{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T05:06:47Z","timestamp":1791090407252,"version":"4.1.0"},"reference-count":134,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,4,25]],"date-time":"2023-04-25T00:00:00Z","timestamp":1682380800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Maj Institute of Pharmacology at the Polish Academy of Sciences in Krak\u00f3w Poland"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>The purpose of this paper was to discuss the achievements of in vitro modeling in terms of the blood\u2013brain barrier [BBB] and to create a clear overview of this research area, which is useful in research planning. The text was divided into three main parts. The first part describes the BBB as a functional structure, its constitution, cellular and noncellular components, mechanisms of functioning and importance for the central nervous system, in terms of both protection and nourishment. The second part is an overview of parameters important in terms of establishing and maintaining a barrier phenotype that allows for formulating criteria of evaluation of the BBB in vitro models. The third and last part discusses certain techniques for developing the BBB in vitro models. It describes subsequent research approaches and models, as they underwent change alongside technological advancement. On the one hand, we discuss possibilities and limitations of different research approaches: primary cultures vs. cell lines and monocultures vs. multicultures. On the other hand, we review advantages and disadvantages of specific models, such as models-on-a-chip, 3D models or microfluidic models. We not only attempt to state the usefulness of specific models in different kinds of research on the BBB but also emphasize the significance of this area of research for advancement of neuroscience and the pharmaceutical industry.<\/jats:p>","DOI":"10.3390\/bioengineering10050519","type":"journal-article","created":{"date-parts":[[2023,4,26]],"date-time":"2023-04-26T01:44:42Z","timestamp":1682473482000},"page":"519","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Experimental Models to Study the Functions of the Blood\u2013Brain Barrier"],"prefix":"10.3390","volume":"10","author":[{"given":"Andrzej","family":"\u0141ach","sequence":"first","affiliation":[{"name":"Laboratory of Neuropharmacology and Epigenetics, Department of Pharmacology, Maj Institute of Pharmacology, Polish Academy of Sciences, 31-343 Krak\u00f3w, Poland"},{"name":"Department of Nutrition, Animal Biotechnology and Fisheries, Faculty of Animal Sciences, University of Agriculture, 30-059 Krak\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3620-3902","authenticated-orcid":false,"given":"Agnieszka","family":"Wnuk","sequence":"additional","affiliation":[{"name":"Laboratory of Neuropharmacology and Epigenetics, Department of Pharmacology, Maj Institute of Pharmacology, Polish Academy of Sciences, 31-343 Krak\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna Katarzyna","family":"W\u00f3jtowicz","sequence":"additional","affiliation":[{"name":"Department of Nutrition, Animal Biotechnology and Fisheries, Faculty of Animal Sciences, University of Agriculture, 30-059 Krak\u00f3w, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/nrn1824","article-title":"Astrocyte-endothelial interactions at the blood-brain barrier","volume":"7","author":"Abbott","year":"2006","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1047778","DOI":"10.3389\/fnins.2023.1047778","article-title":"Blood-brain barrier endothelial cells in neurodegenerative diseases: Signals from the \u201cbarrier\u201d","volume":"24","author":"Yuan","year":"2023","journal-title":"Front. Neurosci."},{"key":"ref_3","first-page":"748","article-title":"3D brain angiogenesis model to reconstitute functional human blood-brain barrier in vitro","volume":"10","author":"Lee","year":"2019","journal-title":"Biotechnol. Bioeng."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Memis, I., Mittal, R., Furar, E., White, I., Eshraghi, R.S., Mittal, J., and Eshraghi, A.A. (2022). Altered Blood Brain Barrier Permeability and Oxidative Stress in Cntnap2 Knockout Rat Model. J. Clin. Med., 11.","DOI":"10.3390\/jcm11102725"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"83","DOI":"10.3389\/fpsyt.2017.00083","article-title":"Neurovascular Unit Dysfunction and Blood-Brain Barrier Hyperpermeability Contribute to Schizophrenia Neurobiology: A Theoretical Integration of Clinical and Experimental Evidence","volume":"8","author":"Najjar","year":"2017","journal-title":"Front. Psychiatry"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Marx, U., and Sandig, V. (2007). Drug Testing In Vitro Breakthroughs and Trends in Cell Culture Technology, Wiley-VCH. [1st ed.].","DOI":"10.1002\/9783527609611"},{"key":"ref_7","first-page":"200","article-title":"Rola bariery krew-p\u0142yn m\u00f3zgowo-rdzeniowy i bariery krew m\u00f3zg w utrzymaniu homeostazy o\u015brodkowego uk\u0142adu nerwowego","volume":"14","author":"Lewoniewska","year":"2018","journal-title":"Pol. Przegl. Neurol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Barichello, T. (2019). Blood Brain Barrier, Humana Press. [1st ed.].","DOI":"10.1007\/978-1-4939-8946-1"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"113","DOI":"10.55782\/ane-2011-1828","article-title":"In vitro models of the blood-brain barrier","volume":"71","author":"Wilhelm","year":"2011","journal-title":"Acta Neurobiol. Exp."