The practice of conventional cell culture techniques affects the quality of biological research in mechanobiology by not replicating the cell’s microenvironment adequately. Microfluidic devices resembling 3D in vivo cell culture offer viable alternatives for mechanobiology studies and other applications including cell screening, cell separation, and point-of-care diagnostics. This review highlights recent advances in additive manufacturing (AM) to fabricate microfluidic and millifluidic devices through fast design iterations and cost reduction compared with conventional device manufacturing. Key advances in AM technologies such as fused deposition modeling (FDM), stereolithography apparatus (SLA), and digital light processing (DLP) have allowed the direct manufacture of complex microfluidic devices with master molds and sacrificial structures using multiple material components. Currently, available 3D printed devices for the precise fabrication of milli- and microfluidic devices, essential to mimicking the cellular microenvironment, while economically reducing production costs by eliminating expensive clean-room environments and reducing material waste are highlighted.
WalkerM, GodinM and PellingAE.A vacuum-actuated microtissue stretcher for long-term exposure to oscillatory strain within a 3D matrix. Biomed Microdevices2018; 20.
2.
SukiB, ParameswaranH, ImsirovicJ, . Regulatory roles of fluctuation-driven mechanotransduction in cell function. Physiology2016; 31: 346–358.
3.
GriffithCM, HuangSA, ChoC, . Microfluidics for the study of mechanotransduction. J Phys D Appl Phys2020; 53: 224004.
4.
SackmannEK, FultonAL and BeebeDJ.The present and future role of microfluidics in biomedical research. Nature2014; 507: 181–189.
5.
HakkinenKM, HarunagaJS, DoyleAD, . Direct comparisons of the morphology, migration, cell adhesions, and actin cytoskeleton of fibroblasts in four different three-dimensional extracellular matrices. Tissue Eng Part A2011; 17: 713–724.
6.
SantosA and LagaresD.Matrix stiffness: the conductor of organ fibrosis. Curr Rheumatol Rep2018; 20: 2–15.
7.
VillegasM, CetinicZ, ShakeriA, . Fabricating smooth PDMS microfluidic channels from low-resolution 3D printed molds using an omniphobic lubricant-infused coating. Anal Chim Acta2018; 1000: 248–255.
FioreVF, StranePW, BryksinAV, . Conformational coupling of integrin and Thy-1 regulates Fyn priming and fibroblast mechanotransduction. J Cell Biol2015; 211: 173–190.
12.
NakaoA, HasegawaY, TsuchiyaY, . Expression of cell adhesion molecules in the lungs of patients with idiopathic pulmonary fibrosis. Chest1995; 108: 233–239.
HerumK, LundeI, McCullochA, . The soft- and hard-heartedness of cardiac fibroblasts: mechanotransduction signaling pathways in fibrosis of the heart. J Clin Med2017; 6: 53.
15.
TrubeljaA and BaoG.Molecular mechanisms of mechanosensing and mechanotransduction in living cells. Ext Mech Lett2018; 20: 91–98.
16.
CastiauxAD, SpenceDM and MartinRS.Review of 3D cell culture with analysis in microfluidic systems. Anal Methods2019; 11: 4220–4232.
17.
LiW, ZhouJ and XuY.Study of the in vitro cytotoxicity testing of medical devices. Biomed Rep2015; 3: 617–620.
18.
HuhD, MatthewsBD, MammotoA, . Reconstituting organ-level lung functions on a chip. Science2010; 328: 1662–1668.
19.
HuhD, KimHJ, FraserJP, . Microfabrication of human organs-on-chips. Nat Protoc2013; 8: 2135–2157.
20.
AhmedI, SullivanK and PriyeA.multi-resin masked stereolithography (MSLA) 3D printing for rapid and inexpensive prototyping of microfluidic chips with integrated functional components. Biosensors2022; 12: 652.
21.
ChenC, TownsendAD, HayterEA, . Insert-based microfluidics for 3D cell culture with analysis. Anal Bioanal Chem2018; 410: 3025–3035.
22.
HeY, WuY, FuJZ, . Developments of 3D printing microfluidics and applications in chemistry and biology: a review. Electroanalysis2016; 28: 1658–1678.
23.
TonyA, BadeaI, YangC, . The additive manufacturing approach to polydimethylsiloxane (PDMS) microfluidic devices: review and future directions. Polymers2023; 15: 1926.
24.
SahaR, SarkarM, ChoudhurySS, . Evolution of 3d printing technology in fabrication of microfluidic devices and biological applications: a comprehensive review. J Micromanuf2024; 7: 110–140.
