bims-ecemfi 2024-06-30 papers

bims-ecemfi

Biomed News

on ECM and fibroblasts

Issue of 2024‒06‒30
twenty papers selected by
Badri Narayanan Narasimhan, University of California, San Diego

The fusion of physics and biology in early mammalian embryogenesis.
Mechanobiology of Adipocytes.
Cancer cell migration depends on adjacent ASC and adipose spheroids in a 3D bioprinted breast cancer model.
Enhancing cell adhesion in synthetic hydrogels via physical confinement of peptide-functionalized polymer clusters.
The extracellular matrix integrates mitochondrial homeostasis.
Matrix stiffness influences response to chemo and targeted therapy in brain metastatic breast cancer cells.
Interplay of Spatial and Topological Defects in Polymer Networks.
Automated Analysis of Extracellular Matrix Invasion of Cancer Cells from Tumor Spheroids.
Effects of coordination and stiffness scale separation in disordered elastic networks.
Side-view optical microscopy-assisted atomic force microscopy for thickness-dependent nanobiomechanics.
Multifunctional hydrogels with spatially controlled light activation with photocaged oligonucleotides.
Mechano-inhibition of endocytosis sensitizes cancer cells to Fas-induced Apoptosis.
Metabolomic Profiling and Characterization of a Novel 3D Culture System for Studying Chondrocyte Mechanotransduction.
Combined effects of matrix stiffness and obesity-associated signaling directs progressive phenotype in PANC-1 pancreatic cancer cells in vitro.
Synergistic Coassembly of Folic Acid-Based Supramolecular Polymer with a Covalent Polymer Toward Fabricating Functional Antibacterial Biomaterials.
Cancer-associated fibroblasts promote proliferation, angiogenesis, metastasis and immunosuppression in gastric cancer.
Modeling 3D Tumor Invasiveness to Modulate Macrophage Phenotype in a Human-based Hydrogel Platform.
Hydrogel morphogenesis induced by force-controlled growth.
Protocol for real-time assessment of mitochondrial and glycolytic ATP production in patient-derived glioma stem-like cells.
A Lifeact-EGFP quail for studying actin dynamics in vivo.

Curr Top Dev Biol. 2024 ;pii: S0070-2153(24)00081-4. [Epub ahead of print]160 31-64

The fusion of physics and biology in early mammalian embryogenesis.

Walter Piszker, Mijo Simunovic.

  Biomechanics in embryogenesis is a dynamic field intertwining the physical forces and biological processes that shape the first days of a mammalian embryo. From the first cell fate bifurcation during blastulation to the complex symmetry breaking and tissue remodeling in gastrulation, mechanical cues appear critical in cell fate decisions and tissue patterning. Recent strides in mouse and human embryo culture, stem cell modeling of mammalian embryos, and biomaterial design have shed light on the role of cellular forces, cell polarization, and the extracellular matrix in influencing cell differentiation and morphogenesis. This chapter highlights the essential functions of biophysical mechanisms in blastocyst formation, embryo implantation, and early gastrulation where the interplay between the cytoskeleton and extracellular matrix stiffness orchestrates the intricacies of embryogenesis and placenta specification. The advancement of in vitro models like blastoids, gastruloids, and other types of embryoids, has begun to faithfully recapitulate human development stages, offering new avenues for exploring the biophysical underpinnings of early development. The integration of synthetic biology and advanced biomaterials is enhancing the precision with which we can mimic and study these processes. Looking ahead, we emphasize the potential of CRISPR-mediated genomic perturbations coupled with live imaging to uncover new mechanosensitive pathways and the application of engineered biomaterials to fine-tune the mechanical conditions conducive to embryonic development. This synthesis not only bridges the gap between experimental models and in vivo conditions to advancing fundamental developmental biology of mammalian embryogenesis, but also sets the stage for leveraging biomechanical insights to inform regenerative medicine.

Keywords:  Blastulation; Embryonic biomechanics; Gastrulation; Implantation; Mechanosignaling; Mechanotransduction; Stem cell embryo models

DOI:  https://doi.org/10.1016/bs.ctdb.2024.05.001
Biology (Basel). 2024 Jun 13. pii: 434. [Epub ahead of print]13(6):

Mechanobiology of Adipocytes.

Sean P Blade, Dylan J Falkowski, Sarah N Bachand, Steven J Pagano, LiKang Chin.

