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1.
Polymers (Basel) ; 16(20)2024 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-39458703

RESUMO

In this investigation, we explored the microrheological characteristics of dilute hydrogels composed exclusively of Poly(vinyl) alcohol (PVA), Phytagel (PHY), and a blend of the two in varying concentrations. Each of these polymers has established applications in the biomedical field, such as drug delivery and lens drops. This study involved varying the sample concentrations from 0.15% to 0.3% (w/w) to assess how the concentration influenced the observed rheological response. Two probe sizes were employed to examine the impact of the size and verify the continuity hypothesis. The use of two polymer blends revealed their immiscibility and tendency to undergo phase separation, as supported by the existing literature. Exploring the microrheological structure is essential for a comprehensive understanding of the molecular scale. Dynamic light scattering (DLS) was chosen due to its wide frequency range and widespread availability. The selected dilute concentration range was hypothesized to fall within the transition from an ergodic to a non-ergodic medium. Properly identifying the sample's nature during an analysis-whether it is ergodic or not-is critical, as highlighted in the literature. The obtained results clearly demonstrate an overlap in the results for the storage (G') and loss moduli (G″) for the different probe particle sizes, confirming the fulfillment of the continuum hypothesis.

2.
Int J Biol Macromol ; 282(Pt 1): 136813, 2024 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-39447801

RESUMO

This work aimed to exploit the effects of pomegranate peel (PP) addition (10 %) and homogenization process (0-12,000 rpm) on the film-forming behavior and interactions of starch film to improve its microstructures and properties. Microrheology results showed that the viscosity of the film suspension was reduced by homogenization, and the amylopectin aggregation was restricted by PP particles under film formation. This helped improve the accessibility of starch and PP, increasing the hydrogen bond energy in the composites from 18.12 to 19.35 kJ/mol and promoting the ester bonds as confirmed by the FTIR results. As a result, the starch film reinforced by PP and homogenization showed a uniform and compact microstructure with reduced water activity, increased UV-blocking capacity (from 67.46 % to 90.11 %), enhanced mechanical resistance, and reduced water vapor permeability (from 4.70 to 2.21 × 10-10·g·m-1·s-1·Pa-1). Additionally, the starch film with 10 % PP also presented superior antimicrobial properties and antioxidant activity with rates of over 86 %, providing an effective strawberry preservation function.

3.
Acta Biomater ; 2024 Oct 29.
Artigo em Inglês | MEDLINE | ID: mdl-39481624

RESUMO

Understanding how polymers deform in response to local stresses and strains, and how strains propagate from a local disturbance, are grand challenges in wide-ranging fields from materials manufacturing to cell mechanics. These dynamics are particularly complex for blends of polymers of distinct topologies, for which several different species-dependent mechanisms may contribute. Here, we use OpTiDDM (Optical-Tweezers-integrating-Differential-Dynamic-Microscopy) to elucidate deformation fields and propagation dynamics of binary blends of linear, ring and supercoiled DNA of varying sizes. We reveal robust non-monotonic dependence of strain alignment and superdiffusive transport with strain rate. However, peak alignment and superdiffusivity are surprisingly decoupled, occurring at different strain rates resonant with the distinct relaxation rates of the different topologies. Despite this universal resonance, we find that strain propagation of ring-linear blends is dictated by entanglements while supercoiled-ring blends are governed by Rouse dynamics. Our results capture critical subtleties in propagation and deformation dynamics of topological blends, shedding new light on the governing physics and offering a route towards decoupled tuning of response features. We anticipate our approach to be broadly generalizable to mapping the deformation dynamics of polymer blends, with an eye towards bottom-up bespoke materials design. STATEMENT OF SIGNIFICANCE: In biology and in manufacturing, biomaterials are often subject to localized and spatially nonuniform strains and stresses. Yet, understanding the extent to which strains are absorbed, distributed, or propagated across different spatiotemporal scales remains a grand challenge. Here, we combine optical tweezers with differential dynamic microscopy to elucidate deformation fields and propagation dynamics of blends of linear, ring and supercoiled DNA, revealing robust non-monotonic trends and decoupling of strain alignment and superdiffusivity, and capturing critical subtleties in propagation and deformation dynamics. Our results, shedding important new physical insight to guide decoupled tuning of response features, may be leveraged to map the deformation dynamics of wide-ranging systems of biopolymers and other macromolecules, with an eye towards bottom-up bespoke biomaterials design.

