bims-ecemfi 2024-11-17 papers

bims-ecemfi

Biomed News

on ECM and fibroblasts

Issue of 2024–11–17
thirteen papers selected by
Badri Narayanan Narasimhan, University of California, San Diego

Cell response to extracellular matrix viscous energy dissipation outweighs high-rigidity sensing.
Viscoelastic High-Molecular-Weight Hyaluronic Acid Hydrogels Support Rapid Glioblastoma Cell Invasion with Leader-Follower Dynamics.
Plasticity of 3D Hydrogels Predicts Cell Biological Behavior.
Viscosity regulates cell spreading and cell-extracellular matrix interactions.
Hydrogel Elastic Energy: A Stressor Triggering an Adaptive Stress-Mediated Cell Response.
The Photocleavable Protein PhoCl-Based Dynamic Hydrogels.
Experimentally-driven mathematical model to understand the effects of matrix deprivation in breast cancer metastasis.
Leveraging Cell Migration Dynamics to Discriminate Between Senescent and Presenescent Human Mesenchymal Stem Cells.
Cell shape affects bacterial colony growth under physical confinement.
Photoinitiator-free light-mediated crosslinking of dynamic polymer and pristine protein networks.
Monophasic hyaluronic acid-silica hybrid hydrogels for articular cartilage applications.
Magnetoactive, Kirigami-Inspired Hammocks to Probe Lung Epithelial Cell Function.
Mechano-gradients drive morphogen-noise correction to ensure robust patterning.

Sci Adv. 2024 Nov 15. 10(46): eadf9758

Cell response to extracellular matrix viscous energy dissipation outweighs high-rigidity sensing.

Carla Huerta-López, Alejandro Clemente-Manteca, Diana Velázquez-Carreras, Francisco M Espinosa, Juan G Sanchez, Álvaro Martínez-Del-Pozo, María García-García, Sara Martín-Colomo, Andrea Rodríguez-Blanco, Ricardo Esteban-González, Francisco M Martín-Zamora, Luis I Gutierrez-Rus, Ricardo Garcia, Pere Roca-Cusachs, Alberto Elosegui-Artola, Miguel A Del Pozo, Elías Herrero-Galán, Pablo Sáez, Gustavo R Plaza, Jorge Alegre-Cebollada.

The mechanics of the extracellular matrix (ECM) determine cell activity and fate through mechanoresponsive proteins including Yes-associated protein 1 (YAP). Rigidity and viscous relaxation have emerged as the main mechanical properties of the ECM steering cell behavior. However, how cells integrate coexisting ECM rigidity and viscosity cues remains poorly understood, particularly in the high-stiffness regime. Here, we have exploited engineered stiff viscoelastic protein hydrogels to show that, contrary to current models of cell-ECM interaction, substrate viscous energy dissipation attenuates mechanosensing even when cells are exposed to higher effective rigidity. This unexpected behavior is however readily captured by a pull-and-hold model of molecular clutch-based cell mechanosensing, which also recapitulates opposite cellular response at low rigidities. Consistent with predictions of the pull-and-hold model, we find that myosin inhibition can boost mechanosensing on cells cultured on dissipative matrices. Together, our work provides general mechanistic understanding on how cells respond to the viscoelastic properties of the ECM.

DOI: https://doi.org/10.1126/sciadv.adf9758
Adv Mater. 2024 Nov 07. e2404885

Viscoelastic High-Molecular-Weight Hyaluronic Acid Hydrogels Support Rapid Glioblastoma Cell Invasion with Leader-Follower Dynamics.

Emily M Carvalho, Erika A Ding, Atul Saha, Diana Cruz Garcia, Anna Weldy, Peter-James H Zushin, Andreas Stahl, Manish K Aghi, Sanjay Kumar.

