Hydrogel mechanics regulate fibroblast DNA methylation and chromatin condensation

Sumey, Jenna L.; Johnston, Peyton C; Harrell, Abigail M; Caliari, Steven R.

doi:10.1039/d2bm02058k

Cited by 8 publications

(6 citation statements)

References 72 publications

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“…Hyaluronic acid (HA) hydrogels have seen widespread use for cell culture due to their biological relevance and their ability to be highly modified with various functional groups, including a range of cross-linking handles and bioactive moieties . For example, various modes of HA cross-linking have enabled orthogonal control over mechanical properties ,− as well as in situ stiffening. , Further, the independent control of bioactive cues has enabled investigation into the distinct roles of integrin engagement and mechanics on cell behavior. , Given that HA hydrogels permit a high degree of tunability, there is a need for hydrogel fabrication approaches that allow for combinatorial testing of the many independent cell-instructive variables that may be achieved with increased throughput.…”

Section: Resultsmentioning

confidence: 99%

“… 20 For example, various modes of HA cross-linking have enabled orthogonal control over mechanical properties 18 , 21 − 23 as well as in situ stiffening. 24 , 25 Further, the independent control of bioactive cues has enabled investigation into the distinct roles of integrin engagement and mechanics on cell behavior. 6 , 26 Given that HA hydrogels permit a high degree of tunability, there is a need for hydrogel fabrication approaches that allow for combinatorial testing of the many independent cell-instructive variables that may be achieved with increased throughput.…”

Section: Resultsmentioning

confidence: 99%

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Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

Skelton,

Gentry,

Astrab

et al. 2024

ACS Biomater. Sci. Eng.

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Hydrogels have gained significant popularity as model platforms to study reciprocal interactions between cells and their microenvironment. While hydrogel tools to probe many characteristics of the extracellular space have been developed, fabrication approaches remain challenging and time-consuming, limiting multiplexing or widespread adoption. Thus, we have developed a modular fabrication approach to generate distinct hydrogel microenvironments within the same 96-well plate for increased throughput of fabrication as well as integration with existing high-throughput assay technologies. This approach enables in situ hydrogel mechanical characterization and is used to generate both elastic and viscoelastic hydrogels across a range of stiffnesses. Additionally, this fabrication method enabled a 3-fold reduction in polymer and up to an 8-fold reduction in fabrication time required per hydrogel replicate. The feasibility of this platform for two-dimensional (2D) cell culture applications was demonstrated by measuring both population-level and single-cell-level metrics via microplate reader and high-content imaging. Finally, a 96-well hydrogel array was utilized for three-dimensional (3D) cell culture, demonstrating the ability to support high cell viability. Together, this work demonstrates a versatile and easily adaptable fabrication approach that can support the ever-expanding tool kit of hydrogel technologies for cell culture applications.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

Skelton,

Gentry,

Astrab

et al. 2024

ACS Biomater. Sci. Eng.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Hyaluronic acid (HA) hydrogels have seen widespread use for cell culture due to their biological relevance and their ability to be highly modified with various functional groups, including a range of crosslinking handle s and bioactive moieties 16 . For example, various modes of HA crosslinking have enabled orthogonal control over mechanical properties 14,17–19 as well as in situ stiffening 20,21 . Further, the independent control of bioactive cues has enabled investigation into the distinct roles of integrin engagement and mechanics on cell behavior 6,22 .…”

Section: Resultsmentioning

confidence: 99%

Modular multiwell viscoelastic hydrogel platform for two- and three-dimensional cell culture applications

Skelton,

Gentry,

Astrab

et al. 2023

Preprint

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Hydrogels have gained significant popularity as model platforms to study the reciprocal interactions between cells and their microenvironment. While hydrogel tools to probe many characteristics of the extracellular space have been developed, fabrication approaches remain challenging and time-consuming, limiting multiplexing or widespread adoption. Thus, we have developed a modular fabrication approach to generate distinct hydrogel microenvironments within 96-well plates for increased throughput of fabrication as well as integration with existing high-throughput assay technologies. This approach enablesin situhydrogel mechanical characterization and was used to generate both elastic and viscoelastic hydrogels across a range of stiffnesses. Additionally, this fabrication method enabled a 3-fold reduction in polymer and up to an 8-fold reduction in fabrication time required per hydrogel replicate. The feasibility of this platform for cell culture applications was demonstrated by measuring both population-level and single cell-level metrics via microplate reader and high-content imaging. Finally, the 96-well hydrogel array was utilized for 3D cell culture, demonstrating the ability to support high cell viability. Together, this work demonstrates a versatile and easily adoptable fabrication approach that can support the ever-expanding tool kit of hydrogel technologies for cell culture applications.