4.
J Biophotonics ; : e202400379, 2024 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-39389583

RESUMO

The blood rheology in vitro in glass or plastic microfluidic chips is different from that in vivo in blood vessels with similar geometry. Absence of vascular endothelium is suggested to cause these discrepancies. This work aims to perform in vitro measurements of blood microrheologic parameters in a slit microfluidic channel covered with endothelial cells (HUVEC). The laser aggregometry was employed to measure the intensity of laser light, backscattered from the blood flow, as a function of shear stress to evaluate the hydrodynamic strength of red blood cells (RBC) aggregates in terms of critical shear stress (CSS). The results demonstrated a decrease in CSS accompanied by an increase in the accuracy of its measurement at similar shear stresses when endothelial cells were present in the channel. The findings hold valuable implications for advanced approaches for endothelization of microfluidic devices, facilitating the study of blood flow dynamics in physiologically more relevant environment.

5.
Polymers (Basel) ; 16(18)2024 Sep 16.
Artigo em Inglês | MEDLINE | ID: mdl-39339082

RESUMO

This work investigated the gelation kinetics and mechanical properties of agarose hydrogels studied at different concentrations (in the range 1-5 g/L) and temperatures. Rheological measurements were performed by diffusing wave spectroscopy (DWS) using polystyrene and titanium dioxide particles as probes. The study emphasized the influence of gelation kinetics on the mechanical behavior of the hydrogels. The results showed that the gel properties were closely related to the thermal history and aging time of the samples. The insights gained from this study are critical for optimizing the performance of agarose hydrogels in specific applications and highlight the importance of controlling the concentration and thermal conditions during hydrogel preparation.

6.
ACS Biomater Sci Eng ; 10(9): 5701-5713, 2024 Sep 09.
Artigo em Inglês | MEDLINE | ID: mdl-39171932

RESUMO

Covalent adaptable networks (CANs) are polymeric networks with cross-links that can break and reform in response to external stimuli, including pH, shear, and temperature, making them potential materials for use as injectable cell delivery vehicles. In the native niche, cells rearrange the extracellular matrix (ECM) to undergo basic functions including migration, spreading, and proliferation. Bond rearrangement enables these hydrogels to mimic viscoelastic properties of the native ECM which promote migration and delivery from the material to the native tissue. In this work, we characterize thioester CANs to inform their design as effective cell delivery vehicles. Using bulk rheology, we characterize the rearrangement of these networks when they are subjected to strain, which mimics the strain applied by a syringe, and using multiple particle tracking microrheology (MPT) we measure cell-mediated remodeling of the pericellular region. Thioester networks are formed by photopolymerizing 8-arm poly(ethylene glycol) (PEG)-thiol and PEG-thioester norbornene. Bulk rheology measures scaffold properties during low and high strain and demonstrates that thioester scaffolds can recover rheological properties after high strain is applied. We then 3D encapsulated human mesenchymal stem cells (hMSCs) in thioester scaffolds. Using MPT, we characterize degradation in the pericellular region. Encapsulated hMSCs degrade these scaffolds within ≈4 days post-encapsulation. We hypothesize that this degradation is mainly due to cytoskeletal tension that cells apply to the matrix, causing adaptable thioester bonds to rearrange, leading to degradation. To verify this, we inhibited cytoskeletal tension using blebbistatin, a myosin-II inhibitor. Blebbistatin-treated cells can degrade these networks only by secreting enzymes including esterases. Esterases hydrolyze thioester bonds, which generate free thiols, leading to bond exchange. Around treated cells, we measure a decrease in the extent of pericellular degradation. We also compare cell area, eccentricity, and speed of untreated and treated cells. Inhibiting cytoskeletal tension results in significantly smaller cell area, more rounded cells, and lower cell speeds when compared to untreated cells. Overall, this work shows that cytoskeletal tension plays a major role in hMSC-mediated degradation of thioester networks. Cytoskeletal tension is also important for the spreading and motility of hMSCs in these networks. This work informs the design of thioester scaffolds for tissue regeneration and cell delivery.


