Application of Membrane Permeability Evaluated in In Vitro Analyses to Estimate Blood–Retinal Barrier Permeability

2012 ◽  
Vol 101 (7) ◽  
pp. 2596-2605 ◽  
Author(s):  
Yoshiyuki Kubo ◽  
Eri Fukui ◽  
Shin-Ichi Akanuma ◽  
Masanori Tachikawa ◽  
Ken-Ichi Hosoya
Keyword(s):  
Membrane Permeability ◽  
Blood Retinal Barrier ◽  
Barrier Permeability

scholarly journals Assessment of Ocriplasmin Effects on the Vitreoretinal Compartment in Porcine and Human Model Systems

2017 ◽  
Vol 2017 ◽  
pp. 1-12 ◽  
Author(s):  
Bart Jonckx ◽  
Michael Porcu ◽  
Aurelie Candi ◽  
Isabelle Etienne ◽  
Philippe Barbeaux ◽  
...  
Keyword(s):  
Posterior Vitreous Detachment ◽  
Subretinal Fluid ◽  
Model Systems ◽  
Human Model ◽  
Blood Retinal Barrier ◽  
Barrier Permeability ◽  
Cell Type Specific ◽  
Insight Into

Ocriplasmin (Jetrea®) is a recombinant protease used to treat vitreomacular traction. To gain insight into vitreoretinal observations reported after ocriplasmin treatment, we have developed anin vivoporcine ocriplasmin-induced posterior vitreous detachment (PVD) model in which we investigated vitreoretinal tissues by optical coherence tomography, histology, and cytokine profiling. Eight weeks postinjection, ocriplasmin yielded PVD in 82% of eyes. Subretinal fluid (85%) and vitreous hyperreflective spots (45%) were resolved by week 3. Histological analysis of extracellular matrix (ECM) proteins such as laminin, fibronectin, and collagen IV indicated no retinal ocriplasmin-induced ECM distribution changes. Retinal morphology was unaffected in all eyes. Cytokine profiles of ocriplasmin-treated eyes were not different from vehicle. In cell-based electrical resistance assays, blood-retinal barrier permeability was altered by ocriplasmin concentrations of 6 μg/mL and higher, with all effects being nontoxic, cell-type specific, and reversible. Ocriplasmin was actively taken up by RPE and Müller cells, and our data suggest both lysosomal and transcellular clearance routes for ocriplasmin. In conclusion, transient hyperreflective spots and fluid in a porcine ocriplasmin-induced PVD model did not correlate with retinal ECM rearrangement nor inflammation. Reversiblein vitroeffects on blood-retinal barrier permeability provide grounds for a hypothesis on the mechanisms behind transient subretinal fluid observed in ocriplasmin-treated patients.

scholarly journals Influences of advanced glycosylation end products on the inner blood–retinal barrier in a co-culture cell model in vitro

2020 ◽  
Vol 15 (1) ◽  
pp. 619-628
Author(s):  
Chen Yuan ◽  
Ya Mo ◽  
Jie Yang ◽  
Mei Zhang ◽  
Xuejun Xie
Keyword(s):  
Electrical Resistance ◽  
Cell Model ◽  
Polyclonal Antibodies ◽  
Time Dependent ◽  
Dependent Manner ◽  
Blood Retinal Barrier ◽  
Advanced Glycosylation End Products ◽  
End Products ◽  
Advanced Glycosylation

AbstractAdvanced glycosylation end products (AGEs) are harmful factors that can damage the inner blood–retinal barrier (iBRB). Rat retinal microvascular endothelial cells (RMECs) were isolated and cultured, and identified by anti-CD31 and von Willebrand factor polyclonal antibodies. Similarly, rat retinal Müller glial cells (RMGCs) were identified by H&E staining and with antibodies of glial fibrillary acidic protein and glutamine synthetase. The transepithelial electrical resistance (TEER) value was measured with a Millicell electrical resistance system to observe the leakage of the barrier. Transwell cell plates for co-culturing RMECs with RMGCs were used to construct an iBRB model, which was then tested with the addition of AGEs at final concentrations of 50 and 100 mg/L for 24, 48, and 72 h. AGEs in the in vitro iBRB model constructed by RMEC and RMGC co-culture led to the imbalance of the vascular endothelial growth factor (VEGF) and pigment epithelial derivative factor (PEDF), and the permeability of the RMEC layer increased because the TEER decreased in a dose- and time-dependent manner. AGEs increased VEGF but lowered PEDF in a dose- and time-dependent manner. The intervention with AGEs led to the change of the transendothelial resistance of the RMEC layer likely caused by the increased ratio of VEGF/PEDF.

scholarly journals Bacillus cereus Induces Permeability of an In Vitro Blood-Retina Barrier

2008 ◽  
Vol 76 (4) ◽  
pp. 1358-1367 ◽  
Author(s):  
A. L. Moyer ◽  
R. T. Ramadan ◽  
J. Thurman ◽  
A. Burroughs ◽  
M. C. Callegan
Keyword(s):  
Quorum Sensing ◽  
Bacillus Cereus ◽  
Barrier Function ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Toxin Production ◽  
Global Regulator ◽  
Barrier Permeability ◽  
Cell Monolayers

