Microextrusion printing cell-laden networks of type I collagen with patterned fiber alignment and geometry

Nerger, Bryan A.; Brun, Pierre-Thomas; Nelson, Celeste M.

doi:10.1039/c8sm02605j

Cited by 90 publications

(95 citation statements)

References 45 publications

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“…More prescriptive tissue engineering scaffolds built through 3D bioprinting of cells and ECM, or subtractive hollowing of hydrogels can potentially impose cues for long‐range tissue organization. [ 6–9 ] However, such scaffolds do not yet achieve the progressive elaboration of cell and ECM position, density, and composition over time that integrates tissue structure from the cellular to organ scales. The ability to mimic this could bring powerful advances in tissue engineering.…”

Section: Figurementioning

confidence: 99%

Guiding Cell Network Assembly using Shape‐Morphing Hydrogels

et al. 2020

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Forces and relative movement between cells and extracellular matrix (ECM) are crucial to the self‐organization of tissues during development. However, the spatial range over which these dynamics can be controlled in engineering approaches is limited, impeding progress toward the construction of large, structurally mature tissues. Herein, shape‐morphing materials called “kinomorphs” that rationally control the shape and size of multicellular networks are described. Kinomorphs are sheets of ECM that change their shape, size, and density depending on patterns of cell contractility within them. It is shown that these changes can manipulate structure‐forming behaviors of epithelial cells in many spatial locations at once. Kinomorphs are built using a new photolithographic technology to pattern single cells into ECM sheets that are >10× larger than previously described. These patterns are designed to partially mimic the branch geometry of the embryonic kidney epithelial network. Origami‐inspired simulations are then used to predict changes in kinomorph shapes. Last, kinomorph dynamics are shown to provide a centimeter‐scale program that sets specific spatial locations in which ≈50 µm‐diameter epithelial tubules form by cell coalescence and structural maturation. The kinomorphs may significantly advance organ‐scale tissue construction by extending the spatial range of cell self‐organization in emerging model systems such as organoids.

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Section: Figurementioning

confidence: 99%

Guiding Cell Network Assembly using Shape‐Morphing Hydrogels

et al. 2020

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“…Therefore, the low-grade enzymatic crosslinking of the THA preserved at least in part the capability of the fibrils to comply with the shear stimulus when extruded. Shear-induced alignment of fibrillar structures has been reported by different groups [34,35]. Kim et al controlled the fiber alignment in dependence of the needle diameter during printing showing a higher degree of alignment when using 30G compared to 20G needles [34].…”

Section: Discussionmentioning

confidence: 92%

Tissue mimetic hyaluronan bioink containing collagen fibers with controlled orientation modulating cell morphology and alignment

Schwab

Hélary

Richards

et al. 2020

Preprint

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Biofabrication is providing scientists and clinicians the ability to produce engineered tissues with desired shapes, chemical and biological gradients. Typical resolutions achieved with extrusion-based bioprinting are at the macroscopic level. However, for capturing the fibrillar nature of the extracellular matrix (ECM), it is necessary to arrange ECM components at smaller scales, down to the sub-micron and the molecular level. In this study, we introduce a (bio)ink containing hyaluronan (HA) as tyramine derivative (THA) and collagen (Col). Similarly to other connective tissues, in this (bio)ink Col is present in fibrillar form and HA as viscoelastic space filler. THA was enzymatically crosslinked under mild conditions allowing simultaneous Col fibrillogenesis, thus achieving a homogeneous distribution of Col fibrils within the viscoelastic HA-based matrix. THA-Col composite displayed synergistic properties in terms of storage modulus and shear-thinning, translating into good printability. Shear-induced alignment of the Col fibrils along the printing direction was achieved and quantified via immunofluorescence and second harmonic generation. Cell-free and cell-laden constructs were printed and characterized, analyzing the influence of the controlled microscopic anisotropy on cell behavior and chondrogenic differentiation. THA-Col showed cell instructive properties modulating hMSC adhesion, morphology and sprouting from spheroids stimulated by the presence and the orientation of Col fibers. Actin filament staining showed that hMSCs embedded into aligned constructs displayed increased cytoskeleton alignment along the fibril direction. Based on gene expression of cartilage/bone markers and matrix production, hMSCs embedded into the bioink displayed chondrogenic differentiation comparable or superior to standard pellet culture by means of proteoglycan production (Safranin O staining and proteoglycan quantification) as well as increase in cartilage related genes. The possibility of printing matrix components with control over microscopic alignment brings biofabrication one step closer to capturing the complexity of native tissues.

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“…), contact guiding 53,104,105 (Based on the adsorption capacity of collagen on the surface of substrate, oriented fibers were prepared), wet spinning, 106 and liquid-crystal mechanism preparation. 107 It is worth mentioning that, it has gradually been applied in the preparation of oriented fibers, [108][109][110] with the development of 3D printing technology; however, the degree of fiber orientation needs to be further improved. Compared with mechanical method, nonmechanical method, such as electrospinning, magnetic field, and so on, can set all kinds of parameters accurately and conveniently, to control the change of fiber orientation.…”

Section: Nonmechanical Factorsmentioning

confidence: 99%

Mechanical Roles in Formation of Oriented Collagen Fibers

Lin

Shi

Men

et al. 2020

Tissue Engineering Part B: Reviews

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Collagen is a structural protein that is widely present in vertebrates, being usually distributed in tissues in the form of fibers. In living organisms, fibers are organized in different orientations in various tissues. As the structural base in connective tissue and load-bearing tissue, the orientation of collagen fibers plays an extremely important role in the mechanical properties and physiological and biochemical functions. The study on mechanics role in formation of oriented collagen fibers enables us to understand how discrete cells use limited molecular materials to create tissues with different structures, thereby promoting our understanding of the mechanism of tissue formation from scratch, from invisible to tangible. However, the current understanding of the mechanism of fiber orientation is still insufficient. In addition, existing fabrication methods of oriented fibers are varied and involve interdisciplinary study, and the achievements of each experiment are favorable to the construction and improvement of the fiber orientation theory. To this end, this review focuses on the preparation methods of oriented fibers and proposes a model explaining the formation process of oriented fibers in tendons based on the existing fiber theory.

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Microextrusion printing cell-laden networks of type I collagen with patterned fiber alignment and geometry

Abstract: Cell-laden networks of aligned type I collagen fibers are fabricated using 3D microextrusion printing of collagen-Matrigel inks.

Cited by 90 publications

References 45 publications

Guiding Cell Network Assembly using Shape‐Morphing Hydrogels

Guiding Cell Network Assembly using Shape‐Morphing Hydrogels

Tissue mimetic hyaluronan bioink containing collagen fibers with controlled orientation modulating cell morphology and alignment

Mechanical Roles in Formation of Oriented Collagen Fibers

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