Assuntos
Hidrogéis , Células-Tronco Mesenquimais , Reologia , Compostos de Sulfidrila , Hidrogéis/química , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Compostos de Sulfidrila/química , Polietilenoglicóis/química , Matriz Extracelular/metabolismo , Matriz Extracelular/química , Ésteres/química , Alicerces Teciduais/química
7.
Biochem Biophys Res Commun ; 736: 150496, 2024 Aug 03.
Artigo em Inglês | MEDLINE | ID: mdl-39128264

RESUMO

The pancreatic ductal adenocarcinoma (PDAC) tumor microenvironment is distinguished by a high degree of fibrosis and inflammation, known as desmoplasia. Desmoplasia increases the stromal deposition and extracellular matrix (ECM) stiffness observed in the tumor microenvironment, contributing to the dampened penetration of pharmacological agents. The molecular and biophysical composition of the ECM during the earliest cellular changes in the development of PDAC, i.e. acinar ductal metaplasia (ADM), has not been extensively explored. We report that the mRNA expression of key protein components of the ECM increases during ADM in p48Cre/+;LSL-KrasG12D (KC) mouse acinar organoids cultured in Matrigel. Treatment of the organoids with small molecular weight epigenetic modulating compounds that inhibit or reverse ADM (largazole, FK228 and chaetocin) dramatically reduced the tissue mRNA expression of collagens, hyaluronan synthase, laminin and fibronectin. The storage moduli, determined by video tracking of fluorescent nanoparticles embedded into the Matrigel, increased during ADM and was reduced following treatment with the epigenetic modulating compounds. We report that the ECM of mouse organoids stiffens during ADM and is further enhanced by the presence of mutant Kras. Moreover, select HDAC and HMT inhibitors reduced the mRNA expression of ECM components and ECM stiffness during inhibition and reversal of ADM, suggesting that these compounds may be useful as adjuvants to enhance the tumor penetration of agents used to treat PDAC.

8.
Int J Biol Macromol ; 277(Pt 3): 134378, 2024 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-39097048

RESUMO

The soy hull polysaccharide (SHP) exhibits excellent interfacial activity and holds potential as an emulsifier for emulsions. To reveal the behavior of SHP at the water/oil (W/O) interface in situ, molecular dynamics (MD) simulations and particle tracking microrheology were used in this study. The results of MD reveal that SHP molecular spontaneously move toward the interface and rhamnogalacturonan-I initiates this movement, while its galacturonic acids on it act as anchors to immobilize the SHP molecules at the W/O interface. Microrheology results suggest that SHP forms microgels at the W/O interface, with the lattices of the microgels continually undergoing dynamic changes. At low concentrations of SHP and short interfacial formation time, the network of the microgels is weak and dominated by viscous properties. However, when SHP reaches 0.75 % and the interfacial formation time is about 60 min, the microgels show perfect elasticity, which is beneficial for stabilizing emulsions.


Assuntos
Emulsões , Glycine max , Simulação de Dinâmica Molecular , Polissacarídeos , Reologia , Água , Água/química , Glycine max/química , Polissacarídeos/química , Emulsões/química , Óleos/química , Viscosidade , Pectinas/química , Microgéis/química
9.
Proc Natl Acad Sci U S A ; 121(33): e2403740121, 2024 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-39102540

RESUMO

The formation of macrophage-derived foam cells has been recognized as the pathological hallmark of atherosclerotic diseases. However, the pathological evolution dynamics and underlying regulatory mechanisms remain largely unknown. Herein, we introduce a single-particle rotational microrheology method for pathological staging of macrophage foaming and antiatherosclerotic explorations by probing the dynamic changes of lysosomal viscous feature over the pathological evolution progression. The principle of this method involves continuous monitoring of out-of-plane rotation-caused scattering brightness fluctuations of the gold nanorod (AuNR) probe-based microrheometer and subsequent determination of rotational relaxation time to analyze the viscous feature in macrophage lysosomes. With this method, we demonstrated the lysosomal viscous feature as a robust pathological reporter and uncovered three distinct pathological stages underlying the evolution dynamics, which are highly correlated with a pathological stage-dependent activation of the NLRP3 inflammasome-involved positive feedback loop. We also validated the potential of this positive feedback loop as a promising therapeutic target and revealed the time window-dependent efficacy of NLRP3 inflammasome-targeted drugs against atherosclerotic diseases. To our knowledge, the pathological staging of macrophage foaming and the pathological stage-dependent activation of the NLRP3 inflammasome-involved positive feedback mechanism have not yet been reported. These findings provide insights into in-depth understanding of evolutionary features and regulatory mechanisms of macrophage foaming, which can benefit the analysis of effective therapeutical drugs as well as the time window of drug treatment against atherosclerotic diseases in preclinical studies.