  The growing obesity epidemic necessitates increased research on adipocyte and adipose tissue function and disease mechanisms that progress obesity. Historically, adipocytes were viewed simply as storage for excess energy. However, recent studies have demonstrated that adipocytes play a critical role in whole-body homeostasis, are involved in cell communication, experience forces in vivo, and respond to mechanical stimuli. Changes to the adipocyte mechanical microenvironment can affect function and, in some cases, contribute to disease. The aim of this review is to summarize the current literature on the mechanobiology of adipocytes. We reviewed over 100 papers on how mechanical stress is sensed by the adipocyte, the effects on cell behavior, and the use of cell culture scaffolds, particularly those with tunable stiffness, to study adipocyte behavior, adipose cell and tissue mechanical properties, and computational models. From our review, we conclude that adipocytes are responsive to mechanical stimuli, cell function and adipogenesis can be dictated by the mechanical environment, the measurement of mechanical properties is highly dependent on testing methods, and current modeling practices use many different approaches to recapitulate the complex behavior of adipocytes and adipose tissue. This review is intended to aid future studies by summarizing the current literature on adipocyte mechanobiology.

Keywords:  adipocyte biology; finite element analysis; mechanobiology; obesity

DOI:  https://doi.org/10.3390/biology13060434
Biofabrication. 2024 Jun 27. 16(3):

Cancer cell migration depends on adjacent ASC and adipose spheroids in a 3D bioprinted breast cancer model.

Hannes Horder, David Böhringer, Nadine Endrizzi, Laura S Hildebrand, Alessandro Cianciosi, Sabrina Stecher, Franziska Dusi, Sophie Schweinitzer, Martin Watzling, Jürgen Groll, Tomasz Jüngst, Jörg Teßmar, Petra Bauer-Kreisel, Ben Fabry, Torsten Blunk.

  Breast cancer develops in close proximity to mammary adipose tissue and interactions with the local adipose environment have been shown to drive tumor progression. The specific role, however, of this complex tumor microenvironment in cancer cell migration still needs to be elucidated. Therefore, in this study, a 3D bioprinted breast cancer model was developed that allows for a comprehensive analysis of individual tumor cell migration parameters in dependence of adjacent adipose stroma. In this co-culture model, a breast cancer compartment with MDA-MB-231 breast cancer cells embedded in collagen is surrounded by an adipose tissue compartment consisting of adipose-derived stromal cell (ASC) or adipose spheroids in a printable bioink based on thiolated hyaluronic acid. Printing parameters were optimized for adipose spheroids to ensure viability and integrity of the fragile lipid-laden cells. Preservation of the adipogenic phenotype after printing was demonstrated by quantification of lipid content, expression of adipogenic marker genes, the presence of a coherent adipo-specific extracellular matrix, and cytokine secretion. The migration of tumor cells as a function of paracrine signaling of the surrounding adipose compartment was then analyzed using live-cell imaging. The presence of ASC or adipose spheroids substantially increased key migration parameters of MDA-MB-231 cells, namely motile fraction, persistence, invasion distance, and speed. These findings shed new light on the role of adipose tissue in cancer cell migration. They highlight the potential of our 3D printed breast cancer-stroma model to elucidate mechanisms of stroma-induced cancer cell migration and to serve as a screening platform for novel anti-cancer drugs targeting cancer cell dissemination.

Keywords:  3D bioprinting; adipose tissue; adipose-derived stromal cells; breast cancer model; live-cell imaging; migration; spheroids

DOI:  https://doi.org/10.1088/1758-5090/ad57f7
J Mater Chem B. 2024 Jun 26.

Enhancing cell adhesion in synthetic hydrogels via physical confinement of peptide-functionalized polymer clusters.

Shohei Ishikawa, Hiroyuki Kamata, Takamasa Sakai.