25.
OrrAW, HelmkeBP, BlackmanBR, . Mechanisms of mechanotransduction. Dev Cell2006; 10: 11–20.
HeY, MaoT, GuY, . A simplified yet enhanced and versatile microfluidic platform for cyclic cell stretching on an elastic polymer. Biofabrication2020; 12: 045032.
28.
StottlemireBJ, ChakravartiAR, WhitlowJW, . Remote-controlled 3D Porous magnetic interface toward high-throughput dynamic 3D cell culture. ACS Biomater Sci Eng2021; 9: 4535–4544.
BhardwajG AND WebsterTJ.Increased NIH 3T3 fibroblast functions on cell culture dishes which mimic the nanometer fibers of natural tissues. Int J Nanomed2015; 10: 5293–5299.
31.
LeeJN, JiangX, RyanD, . Compatibility of mammalian cells on surfaces of poly(dimethylsiloxane). Langmuir2004; 20: 11684–11691.
32.
XiaY and WhitesidesGM.Soft lithography. Angew Chem Int Ed1998; 37: 550–575.
ChenJ, LiuCY, WangX, . 3D printed microfluidic devices for circulating tumor cells (CTCs) isolation. Biosens Bioelectron2020; 150: 111900.
41.
SmoakMM, PearceHA and MikosAG.Microfluidic devices for disease modeling in muscle tissue. Biomaterials2019; 198: 250–258.
42.
KreutzerJ, ViehrigM, PölönenRP, . Pneumatic unidirectional cell stretching device for mechanobiological studies of cardiomyocytes. Biomech Model Mechanobiol2020; 19: 291–303.
43.
ShaoY, TanX, NovitskiR, . Uniaxial cell stretching device for live-cell imaging of mechanosensitive cellular functions. Rev Sci Instrum2013; 84: 114304.
44.
HelmkeBP and DaviesPF.Imaging Live Cells Under Mechanical Stress, http://repository.upenn.edu/ime_papershttp://repository.upenn.edu/ime_papers/10 (2003).
45.
HuangL, MathieuPS and HelmkeBP.A stretching device for high-resolution live-cell imaging. Ann Biomed Eng2010; 38: 1728–1740.
46.
NielsenAV, BeauchampMJ, NordinGP, . 3D printed microfluidics. Annu Rev Anal Chem2020; 13: 45–61.
47.
HoCMB, NgSH, LiKHH, . 3D printed microfluidics for biological applications. Lab Chip2015; 15: 3627–3637.
48.
RichmondT and TompkinsN.3D microfluidics in PDMS: manufacturing with 3D molding. Microfluid Nanofluid2021; 25: 76.
49.
MohamedMGA, KumarH, WangZ, . Rapid and inexpensive fabrication of multi-depth microfluidic device using high-resolution LCD stereolithographic 3D printing. J Manuf Mater Process2019; 3: 26–37.
50.
ShanklesPG, MilletLJ, AufrechtJA, . Accessing microfluidics through feature-based design software for 3D printing. PLoS ONE2018; 13: e0192752.
51.
Razavi BazazS, RouhiO, RaoufiMA, . 3D printing of inertial microfluidic devices. Sci Rep2020; 10: 5929.
52.
ShresthaJ, GhadiriM, ShanmugavelM, . A rapidly prototyped lung-on-a-chip model using 3D-printed molds. Organs-on-a-Chip2019; 1: 100001.
53.
LeeJM, ZhangM and YeongWY.Characterization and evaluation of 3D printed microfluidic chip for cell processing. Microfluid Nanofluid2016; 20: 1–15.
54.
OngLJY, IslamA, DasguptaR, . A 3D printed microfluidic perfusion device for multicellular spheroid cultures. Biofabrication2017; 9: 045005.
55.
BeckwithAL, BorensteinJT and Velasquez-GarciaLF.Monolithic, 3D-Printed microfluidic platform for recapitulation of dynamic tumor microenvironments. J Microelectromech Syst2018; 27: 1009–1022.
56.
Hernández-VarasP, BergeU, LockJG, . A plastic relationship between vinculin-mediated tension and adhesion complex area defines adhesion size and lifetime. Nat Commun2015; 6: 7524.
57.
ChanHN, ChenY, ShuY, . Direct, one-step molding of 3D-printed structures for convenient fabrication of truly 3D PDMS microfluidic chips. Microfluid Nanofluid2015; 19: 9–18.
58.