Assuntos
Aterosclerose , Células Espumosas , Ouro , Proteína 3 que Contém Domínio de Pirina da Família NLR , Aterosclerose/patologia , Animais , Ouro/química , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Células Espumosas/patologia , Células Espumosas/metabolismo , Macrófagos/patologia , Macrófagos/metabolismo , Humanos , Lisossomos/metabolismo , Inflamassomos/metabolismo , Nanotubos/química , Reologia
10.
Mol Cell ; 84(14): 2698-2716.e9, 2024 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-39059370

RESUMO

The cell interior is packed with macromolecules of mesoscale size, and this crowded milieu significantly influences cellular physiology. Cellular stress responses almost universally lead to inhibition of translation, resulting in polysome collapse and release of mRNA. The released mRNA molecules condense with RNA-binding proteins to form ribonucleoprotein (RNP) condensates known as processing bodies and stress granules. Here, we show that polysome collapse and condensation of RNA transiently fluidize the cytoplasm, and coarse-grained molecular dynamic simulations support this as a minimal mechanism for the observed biophysical changes. Increased mesoscale diffusivity correlates with the efficient formation of quality control bodies (Q-bodies), membraneless organelles that compartmentalize misfolded peptides during stress. Synthetic, light-induced RNA condensation also fluidizes the cytoplasm. Together, our study reveals a functional role for stress-induced translation inhibition and formation of RNP condensates in modulating the physical properties of the cytoplasm to enable efficient response of cells to stress conditions.


Assuntos
Citoplasma , Polirribossomos , Ribonucleoproteínas , Polirribossomos/metabolismo , Citoplasma/metabolismo , Humanos , Ribonucleoproteínas/metabolismo , Ribonucleoproteínas/genética , Simulação de Dinâmica Molecular , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , Biossíntese de Proteínas , Proteínas de Ligação a RNA/metabolismo , Proteínas de Ligação a RNA/genética , Condensados Biomoleculares/metabolismo , Grânulos de Estresse/metabolismo , Grânulos de Estresse/genética
11.
Micromachines (Basel) ; 15(7)2024 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-39064399

RESUMO

Microrheology, the study of material flow at micron scales, has advanced significantly since Robert Brown's discovery of Brownian motion in 1827. Mason and Weitz's seminal work in 1995 established the foundation for microrheology techniques, enabling the measurement of viscoelastic properties of complex fluids using light-scattering particles. However, existing techniques face limitations in exploring very slow dynamics, crucial for understanding biological systems. Here, we present a proof of concept for a novel microrheology technique called "Optical Halo", which utilises a ring-shaped Bessel beam created by optical tweezers to overcome existing limitations. Through numerical simulations and theoretical analysis, we demonstrate the efficacy of the Optical Halo in probing viscoelastic properties across a wide frequency range, including low-frequency regimes inaccessible to conventional methods. This innovative approach holds promise for elucidating the mechanical behaviour of complex biological fluids.

