Patterned photocrosslinking to establish stiffness anisotropies in fibrous 3D hydrogels

Jagiełło, Alicja; Hu, Qingda; Castillo, Ulysses; Botvinick, Elliot L.

doi:10.1016/j.actbio.2021.12.028

Cited by 13 publications

(14 citation statements)

References 46 publications

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“…To really assess cell sensitivity to these micron-scaled stiffness heterogeneities, one major concern is to decouple the surface density of the adhesive ligands on the substrate from the stiffness pattern. This decoupling was not addressed in former studies where the materials were either permissive to cell adhesion [21,23,25] or not investigated [24]. We thus propose an original protocol for the surface functionalization of polyacrylamide hydrogels that allow to robustly control the uniformity of the adhesive coating in the presence of a rigidity pattern.…”

Section: Introductionmentioning

confidence: 95%

“…Complex nanoand micro-scale patterns of rigidity, with very sharp kilopascals per micrometers (kPa/µm), and gradients at the subcellular scales, have been evidenced in human tissues [19,20], as it is expected from the composition of an extracellular environment that consists of an entanglement of different molecular components with different mechanical properties [5]. Recently, few technologies have addressed the design of theses sharp gradients and their impact on cells, using appositions of materials with distinct mechanical properties [21,22], photodegradation of chemical bonds [23], electron-beam induced reticulation [24], or a posteriori photoreticulation of regions in a well formed 3D scaffold [25]. From these studies, stiffness patterning appears to influence cell behavior.…”

Section: Introductionmentioning

confidence: 99%

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Cells on Hydrogels with Micron-Scaled Stiffness Patterns Demonstrate Local Stiffness Sensing

et al. 2022

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Cell rigidity sensing—a basic cellular process allowing cells to adapt to mechanical cues—involves cell capabilities exerting force on the extracellular environment. In vivo, cells are exposed to multi-scaled heterogeneities in the mechanical properties of the surroundings. Here, we investigate whether cells are able to sense micron-scaled stiffness textures by measuring the forces they transmit to the extracellular matrix. To this end, we propose an efficient photochemistry of polyacrylamide hydrogels to design micron-scale stiffness patterns with kPa/µm gradients. Additionally, we propose an original protocol for the surface coating of adhesion proteins, which allows tuning the surface density from fully coupled to fully independent of the stiffness pattern. This evidences that cells pull on their surroundings by adjusting the level of stress to the micron-scaled stiffness. This conclusion was achieved through improvements in the traction force microscopy technique, e.g., adapting to substrates with a non-uniform stiffness and achieving a submicron resolution thanks to the implementation of a pyramidal optical flow algorithm. These developments provide tools for enhancing the current understanding of the contribution of stiffness alterations in many pathologies, including cancer.

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Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Cells on Hydrogels with Micron-Scaled Stiffness Patterns Demonstrate Local Stiffness Sensing

et al. 2022

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“…It was found that the cytoskeleton and nucleus align along the fibril axes and that transforming growth factor beta 1 (TGF-β1) is upregulated in the cells cultured on aligned fibrils: these cells also underwent EMT in response to aligned fibril in the polymer scaffold [85]. Besides, Jagiello et al [86] presented a 3D fibrous hydrogel with tunable local stiffness and fibril anisotropy to investigate the response of cancer cells to mechanical cues by using a cell-safe method of patterned photocrosslinking which is termed rutheniumcatalyzed photocrosslinking (RCP), and assessed the relationships between fibril alignment and stiffness by using multi-axes optical tweezers active microrheology (AMR) [87].…”

Section: 14mentioning

confidence: 99%

“…Besides, Jagiello et al . [86] presented a 3D fibrous hydrogel with tunable local stiffness and fibril anisotropy to investigate the response of cancer cells to mechanical cues by using a cell‐safe method of patterned photocrosslinking which is termed ruthenium‐catalyzed photocrosslinking (RCP), and assessed the relationships between fibril alignment and stiffness by using multi‐axes optical tweezers active microrheology (AMR) [87].…”