  Hyaluronic acid (HA), the primary component of brain extracellular matrix, is increasingly used to model neuropathological processes, including glioblastoma (GBM) tumor invasion. While elastic hydrogels based on crosslinked low-molecular-weight (LMW) HA are widely exploited for this purpose and have proven valuable for discovery and screening, brain tissue is both viscoelastic and rich in high-MW (HMW) HA, and it remains unclear how these differences influence invasion. To address this question, hydrogels comprised of either HMW (1.5 MDa) or LMW (60 kDa) HA are introduced, characterized, and applied in GBM invasion studies. Unlike LMW HA hydrogels, HMW HA hydrogels relax stresses quickly, to a similar extent as brain tissue, and to a greater extent than many conventional HA-based scaffolds. GBM cells implanted within HMW HA hydrogels invade much more rapidly than in their LMW HA counterparts and exhibit distinct leader-follower dynamics. Leader cells adopt dendritic morphologies similar to invasive GBM cells observed in vivo. Transcriptomic, pharmacologic, and imaging studies suggest that leader cells exploit hyaluronidase, an enzyme strongly enriched in human GBMs, to prime a path for followers. This study offers new insight into how HA viscoelastic properties drive invasion and argues for the use of highly stress-relaxing materials to model GBM.

Keywords:  glioblastoma (GBM); high‐molecular‐weight (HMW); hyaluronic acid (HA); stress‐relaxing; viscoelastic

DOI:  https://doi.org/10.1002/adma.202404885
Biomacromolecules. 2024 Nov 08.

Plasticity of 3D Hydrogels Predicts Cell Biological Behavior.

Andrea Malandrino, Huijun Zhang, Nico Schwarm, David Böhringer, Delf Kah, Christian Kuster, Aldo R Boccaccini, Ben Fabry.

Under 3D culture conditions, cells tend to spread, migrate, and proliferate better in more viscoelastic and plastic hydrogels. Here, we present evidence that the improved cell behavior is facilitated by the lower steric hindrance of a more viscoelastic and plastic matrix with weaker intermolecular bonds. To determine intermolecular bond stability, we slowly insert semispherical tipped needles (100-700 μm diameter) into alginate dialdehyde-gelatin hydrogels and measure stiffness, yield strength, plasticity, and the force at which the surface ruptures (puncture force). To tune these material properties without affecting matrix stiffness, we precross-link the hydrogels with CaCl2 droplets prior to mixing in NIH/3T3 fibroblasts and final cross-linking with CaCl2. Precross-linking introduces microscopic weak spots in the hydrogel, increases plasticity, and decreases puncture force and yield strength. Fibroblasts spread and migrate better in precross-linked hydrogels, demonstrating that intermolecular bond stability is a critical determinant of cell behavior under 3D culture conditions.

DOI: https://doi.org/10.1021/acs.biomac.4c00765
FEBS J. 2024 Nov 11.

Viscosity regulates cell spreading and cell-extracellular matrix interactions.

Hugh Xiao, Xiangyu Gong, Seyma Nayir Jordan, Zixie Liang, Michael Mak.

  Fluid viscosity and osmolarity are among some of the underappreciated mechanical stimuli that cells can detect. Abnormal changes of multiple fluidic factors such as viscosity and osmolarity have been linked with diseases such as cystic fibrosis, cancer, and coronary heart disease. Changes in viscosity have been recently suggested as a regulator of cell locomotion. These novel studies focus on cell migration and spreading on glass substrates and through microchannels, and it remains a question whether viscosity impacts the cellular remodeling of extracellular matrices (ECMs). Here, we demonstrate that elevated viscosity induces cellular remodeling of collagen substrates and enhances cell spreading on ECM-mimetic substrates. Our results expand on recent work showing that viscosity induces increased cellular forces and demonstrates that viscosity can drive local ECM densification. Our data further show that microtubules, Ras-related C3 botulinum toxin substrate 1 (Rac1), actin-related protein 2/3 (Arp2/3) complex, Rho-associated protein kinase 1 (ROCK), and myosin are important regulators of viscosity-induced ECM remodeling. In the context of viscosity-induced cell spreading, cells cultured on glass and collagen substrates exhibit markedly different responses to pharmacological treatments, indicating that microtubules, Rac1, and Arp2/3 play distinct roles in regulating cellular spreading depending on the substrate. In addition, our results demonstrate that high osmotic pressures override viscosity-induced cell spreading by suppressing membrane ruffling. Our results demonstrate viscosity as a regulator of ECM remodeling and cell spreading in a fibrillar microenvironment. We also reveal a complex interplay between viscosity and osmolarity. We anticipate that our research can pave the way for future investigations into the crucial roles played by viscosity in both physiological and pathological conditions.