Artificially synthesized poly(ethylene glycol) (PEG)-based hydrogels are extensively utilized as biomaterials for tissue scaffolds and cell culture matrices due to their non-protein adsorbing properties. Although these hydrogels are inherently non-cell-adhesive, advancements in modifying polymer networks with functional peptides have led to PEG hydrogels with diverse functionalities, such as cell adhesion and angiogenesis. However, traditional methods of incorporating additives into hydrogel networks often result in the capping of crosslinking points with heterogeneous substances, potentially impairing mechanical properties and obscuring the causal relationships of biological functions. This study introduces polymer additives designed to resist prolonged elution from hydrogels, providing a novel approach to facilitate cell culture on non-adhesive surfaces. By clustering tetra-branched PEG to form ultra-high molecular weight hyper-branched structures and functionalizing their termini with cell-adhesive peptides, we successfully entrapped these clusters within the hydrogel matrix without compromising mechanical strength. This method has enabled successful cell culture on inherently non-adhesive PEG hydrogel surfaces at high peptide densities, a feat challenging to achieve with conventional means. The approach proposed in this study not only paves the way for new possibilities with polymer additives but also serves as a new design paradigm for cell culturing on non-cell-adhesive hydrogels.

DOI: https://doi.org/10.1039/d4tb00761a
Cell. 2024 Jun 24. pii: S0092-8674(24)00638-X. [Epub ahead of print]

The extracellular matrix integrates mitochondrial homeostasis.

Hanlin Zhang, C Kimberly Tsui, Gilberto Garcia, Larry K Joe, Haolun Wu, Ayane Maruichi, Wudi Fan, Sentibel Pandovski, Peter H Yoon, Brant M Webster, Jenni Durieux, Phillip A Frankino, Ryo Higuchi-Sanabria, Andrew Dillin.

  Cellular homeostasis is intricately influenced by stimuli from the microenvironment, including signaling molecules, metabolites, and pathogens. Functioning as a signaling hub within the cell, mitochondria integrate information from various intracellular compartments to regulate cellular signaling and metabolism. Multiple studies have shown that mitochondria may respond to various extracellular signaling events. However, it is less clear how changes in the extracellular matrix (ECM) can impact mitochondrial homeostasis to regulate animal physiology. We find that ECM remodeling alters mitochondrial homeostasis in an evolutionarily conserved manner. Mechanistically, ECM remodeling triggers a TGF-β response to induce mitochondrial fission and the unfolded protein response of the mitochondria (UPRMT). At the organismal level, ECM remodeling promotes defense of animals against pathogens through enhanced mitochondrial stress responses. We postulate that this ECM-mitochondria crosstalk represents an ancient immune pathway, which detects infection- or mechanical-stress-induced ECM damage, thereby initiating adaptive mitochondria-based immune and metabolic responses.

Keywords:  TGF-β; TMEM2; extracellular matrix; hyaluronan; immunity; mitochondria

DOI:  https://doi.org/10.1016/j.cell.2024.05.057
Biomater Sci. 2024 Jun 24.

Matrix stiffness influences response to chemo and targeted therapy in brain metastatic breast cancer cells.

Venu Yakati, Lalita A Shevde, Shreyas S Rao.

Breast cancer is the most common malignancy accounting for 12.5% of all newly diagnosed cancer cases across the globe. Breast cancer cells are known to metastasize to distant organs (i.e., brain), wherein they can exhibit a dormant phenotype for extended time periods. These dormant cancer cells exhibit reduced proliferation and therapeutic resistance. However, the mechanisms by which dormant cancer cells exhibit resistance to therapy, in the context of brain metastatic breast cancer (BMBC), is not well understood. Herein, we utilized hyaluronic acid (HA) hydrogels with varying stiffnesses to study drug responsiveness in dormant vs. proliferative BMBC cells. It was found that cells cultured on soft HA hydrogels (∼0.4 kPa) that showed a non-proliferative (dormant) phenotype exhibited resistance to Paclitaxel or Lapatinib. In contrast, cells cultured on stiff HA hydrogels (∼4.5 kPa) that showed a proliferative phenotype exhibited responsiveness to Paclitaxel or Lapatinib. Moreover, dormancy-associated resistance was found to be due to upregulation of the serum/glucocorticoid regulated kinase 1 (SGK1) gene which was mediated, in part, by the p38 signaling pathway. Accordingly, SGK1 inhibition resulted in a dormant-to-proliferative switch and response to therapy. Overall, our study demonstrates that matrix stiffness influences dormancy-associated therapy response mediated, in part, via the p38/SGK1 axis.

DOI: https://doi.org/10.1039/d4bm00342j
ACS Eng Au. 2024 Jun 19. 4(3): 351-358

Interplay of Spatial and Topological Defects in Polymer Networks.

B Ruşen Argun, Antonia Statt.

