scholarly journals Posttraumatic Invasion of Monocytes across the Blood—Cerebrospinal Fluid Barrier

2011 ◽  
Vol 32 (1) ◽  
pp. 93-104 ◽  
Author(s):  
Joanna Szmydynger-Chodobska ◽  
Nathalie Strazielle ◽  
Jessica R Gandy ◽  
Timothy H Keefe ◽  
Brian J Zink ◽  
...  
Keyword(s):  
Cerebrospinal Fluid ◽  
Inflammatory Cells ◽  
Electron Microscopic ◽  
Injured Brain ◽  
Pathophysiological Role ◽  
Choroidal Epithelium ◽  
Epithelial Barriers ◽  
Vitro Model ◽  
A Site

The invasion of inflammatory cells occurring after ischemic or traumatic brain injury (TBI) has a detrimental effect on neuronal survival and functional recovery after injury. We have recently demonstrated that not only the blood-brain barrier, but also the blood-cerebrospinal fluid (CSF) barrier (BCSFB), has a role in posttraumatic recruitment of neutrophils. Here, we show that TBI results in a rapid increase in synthesis and release into the CSF of a major chemoattractant for monocytes, CCL2, by the choroid plexus epithelium, a site of the BCSFB. Using an in vitro model of the BCSFB, we also show that CCL2 is released across the apical and basolateral membranes of the choroidal epithelium, a pattern of chemokine secretion that promotes leukocyte migration across epithelial barriers. Immunohistochemical and electron microscopic analyses of choroidal tissue provide evidence for the movement of monocytes, sometimes in tandem with neutrophils, along the paracellular pathways between adjacent epithelial cells. These data further support the pathophysiological role of BCSFB in promoting the recruitment of inflammatory cells to the injured brain.

scholarly journals The Role of the Choroid Plexus in Neutrophil Invasion after Traumatic Brain Injury

2009 ◽  
Vol 29 (9) ◽  
pp. 1503-1516 ◽  
Author(s):  
Joanna Szmydynger-Chodobska ◽  
Nathalie Strazielle ◽  
Brian J Zink ◽  
Jean-François Ghersi-Egea ◽  
Adam Chodobski
Keyword(s):  
Traumatic Brain Injury ◽  
Brain Injury ◽  
Choroid Plexus ◽  
Neutrophil Migration ◽  
Inflammatory Cells ◽  
Brain Parenchyma ◽  
Electron Microscopic ◽  
Epithelial Barriers

Traumatic brain injury (TBI) frequently results in neuroinflammation, which includes the invasion of neutrophils. After TBI, neutrophils infiltrate the choroid plexus (CP), a site of the blood—cerebrospinal fluid (CSF) barrier (BCSFB), and accumulate in the CSF space near the injury, from where these inflammatory cells may migrate to brain parenchyma. We have hypothesized that the CP functions as an entry point for neutrophils to invade the injured brain. Using the controlled cortical impact model of TBI in rats and an in vitro model of the BCSFB, we show that the CP produces CXC chemokines, such as cytokine-induced neutrophil chemoattractant (CINC)-1 or CXCL1, CINC-2α or CXCL3, and CINC-3 or CXCL2. These chemokines are secreted both apically and basolaterally from the choroidal epithelium, a prerequisite for neutrophil migration across epithelial barriers. Consistent with these findings, we also provide electron microscopic evidence that neutrophils infiltrate the choroidal stroma and subsequently reach the intercellular space between choroidal epithelial cells. This is the first detailed analysis of the BCSFB function related to neutrophil trafficking. Our observations support the role of this barrier in posttraumatic neutrophil invasion.

Accelerated Non-Muscle Contraction after Subarachnoid Hemorrhage: Cerebrospinal Fluid Testing in a Culture Model

Neurosurgery ◽  
1990 ◽  
Vol 27 (6) ◽  
pp. 921-928 ◽  
Author(s):  
Yoshihiro Yamamoto ◽  
David H. Bernanke ◽  
Robert R. Smith
Keyword(s):  
Cerebrospinal Fluid ◽  
Subarachnoid Hemorrhage ◽  
Collagen Matrix ◽  
Aneurysm Rupture ◽  
Lattice Contraction ◽  
Symptomatic Vasospasm ◽  
Luminal Narrowing ◽  
Chronic Vasospasm ◽  
Vitro Model