ABSTRACT Most Bacillus cereus toxin production is controlled by the quorum-sensing-dependent, pleiotropic global regulator plcR, which contributes to the organism's virulence in the eye. The purpose of this study was to analyze the effects of B. cereus infection and plcR-regulated toxins on the barrier function of retinal pigment epithelium (RPE) cells, the primary cells of the blood-retina barrier. Human ARPE-19 cells were apically inoculated with wild-type or quorum-sensing-deficient B. cereus, and cytotoxicity was analyzed. plcR-regulated toxins were not required for B. cereus-induced RPE cytotoxicity, but these toxins did increase the rate of cell death, primarily by necrosis. B. cereus infection of polarized RPE cell monolayers resulted in increased barrier permeability, independent of plcR-regulated toxins. Loss of both occludin and ZO-1 expression occurred by 8 h postinfection, but alterations in tight junctions appeared to precede cytotoxicity. Of the several proinflammatory cytokines analyzed, only interleukin-6 was produced in response to B. cereus infection. These results demonstrate the deleterious effects of B. cereus infection on RPE barrier function and suggest that plcR-regulated toxins may not contribute significantly to RPE barrier permeability during infection.

scholarly journals TMIC-34. IN VITRO STUDY OF ROLE OF ASTROCYTIC S1P3 IN REGULATING BLOOD-BRAIN/TUMOR BARRIER PERMEABILITY

2017 ◽  
Vol 19 (suppl_6) ◽  
pp. vi250-vi250
Author(s):  
Anurag N Paranjape ◽  
Brunilde Gril ◽  
Stephan Woditschka ◽  
Jeffrey Hanson ◽  
Xiaolin Wu ◽  
...  
Keyword(s):  
Brain Tumor ◽  
In Vitro Study ◽  
Vitro Study ◽  
Barrier Permeability ◽  
Blood Brain

scholarly journals Sporadic Emergence of Klebsiella pneumoniae Strains Resistant to Cefepime and Cefpirome in Greek Hospitals

1998 ◽  
Vol 36 (1) ◽  
pp. 266-268 ◽  
Author(s):  
L. S. Tzouvelekis ◽  
E. Tzelepi ◽  
E. Prinarakis ◽  
M. Gazouli ◽  
A. Katrahoura ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Membrane Permeability ◽  
Outer Membrane ◽  
Outer Membrane Permeability ◽  
Greek Hospitals ◽  
Hydrolysis Of

The sporadic emergence of Klebsiella pneumoniae strains resistant to cefepime and cefpirome was observed in Greek hospitals during 1996. Examination of six epidemiologically distinct strains and clones selected in vitro provided indications that resistance is due to the cooperation of decreased outer membrane permeability and hydrolysis of the cephalosporins by SHV-5 β-lactamase, which was produced in large amounts.

scholarly journals Soluble RAGE attenuates AngII-induced endothelial hyperpermeability by disrupting HMGB1-mediated crosstalk between AT1R and RAGE

2019 ◽  
Vol 51 (9) ◽  
pp. 1-15 ◽  
Author(s):  
Jisu Jeong ◽  
Jiye Lee ◽  
Juyeon Lim ◽  
Soyoung Cho ◽  
Soyoung An ◽  
...  
Keyword(s):  
Endothelial Permeability ◽  
Endothelial Barrier ◽  
Blue Staining ◽  
Ang Ii ◽  
Barrier Permeability ◽  
Soluble Rage ◽  
Apolipoprotein E Knockout Mice ◽  
Endothelial Hyperpermeability

Abstract Increased endothelial permeability, one of the earliest signs of endothelial dysfunction, is associated with the development of cardiovascular diseases such as hypertension and atherosclerosis. Recent studies suggest that the receptor for advanced glycation end products (RAGE) regulates endothelial permeability in inflammation. In the present study, we investigated the regulatory mechanism of RAGE in endothelial hyperpermeability induced by angiotensin II (Ang II), a well-known inflammatory mediator, and the potential therapeutic effect of soluble RAGE (sRAGE), a decoy receptor for RAGE ligands. For in vitro studies, Ang II-treated human umbilical vein endothelial cells (HUVECs) were treated with siRNA specific to either RAGE or sRAGE to disrupt RAGE-mediated signaling. Endothelial permeability was estimated using FITC-labeled dextran 40 and a resistance meter. To evaluate intercellular junction disruption, VE-cadherin expression was examined by western blotting and immunocytochemistry. Ang II increased the expression of the Ang II type 1 receptor (AT1R) and RAGE, and this increase was inhibited by sRAGE. sRAGE prevented Ang II-induced VE-cadherin disruption in HUVECs. For in vivo studies, Ang II-infused, atherosclerosis-prone apolipoprotein E knockout mice were utilized. Endothelial permeability was assessed by Evans blue staining of the aorta. Ang II increased endothelial barrier permeability, and this effect was significantly attenuated by sRAGE. Our data demonstrate that blockade of RAGE signaling using sRAGE attenuates Ang II-induced endothelial barrier permeability in vitro and in vivo and indicate the therapeutic potential of sRAGE in controlling vascular permeability under pathological conditions.

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