Keywords:  ECM remodeling; collagen; cytoskeleton; osmolarity; viscosity

DOI:  https://doi.org/10.1111/febs.17306
Adv Healthc Mater. 2024 Nov 13. e2402400

Hydrogel Elastic Energy: A Stressor Triggering an Adaptive Stress-Mediated Cell Response.

Sara Lipari, Pasquale Sacco, Michela Cok, Francesca Scognamiglio, Maurizio Romano, Francesco Brun, Piero Giulio Giulianini, Eleonora Marsich, Finn L Aachmann, Ivan Donati.

  The crosstalk between the cells and the extracellular matrix (ECM) is bidirectional and consists of a pushing/pulling stretch exerted by the cells and a mechanical resistance counteracted by the surrounding microenvironment. It is widely recognized that the stiffness of the ECM, its viscoelasticity, and its overall deformation are the most important traits influencing the response of the cells. Here these three parameters are combined into a concept of elastic energy, which in biological terms represents the mechanical feedback that cells perceive when the ECM is deformed. It is shown that elastic energy is a stress factor that influences the response of cells in three-dimensional (3D) cultures. Strikingly, the higher the elastic energy of the matrix and thus the mechanical feedback, the higher the stress state of the cells, which correlates with the formation of G3BP-mediated stress granules. This condition is associated with an increase in alkaline phosphatase (ALP) activity but a decrease in gene expression and is mediated by the nuclear translocation of Yes-associated protein (YAP). This work supports the importance of considering the elastic energy as mechano-controller in regulating cellular stress state in 3D cultures.

Keywords:  cellular stress state; extracellular matrix mimic; hydrogel; mechanical feedback; mechanosensing

DOI:  https://doi.org/10.1002/adhm.202402400
ACS Biomater Sci Eng. 2024 Nov 15.

The Photocleavable Protein PhoCl-Based Dynamic Hydrogels.

Jingqi Lei, Hongbin Li.

  Dynamic protein hydrogels have attracted increasing attention owing to their tunable physiochemical and mechanical properties, customized functionality, and biocompatibility. Among the different types of dynamic hydrogels, photoresponsive hydrogels are of particular interest. Here, we report the engineering of a photoresponsive protein hydrogel by using the photocleavable protein PhoCl. We employed the well-developed SpyTag and SpyCatcher chemistry to engineer PhoCl-containing covalently cross-linked hydrogels. In the hydrogel network, PhoCl, which can be cleaved into two fragments upon violet irradiation, is employed as a dynamic structural motif to regulate the cross-linking density of the hydrogel network. The resultant PhoCl-containing hydrogels showed photoresponsive viscoelastic properties. Upon violet irradiation, the PhoCl hydrogels soften, leading to an irreversible reduction in the storage moduli. However, no gel-sol transition was observed. Leveraging this light-induced stiffness change, we employed this hydrogel as a cell culture substrate to investigate the mechanobiological response of NIH-3T3 fibroblast cells. Our results showed that 3T3 cells can change their morphologies in response to the stiffness change of the PhoCl hydrogel substrate dynamically, rendering PhoCl-based hydrogels a useful substrate for other mechanobiological studies.

Keywords:  3T3 cells; PhoCl-containing hydrogel; SpyTag and SpyCatcher chemistry; light-induced stiffness change; photoresponsive hydrogels

DOI:  https://doi.org/10.1021/acsbiomaterials.4c01584
NPJ Syst Biol Appl. 2024 Nov 12. 10(1): 132

Experimentally-driven mathematical model to understand the effects of matrix deprivation in breast cancer metastasis.