Abstract The cause of chronic cerebral vasospasm after subarachnoid hemorrhage has been studied intensively, but it is still controversial whether the observable luminal narrowing should be attributed to the contraction of vascular smooth muscle cells or whether it results from some organic change in the wall. A proliferation of myointimal cells, accompanied by increased deposition of collagen, as well as myonecrosis, have been frequently observed several days after aneurysm rupture. Studies from our laboratory showed that these myointimal cells had characteristics identical to myofibroblasts. In this study, we quantitatively and morphologically examined the effect of cerebrospinal fluid on the ability of myofibroblasts to alter collagen matrices using an in vitro model. Myofibroblasts contract the collagen matrix by rearranging or compacting the framework of collagen fibers. Cerebrospinal fluid obtained from patients with recently ruptured aneurysms significantly accelerated lattice contraction, especially when the patient developed symptomatic vasospasm. This study suggests that myofibroblasts in the spastic artery can produce a contractile force that contributes to chronic vasospasm, tightening the proliferated collagen. Some unknown agent present in bloody cerebrospinal fluid accelerates the rearrangement of the collagen lattice by myofibroblasts, both of which have, until now, been considered non-contractile components.

scholarly journals The transcriptional landscape of the murine middle ear epithelium in vitro

2019 ◽  
Author(s):  
Apoorva Mulay ◽  
Md Miraj K Chowdhury ◽  
Cameron James ◽  
Lynne Bingle ◽  
Colin D Bingle
Keyword(s):  
Gene Expression ◽  
Middle Ear ◽  
In Vitro Model ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Inflammatory Cells ◽  
Differentially Expressed ◽  
Vitro Model ◽  
Transcriptional Landscape

AbstractOtitis media (OM) is the most common paediatric disease and leads to significant morbidity. Although understanding of underlying disease mechanisms is hampered by complex pathophysiology, it is clear that epithelial abnormalities underpin the disease. The mechanisms underpinning epithelial remodelling in OM remain unclear. We recently described a novel in vitro model of mouse middle ear epithelial cells (mMEECs) that undergoes mucociliary differentiation into the varied epithelial cell populations seen in the middle ear cavity. We now describe genome wide gene expression profiles of mMEECs as they undergo differentiation. We compared the gene expression profiles of original (uncultured) middle ear cells, confluent cultures of undifferentiated cells (day 0 of ALI) and cells that had been differentiated for 7 days at an ALI. >5000 genes were differentially expressed among the three groups of cells. Approximately 4000 genes were differentially expressed between the original cells and day 0 of ALI culture. The original cell population was shown to contain a mix of cell types, including contaminating inflammatory cells that were lost on culture. Approximately 500 genes were upregulated during ALI induced differentiation. These included some secretory genes and some enzymes but most were associated with the process of ciliogenesis. Our in vitro model of differentiated murine middle ear epithelium exhibits a transcriptional profile consistent with the mucociliary epithelium seen within the middle ear. Knowledge of the transcriptional landscape of this epithelium will provide a basis for understanding the phenotypic changes seen in murine models of OM.

scholarly journals Betaine attenuates LPS-induced downregulation of Occludin and Claudin-1 and restores intestinal barrier function

2020 ◽  
Author(s):  
Jingtao Wu ◽  
Caimei He ◽  
Jie Bu ◽  
Yue Luo ◽  
Shuyuan Yang ◽  
...  
Keyword(s):  
Barrier Function ◽  
Intestinal Barrier ◽  
Protective Role ◽  
Vital Role ◽  
Inflammatory Factors ◽  
Intestinal Barrier Function ◽  
Epithelial Barrier ◽  
Epithelial Barriers ◽  
Vitro Model

Abstract Background：The intestinal epithelial barrier, which works as the first line of defense between the luminal environment and the host, once destroyed, it will cause serious inflammation or other intestinal diseases. Tight junctions (TJs) play a vital role to maintain the integrity of the epithelial barrier. Lipopolysaccharide (LPS), one of the most important inflammatory factors will downregulate specific TJ proteins including Occludin and Claudin-1 and impair integrity of the epithelial barrier. Betaine has excellent anti-inflammatory activity but whether betaine has any effect on TJ proteins, particularly on LPS-induced dysfunction of epithelial barriers remains unknown. The purpose of this study is to explore the pharmacological effect of betaine on improving intestinal barrier function represented by TJ proteins. Intestinal porcine epithelial cells (IPEC-J2) were used as an in vitro model. Results: The results demonstrated that betaine enhanced the expression of TJ proteins while LPS (1µg/mL) downregulates the expression of these proteins. Furthermore, betaine attenuates LPS-induced decreases of TJ proteins both shown by Western blot (WB) and Reverse transcription- polymerase chain reaction (RT-PCR). The immunofluorescent images consistently revealed that LPS induced the disruption of TJ protein Claudin-1 and reduced its expression while betaine could reverse these alterations. Similar protective role of betaine on intestinal barrier function was observed by transepithelial electrical resistance (TEER) approach. Conclusion: In conclusion, our research demonstrated that betaine attenuated LPS-induced downregulation of Occludin and Claudin-1 and restored the intestinal barrier function.