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0079-6107(02)00037-8","article-title":"Tight junction proteins","volume":"81","author":"Betanzos","year":"2003","journal-title":"Prog. Biophys. Mol. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1777","DOI":"10.1083\/jcb.123.6.1777","article-title":"Occludin: A novel integral membrane protein localizing at tight junctions","volume":"123","author":"Furuse","year":"1993","journal-title":"J. Cell Biol."},{"key":"ref_12","first-page":"2892","article-title":"The tight junction protein occludin modulates blood-brain barrier integrity and neurological function after ischemic stroke in mice","volume":"13","author":"Sugiyama","year":"2023","journal-title":"Sci. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1083\/jcb.142.1.117","article-title":"Junctional adhesion molecule, a novel member of the immunoglobulin superfamily that distributes at intercellular junctions and modulates monocyte transmigration","volume":"142","author":"Lostaglio","year":"1998","journal-title":"J. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"773","DOI":"10.1242\/jcs.104.3.773","article-title":"Immunohistochemical localization of adherens junction components in blood-brain barrier microvessels of the rat","volume":"104","author":"Schulze","year":"1993","journal-title":"J. Cell Sci."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Barichello, T. (2019). Blood Brain Barrier, Humana Press. [1st ed.].","DOI":"10.1007\/978-1-4939-8946-1"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/j.bbamem.2007.07.014","article-title":"Desmosome structure, composition and function","volume":"1778","author":"Garrod","year":"2008","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1523\/JNEUROSCI.22-01-00183.2002","article-title":"Protoplasmic astrocytes in CA1 stratum radiatum occupy separate anatomical domains","volume":"22","author":"Bushong","year":"2002","journal-title":"J. Neurosci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1798","DOI":"10.1111\/j.1471-4159.1990.tb01236.x","article-title":"An easier, reproducible, and mass-production method to study the blood-brain barrier in vitro","volume":"54","author":"Dehouck","year":"1990","journal-title":"J. Neurochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/S0169-409X(98)00085-4","article-title":"P-Glycoprotein, a gatekeeper in the blood-brain barrier","volume":"36","author":"Schinkel","year":"1999","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1007\/s10571-004-1375-x","article-title":"In search of the astrocytic factor(s) modulating blood-brain barrier functions in brain capillary endothelial cells in vitro","volume":"25","author":"Haseloff","year":"2005","journal-title":"Cell. Mol. Neurobiol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Gotoh, M., Miyamoto, Y., and Ikeshima-Kataoka, H. (2023). Astrocytic Neuroimmunological Roles Interacting with Microglial Cells in Neurodegenerative Diseases. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24021599"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1274","DOI":"10.1096\/fj.01-0814fje","article-title":"Angiogenesis of the blood-brain barrier in vitro and the function of cerebral pericytes","volume":"16","author":"Ramsauer","year":"2002","journal-title":"FASEB J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1538","DOI":"10.1523\/JNEUROSCI.21-05-01538.2001","article-title":"Induction of astrocyte differentiation by endothelial cells","volume":"21","author":"Mi","year":"2001","journal-title":"J. Neurosci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/S0014-5793(99)00451-2","article-title":"Astrocytes enhance radical defence in capillary endothelial cells constituting the blood-brain barrier","volume":"449","author":"Schroeter","year":"1999","journal-title":"FEBS Lett."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"78","DOI":"10.3389\/fcvm.2018.00078","article-title":"Tissue Specific Origin, Development, and Pathological Perspectives of Pericytes","volume":"5","author":"Yamazaki","year":"2018","journal-title":"Front. Cardiovasc. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"762","DOI":"10.1074\/jbc.R116.760215","article-title":"The Translational Significance of the Neurovascular Unit","volume":"292","author":"McConnell","year":"2017","journal-title":"J. Biol. