Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 10 de 10
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
1.
Biomater Sci ; 9(8): 3051-3068, 2021 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-33666608

RESUMO

3D-printing technologies, such as biofabrication, capitalize on the homogeneous distribution and growth of cells inside biomaterial hydrogels, ultimately aiming to allow for cell differentiation, matrix remodeling, and functional tissue analogues. However, commonly, only the mechanical properties of the bioinks or matrix materials are assessed, while the detailed influence of cells on the resulting mechanical properties of hydrogels remains insufficiently understood. Here, we investigate the properties of hydrogels containing cells and spherical PAAm microgel beads through multi-modal complex mechanical analyses in the small- and large-strain regimes. We evaluate the individual contributions of different filler concentrations and a non-fibrous oxidized alginate-gelatin hydrogel matrix on the overall mechanical behavior in compression, tension, and shear. Through material modeling, we quantify parameters that describe the highly nonlinear mechanical response of soft composite materials. Our results show that the stiffness significantly drops for cell- and bead concentrations exceeding four million per milliliter hydrogel. In addition, hydrogels with high cell concentrations (≥6 mio ml-1) show more pronounced material nonlinearity for larger strains and faster stress relaxation. Our findings highlight cell concentration as a crucial parameter influencing the final hydrogel mechanics, with implications for microgel bead drug carrier-laden hydrogels, biofabrication, and tissue engineering.


Assuntos
Bioimpressão , Microgéis , Alginatos , Gelatina , Hidrogéis , Engenharia Tecidual , Alicerces Teciduais
2.
Sci Rep ; 9(1): 17031, 2019 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-31745109

RESUMO

Mechanical stress exerted and experienced by cells during tissue morphogenesis and organ formation plays an important role in embryonic development. While techniques to quantify mechanical stresses in vitro are available, few methods exist for studying stresses in living organisms. Here, we describe and characterize cell-like polyacrylamide (PAAm) bead sensors with well-defined elastic properties and size for in vivo quantification of cell-scale stresses. The beads were injected into developing zebrafish embryos and their deformations were computationally analyzed to delineate spatio-temporal local acting stresses. With this computational analysis-based cell-scale stress sensing (COMPAX) we are able to detect pulsatile pressure propagation in the developing neural rod potentially originating from polarized midline cell divisions and continuous tissue flow. COMPAX is expected to provide novel spatio-temporal insight into developmental processes at the local tissue level and to facilitate quantitative investigation and a better understanding of morphogenetic processes.


Assuntos
Resinas Acrílicas/química , Fenômenos Biomecânicos/fisiologia , Módulo de Elasticidade , Desenvolvimento Embrionário/fisiologia , Peixe-Zebra/embriologia , Animais , Biologia Computacional , Módulo de Elasticidade/fisiologia , Embrião não Mamífero/embriologia , Técnicas Analíticas Microfluídicas , Morfogênese/fisiologia , Nanopartículas , Estresse Mecânico
3.
J Mater Chem B ; 6(39): 6245-6261, 2018 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-32254615

RESUMO

Cell mechanical measurements are gaining increasing interest in biological and biomedical studies. However, there are no standardized calibration particles available that permit the cross-comparison of different measurement techniques operating at different stresses and time-scales. Here we present the rational design, production, and comprehensive characterization of poly-acrylamide (PAAm) microgel beads mimicking size and overall mechanics of biological cells. We produced mono-disperse beads at rates of 20-60 kHz by means of a microfluidic droplet generator, where the pre-gel composition was adjusted to tune the beads' elasticity in the range of cell and tissue relevant mechanical properties. We verified bead homogeneity by optical diffraction tomography and Brillouin microscopy. Consistent elastic behavior of microgel beads at different shear rates was confirmed by AFM-enabled nanoindentation and real-time deformability cytometry (RT-DC). The remaining inherent variability in elastic modulus was rationalized using polymer theory and effectively reduced by sorting based on forward-scattering using conventional flow cytometry. Our results show that PAAm microgel beads can be standardized as mechanical probes, to serve not only for validation and calibration of cell mechanical measurements, but also as cell-scale stress sensors.

4.
ACS Biomater Sci Eng ; 3(11): 2962-2973, 2017 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-33418716

RESUMO

The measurement of cell stiffness is an important part of biological research with diverse applications in biology, biotechnology and medicine. Real-time deformability cytometry (RT-DC) is a new method to probe cell stiffness at high throughput by flushing cells through a microfluidic channel where cell deformation provides an indicator for cell stiffness (Otto et al. Real-time deformability cytometry: on-the-fly cell 725 mechanical phenotyping. Nat. Methods 2015, 12, 199-202). Here, we propose a full numerical model for single cells in a flow channel to quantitatively relate cell deformation to mechanical parameters. Thereby the cell is modeled as a viscoelastic material surrounded by a thin shell cortex, subject to bending stiffness and cortical surface tension. For small deformations our results show good agreement with a previously developed analytical model that neglects the influence of cell deformation on the fluid flow (Mietke et al. Extracting Cell Stiffness from Real-Time Deformability Cytometry: 728 Theory and Experiment. Biophys. J. 2015, 109, 2023-2036). Including linear elasticity as well as neo-Hookean hyperelasticity, our model is valid in a wide range of cell deformations and allows to extract cell stiffness for largely deformed cells. We introduce a new measure for cell deformation that is capable to distinguish between deformation effects stemming from cell cortex and cell bulk elasticity. Finally, we demonstrate the potential of the method to simultaneously quantify multiple mechanical cell parameters by RT-DC.