Polymer networks are widely used in applications, and the formation of a network and its gel point can be predicted. However, the effects of spatial and topological heterogeneity on the resulting network structure and ultimately the mechanical properties, are less understood. To address this challenge, we generate in silico random networks of cross-linked polymer chains with controlled spatial and topological defects. While all fully reacted networks investigated in this study have the same number of end-functionalized polymer strands and cross-linkers, we vary the degree of spatial and topological heterogeneities systematically. We find that spatially heterogeneous cross-linker distributions result in a reduction in the network's primary loops with increased spatial heterogeneity, the opposite trend as observed in homogeneous networks. By performing molecular dynamics simulations, we investigated the mechanical properties of the networks. Even though spatially heterogeneous networks have more elastically active strands and cross-linkers, they break at lower extensions than the homogeneous networks and sustain slightly lower maximum stresses. Their shear moduli are higher, i.e., stiffer, than theoretically predicted, and higher than their homogeneous gel counterparts. Our results highlight that topological loop defects and spatial heterogeneities result in significantly different network structures and, ultimately, different mechanical properties.

DOI: https://doi.org/10.1021/acsengineeringau.3c00072
ACS Meas Sci Au. 2024 Jun 19. 4(3): 260-266

Automated Analysis of Extracellular Matrix Invasion of Cancer Cells from Tumor Spheroids.

Jacob Heiss, Hossein Tavana.

The main cause of mortality among cancer patients is metastatic disease. Metastasis develops from cancer cells that invade the stromal tissue and intravasate the circulatory or lymphatic systems to eventually form new tumors in other organs. Blocking cancer cell invasion can potentially prevent or reduce the metastatic progression of cancers. Testing different chemical compounds against cell invasion in three-dimensional cultures is a common laboratory technique. The efficacy of the treatments is often evaluated from confocal microscopic images of the cells using image processing. However, the analysis approaches are often subject to variations and inconsistencies due to user decisions that must be made while processing each image. To overcome this limitation, we developed a fully automated method to quantify the invasion of cancer cells from a 3D tumor spheroid into the surrounding extracellular matrix. We demonstrated that this method resolves cell invasion from spheroids of different shapes and sizes and from cells that invade as a cluster or individually. We also showed that this approach can help quantify the dose-dependent anti-invasive effects of a commonly used chemotherapy drug. Our automated method significantly reduces the time and increases the consistency and accuracy of cancer cell invasion analysis in three-dimensional cultures.

DOI: https://doi.org/10.1021/acsmeasuresciau.3c00064
Phys Rev E. 2024 May;109(5-1): 054904

Effects of coordination and stiffness scale separation in disordered elastic networks.

Edan Lerner.

Many fibrous materials are modeled as elastic networks featuring a substantial separation between the stiffness scales that characterize different microscopic deformation modes of the network's constituents. This scale separation has been shown to give rise to emergent complexity in these systems' linear and nonlinear mechanical response. Here we study numerically a simple model featuring said stiffness scale separation in two-dimensions and show that its mechanical response is governed by the competition between the characteristic stiffness of collective nonphononic soft modes of the stiff subsystem, and the characteristic stiffness of the soft interactions. We present and rationalize the behavior of the shear modulus of our complex networks across the unjamming transition at which the stiff subsystem alone loses its macroscopic mechanical rigidity. We further establish a relation in the soft-interaction-dominated regime between the shear modulus, the characteristic frequency of nonphononic vibrational modes, and the mesoscopic correlation length that marks the crossover from a disorder-dominated response to local mechanical perturbations in the near field, to a linear, continuumlike response in the far field. The effects of spatial dimension on the observed scaling behavior are discussed, in addition to the interplay between stiffness scales in strain-stiffened networks, which is relevant to understanding the nonlinear mechanics of non-Brownian fibrous biomatter.

DOI: https://doi.org/10.1103/PhysRevE.109.054904
Nanoscale Adv. 2024 Jun 25. 6(13): 3306-3319

Side-view optical microscopy-assisted atomic force microscopy for thickness-dependent nanobiomechanics.

Yanqi Yang, Mi Li.

