Validation of an immortalized human (hBMEC) in vitro blood-brain barrier model

Eigenmann, Daniela Elisabeth; Jähne, Evelyn A.; Smieško, Martin; Hamburger, Matthias; Oufir, Mouhssin

doi:10.1007/s00216-016-9313-6

Cited by 30 publications

(31 citation statements)

References 42 publications

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“…Stock solutions and working solutions of analytes and internal standard (I.S.) were prepared as described elsewhere [15]. For all compounds, calibration standards (calibrators) in the range of 5.00-500 ng/mL (5.00, 25.0, 50.0, 100, 200, 400, and 500 ng/mL) and quality controls (QCs) at low, middle, and high concentration (QCL: 15.0 ng/mL, QCM: 250 ng/mL, and QCH: 400 ng/mL) were prepared in modified RHB (RHB + 0.2% Tween 20) and modified DMEM (DMEM + 10% FBS for piperine, LAU397, and LAU399; DMEM + 10% FBS + 0.2% Tween 20 for the other analogs) by serial dilution of the respective working solutions.…”

Section: Stock Solutions Calibration Standards and Quality Controlsmentioning

confidence: 99%

“…Immortalized in vitro human BBB model: The immortalized human mono-culture in vitro BBB model based on hBMEC cell line [13] was prepared as reported previously [14,15]. Tissue culture inserts (24-well format) were from Greiner Bio-one (0.336 cm 2 , transparent PET membrane, 3.0 µm pore size, Greiner Bio-one, Frickenhausen, Germany), and were coated with rat tail collagen I.…”

Section: In Vitro Bbb Modelsmentioning

confidence: 99%

“…in the apical-to-basolateral (A→B) and basolateral to apical (B→A) direction), with working solutions containing the test compound (2 µM) and Na-F (10 µg/mL) in RHB + 0.1% BSA. Aliquots of donor and receiver compartments were collected after several time points (15,30,60, and 120 min) (one insert per time point).…”

Section: In Vitro Bbb Modelsmentioning

confidence: 99%

“…where CR and VR are the concentration and volume in the receiver compartments, respectively, and CD is the initial concentration in the donor compartment. The mean cleared volume was plotted as a function of time (15,30, 60, and 120 min), and the slope was estimated by linear regression analysis. Permeability-surface area products (PS) and Pe values were subsequently calculated according to the following equations [20,21]:…”

Section: Calculation Of Permeability Coefficientsmentioning

confidence: 99%

“…In the present study, we assessed the BBB permeability of piperine and five selected piperine analogs ( Fig. 1) in three in vitro cell-based BBB models, namely a recently established human model with immortalized hBMEC cells, a human brain-like endothelial cells (BLEC) model, and a primary animal (bovine endothelial/rat astrocytes co-culture) model [13][14][15][16][17]. Permeability coefficients across the cell monolayers were determined by means of UHPLC-MS/MS, whereby quantitative UHPLC-MS/MS assays in the corresponding matrix were developed for each compound.…”

Section: Introductionmentioning

confidence: 99%

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In vitro blood–brain barrier permeability predictions for GABAA receptor modulating piperine analogs

Eigenmann

Dürig

Jähne

et al. 2016

European Journal of Pharmaceutics and Biopharmaceutics

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The alkaloid piperine from black pepper (Piper nigrum L.) and several synthetic piperine analogs were recently identified as positive allosteric modulators of γ-aminobutyric acid type A (GABAA) receptors. In order to reach their target sites of action, these compounds need to enter the brain by crossing the blood-brain barrier (BBB). We here evaluated piperine and five selected analogs (SCT-66, SCT-64, SCT-29, LAU397, and LAU399) regarding their BBB permeability. Data were obtained in three in vitro BBB models, namely a recently established human model with immortalized hBMEC cells, a human brain-like endothelial cells (BLEC) model, and a primary animal (bovine endothelial/rat astrocytes co-culture) model. For each compound, quantitative UHPLC-MS/MS methods in the range of 5.00-500ng/mL in the corresponding matrix were developed, and permeability coefficients in the three BBB models were determined. In vitro predictions from the two human BBB models were in good agreement, while permeability data from the animal model differed to some extent, possibly due to protein binding of the screened compounds. In all three BBB models, piperine and SCT-64 displayed the highest BBB permeation potential. This was corroborated by data from in silico prediction. For the other piperine analogs (SCT-66, SCT-29, LAU397, and LAU399), BBB permeability was low to moderate in the two human BBB models, and moderate to high in the animal BBB model. Efflux ratios (ER) calculated from bidirectional permeability experiments indicated that the compounds were likely not substrates of active efflux transporters.