5.
Artigo em Inglês | MEDLINE | ID: mdl-26736644

RESUMO

The mechanical properties of cells are known to be a label-free, inherent marker of biological function in health and disease. Wide-spread utilization has so far been impeded by the lack of a convenient measurement technique with sufficient throughput. To address this unmet need, we have recently introduced real-time deformability cytometry (RT-DC) for continuous mechanical single-cell classification of heterogeneous cell populations at rates of several hundred cells per second. Cells are driven through the constriction zone of a microfluidic chip leading to cell deformations due to hydrodynamic stresses only. Our custom-built image processing software performs image acquisition, image analysis and data storage on the fly. The ensuing deformations can be quantified and an analytical model enables the derivation of cell material properties. Performing RT-DC we highlight its potential to identify rare objects in heterogeneous suspensions and to track drug-induced changes in cells. In summary, RT-DC enables marker-free, quantitative phenotyping of heterogeneous cell populations with a throughput comparable to standard flow cytometry.


Assuntos
Citofotometria/instrumentação , Citofotometria/métodos , Desenho de Equipamento , Citometria de Fluxo/métodos , Células HL-60 , Ensaios de Triagem em Larga Escala/instrumentação , Ensaios de Triagem em Larga Escala/métodos , Humanos , Hidrodinâmica , Processamento de Imagem Assistida por Computador , Dispositivos Lab-On-A-Chip
6.
Interface Focus ; 4(2): 20130069, 2014 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-24748957

RESUMO

A cell is a complex material whose mechanical properties are essential for its normal functions. Heating can have a dramatic effect on these mechanical properties, similar to its impact on the dynamics of artificial polymer networks. We investigated such mechanical changes by the use of a microfluidic optical stretcher, which allowed us to probe cell mechanics when the cells were subjected to different heating conditions at different time scales. We find that HL60/S4 myeloid precursor cells become mechanically more compliant and fluid-like when subjected to either a sudden laser-induced temperature increase or prolonged exposure to higher ambient temperature. Above a critical temperature of 52 ± 1°C, we observed active cell contraction, which was strongly correlated with calcium influx through temperature-sensitive transient receptor potential vanilloid 2 (TRPV2) ion channels, followed by a subsequent expansion in cell volume. The change from passive to active cellular response can be effectively described by a mechanical model incorporating both active stress and viscoelastic components. Our work highlights the role of TRPV2 in regulating the thermomechanical response of cells. It also offers insights into how cortical tension and osmotic pressure govern cell mechanics and regulate cell-shape changes in response to heat and mechanical stress.

7.
Biophys J ; 81(2): 767-84, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11463624

RESUMO

When a dielectric object is placed between two opposed, nonfocused laser beams, the total force acting on the object is zero but the surface forces are additive, thus leading to a stretching of the object along the axis of the beams. Using this principle, we have constructed a device, called an optical stretcher, that can be used to measure the viscoelastic properties of dielectric materials, including biologic materials such as cells, with the sensitivity necessary to distinguish even between different individual cytoskeletal phenotypes. We have successfully used the optical stretcher to deform human erythrocytes and mouse fibroblasts. In the optical stretcher, no focusing is required, thus radiation damage is minimized and the surface forces are not limited by the light power. The magnitude of the deforming forces in the optical stretcher thus bridges the gap between optical tweezers and atomic force microscopy for the study of biologic materials.


Assuntos
Eritrócitos/citologia , Eritrócitos/efeitos da radiação , Fibroblastos/citologia , Fibroblastos/efeitos da radiação , Lasers , Células 3T3 , Algoritmos , Animais , Tamanho Celular/efeitos da radiação , Citoesqueleto/efeitos da radiação , Elasticidade/efeitos da radiação , Humanos , Camundongos , Microscopia de Contraste de Fase , Microesferas , Dióxido de Silício , Viscosidade/efeitos da radiação
8.
Phys Rev Lett ; 84(23): 5451-4, 2000 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-10990966

RESUMO

Two counterpropagating laser beams were used to significantly stretch soft dielectrics such as cells. The deforming forces act on the surface between the object and the surrounding medium and are considerably higher than the trapping forces on the object. Radiation damage is avoided since a double-beam trap does not require focusing for stable trapping. Ray optics was used to describe the stress profile on the surface of the trapped object. Measuring the total forces and deformations of well-defined elastic objects validated this approach.


Assuntos
Eritrócitos/citologia , Eritrócitos/efeitos da radiação , Lasers , Animais , Tamanho Celular/efeitos da radiação , Elasticidade/efeitos da radiação , Estresse Mecânico
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...