bims-ecemfi 2024-05-05 papers

bims-ecemfi

Biomed News

on ECM and fibroblasts

Issue of 2024–05–05
eleven papers selected by
Badri Narayanan Narasimhan, University of California, San Diego

Micro-Tensile Rheology of Fibrous Gels Quantifies Strain-dependent Anisotropy.
High-viscosity driven modulation of biomechanical properties of human mesenchymal stem cells promotes osteogenic lineage.
Photo-tuning of Hyaluronic-acid-based Hydrogel Properties to Control Network Formation in Human Vascular Endothelial Cells.
Dual-crosslinking gelatin-hyaluronic acid methacrylate based biomimetic PDAC desmoplastic niche enhances tumor-associated macrophages recruitment and M2-like polarization.
Lumen expansion is initially driven by apical actin polymerization followed by osmotic pressure in a human epiblast model.
Engineering cell instructive microenvironments for in vitro replication of functional barrier organs.
Confinement controls the creep rate in soft granular packings.
Fibrillogenesis in collagen hydrogels accelerated by carboxylated microbeads.
i-Rheo-optical assay: Measuring the viscoelastic properties of multicellular spheroids.
Spatially resolved analysis of microenvironmental gradient impact on cancer cell phenotypes.
The amoeboid migration of monocytes in confining channels requires the local remodeling of the cortical actin cytoskeleton by cofilin-1.

Acta Biomater. 2024 Apr 27. pii: S1742-7061(24)00165-X. [Epub ahead of print]

Micro-Tensile Rheology of Fibrous Gels Quantifies Strain-dependent Anisotropy.

Shahar Goren, Bar Ergaz, Daniel Barak, Raya Sorkin, Ayelet Lesman.

  Semiflexible fiber gels such as collagen and fibrin have unique nonlinear mechanical properties that play an important role in tissue morphogenesis, wound healing, and cancer metastasis. Optical tweezers microrheology has greatly contributed to the understanding of the mechanics of fibrous gels at the microscale, including its heterogeneity and anisotropy. However, the explicit relationship between micromechanical properties and gel deformation has been largely overlooked. We introduce a unique gel-stretching apparatus and employ it to study the relationship between microscale strain and stiffening in fibrin and collagen gels, focusing on the development of anisotropy in the gel. We find that gels stretched by as much as 15% stiffen significantly both in parallel and perpendicular to the stretching axis, and that the parallel axis is 2-3 times stiffer than the transverse axis. We also measure the stiffening and anisotropy along bands of aligned fibers created by aggregates of cancer cells, and find similar effects as in gels stretched with the tensile apparatus. Our results illustrate that the extracellular microenvironment is highly sensitive to deformation, with implications for tissue homeostasis and pathology. STATEMENT OF SIGNIFICANCE: The inherent fibrous architecture of the extracellular matrix (ECM) gives rise to unique strain-stiffening mechanics. The micromechanics of fibrous networks has been studied extensively, but the deformations involved in its stiffening at the microscale were not quantified. Here we introduce an apparatus that enables measuring the deformations in the gel as it is being stretched while simultaneously using optical tweezers to measure its microscale anisotropic stiffness. We reveal that fibrin and collagen both stiffen dramatically already at ∼10% deformation, accompanied by the emergence of significant, yet moderate anisotropy. We measure similar stiffening and anisotropy in the matrix remodeled by the tensile apparatus to those found between cancer cell aggregates. Our results emphasize that small strains are enough to introduce substantial stiffening and anisotropy. These have been shown to result in directional cell migration and enhanced force propagation, and possibly control processes like morphogenesis and cancer metastasis.

Keywords:  Anisotropy; Collagen; Deformation; Extra-cellular matrix; Fiber Alignment; Fibrin; Microrheology; Optical Tweezers; Tumor Microenvironment

DOI:  https://doi.org/10.1016/j.actbio.2024.03.028
Mater Today Bio. 2024 Jun;26 101058

High-viscosity driven modulation of biomechanical properties of human mesenchymal stem cells promotes osteogenic lineage.