Sayoni Maiti, Annapoorni Rangarajan, Venkatesh Kareenhalli.

Normal epithelial cells receive proper signals for growth and survival from attachment to the underlying extracellular matrix (ECM). They perceive detachment from the ECM as a stress and die - a phenomenon termed as 'anoikis'. However, metastatic cancer cells acquire anoikis-resistance and circulate through the blood and lymphatics to seed metastasis. Under normal (adherent) growth conditions, the serine-threonine protein kinase Akt stimulates protein synthesis and cell growth, maintaining an anabolic state in the cancer cell. In contrast, previously we showed that the stress due to matrix deprivation is sensed by yet another serine-threonine kinase, AMP-activated protein kinase (AMPK), that inhibits anabolic pathways while promoting catabolic processes. We illustrated a switch from Akthigh/AMPKlow in adherent condition to AMPKhigh/Aktlow in matrix-detached condition, with consequent metabolic switching from an anabolic to a catabolic state, which aids cancer cell stress-survival. In this study, we utilized these experimental data and developed a deterministic ordinary differential equation (ODE)-based mechanistic mathematical model to mimic attachment-detachment signaling network. To do so, we used the framework of insulin-glucagon signaling with consequent metabolic shifts to capture the pathophysiology of matrix-deprived state in breast cancer cells. Using the developed metastatic breast cancer signaling (MBCS) model, we identified perturbation of several signaling proteins such as IRS, PI3K, PKC, GLUT1, IP3, DAG, PKA, cAMP, and PDE3 upon matrix deprivation. Further, in silico molecular perturbations revealed that several feedback/crosstalks like DAG to PKC, PKC to IRS, S6K1 to IRS, cAMP to PKA, and AMPK to Akt are essential for the metabolic switching in matrix-deprived cancer cells. AMPK knockdown simulations identified a crucial role for AMPK in maintaining these adaptive changes. Thus, this mathematical framework provides insights on attachment-detachment signaling with metabolic adaptations that promote cancer metastasis.

DOI: https://doi.org/10.1038/s41540-024-00443-4
Cell Mol Bioeng. 2024 Oct;17(5): 385-399

Leveraging Cell Migration Dynamics to Discriminate Between Senescent and Presenescent Human Mesenchymal Stem Cells.

Farshad Amiri, Panagiotis Mistriotis.

   Purpose: The suboptimal clinical performance of human mesenchymal stem cells (hMSCs) has raised concerns about their therapeutic potential. One major contributing factor to this issue is the heterogeneous nature of hMSCs. Senescent cell accumulation during stem cell expansion is a key driver of MSC heterogeneity. Current methodologies to eradicate senescent hMSCs have either shown limited success or lack clinical relevance. This study leverages the inherent capacity of hMSCs to migrate toward damaged tissues as a means to discern senescent from presenescent stem cells. Given the established deficiency of senescent cells to migrate through physiologically relevant environments, we hypothesized that a microfluidic device, designed to emulate key facets of in vivo cell motility, could serve as a platform for identifying presenescent cells.
Methods: We employed a Y-shaped microchannel assay, which allows fine-tuning of fluid flow rates and the degree of confinement.
Results: Highly migratory hMSCs detected by the device not only demonstrate increased speed, smaller size, and higher proliferative capacity but also manifest reduced DNA damage and senescence compared to non-migratory cells. Additionally, this assay detects presenescent cells in experiments with mixed early and late passage cells. The introduction of fluid flow through the device can further increase the fraction of highly motile stem cells, improving the assay's effectiveness to remove senescent hMSCs.
Conclusions: Collectively, this assay facilitates the detection and isolation of a highly potent stem cell subpopulation. Given the positive correlation between the migratory potential of administered MSCs and the long-term clinical outcome, delivering homogeneous, highly motile presenescent hMSCs may benefit patient outcomes.
Supplementary Information: The online version contains supplementary material available at 10.1007/s12195-024-00807-0.

