A Novel 2.5D Culture Platform to Investigate the Role of Stiffness Gradients on Adhesion-Independent Cell Migration

Pebworth, Mark-Phillip; Cismas, Sabrina A.; Asuri, Prashanth

doi:10.1371/journal.pone.0110453

Cited by 26 publications

(27 citation statements)

References 44 publications

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“…Researchers have also taken advantage of the ability of hydrogels to swell or shrink in response to external stimuli (e.g., pH, temperature) to develop biosensors for the detection of biomolecules [4,5]. Additionally, their highly porous and hydrated polymer structure mimics the extracellular cellular matrix and renders them highly suitable for in vitro cell culture, and several studies have demonstrated the successful use of hydrogels to encapsulate mammalian cells in a 3D physiological-like environment and develop in vitro models of cell proliferation, migration, and differentiation [6][7][8][9][10]. However, hydrogels have poor mechanical strength, which limits their broad applicability for tissue engineering [11].…”

Section: Introductionmentioning

confidence: 99%

Role of Nanoparticle–Polymer Interactions on the Development of Double-Network Hydrogel Nanocomposites with High Mechanical Strength

et al. 2020

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Extensive experimental and theoretical research over the past several decades has pursued strategies to develop hydrogels with high mechanical strength. Our study investigated the effect of combining two approaches, addition of nanoparticles and crosslinking two different polymers (to create double-network hydrogels), on the mechanical properties of hydrogels. Our experimental analyses revealed that these orthogonal approaches may be combined to synthesize hydrogel composites with enhanced mechanical properties. However, the enhancement in double network hydrogel elastic modulus due to incorporation of nanoparticles is limited by the ability of the nanoparticles to strongly interact with the polymers in the network. Moreover, double-network hydrogel nanocomposites prepared using lower monomer concentrations showed higher enhancements in elastic moduli compared to those prepared using high monomer concentrations, thus indicating that the concentration of hydrogel monomers used for the preparation of the nanocomposites had a significant effect on the extent of nanoparticle-mediated enhancements. Collectively, these results demonstrate that the hypotheses previously developed to understand the role of nanoparticles on the mechanical properties of hydrogel nanocomposites may be extended to double-network hydrogel systems and guide the development of next-generation hydrogels with extraordinary mechanical properties through a combination of different approaches.

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Section: Introductionmentioning

confidence: 99%

Role of Nanoparticle–Polymer Interactions on the Development of Double-Network Hydrogel Nanocomposites with High Mechanical Strength

et al. 2020

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“…Meanwhile, 3D encapsulation of cells to preserve their viability during the freeze-thaw cycle opens up the possibility to further increase freezing efficiencies by varying the biophysical and biochemical properties of the scaffold. For instance, recent studies have demonstrated the individual as well as synergistic effects of matrix stiffness and cell-matrix interactions on cell responses to small molecule toxins (Pebworth et al 2014). While it is not clear if the intracellular responses to freeze-thaw and other toxic insults are similar, it could be postulated that biophysical and biochemical properties of the 3D cell culture matrix may also have a significant effect on cell viability during the freeze-thaw process.…”

Section: Comparison With Other Cryoprotectantsmentioning

confidence: 99%

“…(1) various research and bioprocess activities including viral packaging and drug discovery (Dietmair et al 2012), (2) studies where the effects of micro-environmental cues on cell proliferation, transfection and migration are explored (Pebworth et al 2014), (3) in biopharmaceutical production (Bandaranayake and Almo 2014) and (4) in desiccation research for genome mapping against anhydrobiotic organisms to elucidate common protection mechanisms against dehydration at the genetic level (Leprince and Buitink 2010). HEK cells have been cryopreserved using slow freezing in DMSO-based cocktails recommended for cryopreservation (Chaytor et al 2012, Kleman 2008.…”

Section: Introductionmentioning

confidence: 99%

Trehalose effectiveness as a cryoprotectant in 2D and 3D cell cultures of human embryonic kidney cells

Hara

Tottori

Anders

et al. 2016

Artificial Cells, Nanomedicine, and Biotechnology

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Post cryopreservation viability of human embryonic kidney (HEK) cells under two-dimensional (2D) and three-dimensional (3D) culture conditions was studied using trehalose as the sole cryoprotective agent. An L (3) Taguchi design was used to optimize the cryoprotection cocktail seeding process prior to slow-freezing with the specific aim of maximizing cell viability measured 7 days post thaw, using the combinatorial cell viability and in-vitro cytotoxicity WST assay. At low (200 mM) and medium (800 mM) levels of trehalose concentration, encapsulation in alginate offered a greater protection to cryopreservation. However, at the highest trehalose concentration (1200 mM) and in the absence of the pre-incubation step, there was no statistical difference at the 95% CI (p = 0.0212) between the viability of the HEK cells under 2D and 3D culture conditions estimated to be 17.9 ± 4.6% and 14.0 ± 3.6%, respectively. A parallel comparison between cryoprotective agents conducted at the optimal levels of the L study, using trehalose, dimethylsulfoxide and glycerol in alginate microcapsules yielded a viability of 36.0 ± 7.4% for trehalose, in average 75% higher than the results associated with the other two cell membrane-permeating compounds. In summary, the effectiveness of trehalose has been demonstrated by the fact that 3D cell cultures can readily be equilibrated with trehalose before cryopreservation, thus mitigating the cytotoxic effects of glycerol and dimethylsulfoxide.