Yin-Quan Chen, Ming-Chung Wu, Ming-Tzo Wei, Jean-Cheng Kuo, Helen Wenshin Yu, Arthur Chiou.

  Biomechanical cues could effectively govern cell gene expression to direct the differentiation of specific stem cell lineage. Recently, the medium viscosity has emerged as a significant mechanical stimulator that regulates the cellular mechanical properties and various physiological functions. However, whether the medium viscosity can regulate the mechanical properties of human mesenchymal stem cells (hMSCs) to effectively trigger osteogenic differentiation remains uncertain. The mechanism by which cells sense and respond to changes in medium viscosity, and regulate cell mechanical properties to promote osteogenic lineage, remains elusive. In this study, we demonstrated that hMSCs, cultured in a high-viscosity medium, exhibited larger cell spreading area and higher intracellular tension, correlated with elevated formation of actin stress fibers and focal adhesion maturation. Furthermore, these changes observed in hMSCs were associated with activation of TRPV4 (transient receptor potential vanilloid sub-type 4) channels on the cell membrane. This feedback loop among TRPV4 activation, cell spreading and intracellular tension results in calcium influx, which subsequently promotes the nuclear localization of NFATc1 (nuclear factor of activated T cells 1). Concomitantly, the elevated intracellular tension induced nuclear deformation and promoted the nuclear localization of YAP (YES-associated protein). The concurrent activation of NFATc1 and YAP significantly enhanced alkaline phosphatase (ALP) for pre-osteogenic activity. Taken together, these findings provide a more comprehensive view of how viscosity-induced alterations in biomechanical properties of MSCs impact the expression of osteogenesis-related genes, and ultimately promote osteogenic lineage.

Keywords:  Cell mechanical properties; Extracellular fluid viscosity; NFATc; Osteogenesis; TRPV4; YAP

DOI:  https://doi.org/10.1016/j.mtbio.2024.101058
Adv Healthc Mater. 2024 Apr 29. e2303787

Photo-tuning of Hyaluronic-acid-based Hydrogel Properties to Control Network Formation in Human Vascular Endothelial Cells.

Kelum Chamara Manoj Lakmal Elvitigala, Lakshimi Mohan, Wildan Mubarok, Shinji Sakai.

  In vitro network formation by endothelial cells serves as a fundamental model for studies aimed at understanding angiogenesis. The morphogenesis of these cells to form a network is intricately regulated by the mechanical and biochemical properties of the extracellular matrix. This article presents the effects of modulating these properties in hydrogels derived from phenolated hyaluronic acid (HA-Ph) and gelatin (Gelatin-Ph). Visible-light irradiation in the presence of tris(2,2'-bipyridyl)ruthenium(II) chloride hexahydrate and sodium persulfate induces the crosslinking of these polymers, thereby forming a hydrogel formation and degrading HA-Ph. Human vascular endothelial cells form networks on the hydrogel prepared by visible-light irradiation for 45 min (42 W cm-2 at 450 nm) but not on the hydrogels prepared by irradiation for 15, 30, or 60 min. The irradiation-time-dependent degradation of HA-Ph and the changes in the mechanical stiffness of the hydrogels, coupled with the expressions of RhoA and β-actin genes and CD44 receptors in the cells, reveal that the network formation is synergistically influenced by the hydrogel stiffness and HA-Ph degradation. These findings highlight the potential of tailoring HA-based hydrogel properties to modulate human vascular endothelial cell responses, which is critical for advancing their application in vascular tissue engineering. This article is protected by copyright. All rights reserved.

Keywords:  composite hydrogel; degradation; huvec; hyaluronic acid; photocrosslinking

DOI:  https://doi.org/10.1002/adhm.202303787
Int J Biol Macromol. 2024 Apr 26. pii: S0141-8130(24)02631-X. [Epub ahead of print] 131826

Dual-crosslinking gelatin-hyaluronic acid methacrylate based biomimetic PDAC desmoplastic niche enhances tumor-associated macrophages recruitment and M2-like polarization.