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.brainres.2005.01.027","article-title":"Brain pericytes contribute to the induction and up-regulation of blood-brain barrier functions through transforming growth factor-beta production","volume":"1038","author":"Dohgu","year":"2005","journal-title":"Brain Res"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1007\/s12035-012-8244-2","article-title":"Neurovascular unit: A focus on pericytes","volume":"45","author":"Brites","year":"2012","journal-title":"Mol. Neurobiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1002\/jcp.21508","article-title":"Peripheral nerve pericytes originating from the blood-nerve barrier expresses tight junctional molecules and transporters as barrier-forming cells","volume":"217","author":"Shimizu","year":"2008","journal-title":"J. Cell. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"S13","DOI":"10.1161\/STROKEAHA.108.533117","article-title":"Pericyte signaling in the neurovascular unit","volume":"40","author":"Fisher","year":"2009","journal-title":"Stroke"},{"key":"ref_31","first-page":"633","article-title":"Pericyte morphology and function","volume":"36","author":"Yemisci","year":"2021","journal-title":"Histol. Histopathol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Chaulagain, B., Gothwal, A., Lamptey, R.N.L., Trivedi, R., Mahanta, A.K., Layek, B., and Singh, J. (2023). Experimental Models of In Vitro Blood-Brain Barrier for CNS Drug Delivery: An Evolutionary Perspective. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24032710"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1038\/sj.jcbfm.9600272","article-title":"CNS microvascular pericytes exhibit multipotential stem cell activity","volume":"26","author":"Katychev","year":"2006","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1962","DOI":"10.1002\/stem.1977","article-title":"Brain vascular pericytes following ischemia have multipotential stem cell activity to differentiate into neural and vascular lineage cells","volume":"33","author":"Nakagomi","year":"2015","journal-title":"Stem Cells"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1017\/S1431927611012402","article-title":"Endothelial cell-pericyte interactions stimulate basement membrane matrix assembly: Influence on vascular tube remodeling, maturation, and stabilization","volume":"18","author":"Stratman","year":"2012","journal-title":"Microsc. Microanal."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1452","DOI":"10.3389\/fnins.2019.01452","article-title":"The Neurovascular Unit: Effects of Brain Insults During the Perinatal Period","volume":"13","author":"Bell","year":"2020","journal-title":"Front. Neurosci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.2174\/138161209787846766","article-title":"Developmental and pathogenic mechanisms of basement membrane assembly","volume":"15","author":"Yurchenco","year":"2009","journal-title":"Curr. Pharm. Des."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.brainresrev.2010.05.003","article-title":"Looking at the blood-brain barrier: Molecular anatomy and possible investigation approaches","volume":"64","author":"Cardoso","year":"2010","journal-title":"Brain Res. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/0006-8993(91)91628-E","article-title":"Glial cells and neurons induce blood-brain barrier related enzymes in cultured cerebral endothelial cells","volume":"539","author":"Tontsch","year":"1991","journal-title":"Brain Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/0012-1606(81)90382-1","article-title":"Developing nervous tissue induces formation of blood-brain barrier characteristics in invading endothelial cells: A study using quail\u2014Chick transplantation chimeras","volume":"84","author":"Stewart","year":"1981","journal-title":"Dev. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1016\/j.neuron.2020.09.002","article-title":"Neuronal Activity Regulates Blood-Brain Barrier Efflux Transport through Endothelial Circadian Genes","volume":"108","author":"Pulido","year":"2020","journal-title":"Neuron"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1088\/1758-5090\/acb571","article-title":"Advancements in modelling human blood brain-barrier on a chip","volume":"15","author":"Guarino","year":"2023","journal-title":"Biofabrication"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1007\/s00401-015-1524-y","article-title":"Microglia\u2013blood vessel interactions: A double-edged sword in brain pathologies","volume":"131","author":"Stankovic","year":"2016","journal-title":"Acta Neuropathol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"290","DOI":"10.1126\/science.3276004","article-title":"Perivascular microglial cells of the CNS are bone marrow-derived and present antigen in vivo","volume":"239","author":"Hickey","year":"1988","journal-title":"Science"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1073\/pnas.0805453106","article-title":"Selective targeting of perivascular macrophages for clearance of beta-amyloid in cerebral amyloid angiopathy","volume":"106","author":"Hawkes","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/1357-4310(96)88720-X","article-title":"Transporting therapeutics across the blood-brain barrier","volume":"2","author":"Abbott","year":"1996","journal-title":"Mol. Med. Today"},{"key":"ref_47","unstructured":"Kesharwani, P., and Gupta, U. (2018). Nanotechnology-Based Targeted Drug Delivery Systems for Brain Tumors, Academic Press. [1st ed.]