scholarly journals Response of Astrocytes to Blood Exposure due to Shunt Insertion in vitro

2021 ◽  
Author(s):  
Mira Zaranek ◽  
Rooshan Arshad ◽  
Kevin Zheng ◽  
Carolyn Harris
Keyword(s):  
In Vitro Model ◽  
Cell Attachment ◽  
Ventricular Zone ◽  
Total Cell ◽  
Total Cell Count ◽  
Shunt Insertion ◽  
Catheter Obstruction ◽  
Vitro Model ◽  
Number Of Cells

The breakdown of the ventricular zone (VZ) with the presence of blood in cerebrospinal fluid (CSF) has been shown to increase shunt catheter obstruction in the treatment of hydrocephalus, but the mechanisms by which this occurs are generally unknown. Using a custom-built incubation chamber, we immunofluorescently assayed cell attachment and morphology on shunt catheters with and without blood after 14 days. Samples exposed to blood showed significantly increased cell attachment (average total cell count 392.0±317.1 versus control of 94.7±44.5, P<0.0001). Analysis of the glial fibrillary acidic protein (GFAP) expression showed similar trends (854.4±450.7 versus control of 174.3±116.5, P<0.0001). An in vitro model was developed to represent the exposure of astrocytes to blood following an increase in BBB permeability. Exposure of astrocytes to blood increases the number of cells and their spread on the shunt.

scholarly journals Blood Exposure Causes Ventricular Zone Disruption and Glial Activation In Vitro

2018 ◽  
Vol 77 (9) ◽  
pp. 803-813 ◽  
Author(s):  
Leandro Castaneyra-Ruiz ◽  
Diego M Morales ◽  
James P McAllister ◽  
Steven L Brody ◽  
Albert M Isaacs ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Ventricular Zone ◽  
Glial Activation ◽  
Cell Junctions ◽  
Cell Junction ◽  
Ependymal Cells ◽  
Glial Fibrillary Acid Protein ◽  
Newborn Mice ◽  
Vitro Model

Abstract Intraventricular hemorrhage (IVH) is the most common cause of pediatric hydrocephalus in North America but remains poorly understood. Cell junction-mediated ventricular zone (VZ) disruption and astrogliosis are associated with the pathogenesis of congenital, nonhemorrhagic hydrocephalus. Recently, our group demonstrated that VZ disruption is also present in preterm infants with IVH. On the basis of this observation, we hypothesized that blood triggers the loss of VZ cell junction integrity and related cytopathology. In order to test this hypothesis, we developed an in vitro model of IVH by applying syngeneic blood to cultured VZ cells obtained from newborn mice. Following blood treatment, cells were assayed for N-cadherin-dependent adherens junctions, ciliated ependymal cells, and markers of glial activation using immunohistochemistry and immunoblotting. After 24–48 hours of exposure to blood, VZ cell junctions were disrupted as determined by a significant reduction in N-cadherin expression (p < 0.05). This was also associated with significant decrease in multiciliated cells and increase in glial fibrillary acid protein-expressing cells (p < 0.05). These observations suggest that, in vitro, blood triggers VZ cell loss and glial activation in a pattern that mirrors the cytopathology of human IVH and supports the relevance of this in vitro model to define injury mechanisms.

Mechanical contributions to astrocyte adhesion using a novel in vitro model of catheter obstruction

2010 ◽  
Vol 222 (2) ◽  
pp. 204-210 ◽  
Author(s):  
Carolyn A. Harris ◽  
James H. Resau ◽  
Eric A. Hudson ◽  
Richard A. West ◽  
Candice Moon ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Catheter Obstruction ◽  
Vitro Model

scholarly journals Preterm Intraventricular Hemorrhage in vitro: Modeling the Cytopathology of the Ventricular Zone

2020 ◽  
Author(s):  
Leandro Castaneyra-Ruiz ◽  
James P. McAllister ◽  
Diego M. Morales ◽  
Steven L. Brody ◽  
Albert M. Isaacs ◽  
...  
Keyword(s):  
Intraventricular Hemorrhage ◽  
In Vitro Model ◽  
Ependymal Cell ◽  
Ventricular Zone ◽  
High Temporal Resolution ◽  
Neuroepithelial Cells ◽  
Vitro Model ◽  
Developmental Features ◽  
The Impact

Abstract Background: Severe intraventricular hemorrhage (IVH) is one of the most devastating neurological complications in preterm infants, with the majority suffering long-term neurological morbidity and up to 50 percent developing post hemorrhagic hydrocephalus (PHH). Despite the importance of this disease, its cytopathological mechanisms are not well known. An in vitro model of IVH is required to investigate the effects of blood and its components on the developing ventricular zone (VZ) and its stem cell niche. To address this need, we developed a new in vitro model to mimic the cytopathological conditions of IVH in the preterm infant. Methods: Maturing neuroepithelial cells from the VZ were harvested from the entire lateral ventricles of wild type C57BL/6 mice at 1-4 days of age and expanded in proliferation media for 3-5 days. At confluence, cells were re-plated onto 24-well plates in differentiation media to generate ependymal cells (EC). At approximately 3-5 days, which corresponded to the onset of ependymal cell differentiation based on the appearance of multiciliated cells , phosphate-buffered saline for controls or syngeneic whole blood for IVH was added to the ependymal cell surface. The cells were examined for the expression of EC markers of differentiation and maturation to qualitatively and quantitatively assess the effect of blood exposure on VZ transition from neuroepithelial cells to EC. Discussion: This model will allow investigators to test cytopathological mechanisms contributing to the pathology of IVH with high temporal resolution and query the impact of injury to the maturation of the VZ. This technique recapitulates features of normal maturation of the VZ in vitro, offering the capacity to investigate the developmental features of VZ biogenesis.