The mechanical properties of biomaterials play an important role in regulating life processes, and thus accurately delineating the mechanical properties of biomaterials is critical to understand their functionality. Particularly, atomic force microscopy (AFM) has become a powerful and standard tool for characterizing and analyzing the nanomechanical properties of biomaterials, and providing a capability to visualize the thickness of the specimen during AFM-based force spectroscopy experiments benefits the biomedical applications of AFM. Here, we present a study of side-view optical microscopy-assisted AFM based on the integration of AFM and a detachable side-view optical microscopy module, which is able to image in real time the AFM indentation process from the side-view perspective and consequently facilitates the utilization of AFM-based indentation assay to precisely detect the mechanical properties of a specimen by taking its thickness into account. The effectiveness of side-view optical microscopy-assisted AFM was confirmed on four different types of biomaterial systems, including microfabricated structures, hydrogels, living cells, and cell spheroids, and the experimental results significantly show that the mechanical properties of samples at the micro/nanoscale are closely related to their thickness, vividly illustrating side-view optical microscopy-assisted AFM as a promising approach for accurate nanomechanics of biomaterial systems. The study provides additional possibilities for measuring the thickness-dependent nanomechanical properties of biomaterials by AFM, which will enable AFM-based force spectroscopy technology to address more biological issues with enhanced precision and will benefit the field of mechanobiology.

DOI: https://doi.org/10.1039/d4na00153b
Cell Rep Phys Sci. 2024 May 15. pii: 101922. [Epub ahead of print]5(5):

Multifunctional hydrogels with spatially controlled light activation with photocaged oligonucleotides.

Katelyn Mathis, Saanvi Gaddam, Rishi Koneru, Nikhil Sunkavalli, Catherine Wang, Manan Patel, Afia Ibnat Kohon, Brian Meckes.

Recreating tissue environments with precise control over mechanical, biochemical, and cellular organization is essential for next-generation tissue models for drug discovery, development studies, and the replication of disease environments. However, controlling these properties at cell-scale lengths remains challenging. Here, we report the development of printing approaches that leverage polyethylene glycol diacrylate (PEGDA) hydrogels containing photocaged oligonucleotides to spatially program material characteristics with non-destructive, non-ultraviolet light. We further integrate this system with a perfusion chamber to allow us to alter the composition of PEGDA hydrogels while retaining common light-activatable photocaged DNAs. We demonstrate that the hydrogels can capture DNA functionalized materials, including cells coated with complementary oligonucleotides with spatial control using biocompatible wavelengths. Overall, these materials open pathways to orthogonal capture of any DNA functionalized materials while not changing the sequences of the DNA.

DOI: https://doi.org/10.1016/j.xcrp.2024.101922
Cell Death Dis. 2024 Jun 22. 15(6): 440

Mechano-inhibition of endocytosis sensitizes cancer cells to Fas-induced Apoptosis.

Mehmet H Kural, Umidahan Djakbarova, Bilal Cakir, Yoshiaki Tanaka, Emily T Chan, Valeria I Arteaga Muniz, Yasaman Madraki, Hong Qian, Jinkyu Park, Lorenzo R Sewanan, In-Hyun Park, Laura E Niklason, Comert Kural.

The transmembrane death receptor Fas transduces apoptotic signals upon binding its ligand, FasL. Although Fas is highly expressed in cancer cells, insufficient cell surface Fas expression desensitizes cancer cells to Fas-induced apoptosis. Here, we show that the increase in Fas microaggregate formation on the plasma membrane in response to the inhibition of endocytosis sensitizes cancer cells to Fas-induced apoptosis. We used a clinically accessible Rho-kinase inhibitor, fasudil, that reduces endocytosis dynamics by increasing plasma membrane tension. In combination with exogenous soluble FasL (sFasL), fasudil promoted cancer cell apoptosis, but this collaborative effect was substantially weaker in nonmalignant cells. The combination of sFasL and fasudil prevented glioblastoma cell growth in embryonic stem cell-derived brain organoids and induced tumor regression in a xenograft mouse model. Our results demonstrate that sFasL has strong potential for apoptosis-directed cancer therapy when Fas microaggregate formation is augmented by mechano-inhibition of endocytosis.

DOI: https://doi.org/10.1038/s41419-024-06822-3
bioRxiv. 2024 Jun 12. pii: 2024.06.10.598340. [Epub ahead of print]

Metabolomic Profiling and Characterization of a Novel 3D Culture System for Studying Chondrocyte Mechanotransduction.

Priyanka Brahmachary, Ebru Erdogan, Erik Myers, Ronald K June.