TremblayD, Chagnon-LessardS, MirzaeiM, . A microscale anisotropic biaxial cell stretching device for applications in mechanobiology. Biotechnol Lett2014; 36: 657–665.
59.
MataA, FleischmanAJ and RoyS.Characterization of polydimethylsiloxane (PDMS) properties for biomedical micro/nanosystems. Biomed Microdevices2005; 7: 281–293
60.
PalcheskoRN, ZhangL, SunY, . Development of polydimethylsiloxane substrates with tunable elastic modulus to study cell mechanobiology in muscle and nerve. PLoS ONE2012; 7: e51499.
61.
WangZ, VolinskyAA and GallantND.Crosslinking effect on polydimethylsiloxane elastic modulus measured by custom-built compression instrument. J Appl Polym Sci2014; 131.
62.
ChaC, AntoniadouE, LeeM, . Tailoring hydrogel adhesion to polydimethylsiloxane substrates using polysaccharide glue. Angew Chem Int Ed2013; 52: 6949–6952.
63.
ChuahYJ, KohYT, LimK, . Simple surface engineering of polydimethylsiloxane with polydopamine for stabilized mesenchymal stem cell adhesion and multipotency. Sci Rep2015; 5: 18162.
64.
ChaeSK, RyooJH, LeeSH.Thin and large free-standing PDMS membrane by using polystyrene Petri dish. Biochip J2012; 6: 184–190.
65.
Hernández VeraR, O’CallaghanP, Fatsis-KavalopoulosN, . Modular microfluidic systems cast from 3D-printed molds for imaging leukocyte adherence to differentially treated endothelial cultures. Sci Rep2019; 9: 11321.
66.
SaadatM, TaylorM, HughesA, . Rapid prototyping method for 3D PDMS microfluidic devices using a red femtosecond laser. Adv Mech Eng2020; 12: 168781402098271.
67.
CostaPF, AlbersHJ, LinssenJEA, . Mimicking arterial thrombosis in a 3D-printed microfluidic: In vitro vascular model based on computed tomography angiography data. Lab Chip2017; 17: 2785–2792.
68.
TherriaultD, WhiteSR and LewisJA.Chaotic mixing in three-dimensional microvascular networks fabricated by direct-write assembly. Nat Mater2003; 2: 265–271.
69.
LeeH, T-SJang, HanG, . Freeform 3D printing of vascularized tissues: challenges and strategies. J Tissue Eng2021; 12: 204173142110572.
70.
SaggiomoV and VeldersAH.Simple 3D printed scaffold-removal method for the fabrication of intricate microfluidic devices. Adv Sci2015; 2: 1500125.
71.
KangK, OhS, YiH, . Fabrication of truly 3D microfluidic channel using 3D-printed soluble mold. Biomicrofluidics2018; 12: 014105.
72.
GohWH and HashimotoM.Fabrication of 3D microfluidic channels and in-channel features using 3D printed, water-soluble sacrificial mold. Macromol Mater Eng2018; 303: 1700484.
73.
GonzálezG, RoppoloI, PirriCF, . Current and emerging trends in polymeric 3D printed microfluidic devices. Addit Manuf2022; 55: 102867.
74.
NelsonMD, RamkumarN and GaleBK.Flexible, transparent, sub-100 µm microfluidic channels with fused deposition modeling 3D-printed thermoplastic polyurethane. J Micromech Microeng2019; 29095010.
75.
UrriosA, Parra-CabreraC, BhattacharjeeN, . 3D-printing of transparent bio-microfluidic devices in PEG-DA. Lab Chip2016; 16: 2287–2294.
AlessandriK, FeyeuxM, GurchenkovB, . A 3D printed microfluidic device for production of functionalized hydrogel microcapsules for culture and differentiation of human Neuronal Stem Cells (hNSC). Lab Chip2016; 16: 1593–1604.
78.
HoKKY, WangYL, WuJ, . Advanced microfluidic device designed for cyclic compression of single adherent cells. Front Bioeng Biotechnol2018; 6:148.
79.
KreutzerJ, ViehrigM, A-JMaki, . Pneumatically actuated elastomeric device for simultaneous mechanobiological studies and live-cell fluorescent microscopy. In: 2017 International Conference on Manipulation, Automation and Robotics at Small Scales (MARSS). IEEE, pp. 1–5.
80.
KimHJ, HuhD, HamiltonG, . Human gut-on-a-chip inhabited by microbial flora that experiences intestinal peristalsis-like motions and flow. Lab Chip2012; 12: 2165–2174.
81.
