Fabrication of biomimetic placental barrier structures within a microfluidic device utilizing two-photon polymerization

Denise Mandt, Peter Gruber, Marica Markovic, Maximillian Tromayer, Mario Rothbauer, Sebastian Rudi Adam Krayz, Faheem Ali, Jasper van Hoorick, Wolfgang Holnthoner, Severin Mühleder, Peter Dubruel, Sandra Van Vlierberghe, Peter Ertl, Robert Liska, Aleksandr Ovsianikov

Article ID: 144
Vol 4, Issue 2, 2018, Article identifier:144

VIEWS - 7541 (Abstract) 952 (PDF)


The placenta is a transient organ, essential for development and survival of the unborn fetus. It interfaces the body of the pregnant woman with the unborn child and secures transport of endogenous and exogenous substances. Maternal and fetal blood are thereby separated at any time, by the so-called placental barrier. Current in vitro approaches fail to model this multifaceted structure, therefore research in the field of placental biology is particularly challenging. The present study aimed at establishing a novel model, simulating placental transport and its implications on development, in a versatile but reproducible way. The basal membrane was replicated using a gelatin-based material, closely mimicking the composition and properties of the natural extracellular matrix. The microstructure was produced by using a high-resolution 3D printing method – the two-photon polymerization (2PP). In order to structure gelatin by 2PP, its primary amines and carboxylic acids are modified with methacrylamides and methacrylates (GelMOD-AEMA), respectively. High-resolution structures in the range of a few micrometers were produced within the intersection of a customized microfluidic device, separating the x-shaped chamber into two isolated cell culture compartments. Human umbilical-vein endothelial cells (HUVEC) seeded on one side of this membrane simulate the fetal compartment while human choriocarcinoma cells, isolated from placental tissue (BeWo B30) mimic the maternal syncytium. This barrier model in combination with native flow profiles can be used to mimic the microenvironment of the placenta, investigating different pharmaceutical, clinical and biological scenarios. As proof-of-principle, this bioengineered placental barrier was used for the investigation of transcellular transport processes. While high molecular weight substances did not permeate, smaller molecules in the size of glucose were able to diffuse through the barrier in a time-depended manner. We envision to apply this bioengineered placental barrier for pathophysiological research, where altered nutrient transport is associated with health risks for the fetus.


high resolution 3D printing; placental barrier; model; microstructure; two-photon polymerization

Full Text:

Download PDF

Included Database


Lee J S, Romero R, Han Y M, et al., 2015, Placenta-ona-chip: A novel platform to study the biology of the human placenta. J Matern Neonatal Med, 29(7): 1046–1054. http://dx.doi.org/10.3109/14767058.2015.1038518

Ren K, Zhou J, Wu H, 2013, Materials for microfluidic chip fabrication. Acc Chem Res, 46(11): 2396–2406. http://dx.doi.org/10.1021/ar300314s

Blundell C, Tess E R, Schanzer A S R, et al., 2016, A microphysiological model of the human placental barrier. Lab Chip, 16(16): 3065–3073. http://dx.doi.org/10.1039/c6lc00259e

Sakolish C M, Esch M B, Hickman J J, et al., 2016, Modeling barrier tissues in vitro: Methods, achievements, and challenges. EBioMedicine, 5(C): 30–39. http://dx.doi.org/10.1016/j.ebiom.2016.02.023

Djagny K B, Wang Z, Xu S, et al., 2001, Gelatin: A valuable protein for food and pharmaceutical industries. Crit Rev Food Sci Nutr, 41(6): 481–492. http://dx.doi.org/10.1080/20014091091904

Peinemann K V, Nunes S P, 2007, Application of membranes in tissue engineering and biohybrid organ technology. Membrane technology: Membranes for life sciences, 1st edition, pp. 343, 2007. http://dx.doi.org/10.1002/9783527631360.ch8

