Combined integrin αvβ3 and lactoferrin receptor targeted docetaxel liposomes enhance the brain targeting effect and anti-glioma effect

AbstractThe integrin αvβ3 receptor and Lactoferrin receptor (LfR) are over-expressed in both cerebral microvascular endothelial cells and glioma cells. RGD tripeptide and Lf can specifically bind with integrin αvβ3 receptor and LfR, respectively. In our study, RGD and Lf dual-modified liposomes loaded with docetaxel (DTX) were designed to enhance the brain targeting effect and treatment of glioma. Our in vitro studies have shown that RGD-Lf-LP can significantly enhance the cellular uptake of U87 MG cells and human cerebral microvascular endothelial cells (hCMEC/D3) when compared to RGD modified liposomes (RGD-LP) and Lf modified liposomes (Lf-LP). Free RGD and Lf competitively reduced the cellular uptake of RGD-Lf-LP, in particular, free RGD played a main inhibitory effect on cellular uptake of RGD-Lf-LP in U87 MG cells, yet free Lf played a main inhibitory effect on cellular uptake of RGD-Lf-LP in hCMEC/D3 cells. RGD-Lf-LP can also significantly increase penetration of U87 MG tumor spheroids, and RGD modification plays a dominating role on promoting the penetration of U87 MG tumor spheroids. The results of in vitro BBB model were shown that RGD-Lf-LP-C6 obviously increased the transport of hCMEC/D3 cell monolayers, and Lf modification plays a dominating role on increasing the transport of hCMEC/D3 cell monolayers. In vivo imaging proved that RGD-Lf-LP shows stronger targeting effects for brain orthotopic gliomas than that of RGD-LP and Lf-LP. The result of tissue distribution confirmed that RGD-LF-LP-DTX could significantly increase brain targeting after intravenous injection. Furthermore, RGD-LF-LP-DTX (a dose of 5 mg kg−1 DTX) could significantly prolong the survival time of orthotopic glioma-bearing mice. In summary, RGD and LF dual modification are good combination for brain targeting delivery, RGD-Lf-LP-DTX could enhance brain targeting effects, and is thus a promising chemotherapeutic drug delivery system for treatment of glioma. Graphical abstract

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Anthropogenic Iron Oxide Nanoparticles Induce Damage to Brain Microvascular Endothelial Cells Forming the Blood-Brain Barrier

Advances in Alzheimer’s Disease - Alzheimer’s Disease and Air Pollution ◽

10.3233/aiad210010 ◽

2021 ◽

Author(s):

Lorena Gárate-Vélez ◽

Claudia Escudero-Lourdes ◽

Daniela Salado-Leza ◽

Armando González-Sánchez ◽

Ildemar Alvarado-Morales ◽

...

Keyword(s):

Endothelial Cells ◽

Blood Brain Barrier ◽

Brain Barrier ◽

Microvascular Endothelial Cells ◽

Brain Microvascular Endothelial Cells ◽

Blood Brain ◽

Fe3o4 Nps ◽

Microvascular Endothelial ◽

The Brain

Background: Iron nanoparticles, mainly in magnetite phase (Fe3O4 NPs), are released to the environment in areas with high traffic density and braking frequency. Fe3O4 NPs were found in postmortem human brains and are assumed to get directly into the brain through the olfactory nerve. However, these pollution-derived NPs may also translocate from the lungs to the bloodstream and then, through the blood-brain barrier (BBB), into the brain inducing oxidative and inflammatory responses that contribute to neurodegeneration. Objective: To describe the interaction and toxicity of pollution-derived Fe3O4 NPs on primary rat brain microvascular endothelial cells (rBMECs), main constituents of in vitro BBB models. Methods: Synthetic bare Fe3O4 NPs that mimic the environmental ones (miFe3O4) were synthesized by co-precipitation and characterized using complementary techniques. The rBMECs were cultured in Transwell® plates. The NPs-cell interaction was evaluated through transmission electron microscopy and standard colorimetric in vitro assays. Results: The miFe3O4 NPs, with a mean diameter of 8.45 ± 0.14 nm, presented both magnetite and maghemite phases, and showed super-paramagnetic properties. Results suggest that miFe3O4 NPs are internalized by rBMECs through endocytosis and that they are able to cross the cells monolayer. The lowest miFe3O4 NPs concentration tested induced mid cytotoxicity in terms of 1) membrane integrity (LDH release) and 2) metabolic activity (MTS transformation). Conclusion: Pollution-derived Fe3O4 NPs may interact and cross the microvascular endothelial cells forming the BBB and cause biological damage.