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“…To determine whether the RGD-binding site on integrin is required for aMPV/B F protein-mediated membrane fusion, we incubated Vero and BHK-21 cells with RGD peptides that can block integrin as previously described (27,40). As shown in Fig.…”

Section: Peptide Blockade Of Integrin Reduces the Fusogenicity Of Thementioning

confidence: 99%

Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection

Yun

Guan

Liu

et al. 2016

Journal of Biological Chemistry

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Avian metapneumovirus (aMPV) fusion (F) protein mediates virus-cell membrane fusion to initiate viral infection, which requires F protein binding to its receptor(s) on the host cell surface. However, the receptor(s) for aMPV F protein is still not identified. All known subtype B aMPV (aMPV/B) F proteins contain a conserved Arg-Asp-Asp (RDD) motif, suggesting that the aMPV/B F protein may mediate membrane fusion via the binding of RDD to integrin. When blocked with integrin-specific peptides, aMPV/B F protein fusogenicity and viral replication were significantly reduced. Specifically we identified integrin ␣v and/or ␤1-mediated F protein fusogenicity and viral replication using antibody blocking, small interfering RNAs (siRNAs) knockdown, and overexpression. Additionally, overexpression of integrin ␣v and ␤1 in aMPV/B non-permissive cells conferred aMPV/B F protein binding and aMPV/B infection. When RDD was altered to RAE (Arg-Ala-Glu), aMPV/B F protein binding and fusogenic activity were profoundly impaired. These results suggest that integrin ␣v␤1 is a functional receptor for aMPV/B F protein-mediated membrane fusion and virus infection, which will provide new insights on the fusogenic mechanism and pathogenesis of aMPV. Avian metapneumovirus (aMPV)3 belongs to the genus Metapneumovirus in the subfamily Pneumovirinae of paramyxoviridae (1, 2). aMPV is a major cause of acute rhinotracheitis in turkeys and is associated with swollen head syndrome in chickens. Thus, aMPV is considered a major threat to the poultry industry (1, 2). Based on phylogenetic analysis, aMPV has been divided into four subtypes: aMPV/A, aMPV/B, aMPV/C, and aMPV/D (3).The entry of a paramyxovirus into the host cell initially requires fusion of the viral envelope and cellular membrane, which is mediated by the glycoproteins on the viral envelope (4 -6). For viruses of the Paramyxovirinae subfamily, such as Newcastle disease virus, fusion (F) protein-mediated membrane fusion requires the assistance of the hemagglutininneuraminidase (HN) protein (7-10). However, viruses in the Pneumovirinae subfamily including human respiratory syncytial virus (hRSV), human metapneumovirus (hMPV), and aMPV, the F protein alone in the absence of the small hydrophobic and attachment (G) proteins can mediate membrane fusion and viral infection (11-15), suggesting that the mechanism by which hRSV, hMPV, and aMPV mediate virus-cell membrane fusion might be unique among the Paramyxovirinae subfamily.The F protein of Paramyxovirinae viruses is activated by a non-F attachment glycoprotein bound to its receptor(s) on the host cell (10, 16). The F protein then undergoes a coordinated series of conformational changes to its most stable form to promote membrane fusion (17-20). Thus, binding of the attachment glycoprotein to the cellular receptor(s) is critical for triggering the fusion process (21,22). For example, Newcastle disease virus HN protein attaches to sialic acid-containing receptors on the host cell surface and then activates the F protein to induce membrane fu...

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A Novel 2.5D Culture Platform to Investigate the Role of Stiffness Gradients on Adhesion-Independent Cell Migration

Cited by 26 publications

References 44 publications

Role of Nanoparticle–Polymer Interactions on the Development of Double-Network Hydrogel Nanocomposites with High Mechanical Strength

Role of Nanoparticle–Polymer Interactions on the Development of Double-Network Hydrogel Nanocomposites with High Mechanical Strength

Trehalose effectiveness as a cryoprotectant in 2D and 3D cell cultures of human embryonic kidney cells

Integrin αvβ1 Modulation Affects Subtype B Avian Metapneumovirus Fusion Protein-mediated Cell-Cell Fusion and Virus Infection

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