."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"E836","DOI":"10.1152\/ajpendo.00496.2009","article-title":"Will the original glucose transporter isoform please stand up!","volume":"297","author":"Carruthers","year":"2009","journal-title":"Am. J. Physiol. Endocrinol. Metab."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"837","DOI":"10.1111\/j.1471-4159.1975.tb04415.x","article-title":"Synthetic amino acids and the nature of L-DOPA transport at the blood-brain barrier","volume":"25","author":"Wade","year":"1975","journal-title":"J. Neurochem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1317","DOI":"10.1208\/s12248-017-0110-8","article-title":"SLC and ABC Transporters: Expression, Localization, and Species Differences at the Blood-Brain and the Blood-Cerebrospinal Fluid Barriers","volume":"19","author":"Morris","year":"2017","journal-title":"AAPS J."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S0169-409X(00)00139-3","article-title":"Delivery of peptides and proteins through the blood-brain barrier","volume":"46","author":"Bickel","year":"2001","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"632","DOI":"10.1073\/pnas.85.2.632","article-title":"Transcytotic pathway for blood-borne protein through the blood-brain barrier","volume":"85","author":"Broadwell","year":"1988","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1007\/BF01190839","article-title":"Ultracytochemical characterization of anionic sites in the wall of brain capillaries","volume":"18","author":"Vorbrodt","year":"1989","journal-title":"J. Neurocytol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"17867","DOI":"10.1073\/pnas.0408191101","article-title":"Tumor imaging by means of proteolytic activation of cell-penetrating peptides","volume":"101","author":"Jiang","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"46","DOI":"10.29245\/2572-9411\/2016\/3.1026","article-title":"Transepithelial\/endothelial Electrical Resistance (TEER) theory and applications for microfluidic body-on-a-chip devices","volume":"1","author":"Elbrecht","year":"2016","journal-title":"J. Rare Dis. Res. Treat."},{"key":"ref_56","first-page":"51300","article-title":"Electric cell-substrate impedance sensing for the quantification of endothelial proliferation, barrier function, and motility","volume":"85","author":"Szulcek","year":"2014","journal-title":"J. Vis. Exp."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"228","DOI":"10.1016\/S1046-2023(03)00029-X","article-title":"Functional analysis of tight junctions","volume":"30","author":"Matter","year":"2003","journal-title":"Methods"},{"key":"ref_58","first-page":"19","article-title":"Temperature corrected transepithelial electrical resistance (TEER) measurement to quantify rapid changes in paracellular permeability","volume":"65","author":"Blume","year":"2010","journal-title":"Pharmazie"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1021\/js950269u","article-title":"Transport properties are not altered across Caco-2 cells with heightened TEER despite underlying physiological and ultrastructural changes","volume":"85","author":"Lu","year":"1996","journal-title":"J. Pharm. Sci."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1725","DOI":"10.1083\/jcb.115.6.1725","article-title":"A cell culture model of the blood-brain barrier","volume":"115","author":"Rubin","year":"1991","journal-title":"J. Cell Biol."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"2011","DOI":"10.4103\/1673-5374.172320","article-title":"In vitro model of the blood-brain barrier established by co-culture of primary cerebral microvascular endothelial and astrocyte cells","volume":"10","author":"Wang","year":"2015","journal-title":"Neural Regen. Res."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1089\/scd.2017.0003","article-title":"Paving the Way Toward Complex Blood-Brain Barrier Models Using Pluripotent Stem Cells","volume":"26","author":"Lauschke","year":"2017","journal-title":"Stem Cells Dev."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1177\/2211068214561025","article-title":"TEER measurement techniques for in vitro barrier model systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J. Lab. Autom."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1038\/nprot.2007.55","article-title":"Freeze-fracture electron microscopy","volume":"2","author":"Severs","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1007\/BF01224762","article-title":"Freeze-fracture evidence for a novel restricting junction at the blood-brain barrier of the cuttlefish Sepia officinalis","volume":"21","author":"Lane","year":"1992","journal-title":"J. Neurocytol."