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Abstract P129: The High Blood Pressure-Malaria Protection Hypothesis

Hypertension ◽

10.1161/hyp.68.suppl_1.p129 ◽

2016 ◽

Vol 68 (suppl_1) ◽

Author(s):

Julio Gallego-Delgado ◽

Thomas Walther ◽

Ana Rodriguez

Keyword(s):

Endothelial Cells ◽

Cerebral Malaria ◽

Microvascular Endothelial Cells ◽

Ang Ii ◽

Asian Origin ◽

Microvascular Endothelial ◽

The Brain

For decades, researchers have been fascinated by the idea of a causative connection between hypertension and malaria as the prevalence of hypertension is higher in populations that have been exposed to malaria for long periods. Cerebral malaria is a multi-factorial syndrome involving the interaction between P. falciparum -infected red blood cells ( Pf -iRBC) and host cerebral microvascular endothelial cells. Disruption of the blood-brain-barrier (BBB), ranging from increased permeability to complete loss of inter-endothelial junctions (IEJ) and to petechial hemorrhages in the brain, is a characteristic feature of cerebral malaria. Our in vitro experiments show that Pf -iRBC induce the disruption of IEJ in human brain microvascular endothelial cells (HBMEC) mediated by the activation of β-catenin. Protection from this effect is achieved by blockade of the angiotensin II (Ang II) type 1 receptor (AT1) or stimulation of the type 2 receptor (AT2), which abrogate Pf -iRBC-induced activation of β-catenin and prevent the disruption HBMEC monolayers. Using a mouse model of cerebral malaria, we observed similar effects after treatment with Ang II receptors modulators, leading to protection against cerebral malaria, reduced cerebral hemorrhages (1.98 and 1.17 vs 0.63 for control, AT1 blocker and AT2 agonist respectively; p<0.05) and increased survival. While only 25% survived in controls (4 of 16 mice), 81.7% (13 of 15) and 71.4% (5 of 7) survived under AT1 blockade or AT2 agonist, respectively. In contrast, AT2-deficient mice were more susceptible to cerebral malaria (0% survival; 0 of 9). A causal association between high levels of Ang II and protection from malaria pathogenesis can provide a likely explanation for the increased prevalence in hypertension observed in populations of African and South Asian origin. Furthermore, this potential causative connection might also direct to unique approaches for the effective treatment of cerebral malaria.

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Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro

PLoS ONE ◽

10.1371/journal.pone.0045977 ◽

2012 ◽

Vol 7 (9) ◽

pp. e45977 ◽

Cited By ~ 10

Author(s):

Johanna P. Laakkonen ◽

Tatjana Engler ◽

Ignacio A. Romero ◽

Babette Weksler ◽

Pierre-Olivier Couraud ◽

...

Keyword(s):

Endothelial Cells ◽

Microvascular Endothelial Cells ◽

Brain Microvascular Endothelial Cells ◽

Adenovirus Vectors ◽

Microvascular Endothelial ◽

The Brain

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The Influence of Capsaicin on the Integrity of Microvascular Endothelial Cell Monolayers

International Journal of Molecular Sciences ◽

10.3390/ijms20010122 ◽

2018 ◽

Vol 20 (1) ◽

pp. 122 ◽

Cited By ~ 4

Author(s):

Mathias Kaiser ◽

Malgorzata Burek ◽

Stefan Britz ◽

Frauke Lankamp ◽

Steffi Ketelhut ◽

...

Keyword(s):

Endothelial Cells ◽

Tight Junctions ◽

In Vitro Assays ◽

Microvascular Endothelial Cells ◽

Microvascular Endothelial Cell ◽

Drug Bioavailability ◽

Cell Monolayers ◽

Microvascular Endothelial ◽

Arteries And Veins

Microvascular endothelial cells are an essential part of many biological barriers, such as the blood–brain barrier (BBB) and the endothelium of the arteries and veins. A reversible opening strategy to increase the permeability of drugs across the BBB could lead to improved therapies due to enhanced drug bioavailability. Vanilloids, such as capsaicin, are known to reversibly open tight junctions of epithelial and endothelial cells. In this study, we used several in vitro assays with the murine endothelial capillary brain cells (line cEND) as a BBB model to characterize the interaction between capsaicin and endothelial tight junctions.