Keywords:  Aging; Confinement; Fluid forces; Senescence; Stem cell migration

DOI:  https://doi.org/10.1007/s12195-024-00807-0
Nat Commun. 2024 Nov 08. 15(1): 9561

Cell shape affects bacterial colony growth under physical confinement.

M Sreepadmanabh, Meenakshi Ganesh, Pratibha Sanjenbam, Christina Kurzthaler, Deepa Agashe, Tapomoy Bhattacharjee.

Evidence from homogeneous liquid or flat-plate cultures indicates that biochemical cues are the primary modes of bacterial interaction with their microenvironment. However, these systems fail to capture the effect of physical confinement on bacteria in their natural habitats. Bacterial niches like the pores of soil, mucus, and infected tissues are disordered microenvironments with material properties defined by their internal pore sizes and shear moduli. Here, we use three-dimensional matrices that match the viscoelastic properties of gut mucus to test how altering the physical properties of their microenvironment influences the growth of bacteria under confinement. We find that low aspect ratio (spherical) bacteria form compact, spherical colonies under confinement while high aspect ratio (rod-shaped) bacteria push their progenies further outwards to create elongated colonies with a higher surface area, enabling increased access to nutrients. As a result, the population growth of high aspect ratio bacteria is, under the tested conditions, more robust to increased physical confinement compared to that of low aspect ratio bacteria. Thus, our experimental evidence supports that environmental physical constraints can play a selective role in bacterial growth based on cell shape.

DOI: https://doi.org/10.1038/s41467-024-53989-6
Biomater Sci. 2024 Nov 12.

Photoinitiator-free light-mediated crosslinking of dynamic polymer and pristine protein networks.

Riccardo Rizzo, Dylan M Barber, Jackson K Wilt, Alexander J Ainscough, Jennifer A Lewis.

Light-based patterning of synthetic and biological hydrogels enables precise spatial and temporal control over the formation of chemical bonds. However, photoinitiators are typically used to generate free radicals, which are detrimental to human cells. Here, we report a photoinitiator- and radical-free method based on ortho-nitrobenzyl alcohol (oNBA) photolysis, which gives rise to highly reactive nitroso and benzaldehyde groups. Synthetic hydrogel and pristine protein networks can rapidly form in the presence of these photo-generated reactive species. Thiol -oNBA bonds yield dynamic hydrogel networks (DHNs) via N-semimercaptal linkages that exhibit thixotropy, stress relaxation, and on-demand reversible gel-to-liquid transitions, while amine-oNBA bonds can be exploited to crosslink pristine proteins, such as gelatin and fibrinogen, by targeting their primary amines. Since this approach does not require incorporation of photoreactive moieties along the backbone, the resulting crosslinked proteins are well suited for bioadhesives. Our photoinitiator-free platform provides a versatile approach for rapidly creating synthetic and biological hydrogels for applications ranging from tissue engineering to biomedical devices.

DOI: https://doi.org/10.1039/d4bm00849a
Biomater Adv. 2024 Nov 04. pii: S2772-9508(24)00332-7. [Epub ahead of print]167 214089

Monophasic hyaluronic acid-silica hybrid hydrogels for articular cartilage applications.

Huijun Zhang, Jessica Faber, Silvia Budday, Qingsen Gao, Sonja Kuth, Kai Zheng, Aldo R Boccaccini.

  Hyaluronic acid (HA), an FDA-approved natural polymer and important component of the extracellular matrix (ECM), has been widely used to develop hydrogels for cartilage regeneration. However, HA hydrogels often exhibit poor mechanical properties and unsuitable degradability, limiting their capability to support cell growth in cartilage. To overcome these challenges, this study modifies HA with a silica precursor and the coupling agent (3-Glycidyloxypropyl) trimethoxysilane (GPTMS) to develop a monophasic organic-inorganic hybrid HA-silica hydrogel. In this system, the inorganic silicate and organic HA components interpenetrate and bond covalently at the molecular level. The HA-silica hybrid hydrogel achieves a compressive modulus of 143 kPa at the highest GPTMS/HA molar ratio of 400. Additionally, in vitro cell studies show that these hybrid hydrogels have no cytotoxicity against MC3T3-E1 and ATDC-5 cells. Cell viability and morphology tests further confirm excellent cell adhesion on the hybrid scaffold. These results indicate that the developed HA-silica hybrid hydrogel is a suitable candidate for cartilage regeneration applications.