12.
Gels ; 10(6)2024 Jun 16.
Artigo em Inglês | MEDLINE | ID: mdl-38920947

RESUMO

Food hydrogels, used as delivery systems for bioactive compounds, can be formulated with various food-grade biopolymers. Their industrial utility is largely determined by their physicochemical properties. However, comprehensive data on the properties of pea protein-psyllium binary hydrogels under different pH and ionic strength conditions are limited. The aim of this research was to evaluate the impact of pH (adjusted to 7, 4.5, and 3) and ionic strength (modified by NaCl addition to 0.15 and 0.3 M) on the physical stability, color, texture, microrheological, and viscoelastic properties of these hydrogels. Color differences were most noticeable at lower pH levels. Inducing hydrogels at pH 7 (with or without NaCl) and pH 4.5 and 3 (without NaCl) resulted in complete gel structures with low stability, low elastic and storage moduli, and low complex viscosity, making them easily spreadable. Lower pH inductions (4.5 and 3) in the absence of NaCl resulted in hydrogels with shorter linear viscoelastic regions. Hydrogels induced at pH 4.5 and 3 with NaCl had high structural stability, high G' and G" moduli, complex viscosity, and high spreadability. Among the tested induction conditions, pH 3 with 0.3 M NaCl allowed for obtaining a hydrogel with the highest elastic and storage moduli values. Adjusting pH and ionic strength during hydrogel induction allows for modifying and tailoring their properties for specific industrial applications.

13.
Adv Mater ; 36(32): e2401745, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38815174

RESUMO

Mucus is a dynamic biological hydrogel, composed primarily of the glycoprotein mucin, exhibits unique biophysical properties and forms a barrier protecting cells against a broad-spectrum of viruses. Here, this work develops a polyglycerol sulfate-based dendronized mucin-inspired copolymer (MICP-1) with ≈10% repeating units of activated disulfide as cross-linking sites. Cryo-electron microscopy (Cryo-EM) analysis of MICP-1 reveals an elongated single-chain fiber morphology. MICP-1 shows potential inhibitory activity against many viruses such as herpes simplex virus 1 (HSV-1) and SARS-CoV-2 (including variants such as Delta and Omicron). MICP-1 produces hydrogels with viscoelastic properties similar to healthy human sputum and with tuneable microstructures using linear and branched polyethylene glycol-thiol (PEG-thiol) as cross-linkers. Single particle tracking microrheology, electron paramagnetic resonance (EPR) and cryo-scanning electron microscopy (Cryo-SEM) are used to characterize the network structures. The synthesized hydrogels exhibit self-healing properties, along with viscoelastic properties that are tuneable through reduction. A transwell assay is used to investigate the hydrogel's protective properties against viral infection against HSV-1. Live-cell microscopy confirms that these hydrogels can protect underlying cells from infection by trapping the virus, due to both network morphology and anionic multivalent effects. Overall, this novel mucin-inspired copolymer generates mucus-mimetic hydrogels on a multi-gram scale. These hydrogels can be used as models for disulfide-rich airway mucus research, and as biomaterials.


Assuntos
Herpesvirus Humano 1 , Hidrogéis , Muco , SARS-CoV-2 , Hidrogéis/química , Hidrogéis/farmacologia , Herpesvirus Humano 1/efeitos dos fármacos , Humanos , Muco/metabolismo , SARS-CoV-2/efeitos dos fármacos , Mucinas/química , Mucinas/metabolismo , Antivirais/farmacologia , Antivirais/química , Polímeros/química , Polímeros/farmacologia , Animais , Dissulfetos/química , Polietilenoglicóis/química , Microscopia Crioeletrônica , COVID-19/virologia , Glicerol
14.
Polymers (Basel) ; 16(10)2024 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-38794527

RESUMO

Recently, there has been interest in determining the viscoelastic properties of polymeric liquids and other complex fluids by means of Diffusing Wave Spectroscopy (DWS). In this technique, light-scattering spectroscopy is applied to highly turbid fluids containing optical probe particles. The DWS spectrum is used to infer the time-dependent mean-square displacement and time-dependent diffusion coefficient D of the probes. From D, values for the storage modulus G'(ω) and the loss modulus G''(ω) are obtained. This paper is primarily concerned with the inference of the mean-square displacement from a DWS spectrum. However, in much of the literature, central to the inference that is said to yield D is an invocation g(1)(t)=exp(-2q2X(t)2¯) of the Gaussian Approximation for the field correlation function g(1)(t) of the scattered light in terms of the mean-square displacement X(t)2¯ of a probe particle during time t. Experiment and simulation both show that the Gaussian approximation is invalid for probes in polymeric liquids and other complex fluids. In this paper, we obtain corrections to the Gaussian approximation that will assist in interpreting DWS spectra of probes in polymeric liquids. The corrections reveal that these DWS spectra receive contributions from higher moments X(t)2n¯, n>1, of the probe displacement distribution function.