scholarly journals Design of a Platelet-Mediated Delivery System for Drug-Incorporated Nanospheres to Enhance Anti-Tumor Therapeutic Effect

Pharmaceutics ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 1724
Author(s):  
Jun-ichiro Jo ◽  
Tsubasa Emi ◽  
Yasuhiko Tabata
Keyword(s):  
Tumor Cells ◽  
Delivery System ◽  
Biological Properties ◽  
Poly Vinyl Alcohol ◽  
Electron Microscopic ◽  
Lower Chamber ◽  
Vitro Model ◽  
Polymer Surfactant ◽  
Upper Chamber

The objective of this study is to construct a platelet-mediated delivery system for drug-incorporated nanospheres. Nanospheres of poly(lactic-co-glycolic acid) (PLGA-NS) with different sizes and surface properties were prepared by changing the preparation parameters, such as the type of polymer surfactant, the concentration of polymer surfactant and PLGA, and the stirring rate. When incubated with platelets, PLGA-NS prepared with poly(vinyl alcohol) suppressed the platelet activation. Scanning electron microscopic and flow cytometry examinations revealed that platelets associated with PLGA-NS (platelet hybrids, PH) had a similar appearance and biological properties to those of the original platelets. In addition, the PH with PLGA-NS specifically adhered onto the substrate pre-coated with fibrin to a significantly great extent compared with PLGA-NS alone. When applied in an in vitro model of tumor tissue which was composed of an upper chamber pre-coated with fibrin and a lower chamber culturing tumor cells, the PH with PLGA-NS incorporating an anti-tumor drug were delivered to the tumor cells through the specific adhesion onto the upper chamber and, consequently, drug release from the upper chamber took place, resulting in the growth suppression of tumor cells. It is concluded that the drug delivery system based on PH is promising for tumor treatment.

Chemotaxis of T-cells after infection of human choroid plexus papilloma cells with Echovirus 30 in an in vitro model of the blood–cerebrospinal fluid barrier

2012 ◽  
Vol 170 (1-2) ◽  
pp. 66-74 ◽  
Author(s):  
Henriette Schneider ◽  
Claudia Ellen Weber ◽  
Julia Schoeller ◽  
Ulrike Steinmann ◽  
Julia Borkowski ◽  
...  
Keyword(s):  
T Cells ◽  
Cerebrospinal Fluid ◽  
Choroid Plexus ◽  
In Vitro Model ◽  
Choroid Plexus Papilloma ◽  
Echovirus 30 ◽  
Human Choroid Plexus ◽  
Vitro Model ◽  
Plexus Papilloma

scholarly journals High Mobility Group Box 1 Protein Induction byMycobacterium BovisBCG

2007 ◽  
Vol 2007 ◽  
pp. 1-8 ◽  
Author(s):  
Péter Hofner ◽  
György Seprényi ◽  
András Miczák ◽  
Krisztina Buzás ◽  
Zsófia Gyulai ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
In Vitro Model ◽  
Nuclear Protein ◽  
High Mobility ◽  
High Mobility Group ◽  
Pathophysiological Role ◽  
Vitro Model