Di Wu, Tiancheng Gong, Zhongxiang Sun, Xihao Yao, Dongzhi Wang, Qiyang Chen, Qingsong Guo, Xiaohong Li, Yibing Guo, Yuhua Lu.

  The tumor microenvironment (TME) of pancreatic ductal adenocarcinoma (PDAC) is characterized by deposition of desmoplastic matrix (including collagen and hyaluronic acid). And the interactions between tumor-associated macrophages (TAMs) and tumor cells play a crucial role in progression of PDAC. Hence, the appropriate model of tumor cell-macrophage interaction within the unique PDAC TME is of significantly important. To this end, a 3D tumor niche based on dual-crosslinking gelatin methacrylate and hyaluronic acid methacrylate hydrogels was constructed to simulate the desmoplastic tumor matrix with matching compressive modulus and composition. The bionic 3D tumor niche creates an immunosuppressive microenvironment characterized by the downregulation of M1 markers and upregulation of M2 markers in TAMs. Mechanistically, RNA-seq analysis revealed that the PI3K-AKT signaling pathway might modulate the phenotypic balance and recruitment of macrophages through regulating SELE and VCAM-1. Furthermore, GO and GSEA revealed the biological process of leukocyte migration and the activation of cytokine-associated signaling were involved. Finally, the 3D tumor-macrophage niches with three different ratios were fabricated which displayed increased M2-like polarization and stemness. The utilization of the 3D tumor niche has the potential to provide a more accurate investigation of the interplay between PDAC tumor cells and macrophages within an in vivo setting.

Keywords:  Dual-crosslinking gelatin-hyaluronic acid methacrylate; Macrophage m2-like polarization; Pancreatic ductal adenocarcinoma

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.131826
Cell Stem Cell. 2024 May 02. pii: S1934-5909(24)00097-3. [Epub ahead of print]31(5): 640-656.e8

Lumen expansion is initially driven by apical actin polymerization followed by osmotic pressure in a human epiblast model.

Dhiraj Indana, Andrei Zakharov, Youngbin Lim, Alexander R Dunn, Nidhi Bhutani, Vivek B Shenoy, Ovijit Chaudhuri.

  Post-implantation, the pluripotent epiblast in a human embryo forms a central lumen, paving the way for gastrulation. Osmotic pressure gradients are considered the drivers of lumen expansion across development, but their role in human epiblasts is unknown. Here, we study lumenogenesis in a pluripotent-stem-cell-based epiblast model using engineered hydrogels. We find that leaky junctions prevent osmotic pressure gradients in early epiblasts and, instead, forces from apical actin polymerization drive lumen expansion. Once the lumen reaches a radius of ∼12 μm, tight junctions mature, and osmotic pressure gradients develop to drive further growth. Computational modeling indicates that apical actin polymerization into a stiff network mediates initial lumen expansion and predicts a transition to pressure-driven growth in larger epiblasts to avoid buckling. Human epiblasts show transcriptional signatures consistent with these mechanisms. Thus, actin polymerization drives lumen expansion in the human epiblast and may serve as a general mechanism of early lumenogenesis.

Keywords:  actin; computational modeling; embryogenesis; engineered hydrogels; human epiblast model; induced pluripotent stem cells; lumen; morphogenesis; osmotic pressure; tissue biophysics

DOI:  https://doi.org/10.1016/j.stem.2024.03.016
Adv Healthc Mater. 2024 May 02. e2400357

Engineering cell instructive microenvironments for in vitro replication of functional barrier organs.

Francesco Urciuolo, Giorgia Imparato, Paolo Antonio Netti.