15.
Acta Biomater ; 181: 272-281, 2024 06.
Artigo em Inglês | MEDLINE | ID: mdl-38685460

RESUMO

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.


Assuntos
Fibrina , Géis , Reologia , Anisotropia , Géis/química , Fibrina/química , Humanos , Resistência à Tração , Estresse Mecânico , Colágeno/química , Animais
16.
ACS Nano ; 18(16): 10738-10757, 2024 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-38609349

RESUMO

Biomolecular condensates play important roles in a wide array of fundamental biological processes, such as cellular compartmentalization, cellular regulation, and other biochemical reactions. Since their discovery and first observations, an extensive and expansive library of tools has been developed to investigate various aspects and properties, encompassing structural and compositional information, material properties, and their evolution throughout the life cycle from formation to eventual dissolution. This Review presents an overview of the expanded set of tools and methods that researchers use to probe the properties of biomolecular condensates across diverse scales of length, concentration, stiffness, and time. In particular, we review recent years' exciting development of label-free techniques and methodologies. We broadly organize the set of tools into 3 categories: (1) imaging-based techniques, such as transmitted-light microscopy (TLM) and Brillouin microscopy (BM), (2) force spectroscopy techniques, such as atomic force microscopy (AFM) and the optical tweezer (OT), and (3) microfluidic platforms and emerging technologies. We point out the tools' key opportunities, challenges, and future perspectives and analyze their correlative potential as well as compatibility with other techniques. Additionally, we review emerging techniques, namely, differential dynamic microscopy (DDM) and interferometric scattering microscopy (iSCAT), that have huge potential for future applications in studying biomolecular condensates. Finally, we highlight how some of these techniques can be translated for diagnostics and therapy purposes. We hope this Review serves as a useful guide for new researchers in this field and aids in advancing the development of new biophysical tools to study biomolecular condensates.


Assuntos
Condensados Biomoleculares , Condensados Biomoleculares/química , Condensados Biomoleculares/metabolismo , Microscopia de Força Atômica , Pinças Ópticas , Humanos , Microscopia/métodos
17.
ACS Biomater Sci Eng ; 10(5): 3378-3386, 2024 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-38517700

RESUMO

The presence of colloidal scaffolds composed of proteins and hyaluronic acid engenders unique viscous and elastic properties to the synovial fluid (SF). While the elastic resistance of SF due to the presence of such nanoscale structures provides the load-bearing capacity, the viscous nature enables fluidity of the joints during the movements to minimize the wear and tear of the adjacent muscle, cartilage, or bone tissues. It is well-known that the hypoxic conditions at the bone joints often increase the lactic acid (LA) concentration due to the occurrence of excess anaerobic respiration during either hyperactivity or arthritic conditions. The present study uncovers that in such a scenario, beyond a critical loading of LA, the colloidal nanoscaffolds of SF break down to precipitate higher molecular weight (MW) proteins and hyaluronic acid (HA). Subsequently, the viscosity and elasticity of SF reduce drastically to manifest a fluid that has reduced load bearing and wear and tear resistance capacity. Interestingly, the study also suggests that a heathy SF is a viscoelastic fluid with a mild Hookean elasticity and non-Newtonian fluidity, which eventually transforms into a viscous watery liquid in the presence of a higher loading of LA. We employ this knowledge to biosynthesize an artificial SF that emulates the characteristics of the real one. Remarkably, the spatiotemporal microscopic images uncover that even for the artificial SF, a dynamic cross-linking of the high MW proteins and HA takes place before precipitating out of the same from the artificial SF matrix, emulating the real one. Control experiments suggest that this phenomenon is absent in the case when LA is mixed with either pure HA or proteins. The experiments unfold the specific role of LA in the destruction of colloidal nanoscaffolds of synovia, which is an extremely important requirement for the biosynthesis and translation of artificial synovial fluid.


