB-45 (1:1) (Enzo Diag. Co) overnight at 4°C. After extensive washing with PBS, grids were treated with Goat-anti-mouse-IgG-Gold (5nm) (1:10) (Amersham Life Sci) for 1 hour, at room temperature.

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