scholarly journals In vitro study after exposure to the aqueous extract of Piper amalago L. shows changes of morphology, proliferation, cytoskeleton and molecules of the extracellular matrix

2021 ◽  
Vol 10 (4) ◽  
pp. e0110413289
Author(s):  
Vera Lucia Pereira dos Santos ◽  
Célia Regina Cavichiolo Franco ◽  
Ricardo Wagner ◽  
Caroline Dadalt Silva ◽  
Célia Regina Cavichiolo Franco ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cell Viability ◽  
Cell Body ◽  
In Vitro Model ◽  
In Vitro Study ◽  
Healing Agent ◽  
Vitro Model ◽  
Insect Bites ◽  
Number Of Cells

Piper amalago L. is a medicinal plant traditionally used as a healing agent for wounds, burns, abscesses, boils, and insect bites. The current study aimed to evaluate the possible effects of the aqueous crude extract obtained from P. amalago leaves, in different concentrations and in different incubation times, using the in vitro model of mouse fibroblasts (3T3). The extract was tested in different concentrations at the 24 h incubation time for analysis of cell viability, cytotoxicity, proliferation, cell morphology, immunostaining, adhesion and cell spreading assays, as well as to determine the hydroxyproline concentration and activity of the metalloproteinase MMP2. Morphologically, after exposure to the concentrations of 15 and 150 µg/mL, the cells maintained the morphology, yet a greater number of cells with more expansions of the cell body and larger than the control cells were observed. The treated cell culture also showed a greater number of cells, larger cells, a greater expansion of the cell body, adherent cells spread over the substrate, and a more juxtaposed, central and spherical nucleus. The treatment induced greater cell adhesion to the polymer, fibronectin, and collagen I. Biochemical results showed a significant increase in the hydroxyproline amino acid after exposure for 96 h. The extract did not induce loss of cell viability until the concentration reached 150 µg/mL, positively modulating proliferation, morphology, adhesion, degree of spreading, and organization of microfilaments. The extract also promoted a significant increase in the hydroxyproline amino acid.

Growth of yeast cultures as in vitro model for investigating homœopathic medicines

1985 ◽  
Vol 74 (03) ◽  
pp. 132-140 ◽  
Author(s):  
William A. Steffen
Keyword(s):  
Cell Cultures ◽  
In Vitro Model ◽  
Total Cell ◽  
Yeast Cells ◽  
Exponential Growth Phase ◽  
Malt Extract ◽  
Remedial Action ◽  
Yeast Cultures ◽  
Vitro Model

AbstractThe question of a sensitive response in the growth of yeast cell cultures to treatments with homœopathic dilutions (potencies), has been raised in a number of recent publications,1,2,3. A further set of growth trials with this model performed at our Institute are reported here.The actions of 8x to 16x decimal potencies of AgNO3, CuSO4, HgCl2, and NaCl on the growth of cell cultures of Schizosaccharomyces pombe were investigated. This included a set of trials with experimental poisoning of the cells by toxic levels of CuSO4 and subsequent study of the remedial action by potencies of the same substance.Yeast cells were grown in batch culture in a glucose/malt extract medium. Growth was interrupted shortly before the end of the exponential growth phase and total cell concentrations were assessed by means of a Coulter Counter. The obtained data were subjected to statistical analysis. Each treatment range was tested in three separate trials.The results obtained failed to indicate any response in the growth rate of Sch. pombe to the potency treatments.

scholarly journals Preterm intraventricular hemorrhage in vitro: modeling the cytopathology of the ventricular zone

2020 ◽  
Author(s):  
Leandro Castaneyra-Ruiz ◽  
James P. McAllister ◽  
Diego M. Morales ◽  
Steven L. Brody ◽  
Albert M. Isaacs ◽  
...  
Keyword(s):  
Intraventricular Hemorrhage ◽  
In Vitro Model ◽  
Ventricular Zone ◽  
Ependymal Cells ◽  
High Temporal Resolution ◽  
Neuroepithelial Cells ◽  
Vitro Model ◽  
Developmental Features ◽  
The Impact