Assuntos
Coloides , Ácido Hialurônico , Ácido Láctico , Reologia , Líquido Sinovial , Líquido Sinovial/química , Líquido Sinovial/metabolismo , Coloides/química , Viscosidade , Ácido Hialurônico/química , Ácido Láctico/química , Ácido Láctico/metabolismo , Humanos , Elasticidade
18.
Rheol Acta ; 63(3): 205-217, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38440195

RESUMO

Microrheology with optical tweezers (MOT) is an all-optical technique that allows the user to investigate a materials' viscoelastic properties at microscopic scales, and is particularly useful for those materials that feature complex microstructures, such as biological samples. MOT is increasingly being employed alongside 3D imaging systems and particle tracking methods to generate maps showing not only how properties may vary between different points in a sample but also how at a single point the viscoelastic properties may vary with direction. However, due to the diffraction limited shape of focussed beams, optical traps are inherently anisotropic in 3D. This can result in a significant overestimation of the fluids' viscosity in certain directions. As such, the rheological properties can only be accurately probed along directions parallel or perpendicular to the axis of trap beam propagation. In this work, a new analytical method is demonstrated to overcome this potential artefact. This is achieved by performing principal component analysis on 3D MOT data to characterise the trap, and then identify the frequency range over which trap anisotropy influences the data. This approach is initially applied to simulated data for a Newtonian fluid where the trap anisotropy induced maximum error in viscosity is reduced from ~ 150% to less than 6%. The effectiveness of the method is corroborated by experimental MOT measurements performed with water and gelatine solutions, thus confirming that the microrheology of a fluid can be extracted reliably across a wide frequency range and in any arbitrary direction. This work opens the door to fully spatially and angularly resolved 3D mapping of the rheological properties of soft materials over a broad frequency range.

19.
Gels ; 10(3)2024 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-38534589

RESUMO

As a soluble fiber, inulin is present in many plants and has many applications in food and non-food products. In this work, we investigated the rheological properties of inulin dispersions at seven concentrations. The linear viscoelastic properties of inulin were determined using a conventional mechanical rheometer. At 25 wt%, inulin exhibited fluid-like viscoelastic liquid behavior. However, when concentrations were ≥27.5 wt%, inulin exhibited gel-like viscoelastic properties. The viscoelastic properties (moduli and viscosities) increased with increasing inulin concentration. The high-frequency linear rheological properties of inulin were also investigated using the modern light scattering technique, diffusion wave spectroscopy (DWS). The diffusion wave spectroscopy (DWS) measurements showed the amplitude of complex moduli (|G*(ω)|) of inulin gels (≥27.5 wt%) to be proportional to ½ power law of the frequency, which suggests inulin gels behave similarly to flexible polymers. The non-linear steady shear experiments demonstrated that inulin exhibited shear-thinning behavior that was well fitted by a power law constitutive model. The trend of the power law exponent from the experiments indicated that the shear-thinning extent for inulin was greater as the inulin concentration increased. The results of this work indicated that the properties of inulin gel can be manipulated by altering its concentration. Therefore, the desired inulin product can be designed accordingly. These results can be used to direct further food and non-food applications, such as wound healing materials for inulin gels.

20.
Cell Rep ; 43(3): 113917, 2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38446665

RESUMO

Embryonic genome activation (EGA) marks the transition from dependence on maternal transcripts to an embryonic transcriptional program. The precise temporal regulation of gene expression, specifically the silencing of the Dux/murine endogenous retrovirus type L (MERVL) program during late 2-cell interphase, is crucial for developmental progression in mouse embryos. How this finely tuned regulation is achieved within this specific window is poorly understood. Here, using particle-tracking microrheology throughout the mouse oocyte-to-embryo transition, we identify a surge in cytoplasmic viscosity specific to late 2-cell interphase brought about by high microtubule and endomembrane density. Importantly, preventing the rise in 2-cell viscosity severely impairs nuclear reorganization, resulting in a persistently open chromatin configuration and failure to silence Dux/MERVL. This, in turn, derails embryo development beyond the 2- and 4-cell stages. Our findings reveal a mechanical role of the cytoplasm in regulating Dux/MERVL repression via nuclear remodeling during a temporally confined period in late 2-cell interphase.


Assuntos
Desenvolvimento Embrionário , Retrovirus Endógenos , Camundongos , Animais , Viscosidade , Desenvolvimento Embrionário/genética , Cromatina , Citoplasma , Regulação da Expressão Gênica no Desenvolvimento
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