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Effects of Dipyrone on Prostaglandin Production by Human Platelets and Cultured Bovine Aortic Endothelial Cells

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657338 ◽

1983 ◽

Vol 49 (02) ◽

pp. 132-137 ◽

Cited By ~ 16

Author(s):

A Eldor ◽

G Polliack ◽

I Vlodavsky ◽

M Levy

Keyword(s):

Endothelial Cells ◽

Arachidonic Acid ◽

Competitive Inhibition ◽

Inhibitory Effect ◽

Human Platelets ◽

Ionophore A23187 ◽

Aortic Endothelial Cells ◽

Bovine Aortic Endothelial Cells ◽

Aortic Endothelial

SummaryDipyrone and its metabolites 4-methylaminoantipyrine, 4-aminoantipyrine, 4-acetylaminoantipyrine and 4-formylaminoan- tipyrine inhibited the formation of thromboxane A2 (TXA2) during in vitro platelet aggregation induced by ADP, epinephrine, collagen, ionophore A23187 and arachidonic acid. Inhibition occurred after a short incubation (30–40 sec) and depended on the concentration of the drug or its metabolites and the aggregating agents. The minimal inhibitory concentration of dipyrone needed to completely block aggregation varied between individual donors, and related directly to the inherent capacity of their platelets to synthesize TXA2.Incubation of dipyrone with cultured bovine aortic endothelial cells resulted in a time and dose dependent inhibition of the release of prostacyclin (PGI2) into the culture medium. However, inhibition was abolished when the drug was removed from the culture, or when the cells were stimulated to produce PGI2 with either arachidonic acid or ionophore A23187.These results indicate that dipyrone exerts its inhibitory effect on prostaglandins synthesis by platelets or endothelial cells through a competitive inhibition of the cyclooxygenase system.

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Radiobiological Studies of Microvascular Damage through In Vitro Models: A Methodological Perspective

Cancers ◽

10.3390/cancers13051182 ◽

2021 ◽

Vol 13 (5) ◽

pp. 1182

Author(s):

Luca Possenti ◽

Laura Mecchi ◽

Andrea Rossoni ◽

Veronica Sangalli ◽

Simone Bersini ◽

...

Keyword(s):

Endothelial Cells ◽

In Vitro Models ◽

Microvascular Endothelial Cells ◽

Microvascular Damage ◽

Essential Information ◽

Reliable Model ◽

Critical Issues ◽

Irradiation Scheme ◽

In Vitro Effects

Ionizing radiation (IR) is used in radiotherapy as a treatment to destroy cancer. Such treatment also affects other tissues, resulting in the so-called normal tissue complications. Endothelial cells (ECs) composing the microvasculature have essential roles in the microenvironment’s homeostasis (ME). Thus, detrimental effects induced by irradiation on ECs can influence both the tumor and healthy tissue. In-vitro models can be advantageous to study these phenomena. In this systematic review, we analyzed in-vitro models of ECs subjected to IR. We highlighted the critical issues involved in the production, irradiation, and analysis of such radiobiological in-vitro models to study microvascular endothelial cells damage. For each step, we analyzed common methodologies and critical points required to obtain a reliable model. We identified the generation of a 3D environment for model production and the inclusion of heterogeneous cell populations for a reliable ME recapitulation. Additionally, we highlighted how essential information on the irradiation scheme, crucial to correlate better observed in vitro effects to the clinical scenario, are often neglected in the analyzed studies, limiting the translation of achieved results.

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Age influences susceptibility of brain capillary endothelial cells to La Crosse virus infection and cell death

Journal of Neuroinflammation ◽

10.1186/s12974-021-02173-4 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Rahul Basu ◽

Vinod Nair ◽

Clayton W. Winkler ◽

Tyson A. Woods ◽

Iain D. C. Fraser ◽

...