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"943","DOI":"10.1002\/cne.22273","article-title":"Differential distribution of tight junction proteins suggests a role for tanycytes in blood-hypothalamus barrier regulation in the adult mouse brain","volume":"518","author":"Mullier","year":"2010","journal-title":"J. Comp. Neurol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1016\/0006-8993(90)91425-G","article-title":"Blood-brain barrier characteristic enzymatic properties in cultured brain capillary endothelial cells","volume":"514","author":"Meyer","year":"1990","journal-title":"Brain Res."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"3229","DOI":"10.1007\/s11095-014-1464-6","article-title":"In vitro cerebrovascular modeling in the 21st century: Current and prospective technologies","volume":"31","author":"Palmiotti","year":"2014","journal-title":"Pharm. Res."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1949","DOI":"10.1021\/mp500046f","article-title":"In vitro models of the blood-brain barrier for the study of drug delivery to the brain","volume":"11","author":"Wilhelm","year":"2014","journal-title":"Mol. Pharm."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1021\/jm00004a004","article-title":"IAM chromatography: An in vitro screen for predicting drug membrane permeability","volume":"38","author":"Pidgeon","year":"1995","journal-title":"J. Med. Chem."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1021\/jm970530e","article-title":"Physicochemical high throughput screening: Parallel artificial membrane permeation assay in the description of passive absorption processes","volume":"41","author":"Kansy","year":"1998","journal-title":"J. Med. Chem."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1517\/17425255.1.2.325","article-title":"The rise of PAMPA","volume":"1","author":"Avdeef","year":"2005","journal-title":"Expert Opin. Drug Metab. Toxicol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"1617","DOI":"10.2174\/0929867043365071","article-title":"In silico prediction models for blood-brain barrier permeation","volume":"11","author":"Ecker","year":"2004","journal-title":"Curr. Med. Chem."},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Nag, S. (2003). The Blood\u2013Brain Barrier: Biology and Research Protocols, Humana Press. [1st ed.].","DOI":"10.1385\/1592594190"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1517\/17425255.4.6.721","article-title":"Utilization of membrane vesicle preparations to study drug-ABC transporter interactions","volume":"4","author":"Glavinas","year":"2008","journal-title":"Expert Opin. Drug Metab. Toxicol."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"3591","DOI":"10.2147\/DDDT.S218708","article-title":"In-vitro blood-brain barrier models for drug screening and permeation studies: An overview","volume":"13","author":"Bagchi","year":"2019","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"340","DOI":"10.1016\/j.ejpb.2012.07.020","article-title":"Comparison of brain capillary endothelial cell-based and epithelial (MDCK-MDR1, Caco-2, and VB-Caco-2) cell-based surrogate blood-brain barrier penetration models","volume":"82","author":"Hellinger","year":"2012","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1023\/A:1016462205267","article-title":"Prediction of drug transport through the blood-brain barrier in vivo: A comparison between two in vitro cell models","volume":"19","author":"Lundquist","year":"2002","journal-title":"Pharm. Res."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/j.ijpharm.2004.10.007","article-title":"Evaluation of the MDR-MDCK cell line as a permeability screen for the blood-brain barrier","volume":"288","author":"Wang","year":"2005","journal-title":"Int. J. Pharm."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"2331","DOI":"10.2174\/138161210791920432","article-title":"Nanoparticulate formulations for paclitaxel delivery across MDCK cell monolayer","volume":"16","author":"Xie","year":"2010","journal-title":"Curr. Pharm. Des."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"384982","DOI":"10.1155\/2008\/384982","article-title":"Human brain microvascular endothelial cells and umbilical vein endothelial cells differentially facilitate leukocyte recruitment and utilize chemokines for T cell migration","volume":"2008","author":"Man","year":"2008","journal-title":"Clin. Dev. Immunol."},{"key":"ref_82","doi-asserted-by":"crossref","unstructured":"Langford, D., Hurford, R., Hashimoto, M., Digicaylioglu, M., and Masliah, E. (2005). Signalling crosstalk in FGF2-mediated protection of endothelial cells from HIV-gp120. BMC Neurosci., 6.","DOI":"10.1186\/1471-2202-6-8"},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1634\/stemcells.2008-0922","article-title":"Hematopoietic and endothelial differentiation of human induced pluripotent stem cells","volume":"27","author":"Choi","year":"2009","journal-title":"Stem Cells"},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1038\/nbt.2247","article-title":"Derivation of blood-brain barrier endothelial cells from human pluripotent stem cells","volume":"30","author":"Lippmann","year":"2012","journal-title":"Nat. Biotechnol."