  Multicellular organisms exhibit synergistic effects among their components, giving rise to emergent properties crucial for their genesis and overall functionality and survival. Morphogenesis, the formation of an organism's shape, involves and relies upon intricate and biunivocal interactions among cells and their environment, i.e., the extracellular matrix (ECM). Cells secrete their own ECM which, in turn regulates their morphogenetic program by controlling time and space presentation of matricellular signals. The ECM, once perceived as a passive structure, is now acknowledged as an informative space where both biochemical (eg, growth factors, cytokines, matricellular cues) and biophysical (eg, morphophysical, mechanical) signals are tightly orchestrated. Replicating this sophisticate and highly interconnected informative media in a synthetic scaffold for tissue engineering is unattainable with our current technology and this limits our capability to engineering functional human tissues/organs in vitro and in vivo. This review explores current limitation to in vitro organ morphogenesis, emphasizing the interplay of gene regulatory networks, mechanical factors, and tissue microenvironment cues. In vitro efforts to replicate biological processes, particularly for barrier organs like the lung and intestine, are examined. The importance of maintaining cells within their native microenvironmental context is highlighted to accurately replicate organ-specific morphogenic properties. The review underscores the necessity for microphysiological systems that faithfully reproduce cell-native interactions, emphasizing spatial and temporal signaling cues between cells and the ECM. This approach is vital to improve such microphysiological systems used to for advancing our understanding of developmental disorders and disease progression. This article is protected by copyright. All rights reserved.

Keywords:  Cell and Tissue microenvironment; cell morphogenetic program; gut‐on‐chip, lung‐on‐chipTissue Engineering

DOI:  https://doi.org/10.1002/adhm.202400357
Soft Matter. 2024 May 01.

Confinement controls the creep rate in soft granular packings.

Joshua A Dijksman, Tom Mullin.

Flow in soft materials encompasses a wide range of viscous, plastic and elastic phenomena which provide challenges to modelling at the microscopic level. To create a controlled flow, we perform falling ball viscometry tests on packings of soft, frictionless hydrogel spheres. Systematic creep flow is found when a controlled driving stress is applied to a sinking sphere embedded in a packing. Here, we take the novel approach of applying an additional global confinement stress to the packing using an external load. This has enabled us to identify two distinct creep regimes. When confinement stress is small, the creep rate is independent of the load imposed. For larger confinement stresses, we find that the creep rate is set by the mechanical load acting on the packing. In the latter regime, the creep rate depends exponentially on the imposed stress. We can combine the two regimes via a rescaling onto a master curve, capturing the creep rate over five orders of magnitude. Our results indicate that bulk creep phenomena in these soft materials can be subtly controlled using an external mechanical force.

DOI: https://doi.org/10.1039/d3sm01755a
Biomed Mater. 2024 Apr 30.

Fibrillogenesis in collagen hydrogels accelerated by carboxylated microbeads.

Laura Rodríguez-Mandujano, Reinher Pimentel-Domínguez, Elisa Tamariz, Edgar Campos-Puente, Astrid Lorena Giraldo-Betancur, Remy Avila.

  Collagen type I is a material widely used for 3D cell culture and tissue engineering. Different architectures, such as gels, sponges, membranes, and nanofibers, can be fabricated with it. In collagen hydrogels, the formation of fibrils and fibers depends on various parameters, such as the source of collagen, pH, temperature, concentration, age, etc. In this work, we study the fibrillogenesis process in collagen type I hydrogels with different types of microbeads embedded, using optical techniques such as turbidity assay and confocal reflectance microscopy. We observe that microbeads embedded in the collagen matrix hydrogels modify the fibrillogenesis. Our results show that carboxylated fluorescent microbeads accelerate 3.6 times the gelation, while silica microbeads slow down the formation of collagen fibrils by a factor of 1.9, both compared to pure collagen hydrogels. Our observations suggest that carboxylate microbeads act as nucleation sites and the early collagen fibrils bind to the microbeads.&#xD.

Keywords:  Carboxylated beads; Collagen hydrogels; Confocal reflectance microscopy; Fibrillogenesis; Turbidimetry

DOI:  https://doi.org/10.1088/1748-605X/ad459a
Mater Today Bio. 2024 Jun;26 101066

i-Rheo-optical assay: Measuring the viscoelastic properties of multicellular spheroids.

Rosalia Ferraro, Stefano Guido, Sergio Caserta, Manlio Tassieri.