Articular chondrocytes synthesize and maintain the avascular and aneural articular cartilage. In vivo these cells are surrounded by a 3D pericellular matrix (PCM) containing predominantly collagen VI. The PCM protects chondrocytes and facilitates mechanotransduction, and PCM stiffness is critical in transmitting biomechanical signals to chondrocytes. Various culture systems with different hydrogels have been used to encapsulate chondrocytes for 3D culture, but many lack either the PCM or the in vivo stiffness of the cartilage matrix. Here, we demonstrate that primary chondrocytes cultured in alginate will form a pericellular matrix and display a phenotype similar to in vivo conditions. We found that primary human and bovine chondrocytes, when cultured in alginate beads with addition of sodium L-ascorbate for 7 days, had a pronounced PCM, retained their phenotype, and synthesized both collagens VI and II. This novel culture system enables alginate-encapsulated chondrocytes to develop a robust PCM thereby creating a model system to study mechanotransduction. We also observed distinct compression-induced changes in metabolomic profiles between the monolayer-agarose and alginate-released agarose-embedded chondrocytes indicating physiological changes in cell metabolism. Our data suggest that 3D preculture of chondrocytes in alginate before encapsulation in physiologically-stiff agarose leads to a pronounced development of pericellular matrix that is sustained in the presence of ascorbate. This novel model can be useful in studying the mechanism by which chondrocytes respond to cyclical compression and other types of loading simulating in vivo physiological conditions.

DOI: https://doi.org/10.1101/2024.06.10.598340
bioRxiv. 2024 Jun 14. pii: 2024.06.11.598541. [Epub ahead of print]

Combined effects of matrix stiffness and obesity-associated signaling directs progressive phenotype in PANC-1 pancreatic cancer cells in vitro.

A E Jones, J F Netto, T L Foote, B N K Ruliffson, C F Whittington.

Obesity is a leading risk factor of pancreatic ductal adenocarcinoma (PDAC) that contributes to poor disease prognosis and outcomes. Retrospective studies have identified this link, but interactions surrounding obesity and PDAC are still unclear. Research has shifted to contributions of fibrosis (desmoplasia) on malignancy, which involves increased deposition of collagens and other extracellular matrix (ECM) molecules and increased ECM crosslinking, all of which contribute to increased tissue stiffening. However, fibrotic stiffening is underrepresented as a model feature in current PDAC models. Fibrosis is shared between PDAC and obesity, and can be leveraged for in vitro model design, as current animal obesity models of PDAC are limited in their ability to isolate individual components of fibrosis to study cell behavior. In the current study, methacrylated type I collagen (PhotoCol®) was photo-crosslinked to pathological stiffness levels to recapitulate fibrotic ECM stiffening. PANC-1 cells were encapsulated within PhotoCol®, and the tumor-tissue constructs were prepared to represent normal (healthy) (∼600 Pa) and pathological (∼2000 Pa) tissues. Separately, human mesenchymal stem cells were differentiated into adipocytes representing lean (2D differentiation) and obese fat tissue (3D collagen matrix differentiation), and conditioned media was applied to PANC-1 tumor-tissue constructs. Conditioned media from obese adipocytes showed increased vimentin expression, a hallmark of invasiveness and progression, that was not seen after exposure to media from lean adipocytes or control media. Characterization of the obese adipocyte secretome suggested that some PANC-1 differences may arise from increased interleukin-8 and -10 compared to lean adipocytes. Additionally, high matrix stiffness associated induced an amoeboid morphology in PANC-1 cells that was not present at low stiffness. Amoeboid morphology is an accessory to epithelial-to-mesenchymal transition and is used to navigate complex ECM environments. This plasticity has greater implications for treatment efficacy of metastatic cancers. Overall, this work 1) highlights the importance of investigating PDAC-obesity interactions to study the effects on disease progression and persistence, 2) establishes PhotoCol® as a matrix material that can be leveraged to study amoeboid morphology and invasion in PDAC, and 3) emphasizes the importance of integrating both biophysical and biochemical interactions associated within both pathologies for in vitro PDAC models.

DOI: https://doi.org/10.1101/2024.06.11.598541
ACS Appl Mater Interfaces. 2024 Jun 24.

Synergistic Coassembly of Folic Acid-Based Supramolecular Polymer with a Covalent Polymer Toward Fabricating Functional Antibacterial Biomaterials.

Ipsita Sahu, Jaya Verma, Ashis Kumar Bera, Shreya Pande, Aashwini Bhavsar, Falguni Pati, Priyadarshi Chakraborty.