},{"key":"ref_85","first-page":"169","article-title":"Cell-based in vitro models for studying blood\u2013brain barrier (BBB) permeability","volume":"Volume 1","author":"Sarmento","year":"2016","journal-title":"Concepts and Models for Drug Permeability Studies"},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"862","DOI":"10.1177\/0271678X16630991","article-title":"In vitro models of the blood-brain barrier: An overview of commonly used brain endothelial cell culture models and guidelines for their use","volume":"36","author":"Helms","year":"2016","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1007\/BF00166997","article-title":"Isolation and characterization of microvessels from normal brain and brain tumors","volume":"11","author":"Silbergeld","year":"1991","journal-title":"J. Neurooncol."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"R191","DOI":"10.1152\/ajpregu.00305.2001","article-title":"Xenobiotic efflux pumps in isolated fish brain capillaries","volume":"282","author":"Miller","year":"2002","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_89","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1006\/bbrc.1996.0239","article-title":"Endothelin stimulates protein phosphorylation in blood-brain barrier","volume":"219","year":"1996","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"7091","DOI":"10.1523\/JNEUROSCI.2711-13.2014","article-title":"1\u03b1,25-Dihydroxyvitamin D3 reduces cerebral amyloid-\u03b2 accumulation and improves cognition in mouse models of Alzheimer\u2019s disease","volume":"34","author":"Durk","year":"2014","journal-title":"J. Neurosci."},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"Salvador, E., K\u00f6ppl, T., H\u00f6rmann, J., Sch\u00f6nh\u00e4rl, S., Bugaeva, P., Kessler, A.F., Burek, M., Ernestus, R.-I., L\u00f6hr, M., and Hagemann, C. (2023). Tumor Treating Fields (TTFields) Induce Cell Junction Alterations in a Human 3D In Vitro Model of the Blood-Brain Barrier. Pharmaceutics, 15.","DOI":"10.3390\/pharmaceutics15010185"},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"107","DOI":"10.2174\/157488911795933910","article-title":"Patented in vitro blood-brain barrier models in CNS drug discovery","volume":"6","author":"Veszelka","year":"2011","journal-title":"Recent Pat. CNS Drug Discov."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"1872","DOI":"10.1096\/fj.04-3458fje","article-title":"Blood-brain barrier-specific properties of a human adult brain endothelial cell line","volume":"19","author":"Weksler","year":"2005","journal-title":"FASEB J."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1080\/17460441.2017.1253676","article-title":"New experimental models of the blood-brain barrier for CNS drug discovery","volume":"12","author":"Kaisar","year":"2017","journal-title":"Expert Opin. Drug Discov."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1186\/s12987-020-00222-3","article-title":"Commentary on human pluripotent stem cell-based blood-brain barrier models","volume":"17","author":"Lippmann","year":"2020","journal-title":"Fluids Barriers CNS"},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"e22770","DOI":"10.1096\/fj.202201422R","article-title":"High and low permeability of human pluripotent stem cell-derived blood-brain barrier models depend on epithelial or endothelial features","volume":"37","author":"Girard","year":"2023","journal-title":"FASEB J."},{"key":"ref_97","doi-asserted-by":"crossref","unstructured":"Singh, N.R., Gromnicova, R., Brachner, A., Kraev, I., Romero, I.A., Neuhaus, W., and Male, D. (2023). A hydrogel model of the human blood-brain barrier using differentiated stem cells. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0283954"},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"2755","DOI":"10.2174\/138161211797440159","article-title":"in vitro modeling of the blood-brain barrier: Simplicity versus complexity","volume":"17","author":"Ogunshola","year":"2011","journal-title":"Curr. Pharm. Des."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.brainres.2012.06.057","article-title":"Establishment of a simplified in vitro porcine blood-brain barrier model with high transendothelial electrical resistance","volume":"1521","author":"Patabendige","year":"2012","journal-title":"Brain Res."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.brainres.2012.08.031","article-title":"An immortalised astrocyte cell line maintains the in vivo phenotype of a primary porcine in vitro blood-brain barrier model","volume":"1479","author":"Cantrill","year":"2012","journal-title":"Brain Res."},{"key":"ref_101","doi-asserted-by":"crossref","unstructured":"Barichello, T. (2019). Blood Brain Barrier, Humana Press. [1st ed.].","DOI":"10.1007\/978-1-4939-8946-1"},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1080\/10611860701288539","article-title":"Primary porcine brain microvascular endothelial cells: Biochemical and functional characterisation as a model for drug transport and targeting","volume":"15","author":"Smith","year":"2007","journal-title":"J. Drug Target."