Keywords:  Cartilage regeneration; Hyaluronic acid; Hybrid hydrogel; Silica

DOI:  https://doi.org/10.1016/j.bioadv.2024.214089
Cell Mol Bioeng. 2024 Oct;17(5): 317-327

Magnetoactive, Kirigami-Inspired Hammocks to Probe Lung Epithelial Cell Function.

Katherine Wei, Avinava Roy, Sonia Ejike, Madeline K Eiken, Eleanor M Plaster, Alan Shi, Max Shtein, Claudia Loebel.

   Introduction: Mechanical forces provide critical biological signals to cells. Within the distal lung, tensile forces act across the basement membrane and epithelial cells atop. Stretching devices have supported studies of mechanical forces in distal lung epithelium to gain mechanistic insights into pulmonary diseases. However, the integration of curvature into devices applying mechanical forces onto lung epithelial cell monolayers has remained challenging. To address this, we developed a hammock-shaped platform that offers desired curvature and mechanical forces to lung epithelial monolayers.
Methods: We developed hammocks using polyethylene terephthalate (PET)-based membranes and magnetic-particle modified silicone elastomer films within a 48-well plate that mimic the alveolar curvature and tensile forces during breathing. These hammocks were engineered and characterized for mechanical and cell-adhesive properties to facilitate cell culture. Using human small airway epithelial cells (SAECs), we measured monolayer formation and mechanosensing using F-Actin staining and immunofluorescence for cytokeratin to visualize intermediate filaments.
Results: We demonstrate a multi-functional design that facilitates a range of curvatures along with the incorporation of magnetic elements for dynamic actuation to induce mechanical forces. Using this system, we then showed that SAECs remain viable, proliferate, and form an epithelial cell monolayer across the entire hammock. By further applying mechanical stimulation via magnetic actuation, we observed an increase in proliferation and strengthening of the cytoskeleton, suggesting an increase in mechanosensing.
Conclusion: This hammock strategy provides an easily accessible and tunable cell culture platform for mimicking distal lung mechanical forces in vitro. We anticipate the promise of this culture platform for mechanistic studies, multi-modal stimulation, and drug or small molecule testing, extendable to other cell types and organ systems.
Supplementary Information: The online version contains supplementary material available at 10.1007/s12195-024-00808-z.

Keywords:  Dynamic culture; Lung epithelial cells; Magnetic actuation Kirigami; Mechanotransduction

DOI:  https://doi.org/10.1007/s12195-024-00808-z
Sci Adv. 2024 Nov 15. 10(46): eadp2357

Mechano-gradients drive morphogen-noise correction to ensure robust patterning.

Kana Aoki, Taiki Higuchi, Yuki Akieda, Kotone Matsubara, Yasuyuki Ohkawa, Tohru Ishitani.

Morphogen gradients instruct cells to pattern tissues. Although the mechanisms by which morphogens transduce chemical signals have been extensively studied, the roles and regulation of the physical communication between morphogen-receiver cells remain unclear. Here, we show that the Wnt/β-catenin-morphogen gradient, which patterns the embryonic anterior-posterior (AP) axis, generates intercellular tension gradients along the AP axis by controlling membrane cadherin levels in zebrafish embryos. This "mechano-gradient" is used for the cell competition-driven correction of noisy morphogen gradients. Naturally and artificially generated unfit cells, producing noisy Wnt/β-catenin gradients, induce local deformation of the mechano-gradients that activate mechanosensitive calcium channels in the neighboring fit cells, which then secrete annexin A1a to kill unfit cells. Thus, chemo-mechanical interconversion-mediated competitive communication between the morphogen-receiver cells ensures precise tissue patterning.

DOI: https://doi.org/10.1126/sciadv.adp2357