  Supramolecular biomaterials can recapitulate the structural and functional facets of the native extracellular matrix and react to biochemical cues, leveraging the unique attributes of noncovalent interactions, including reversibility and tunability. However, the low mechanical properties of supramolecular biomaterials can restrict their utilization in specific applications. Combining the advantages of supramolecular polymers with covalent polymers can lead to the fabrication of tailor-made biomaterials with enhanced mechanical properties/degradability. Herein, we demonstrate a synergistic coassembled self-healing gel as a multifunctional supramolecular material. As the supramolecular polymer component, we chose folic acid (vitamin B9), an important biomolecule that forms a gel comprising one-dimensional (1D) supramolecular polymers. Integrating polyvinyl alcohol (PVA) into this supramolecular gel alters its ultrastructure and augments its mechanical properties. A drastic improvement of complex modulus (G*) (∼3674 times) was observed in the folic acid-PVA gel with 15% w/v PVA (33215 Pa) compared with the folic acid gel (9.04 Pa). The coassembled hydrogels possessed self-healing and injectable/thixotropic attributes and could be printed into specific three-dimensional (3D) shapes. Synergistically, the supramolecular polymers of folic acid also improve the toughness, durability, and ductility of the PVA films. A nanocomposite of the gels with silver nanoparticles exhibited excellent catalytic efficiency and antibacterial activity. The folic acid-PVA coassembled gels and films also possessed high cytocompatibility, substantiated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and live-dead assays. Taken together, the antibacterial and cell-adhesive attributes suggest potential applications of these coassembled biomaterials for tissue engineering and wound healing.

Keywords:  antibacterial property; catalysis; cytocompatibility; free-standing films; gel nanocomposites; self-healing property; supramolecular gel

DOI:  https://doi.org/10.1021/acsami.4c06785
Matrix Biol. 2024 Jun 25. pii: S0945-053X(24)00088-X. [Epub ahead of print]

Cancer-associated fibroblasts promote proliferation, angiogenesis, metastasis and immunosuppression in gastric cancer.

Peiyuan Li, Huan Zhang, Tao Chen, Yajing Zhou, Jiaoyang Yang, Jin Zhou.

  Despite advances in surgery, radiotherapy and immunotherapy, the mortality rate for gastric cancer remains one of the highest in the world. A large body of evidence has demonstrated that cancer-associated fibroblasts (CAFs), as core members of the stroma, can secrete cytokines, proteins and exosomes to create a tumour microenvironment that is conducive to cancer cell survival. CAFs can also interact with cancer cells to form a complex signalling network, enabling cancer cells to more easily metastasise to other organs and tissues in the body and develop metastatic foci. In this review, we provide an overview of the CAFs concept and activators. We focus on elucidating their effects on immune cells, intratumoural vasculature, extracellular matrix, as well as cancer cell activity, metastatic power and metabolism, and on enhancing the metastatic ability of cancer cells through activation of JAK/STAT, NF/κB and CXCL12/CXCR4. Various therapeutic agents targeting CAFs are also under development and are expected to improve the prognosis of gastric cancer in combination with existing treatment options.

Keywords:  Cancer-associated fibroblasts; Cytokines; Gastric cancer; Metastasis; Therapeutic target

DOI:  https://doi.org/10.1016/j.matbio.2024.06.004
Macromol Biosci. 2024 Jun 28. e2400227

Modeling 3D Tumor Invasiveness to Modulate Macrophage Phenotype in a Human-based Hydrogel Platform.

Cátia F Monteiro, Catarina R Almeida, Catarina A Custódio, João F Mano.

  The immune system is a pivotal player in determining tumor fate, contributing to the immunosuppressive microenvironment that supports tumor progression. Considering the emergence of biomaterials as promising platforms to mimic the tumor microenvironment, human platelet lysate(PLMA)-based hydrogel beads are proposed as 3D platforms to recapitulate the tumor milieu and recreate the synergistic tumor-macrophage communication. Having characterized the biomaterial-mediated pro-regenerative macrophage phenotype, an osteosarcoma spheroid encapsulated into a PLMA hydrogel bead was explored to study macrophage immunomodulation through paracrine signaling. The culture of PLMA-Tumor beads on the top of a 2D monolayer of macrophages revealed that tumor cells triggered morphologic and metabolic adaptations in macrophages. The cytokine profile, coupled with the upregulation of gene and protein anti-inflammatory biomarkers clearly indicated macrophage polarization toward an M2-like phenotype. Moreover, the increased gene expression of chemokines identified as pro-tumoral environmental regulators suggested a tumor-associated macrophage phenotype, exclusively stimulated by tumor cells. This pro-tumoral microenvironment was also found to enhance tumor invasiveness ability and proliferation. Besides providing a robust in vitro immunomodulatory tumor model that faithfully recreates the tumor-macrophage interplay, this human-based platform has the potential to provide fundamental insights into immunosuppressive signaling and predict immune-targeted response. This article is protected by copyright. All rights reserved.