},{"key":"ref_103","doi-asserted-by":"crossref","unstructured":"Park, J.S., Choe, K., Khan, A., Jo, M.H., Park, H.Y., Kang, M.H., Park, T.J., and Kim, M.O. (2023). Establishing Co-Culture Blood-Brain Barrier Models for Different Neurodegeneration Conditions to Understand Its Effect on BBB Integrity. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24065283"},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1111\/j.1582-4934.2005.tb00362.x","article-title":"Permeability properties of a three-cell type in vitro model of blood-brain barrier","volume":"9","author":"Schiera","year":"2005","journal-title":"J. Cell. Mol. Med."},{"key":"ref_105","doi-asserted-by":"crossref","unstructured":"Boghdeh, N.A., Risner, K.H., Barrera, M.D., Britt, C.M., Schaffer, D.K., Alem, F., Brown, J.A., Wikswo, J.P., and Narayanan, A. (2022). Application of a Human Blood Brain Barrier Organ-on-a-Chip Model to Evaluate Small Molecule Effectiveness against Venezuelan Equine Encephalitis Virus. Viruses, 14.","DOI":"10.3390\/v14122799"},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1002\/glia.20189","article-title":"Induction of blood-brain barrier properties in cultured brain capillary endothelial cells: Comparison between primary glial cells and C6 cell line","volume":"51","author":"Boveri","year":"2005","journal-title":"Glia"},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1242\/jcs.00281","article-title":"Tumour-endothelium interactions in co-culture: Coordinated changes of gene expression profiles and phenotypic properties of endothelial cells","volume":"116","author":"Khodarev","year":"2003","journal-title":"J. Cell. Sci."},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1021\/acsabm.8b00654","article-title":"Comparison of Blood-Brain Barrier Models for in vitro Biological Analysis: One Cell Type vs Three Cell Types","volume":"2","author":"Song","year":"2019","journal-title":"ACS Appl. Bio Mater."},{"key":"ref_109","doi-asserted-by":"crossref","unstructured":"Barichello, T. (2019). Blood Brain Barrier, Humana Press. [1st ed.].","DOI":"10.1007\/978-1-4939-8946-1"},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1093\/cvr\/cvq146","article-title":"Shear stress and the endothelial transport barrier","volume":"87","author":"Tarbell","year":"2010","journal-title":"Cardiovasc. Res."},{"key":"ref_111","doi-asserted-by":"crossref","unstructured":"Cucullo, L., Hossain, M., Puvenna, V., Marchi, N., and Janigro, D. (2011). The role of shear stress in Blood-Brain Barrier endothelial physiology. BMC Neurosci., 12.","DOI":"10.1186\/1471-2202-12-40"},{"key":"ref_112","doi-asserted-by":"crossref","first-page":"1851","DOI":"10.1063\/1.1136909","article-title":"Apparatus for subjecting living cells to fluid shear stress","volume":"53","author":"Bussolari","year":"1982","journal-title":"Rev. Sci. Instrum."},{"key":"ref_113","doi-asserted-by":"crossref","first-page":"119ra14","DOI":"10.1126\/scitranslmed.3003197","article-title":"CXCL12-induced monocyte-endothelial interactions promote lymphocyte transmigration across an in vitro blood-brain barrier","volume":"4","author":"Man","year":"2012","journal-title":"Sci. Transl. Med."},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/s12195-015-0424-5","article-title":"A Parallel-Plate Flow Chamber for Mechanical Characterization of Endothelial Cells Exposed to Laminar Shear Stress","volume":"9","author":"Wong","year":"2016","journal-title":"Cell. Mol. Bioeng."},{"key":"ref_115","doi-asserted-by":"crossref","unstructured":"Kemeny, S.F., Figueroa, D.S., and Clyne, A.M. (2013). Hypo- and hyperglycemia impair endothelial cell actin alignment and nitric oxide synthase activation in response to shear stress. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0066176"},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/S0006-8993(03)02980-9","article-title":"Use of a three-dimensional in vitro model of the rat blood-brain barrier to assay nucleoside efflux from brain","volume":"980","author":"Parkinson","year":"2003","journal-title":"Brain Res."},{"key":"ref_117","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1002\/jps.23022","article-title":"In vitro blood-brain barrier models: Current and perspective technologies","volume":"101","author":"Naik","year":"2012","journal-title":"J. Pharm. Sci."},{"key":"ref_118","doi-asserted-by":"crossref","unstructured":"Barichello, T. (2019). Blood Brain Barrier, Humana Press. [1st ed.].","DOI":"10.1007\/978-1-4939-8946-1"},{"key":"ref_119","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1038\/sj.jcbfm.9600525","article-title":"Immortalized human brain endothelial cells and flow-based vascular modeling: A marriage of convenience for rational neurovascular studies","volume":"28","author":"Cucullo","year":"2008","journal-title":"J. Cereb. Blood Flow Metab."