Van Den Bulcke A I, Bogdanov B, De Rooze N, et al., 2000, Structural and rheological properties of methacrylamide modified gelatin hydrogels. Biomacromolecules, 1(1): 31–38. http://dx.doi.org/10.1021/bm990017d

Ovsianikov A, Mironov V, Stampfl J, et al., 2012, Engineering 3D cell-culture matrices: Multiphoton pro­cessing technologies for biological & tissue engineering applications. Expert Rev Med Devices, 9(6): 613–633. http://dx.doi.org/10.1586/erd.12.48

Hölzl K, Lin S, Tytgat L, et al, 2016, Bioink properties before, during and after 3D bioprinting. Biofabrication, 8(3): 032002. http://dx.doi.org/10.1088/1758-5090/8/3/032002

Van Hoorick J, Gruber P, Markovic M, et al., 2017, Cross-linkable gelatins with superior mechanical properties through carboxylic acid modification: Increasing the two-photon polymerization potential. Biomacromolecules, 18(10): 3260–3272. http://dx.doi.org/10.1021/acs.biomac.7b00905

Tayalia P, Mendonca C R, Baldacchini T, et al., 2008, 3D cell-migration studies using two-photon engineered polymer scaffolds. Adv Mater, 20(23): 4494–4498. http://dx.doi.org/10.1002/adma.200801319

Paz V F, Emons M, Obata K, et al., 2012, Development of functional sub-100 nm structures with 3D two-photon polymerization technique and optical methods for characterization. J Laser Appl, 24(4): 293–301. http://dx.doi.org/10.2351/1.4712151

Stampfl J, Liska R, Ovsinikov A, 2016, Multiphoton lithography: Techniques, materials, and applications. in Stampfl J, Liska R, Ovsinikov A, (Eds.) John Wiley & Sons, ISBN: 978-3-527-33717-0

Markovic M, Van Hoorick J, Hölzl K, et al., 2015, Hybrid tissue engineering scaffolds by combination of three-dimensional printing and cell photoencapsulation. J Nanotechnol Eng Med, 6(2): 0210011–210017. http://dx.doi.org/10.1115/1.4031466

Ovsianikov A, Muehleder S, Torgersen T, et al., 2014, Laser photofabrication of cell-containing hydrogel constructs. Langmuir, 30(13): 3787–3794. http://dx.doi.org/10.1021/la402346z

Faller A, Schünke M, Schünke G, et al., 2012, Fortpflanzung, Entwikclung und Geburt [in German]. Reproduction, development and birth. in Der Körper des Menschen, Stuttgart: Georg Thieme Verlag, 16th edition, pp. 752ff, 2012.

Desoye G, Gauster M, Wadsack C, et al., 2011, Placental transport in pregnancy pathologies. Am J Clin Nutr, 94(6): 1896–1902. http://dx.doi.org/10.3945/ajcn.110.000851

Gallo L A, Barrett H L, Dekker N M, 2016, Review: Placental transport and metabolism of energy substrates in maternal obesity and diabetes. Placenta, 54: 59–67. http://dx.doi.org/10.1016/j.placenta.2016.12.006

Gaccioli F, Lager S, Powell T L, et al., 2012, Placental transport in response to altered maternal nutrition. J Dev Orig Health Dis, 4(2): 1–15. http://dx.doi.org/10.1017/S2040174412000529

Gaither K, Quraishi A N, Illsley N P, 2016, Diabetes alters the expression and activity of the human placental GLUT1 glucose transporter. J Clin Endocrinol Metab, 84(2): 695–701. http://dx.doi.org/10.1210/jcem.84.2.5438

Jansson T, Ekstrand Y, Wennergren M, et al., 2001, Placental glucose transport in gestational diabetes mellitus. Am J Obstet Gynecol, 184(2): 111–116. http://dx.doi.org/10.1067/mob.2001.108075

Miura S, Sato K, Kato-Negishi M, et al., 2015, Fluid shear triggers microvilli formation via mechanosensitive activation of TRPV6. Nat Commun, 6(12): 8871. http://dx.doi.org/10.1038/ncomms9871

Caplin J D, 2016, Utilizing microfluidic technology to replicate placental functions in a drug testing model. 2016. Global Congress on NanoEngineering for Medicine and Biology.