Keywords:  3D tumor model; human protein‐derived hydrogel; immunomodulation; macrophage polarization

DOI:  https://doi.org/10.1002/mabi.202400227
Proc Natl Acad Sci U S A. 2024 Jul 02. 121(27): e2402587121

Hydrogel morphogenesis induced by force-controlled growth.

Zhi Jian Wang, Ji Lin, Tasuku Nakajima, Jian Ping Gong.

  Morphogenesis is one of the most marvelous natural phenomena. The morphological characteristics of biological organs develop through growth, which is often triggered by mechanical force. In this study, we propose a bioinspired strategy for hydrogel morphogenesis through force-controlled chemical reaction and growth under isothermal conditions. We adopted a double network (DN) hydrogel with sacrificial bonds. Applying mechanical force to the gel caused deformation and sacrificial bond rupture. By supplying monomers to the gel, the radicals generated by the bond rupture triggered the formation of a new network inside the deformed gel. This new network conferred plasticity to the elastic gel, allowing it to maintain its deformed shape, along with increased volume and strength. We demonstrated that sheet-shaped DN hydrogels rapidly adopted various three-dimensional shapes at ambient temperature when subjected to forces such as drawing and blowing. This mechanism enables morphogenesis of elastic hydrogels and will promote the application of these materials in biomedical fields and soft machines.

Keywords:  blow molding; double network hydrogel; growth-induced-plasticity; mechanochemistry; shape morphing

DOI:  https://doi.org/10.1073/pnas.2402587121
STAR Protoc. 2024 Jun 27. pii: S2666-1667(24)00324-1. [Epub ahead of print]5(3): 103159

Protocol for real-time assessment of mitochondrial and glycolytic ATP production in patient-derived glioma stem-like cells.

Pratibha Sharma, Vinay K Puduvalli.

  Glioma cells switch between energetic pathways to adapt and resist therapies. We present a protocol for measuring mitochondrial and glycolytic ATP rates in patient-derived glioma stem-like cells using a Seahorse XF ATP rate assay. We describe steps for growing 3D glioma stem-like cells, attaching cells to the assay plate, preparing drugs, and running the ATP rate assay. We also detail procedures for imaging viable cell numbers and normalization, with tips to overcome pitfalls in Agilent Seahorse assays.

Keywords:  Cancer; Cell Biology; Cell culture; Cell-based Assays; Health Sciences; Metabolism; Neuroscience

DOI:  https://doi.org/10.1016/j.xpro.2024.103159
J Cell Biol. 2024 Sep 02. pii: e202404066. [Epub ahead of print]223(9):

A Lifeact-EGFP quail for studying actin dynamics in vivo.

Yanina D Alvarez, Marise van der Spuy, Jian Xiong Wang, Ivar Noordstra, Siew Zhuan Tan, Murron Carroll, Alpha S Yap, Olivier Serralbo, Melanie D White.

Here, we report the generation of a transgenic Lifeact-EGFP quail line for the investigation of actin organization and dynamics during morphogenesis in vivo. This transgenic avian line allows for the high-resolution visualization of actin structures within the living embryo, from the subcellular filaments that guide cell shape to the supracellular assemblies that coordinate movements across tissues. The unique suitability of avian embryos to live imaging facilitates the investigation of previously intractable processes during embryogenesis. Using high-resolution live imaging approaches, we present the dynamic behaviors and morphologies of cellular protrusions in different tissue contexts. Furthermore, through the integration of live imaging with computational segmentation, we visualize cells undergoing apical constriction and large-scale actin structures such as multicellular rosettes within the neuroepithelium. These findings not only enhance our understanding of tissue morphogenesis but also demonstrate the utility of the Lifeact-EGFP transgenic quail as a new model system for live in vivo investigations of the actin cytoskeleton.

DOI: https://doi.org/10.1083/jcb.202404066