Keyword(s):

Endothelial Cells ◽

Cell Death ◽

Virus Infection ◽

La Crosse Virus ◽

Brain Capillary ◽

Adult Mice ◽

Age Related ◽

Capillary Endothelial Cells ◽

The Brain

Abstract Background A key factor in the development of viral encephalitis is a virus crossing the blood-brain barrier (BBB). We have previously shown that age-related susceptibility of mice to the La Crosse virus (LACV), the leading cause of pediatric arbovirus encephalitis in the USA, was associated with the ability of the virus to cross the BBB. LACV infection in weanling mice (aged around 3 weeks) results in vascular leakage in the olfactory bulb/tract (OB/OT) region of the brain, which is not observed in adult mice aged > 6–8 weeks. Thus, we studied age-specific differences in the response of brain capillary endothelial cells (BCECs) to LACV infection. Methods To examine mechanisms of LACV-induced BBB breakdown and infection of the CNS, we analyzed BCECs directly isolated from weanling and adult mice as well as established a model where these cells were infected in vitro and cultured for a short period to determine susceptibility to virus infection and cell death. Additionally, we utilized correlative light electron microscopy (CLEM) to examine whether changes in cell morphology and function were also observed in BCECs in vivo. Results BCECs from weanling, but not adult mice, had detectable infection after several days in culture when taken ex vivo from infected mice suggesting that these cells could be infected in vitro. Further analysis of BCECs from uninfected mice, infected in vitro, showed that weanling BCECs were more susceptible to virus infection than adult BCECs, with higher levels of infected cells, released virus as well as cytopathic effects (CPE) and cell death. Although direct LACV infection is not detected in the weanling BCECs, CLEM analysis of brain tissue from weanling mice indicated that LACV infection induced significant cerebrovascular damage which allowed virus-sized particles to enter the brain parenchyma. Conclusions These findings indicate that BCECs isolated from adult and weanling mice have differential viral load, infectivity, and susceptibility to LACV. These age-related differences in susceptibility may strongly influence LACV-induced BBB leakage and neurovascular damage allowing virus invasion of the CNS and the development of neurological disease.

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Isookanin Inhibits PGE2-Mediated Angiogenesis by Inducing Cell Arrest through Inhibiting the Phosphorylation of ERK1/2 and CREB in HMEC-1 Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22126466 ◽

2021 ◽

Vol 22 (12) ◽

pp. 6466

Author(s):

Yingji Xin ◽

Kyungbaeg Roh ◽

Eunae Cho ◽

Deokhoon Park ◽

Wankyunn Whang ◽

...

Keyword(s):

Endothelial Cells ◽

Inflammatory Diseases ◽

Inhibitory Effect ◽

Biological Properties ◽

Tube Formation ◽

Microvascular Endothelial Cells ◽

Potential Candidate ◽

Creb Phosphorylation ◽

Microvascular Endothelial ◽

Cell Arrest

Inflammation is increasingly recognized as a critical mediator of angiogenesis, and unregulated angiogenic responses often involve human diseases. The importance of regulating angiogenesis in inflammatory diseases has been demonstrated through some successful cases of anti-angiogenesis therapies in related diseases, including arthritis, but it has been reported that some synthetic types of antiangiogenic drugs have potential side effects. In recent years, the importance of finding alternative strategies for regulating angiogenesis has begun to attract the attention of researchers. Therefore, identification of natural ingredients used to prevent or treat angiogenesis-related diseases will play a greater role. Isookanin is a phenolic flavonoid presented in Bidens extract, and it has been reported that isookanin possesses some biological properties, including antioxidative and anti-inflammatory effects, anti-diabetic properties, and an ability to inhibit α-amylase. However, its antiangiogenic effects and mechanism thereof have not been studied yet. In this study, our results indicate that isookanin has an effective inhibitory effect on the angiogenic properties of microvascular endothelial cells. Isookanin shows inhibitory effects in multiple stages of PGE2-induced angiogenesis, including the growth, proliferation, migration, and tube formation of microvascular endothelial cells. In addition, isookanin induces cell cycle arrest in S phase, which is also the reason for subsequent inhibition of cell proliferation. The mechanism of inhibiting angiogenesis by isookanin is related to the inhibition of PGE2-mediated ERK1/2 and CREB phosphorylation. These findings make isookanin a potential candidate for the treatment of angiogenesis-related diseases.

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