Functional characterization of 3D contractile smooth muscle tissues generated using a unique microfluidic 3D bioprinting technology

Abstract

Conditions such as asthma and inflammatory bowel disease are characterized by aberrant smooth muscle contraction. It has proven difficult to develop human cellbased models that mimic acute muscle contraction in 2D in vitro cultures due to the nonphysiological chemical and mechanical properties of lab plastics that do not allow for muscle cell contraction. To enhance the relevance of in vitro models for human disease, we describe how functional 3D smooth muscle tissue that exhibits physiological and pharmacologically relevant acute contraction and relaxation responses can be reproducibly fabricated using a unique microfluidic 3D bioprinting technology. Primary human airway and intestinal smooth muscle cells were printed into rings of muscle tissue at high density and viability. Printed tissues contracted to physiological concentrations of histamine (0.01-100 μM) and relaxed to salbutamol, a pharmacological compound used to relieve asthmatic exacerbations. The addition of TGFβ to airway muscle rings induced an increase in unstimulated muscle shortening and a decreased response to salbutamol, a phenomenon which also occurs in chronic lung diseases. Results indicate that the 3D bioprinted smooth muscle is a physiologically relevant in vitro model that can be utilized to study disease pathways and the effects of novel therapeutics on acute contraction and chronic tissue stenosis.

Cell-Seeded Biomaterial Scaffolds: The Urgent Need for Unanswered Accelerated Angiogenesis

“…In this section, we review the different types of implanted cells within scaffolds that can achieve vascularization. To the best of our knowledge, the co-culture of primary basal-like cells with HUVECs 141 has been reported to be a potent cell source for vasculature branching. The results achieved by Ren et al related that seeding a lung scaffold with endothelial and perivascular cells derived from induced pluripotent stem cells can lead to ~75% endothelial coverage.…”

Microvascular Tissue Engineering—A Review

“…On the other hand, the main limitation of extrusion-based bioprinting is the spatial resolution, which is defined by the mechanical translation and the nozzle diameter, which typically lies in the 100 µm range [ 5 , 6 , 107 , 108 , 109 ]. Through the use of multiple printheads, and especially with the introduction of microfluidic and multicomponent extrusion systems, extrusion-based bioprinting allows spatial control over the chemical composition of scaffolds [ 111 , 112 , 113 , 114 ].…”

“…With the development of new materials and process optimization, it has become possible to 3D print complex structures using live cells [ 132 ]. However, to date, SLA remains inferior to extrusion-based in terms of material range, especially internal chemical gradients [ 111 , 112 , 113 , 114 , 132 ].…”

Tools, techniques, and future opportunities for characterizing the mechanobiology of uterine myometrium

“…99,104 3-D muscle tissues have been generated by casting cell-hydrogel mixtures into custom molds 105–107 or by leveraging new advances in bioprinting. 108–110 The contractility of these 3-D tissues has been measured by attaching tissues to a force transducer 105 or by monitoring the deflection of tissue anchor points. 111,112 Although these approaches for patterning tissues and quantifying contractility have yet to be translated to myometrial smooth muscle cells, they are likely compatible and could be implemented as new paradigms for engineering in vitro models of human myometrium that recapitulate key features of native tissue structure, such as alignment and matrix rigidity, while also facilitating rigorous functional measurements at scale.…”