  This study introduces a novel mechanobiology assay, named "i-Rheo-optical assay", that integrates rheology with optical microscopy for analysing the viscoelastic properties of multicellular spheroids. These spheroids serve as three-dimensional models resembling tissue structures. The innovative technique enables real-time observation and quantification of morphological responses to applied stress using a cost-effective microscope coverslip for constant compression force application. By bridging a knowledge gap in biophysical research, which has predominantly focused on the elastic properties while only minimally exploring the viscoelastic nature in multicellular systems, the i-Rheo-optical assay emerges as an effective tool. It facilitates the measurement of broadband viscoelastic compressional moduli in spheroids, here derived from cancer (PANC-1) and non-tumoral (NIH/3T3) cell lines during compression tests. This approach plays a crucial role in elucidating the mechanical properties of spheroids and holds potential for identifying biomarkers to discriminate between healthy tissues and their pathological counterparts. Offering comprehensive insights into the biomechanical behaviour of biological systems, i-Rheo-optical assay marks a significant advancement in tissue engineering, cancer research, and therapeutic development.

Keywords:  Cancer research; Mechanobiology; Multicellular spheroids; Optical assay; Rheology

DOI:  https://doi.org/10.1016/j.mtbio.2024.101066
Sci Adv. 2024 May 03. 10(18): eadn3448

Spatially resolved analysis of microenvironmental gradient impact on cancer cell phenotypes.

Jamie Auxillos, Roxane Crouigneau, Yan-Fang Li, Yifan Dai, Arnaud Stigliani, Isabella Tavernaro, Ute Resch-Genger, Albin Sandelin, Rodolphe Marie, Stine F Pedersen.

Despite the physiological and pathophysiological significance of microenvironmental gradients, e.g., for diseases such as cancer, tools for generating such gradients and analyzing their impact are lacking. Here, we present an integrated microfluidic-based workflow that mimics extracellular pH gradients characteristic of solid tumors while enabling high-resolution live imaging of, e.g., cell motility and chemotaxis, and preserving the capacity to capture the spatial transcriptome. Our microfluidic device generates a pH gradient that can be rapidly controlled to mimic spatiotemporal microenvironmental changes over cancer cells embedded in a 3D matrix. The device can be reopened allowing immunofluorescence analysis of selected phenotypes, as well as the transfer of cells and matrix to a Visium slide for spatially resolved analysis of transcriptional changes across the pH gradient. This workflow is easily adaptable to other gradients and multiple cell types and can therefore prove invaluable for integrated analysis of roles of microenvironmental gradients in biology.

DOI: https://doi.org/10.1126/sciadv.adn3448
Sci Rep. 2024 May 03. 14(1): 10241

The amoeboid migration of monocytes in confining channels requires the local remodeling of the cortical actin cytoskeleton by cofilin-1.

Maria F Ullo, Anna E D'Amico, Sandrine B Lavenus, Jeremy S Logue.

  Within the bloodstream, monocytes must traverse the microvasculature to prevent leukostasis, which is the entrapment of monocytes within the confines of the microvasculature. Using the model cell line, THP-1, and VCAM-1 coated channels to simulate the microvasculature surface, we demonstrate that monocytes predominantly adopt an amoeboid phenotype, which is characterized by the formation of blebs. As opposed to cortical actin flow in leader blebs, cell movement is correlated with myosin contraction at the cell rear. It was previously documented that cofilin-1 promotes cortical actin turnover at leader bleb necks in melanoma cells. In monocytes, our data suggest that cofilin-1 promotes the local upregulation of myosin contractility through actin cytoskeleton remodeling. In support of this concept, cofilin-1 is found to localize to a single cell edge. Moreover, the widespread upregulation of myosin contractility was found to inhibit migration. Thus, monocytes within the microvasculature may avoid entrapment by adopting an amoeboid mode of migration.

Keywords:  Amoeboid; Bleb; Cofilin-1; Leukemia; Monocytes

DOI:  https://doi.org/10.1038/s41598-024-60971-1