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“…Hydrogels require crosslinking for assembly, which can be mediated by either covalent or physical interactions. In addition to naturally occurring physical hydrogels like gelatin, collagen, and alginate, synthetic and natural polymers have been grafted with moieties that physically interact to create robust hydrogels [Wong Po Foo et al, 2009;Rodell et al, 2013;Dånmark et al, 2016;Clarke et al, 2017;Mann et al, 2017;Ding et al, 2018;Sumey et al, 2023]. Another physical gelation mechanism builds on supramolecular assembly of small peptide molecules, in some cases only 2-5 amino acids long, that create fiber networks held together by physical interactions such as hydrogen bonding and pipi stacking [Hartgerink et al, 2001;Schneider et al, 2002;Jayawarna et al, 2006;Haines-Butterick et al, 2007;Smith et al, 2008;Tang et al, 2019a].…”

Section: Introductionmentioning

confidence: 99%

Effects of Cell-Adhesive Ligand Presentation on Pentapeptide Supramolecular Assembly and Gelation: Simulations and Experiments

Thede,

Tang,

Cocker

et al. 2023

Cells Tissues Organs

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The extracellular matrix (ECM) is a complex, hierarchical material containing structural and bioactive components. This complexity makes decoupling the effects of biomechanical properties and cell-matrix interactions difficult, especially when studying cellular processes in a 3D environment. Matrix mechanics and cell adhesion are both known regulators of specific cellular processes such as stem cell proliferation and differentiation. However, more information is required about how such variables impact various neural lineages that could, upon transplantation, therapeutically improve neural function after central nervous system (CNS) injury or disease. Rapidly Assembling Pentapeptides for Injectable Delivery (RAPID) hydrogels are one biomaterial approach to meet these goals, consisting of a family of peptide sequences that assemble into physical hydrogels in physiological media. In this study, we studied our previously reported supramolecularly-assembling RAPID hydrogels functionalized with the ECM-derived cell adhesive peptide ligands RGD, IKVAV, and YIGSR. Using molecular dynamics simulations and experimental rheology we demonstrated that these integrin-binding ligands at physiological concentrations (3-12 mM) did not impact assembly of the KYFIL peptide system. In simulations, molecular measures of assembly such as hydrogen bonding and pi-pi interactions appeared unaffected by cell-adhesion sequence or concentration. Visualizations of clustering and analysis of solvent accessible surface area (SASA) indicated that the integrin-binding domains remained exposed. KYFIL or AYFIL hydrogels containing 3 mM of integrin-binding domains resulted in mechanical properties consistent with their non-functionalized equivalents. This strategy of doping RAPID gels with cell-adhesion sequences allows for the precise tuning of peptide ligand concentration, independent of the rheological properties. The controllability of the RAPID hydrogel system provides an opportunity to investigate the effect of integrin-binding interactions on encapsulated neural cells to discern how hydrogel microenvironment impacts growth, maturation, or differentiation.

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Hydrogel mechanics regulate fibroblast DNA methylation and chromatin condensation

Abstract: Cellular mechanotransduction plays a central role in fibroblast activation during fibrotic disease progression, leading to increased tissue stiffness and reduced organ function. While the role of epigenetics in disease mechanotransduction...

Cited by 8 publications

References 72 publications

Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

Modular multiwell viscoelastic hydrogel platform for two- and three-dimensional cell culture applications

Effects of Cell-Adhesive Ligand Presentation on Pentapeptide Supramolecular Assembly and Gelation: Simulations and Experiments

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