},{"key":"ref_120","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1517\/17425247.2013.762354","article-title":"Drug targeting to brain: A systematic approach to study the factors, parameters and approaches for prediction of permeability of drugs across BBB","volume":"10","author":"Nagpal","year":"2013","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_121","doi-asserted-by":"crossref","first-page":"2324","DOI":"10.1002\/jbm.a.36981","article-title":"Three-dimensional printing of extracellular matrix (ECM)-mimicking scaffolds: A critical review of the current ECM materials","volume":"108","author":"Kumar","year":"2020","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_122","doi-asserted-by":"crossref","first-page":"1193","DOI":"10.1163\/156856205774269485","article-title":"Proliferation and differentiation of human embryonic germ cell derivatives in bioactive polymeric fibrous scaffold","volume":"16","author":"Yim","year":"2005","journal-title":"J. Biomater. Sci. Polym. Ed."},{"key":"ref_123","doi-asserted-by":"crossref","first-page":"179","DOI":"10.2174\/157015908785777210","article-title":"Brain endothelial cell-cell junctions: How to \u201copen\u201d the blood brain barrier","volume":"6","author":"Stamatovic","year":"2008","journal-title":"Curr. Neuropharmacol."},{"key":"ref_124","doi-asserted-by":"crossref","first-page":"934","DOI":"10.1016\/j.tim.2020.06.005","article-title":"Human Organs-on-Chips for Virology","volume":"28","author":"Tang","year":"2020","journal-title":"Trends Microbiol."},{"key":"ref_125","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1186\/s12951-023-01798-2","article-title":"BBB-on-a-chip with integrated micro-TEER for permeability evaluation of multi-functionalized gold nanorods against Alzheimer\u2019s disease","volume":"21","author":"Kogan","year":"2023","journal-title":"J. Nanobiotechnol."},{"key":"ref_126","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s10544-012-9699-7","article-title":"BBB on chip: Microfluidic platform to mechanically and biochemically modulate blood-brain barrier function","volume":"15","author":"Griep","year":"2013","journal-title":"Biomed. Microdevices"},{"key":"ref_127","doi-asserted-by":"crossref","first-page":"114816","DOI":"10.1016\/j.addr.2023.114816","article-title":"An overview of in vitro 3D models of the blood-brain barrier as a tool to predict the in vivo permeability of nanomedicines","volume":"196","year":"2023","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_128","doi-asserted-by":"crossref","first-page":"995","DOI":"10.1016\/j.stem.2019.05.011","article-title":"Human iPSC-Derived Blood-Brain Barrier Chips Enable Disease Modeling and Personalized Medicine Applications","volume":"24","author":"Vatine","year":"2019","journal-title":"Cell Stem Cell"},{"key":"ref_129","doi-asserted-by":"crossref","unstructured":"Kincses, A., Vigh, J.P., Petrovszki, D., Valkai, S., Kocsis, A.E., Walter, F.R., Lin, H.-Y., Jan, J.-S., Deli, M.A., and D\u00e9r, A. (2023). The Use of Sensors in Blood-Brain Barrier-on-a-Chip Devices: Current Practice and Future Directions. Biosensors, 13.","DOI":"10.3390\/bios13030357"},{"key":"ref_130","doi-asserted-by":"crossref","first-page":"542","DOI":"10.1039\/C2LC41033H","article-title":"A modular approach to create a neurovascular unit-on-a-chip","volume":"13","author":"Achyuta","year":"2013","journal-title":"Lab Chip"},{"key":"ref_131","doi-asserted-by":"crossref","first-page":"865","DOI":"10.1038\/nbt.4226","article-title":"A linked organ-on-chip model of the human neurovascular unit reveals the metabolic coupling of endothelial and neuronal cells","volume":"36","author":"Maoz","year":"2018","journal-title":"Nat. Biotechnol."},{"key":"ref_132","doi-asserted-by":"crossref","first-page":"S18","DOI":"10.1186\/scrt379","article-title":"Neurovascular unit on a chip: Implications for translational applications","volume":"4","author":"Alcendor","year":"2013","journal-title":"Stem Cell Res. Ther."},{"key":"ref_133","doi-asserted-by":"crossref","first-page":"1093","DOI":"10.1039\/c2lc41208j","article-title":"SyM-BBB: A microfluidic Blood Brain Barrier model","volume":"13","author":"Prabhakarpandian","year":"2013","journal-title":"Lab Chip"},{"key":"ref_134","doi-asserted-by":"crossref","first-page":"8083","DOI":"10.1038\/s41598-017-07416-0","article-title":"A Low Permeability Microfluidic Blood-Brain Barrier Platform with Direct Contact between Perfusable Vascular Network and Astrocytes","volume":"7","author":"Bang","year":"2017","journal-title":"Sci. Rep."}],"container-title":["Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-5354\/10\/5\/519\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:23:20Z","timestamp":1760124200000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-5354\/10\/5\/519"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,25]]},"references-count":134,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2023,5]]}},"alternative-id":["bioengineering10050519"],"URL":"https:\/\/doi.org\/10.3390\/bioengineering10050519","relation":{},"ISSN":["2306-5354"],"issn-type":[{"value":"2306-5354","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,25]]}}}