Chen S, Zhang Q, Nakamoto T, et al., 2016, Gelatin scaffolds with controlled pore structure and mechanical property for cartilage tissue engineering. Tissue Eng Part C Methods, 22(3): 189–198.

Gorgieva S, Kokol V, 2011, Biomaterials and their biocompatibility: Review and perspectives. InTech, 1–36.

Markovic M, Van Hoorick J, Hölzl K, et al., 2015, Hybrid tissue engineering scaffolds by combination of three-dimensional printing and cell photoencapsulation. J Nanotechnol Eng Med, 6(2): 1–7. http://dx.doi.org/10.1115/1.4031466

Van Hoorick J, Gruber P, Markovic M, et al., 2018, Highly reactive thiol-norbornene photo-click hydrogels: Toward improved processability. Macromolecular Rapid Commun: 1800181, http://dx.doi.org/10.1002/marc.201800181

Nichol J W, Koshy S T, Bae H, et al., 2010, Cell-laden microengineered gelatin methacrylate hydrogels. Biomaterials, 31(21): 5536–5544. http://dx.doi.org/10.1016/j.biomaterials.2010.03.064

Maquoi E, Noel A, Foidart J M, 1997, Matrix metallo­proteinases in choriocarcinoma cell lines: A potential regulatory role of extracellular matrix components. in Placental Molecules in Hemodynamics, Transport, and Cellular Regulation, T. Hata, M. Takayama, I. Taki, and J.-M. Foidart, pp. 585, 1997.

Ruoslahti E, Pierschbacher M D, 1897, New perspectives in cell adhesion: RGD and integrins. Am Assoc Adv Sci, 238(4826): 491–497. http://dx.doi.org/10.1126/science.2821619

PeproTech, 2014, Endothelial cell media-maintenance media for endothelial cells.

Seeger J M, Klingman N, et al., 1985, Improved endothelial cell seeding with cultured cells and fibronectin-coated grafts. J Surg Res, 38(6): 641–647.

Ruoslahti E, 1984, Fibronectin in cell adhesion and invasion. Cancer Metastasis Rev, 3(1): 43–51.

Wang Q, 2017, Fabrication of photo-mediated biomaterial scaffolds. in Smart Materials for Tissue Engineering: Fundamental Principles, Q. Wang, Ed. 2017.

Ren K, Zhou J, Wu H, 2013, Materials for microfluidic chip fabrication. Acc Chem Res, 46(11): 2396–2406. https://dx.doi.org/10.1021/ar300314s

Dendukuri D, Panda P, Haghgooie R, et al., 2008, Modeling of oxygen-inhibited free radical photopolymerization in a PDMS microfluidic device. Macromolecules, 41(22): 8547–8556.

Altannavch T S, Roubalová K, Era P K U Č, 2004, Effect of high glucose concentrations on expression of ELAM-1, VCAM-1 and ICAM-1 in HUVEC with and without cytokine activation. Physiol Res, 53: 77–82. Avaliable from: http://citeseerx.ist.psu.edu/viewdoc/download?doi=

DOI: http://dx.doi.org/10.18063/ijb.v4i2.144


  • There are currently no refbacks.

Copyright (c) 2018 Denise Mandt, Peter Gruber, Marica Markovic, Maximillian Tromayer, Mario Rothbauer, Sebastian Rudi Adam Krayz, Faheem Ali, Jasper van Hoorick, Wolfgang Holnthoner, Severin Muhleder, Peter Dubruel, Sandra van Vlierberghe, Peter Ertl, Robert Liska, Aleksandr Ovsianikov

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.