High mobility group box 1 protein (HMGB1), a nuclear protein, is a critical cytokine that mediates the response to infection, injury, and inflammation. The aim of our study was to elaborate a reliable in vitro model to investigate whetherMycobacterium bovisBCG is able to induce HMGB1 secretion from the monocytic U-937 cells. Western blot technique was applied for the detection of HMGB1 from supernatants of cells, following induction withMycobacterium bovisBCG. Densitometric analysis revealed higher concentrations of HMGB1 in cell supernatants stimulated with BCG than in the supernatants of the control, nonstimulated cells. Further quantitation of the secreted HMGB1 was performed by ELISA. The BCG strain resulted in a higher amount of secreted HMGB1 (450±44 ng/mL) than that of LPS (84±12 ng/mL) orStaphylococcus aureus(150±14 ng/mL). BCG and Phorbol−12-myristate−13acetate (PMA), added together, resulted in the highest HMGB1 secretion (645±125 ng/mL). The translocation of the HMGB1 towards the cytoplasm following infection of cells with BCG was demonstrated by immunofluorescence examinations. Conclusion: Our pilot experiments draw attention to the HMGB1 inducing ability ofMycobacterium bovis. Assesment of the pathophysiological role of this late cytokine in mycobacterial infections demands further in vitro and in vivo examinations.

scholarly journals Different structures and pathologies of alpha-synuclein fibrils derived from preclinical and postmortem patients of Parkinson's disease

2021 ◽  
Author(s):  
Yun Fan ◽  
Yunpeng Sun ◽  
Wenbo Yu ◽  
Youqi Tao ◽  
Wencheng Xia ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Cerebrospinal Fluid ◽  
Lewy Bodies ◽  
Alpha Synuclein ◽  
Electron Microscopic ◽  
Lewy Neurites ◽  
The Difference ◽  
A Minor

alpha-Synuclein (alpha-syn) fibrillar aggregates are the major component of Lewy bodies and Lewy neurites presenting as the pathology hallmark of Parkinson's disease (PD). Studies have shown that alpha-syn is potential to form different conformational fibrils associated with different synucleinopathies, but whether the conformation of alpha-syn fibrils changes in different phases of related diseases is to be explored. Here, we amplified alpha-syn aggregates from the cerebrospinal fluid (CSF) of preclinical (pre-PD) and late-stage postmortem PD (post-PD) patients. Our results show that compared to the CSF of pre-PD, that of post-PD is markedly stronger in seeding in vitro alpha-syn aggregation, and the amplified fibrils are more potent in inducing endogenous alpha-syn aggregation in neurons. Cryo-electron microscopic structures further reveal that the difference between the pre-PD- and post-PD-derived fibrils lies on a minor polymorph which in the pre-PD fibrils is morphologically straight, while in the post-PD fibrils represents a single protofilament assembled by a distinctive conformation of alpha-syn. Our work demonstrates structural and pathological differences between pre-PD and post-PD alpha-syn aggregation and suggests potential alteration of alpha-syn fibrils during the progression of PD clinical phases.

Development of an in Vitro Vessel Wall Model for Studying Certain Aspects of Platelet-Endothelial Interactions

1975 ◽  
Author(s):  
F. M. Booyse ◽  
S. Bell ◽  
B. Sedlak ◽  
M. E. Rafelson
Keyword(s):  
Endothelial Cells ◽  
Cultured Cells ◽  
Electron Microscopic ◽  
Experimental Conditions ◽  
Aortic Endothelial Cells ◽  
Interaction Interaction ◽  
Pinocytotic Vesicles ◽  
Vitro Model ◽  
Calculated Number

Cultured bovine aortic endothelial cells were characterized by the presence of Weibel-Palade bodies, pinocytotic vesicles, Factor VIII antigen and thrombostenin-like contractile proteins. Longitudinal sections of cells showed the presence of an extensive matrix of 100–110 Å extracellular filaments. Treatment of monolayers of these cells with serotonin, thrombin, trypsin, endotoxin and heat caused contraction and exposure of the 100–110 Å filaments. Cultured cells were grown on plastic cover slips (9 × 35 mm, 3.5–4 × 105 cells). Post confluent cultures (2 weeks) were treated (activated) with one of the above agents and exposed to platelets, laleled with 125I (lactoperoxidase method) or 14C-amino acids, for various periods of time. Cover slips were washed and counted. Initial platelet-endothelial interactions were linear and could be expressed quantitatively by plotting radioactivity (CPM or calculated number of platelets) per 3.5–4×105 endothelial cells vs. exposure time. Non-activated control values were subtracted in each case. PGE1 eAMP, ATP and aspirin inhibit this interaction. Citrate and EDTA decreased the interaction. Interaction was increased by red blood cells and rotational flow rates and unaffected by plasma proteins. Preliminary electron microscopic observations suggest platelet attachment at the sites of 100–110 Å microfilament abundance. The feasibility of developing a quantitative in vitro model for studying platelet-endothelial interactions under defined experimental conditions will be considered further.

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