Section: In Vitro Construction Of the Tumor Mechanical Microenvironmentmentioning

confidence: 99%

Functions and clinical significance of mechanical tumor microenvironment: cancer cell sensing, mechanobiology and metastasis

Zhou

Wang

Zhang

et al. 2022

Cancer Communications

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Dynamic and heterogeneous interaction between tumor cells and the surrounding microenvironment fuels the occurrence, progression, invasion, and metastasis of solid tumors. In this process, the tumor microenvironment (TME) fractures cellular and matrix architecture normality through biochemical and mechanical means, abetting tumorigenesis and treatment resistance. Tumor cells sense and respond to the strength, direction, and duration of mechanical cues in the TME by various mechanotransduction pathways. However, far less understood is the comprehensive perspective of the functions and mechanisms of mechanotransduction. Due to the great therapeutic difficulties brought by the mechanical changes in the TME, emerging studies have focused on targeting the adverse mechanical factors in the TME to attenuate disease rather than conventionally targeting tumor cells themselves, which has been proven to be a potential therapeutic approach. In this review, we discussed the origins and roles of mechanical factors in the TME, cell sensing, mechano‐biological coupling and signal transduction, in vitro construction of the tumor mechanical microenvironment, applications and clinical significance in the TME.

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“…Photopatterning, or photolithography, is another method for patterning the stiffness of hydrogels by spatially varying the light irradiance. Photopatterning can be performed in a straightforward manner by using a physical photomask ( Rape et al, 2015 ; Tomba and Villard, 2015 ; Kim et al, 2020 ; Li and Bratlie, 2021 ; Mgharbel et al, 2022 ), via direct laser writing ( Song et al, 2021 ; Jagiełło et al, 2022 ), or by spatially varying the light source itself ( Norris et al, 2016 ; Oh et al, 2022 ). The latter can be achieved using digital micromirror devices (DMDs), such as those contained in some video projectors.…”

Section: Introductionmentioning

confidence: 99%

Visible-Light Stiffness Patterning of GelMA Hydrogels Towards In Vitro Scar Tissue Models

Chalard

Dixon

Taberner

et al. 2022

Front. Cell Dev. Biol.

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Variations in mechanical properties of the extracellular matrix occurs in various processes, such as tissue fibrosis. The impact of changes in tissue stiffness on cell behaviour are studied in vitro using various types of biomaterials and methods. Stiffness patterning of hydrogel scaffolds, through the use of stiffness gradients for instance, allows the modelling and studying of cellular responses to fibrotic mechanisms. Gelatine methacryloyl (GelMA) has been used extensively in tissue engineering for its inherent biocompatibility and the ability to precisely tune its mechanical properties. Visible light is now increasingly employed for crosslinking GelMA hydrogels as it enables improved cell survival when performing cell encapsulation. We report here, the photopatterning of mechanical properties of GelMA hydrogels with visible light and eosin Y as the photoinitiator using physical photomasks and projection with a digital micromirror device. Using both methods, binary hydrogels with areas of different stiffnesses and hydrogels with stiffness gradients were fabricated. Their mechanical properties were characterised using force indentation with atomic force microscopy, which showed the efficiency of both methods to spatially pattern the elastic modulus of GelMA according to the photomask or the projected pattern. Crosslinking through projection was also used to build constructs with complex shapes. Overall, this work shows the feasibility of patterning the stiffness of GelMA scaffolds, in the range from healthy to pathological stiffness, with visible light. Consequently, this method could be used to build in vitro models of healthy and fibrotic tissue and study the cellular behaviours involved at the interface between the two.

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Patterned photocrosslinking to establish stiffness anisotropies in fibrous 3D hydrogels

Cited by 13 publications

References 46 publications

Cells on Hydrogels with Micron-Scaled Stiffness Patterns Demonstrate Local Stiffness Sensing

Cells on Hydrogels with Micron-Scaled Stiffness Patterns Demonstrate Local Stiffness Sensing

Functions and clinical significance of mechanical tumor microenvironment: cancer cell sensing, mechanobiology and metastasis

Visible-Light Stiffness Patterning of GelMA Hydrogels Towards In Vitro Scar Tissue Models

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