MoraesC, ChenJH, SunY, . Microfabricated arrays for high-throughput screening of cellular response to cyclic substrate deformation. Lab Chip2010; 10: 227–234.
82.
LiuB, KimTJ and WangY.Live cell imaging of mechanotransduction. J R Soc Interface 2010; 7: S365–S375
83.
YeungT, GeorgesPC, FlanaganLA, . Effects of substrate stiffness on cell morphology, cytoskeletal structure, and adhesion. Cell Motil Cytoskel2005; 60: 24–34.
84.
Chagnon-LessardS, Jean-RuelH, GodinM, . Mechanotransduction of strain regulates an invasive phenotype in newly transformed epithelial cells. bioRxiv. Epub ahead of print 15 September 2019. DOI: 10.1101/770487.
85.
WuY, HuangY and GunstSJ.Focal adhesion kinase (FAK) and mechanical stimulation negatively regulate the transition of airway smooth muscle tissues to a synthetic phenotype. Am J Physiol Lung Cell Mol Physiol2016; 311: 893–902.
86.
ProvenzanoPP, InmanDR, EliceiriKW, . Matrix density-induced mechanoregulation of breast cell phenotype, signaling and gene expression through a FAK-ERK linkage. Oncogene2009; 28: 4326–4343.
87.
JungbauerS, GaoH, SpatzJP, . Two characteristic regimes in frequency-dependent dynamic reorientation of fibroblasts on cyclically stretched substrates. Biophys J2008; 95: 3470–3478.
88.
KumarA, PlaconeJK and EnglerAJ.Understanding the extracellular forces that determine cell fate and maintenance. Development (Cambridge)2017; 144: 4261–4270.
89.
UnalM, AlapanY, JiaH, . Micro and nano-scale technologies for cell mechanics. Nanobiomedicine2014; 1: 5.
90.
KanabekovaP, KadyrovaA and KulsharovaG.Microfluidic organ-on-a-chip devices for liver disease modeling in vitro. Micromachines2022; 13: 428.
91.
GuenatOT and BerthiaumeF.Incorporating mechanical strain in organs-on-a-chip: Lung and skin. Biomicrofluidics2018; 12(4): 042207
92.
MiltonLA, ViglioneMS, OngLJY, . Vat photopolymerization 3D printed microfluidic devices for organ-on-a-chip applications. Lab Chip2023; 23: 3537–3560.
KambleH, VadiveluR, BartonM, . An electromagnetically actuated double-sided cell-stretching device for mechanobiology research. Micromachines2017; 8(8): 256.
95.
AroushDRB and WagnerHD.Shear-stress profile along a cell focal adhesion. Adv Mater2006; 18: 1537–1540.
96.
BußmannA, ThalhoferT, HoffmannS, . Microfluidic cell transport with piezoelectric micro diaphragm pumps. Micromachines2021; 12(12): 1459.
97.
PathakA, DeshpandeVS, McMeekingRM, . The simulation of stress fibre and focal adhesion development in cells on patterned substrates. J R Soc Interface2008; 5: 507–524.
98.
BalachandranK, AlfordPW, Wylie-SearsJ, . Cyclic strain induces dual-mode endothelial-mesenchymal transformation of the cardiac valve. Proc Natl Acad Sci2011; 108: 19943–19948.
99.
GoldynAM, RiojaBA, SpatzJP, . Force-induced cell polarisation is linked to RhoA-driven microtubule-independent focal-adhesion sliding. J Cell Sci2009; 122: 3644–3651.
100.
AthirasalaA, HirschN and BuxboimA.Nuclear mechanotransduction: sensing the force from within. Curr Opin Cell Biol2017; 46: 119–127.
101.
GhazizadehH and AravamudhanS.A review of two-dimensional in vitro cell stretch platforms. J Mech Med Biol2017; 8: 1730003.
102.
BabicJ, GriscomL, CramerJ, . An easy-to-build and re-usable microfluidic system for live-cell imaging. BMC Cell Biol2018; 19: 8.
103.
RothenbergKE, NeibartSS, LaCroixAS, . Controlling cell geometry affects the spatial distribution of load across vinculin. Cell Mol Bioeng2015; 8: 364–382.
104.
de Almeida Monteiro Melo FerrazM, NagashimaJB, VenzacB, . 3D printed mold leachates in PDMS microfluidic devices. Sci Rep2020; 10: 994.
105.
YoungOM, XuX, SarkerS, . Direct laser writing-enabled 3D printing strategies for microfluidic applications. Lab Chip2024; 24: 2371–2396.