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Section: Stock Solutions Calibration Standards and Quality Controlsmentioning

confidence: 99%

Section: In Vitro Bbb Modelsmentioning

confidence: 99%

Section: In Vitro Bbb Modelsmentioning

confidence: 99%

Section: Calculation Of Permeability Coefficientsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In vitro blood–brain barrier permeability predictions for GABAA receptor modulating piperine analogs

Eigenmann

Dürig

Jähne

et al. 2016

European Journal of Pharmaceutics and Biopharmaceutics

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Progresses in the establishment, evaluation, and application of in vitro blood–brain barrier models

Yin,

Wang

2024

J of Neuroscience Research

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The blood–brain barrier (BBB) is a barrier between the circulatory system and the central nervous system (CNS), contributing to CNS protection and maintaining the brain homeostasis. Establishment of in vitro BBB models that are closer to the microenvironment of the human brain is helpful for evaluating the potential and efficiency of a drug penetrating BBB and thus the clinical application value of the drug. The in vitro BBB models not only provide great convenience for screening new drugs that can access to CNS but also help people to have a deeper study on the mechanism of substances entering and leaving the brain, which makes people have greater opportunities in the treatment of CNS diseases. Up to now, although much effort has been paid to the researches on the in vitro BBB models and many progresses have been achieved, no unified method has been described for establishing a BBB model and there is much work to do and many challenges to be faced with in the future. This review summarizes the research progresses in the establishment, evaluation, and application of in vitro BBB models.

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Brain‐Penetration and Neuron‐Targeting DNA Nanoflowers Co‐Delivering miR‐124 and Rutin for Synergistic Therapy of Alzheimer's Disease

Ouyang

Zhu

et al. 2022

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tau hyperphosphorylation, mitochondrial dysfunction and oxidative stress, and neuroinflammatory response. [3] Among them, the amyloid theory that focuses on excessive deposition of β-amyloid (Aβ) has been extensively studied with wide acceptance. [4] In this theory, imbalance of Aβ metabolism and catabolism in the central nervous system (CNS) has been regarded as the main reason for AD-like pathology. [5] Aβ is the product of a proteolytic process of a transmembrane protein, amyloid precursor protein (APP), by β-secretase 1 (BACE1, β-site APP cleavage enzyme 1) and the γ-secretase. [6] The overgeneration and abnormal aggregation of Aβ 1-42 can cause the collapse of dendritic spines and neuronal loss, induce oxidative stress, and promote inflammatory by activating microglia, which in turn exacerbates the pathogenesis of AD. [7] Several strategies focus on blocking Aβ aggregation and alleviating Aβ-mediated neurotoxicity have been developed, including aggregation inhibition, copper chelation, Aβ photo-oxidation, fibril thermal dissociation, and Aβ degradation. [8] However, many Aβ targeting pharmacological treatments failed to slow down genitive decline or improve global functioning in phase 3 clinical trials although with definite efficacy at animal model. [9] Besides the pathological discrepancy between real clinical patient and AD animal model, [10] another important reason is that AD is a multifactorial disease and therapy targeting Aβ alone is far more enough. [11] Despite significant Alzheimer disease (AD) is the leading cause of dementia that affects millions of old people. Despite significant advances in the understanding of AD pathobiology, no disease modifying treatment is available. MicroRNA-124 (miR-124) is the most abundant miRNA in the normal brain with great potency to ameliorate AD-like pathology, while it is deficient in AD brain. Herein, the authors develop a DNA nanoflowers (DFs)-based delivery system to realize exogenous supplementation of miR-124 for AD therapy. The DFs with well-controlled size and morphology are prepared, and a miR-124 chimera is attached via hybridization. The DFs are further modified with RVG29 peptide to simultaneously realize brain-blood barrier (BBB) penetration and neuron targeting. Meanwhile, Rutin, a small molecular ancillary drug, is co-loaded into the DFs structure via its intercalation into the double stranded DNA region. Interestingly, Rutin could synergize miR-124 to suppress the expression of both BACE1 and APP, thus achieving a robust inhibition of amyloid β generation. The nanosystem could pro-long miR-124 circulation in vivo, promote its BBB penetration and neuron targeting, resulting in a significant increase of miR-124 in the hippocampus of APP/PS1 mice and robust therapeutic efficacy in vivo. Such a bio-derived therapeutic system shows promise as a biocompatible nanomedicine for AD therapy.

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Validation of an immortalized human (hBMEC) in vitro blood-brain barrier model

Cited by 30 publications

References 42 publications

In vitro blood–brain barrier permeability predictions for GABAA receptor modulating piperine analogs

In vitro blood–brain barrier permeability predictions for GABAA receptor modulating piperine analogs

Progresses in the establishment, evaluation, and application of in vitro blood–brain barrier models

Brain‐Penetration and Neuron‐Targeting DNA Nanoflowers Co‐Delivering miR‐124 and Rutin for Synergistic Therapy of Alzheimer's Disease

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