Abstract Background: Severe intraventricular hemorrhage (IVH) is one of the most devastating neurological complications in preterm infants, with the majority suffering long-term neurological morbidity and up to 50 percent developing post-hemorrhagic hydrocephalus (PHH). Despite the importance of this disease, its cytopathological mechanisms are not well known. An in vitro model of IVH is required to investigate the effects of blood and its components on the developing ventricular zone (VZ) and its stem cell niche. To address this need, we developed a protocol from our accepted in vitro model to mimic the cytopathological conditions of IVH in the preterm infant. Methods: Maturing neuroepithelial cells from the VZ were harvested from the entire lateral ventricles of wild type C57BL/6 mice at 1-4 days of age and expanded in proliferation media for 3-5 days. At confluence, cells were re-plated onto 24-well plates in differentiation media to generate ependymal cells (EC). At approximately 3-5 days, which corresponded to the onset of EC differentiation based on the appearance of multiciliated cells, phosphate-buffered saline for controls or syngeneic whole blood for IVH was added to the EC surface. The cells were examined for the expression of EC markers of differentiation and maturation to qualitatively and quantitatively assess the effect of blood exposure on VZ transition from neuroepithelial cells to EC. Discussion: This protocol will allow investigators to test cytopathological mechanisms contributing to the pathology of IVH with high temporal resolution and query the impact of injury to the maturation of the VZ. This technique recapitulates features of normal maturation of the VZ in vitro, offering the capacity to investigate the developmental features of VZ biogenesis.

scholarly journals Preterm Intraventricular Hemorrhage in vitro: Modeling the Cytopathology of the Ventricular Zone

2020 ◽  
Author(s):  
Leandro Castaneyra-Ruiz ◽  
James P. McAllister ◽  
Diego M. Morales ◽  
Steven L. Brody ◽  
Albert M. Isaacs ◽  
...  
Keyword(s):  
Intraventricular Hemorrhage ◽  
In Vitro Model ◽  
Ependymal Cell ◽  
Ventricular Zone ◽  
High Temporal Resolution ◽  
Neuroepithelial Cells ◽  
Vitro Model ◽  
Developmental Features ◽  
The Impact

Abstract Background: Severe intraventricular hemorrhage (IVH) is one of the most devastating neurological complications in preterm infants, with the majority suffering long-term neurological morbidity and up to 50 percent developing post hemorrhagic hydrocephalus (PHH). Despite the importance of this disease, its cytopathological mechanisms are not well known. An in vitro model of IVH is required to investigate the effects of blood and its components on the developing ventricular zone (VZ) and its stem cell niche. To address this need, we developed a new in vitro model to mimic the cytopathological conditions of IVH in the preterm infant. Methods: Maturing neuroepithelial cells from the VZ were harvested from the entire lateral ventricles of wild type C57BL/6 mice at 1-4 days of age and expanded in proliferation media for 3-5 days. At confluence, cells were re-plated onto 24-well plates in differentiation media to generate ependymal cells (EC). At approximately 3-5 days, which corresponded to the onset of ependymal cell differentiation based on the appearance of multiciliated cells , phosphate-buffered saline for controls or syngeneic whole blood for IVH was added to the ependymal cell surface. The cells were examined for the expression of EC markers of differentiation and maturation to qualitatively and quantitatively assess the effect of blood exposure on VZ transition from neuroepithelial cells to EC. Discussion: This model will allow investigators to test cytopathological mechanisms contributing to the pathology of IVH with high temporal resolution and query the impact of injury to the maturation of the VZ. This technique recapitulates features of normal maturation of the VZ in vitro, offering the capacity to investigate the developmental features of VZ biogenesis.

An in vitro model for investigation of vitamin A effects on wound healing

2021 ◽  
pp. 1-6
Author(s):  
Hoda Keshmiri Neghab ◽  
Mohammad Hasan Soheilifar ◽  
Gholamreza Esmaeeli Djavid
Keyword(s):  
Wound Healing ◽  
Endothelial Cell ◽  
Vitamin A ◽  
In Vitro Model ◽  
Cellular Proliferation ◽  
Endothelial Cell Migration ◽  
Normal Fibroblast ◽  
Anti Inflammatory ◽  
Vitro Model

Abstract. Wound healing consists of a series of highly orderly overlapping processes characterized by hemostasis, inflammation, proliferation, and remodeling. Prolongation or interruption in each phase can lead to delayed wound healing or a non-healing chronic wound. Vitamin A is a crucial nutrient that is most beneficial for the health of the skin. The present study was undertaken to determine the effect of vitamin A on regeneration, angiogenesis, and inflammation characteristics in an in vitro model system during wound healing. For this purpose, mouse skin normal fibroblast (L929), human umbilical vein endothelial cell (HUVEC), and monocyte/macrophage-like cell line (RAW 264.7) were considered to evaluate proliferation, angiogenesis, and anti-inflammatory responses, respectively. Vitamin A (0.1–5 μM) increased cellular proliferation of L929 and HUVEC (p < 0.05). Similarly, it stimulated angiogenesis by promoting endothelial cell migration up to approximately 4 fold and interestingly tube formation up to 8.5 fold (p < 0.01). Furthermore, vitamin A treatment was shown to decrease the level of nitric oxide production in a dose-dependent effect (p < 0.05), exhibiting the anti-inflammatory property of vitamin A in accelerating wound healing. These results may reveal the therapeutic potential of vitamin A in diabetic wound healing by stimulating regeneration, angiogenesis, and anti-inflammation responses.

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