Freezing-Induced Cell-Fluid-Matrix Interaction in Engineered Tissue

Author(s):  
Junkyu Jung ◽  
Ka Yaw Teo ◽  
J. Craig Dutton ◽  
Bumsoo Han

Freezing of biological tissues occurs in cryomedicine applications such as cryosurgery and cryopreservation. Although cellular level biophysics during freezing and thawing (F/T) has been extensively studied, tissue level biophysics is not fully understood yet. Especially, the effects of F/T on the functionalities of tissue are not well understood so that the outcomes of cryomedicine applications are highly tissue-type dependent [1]. Although many of the functionalities are associated with the extracellular matrix (ECM), the effect of F/T on ECM microstructure has been overlooked. Quantitative understanding on the post-thaw ECM structure is rarely available, but it is essential to design and improve cryopresevation and cryotherapy protocols for a wide variety of native and engineered tissues.

Author(s):  
Jun K. Jung ◽  
Ka Yaw Teo ◽  
J. Craig Dutton ◽  
Bumsoo Han

Since biological tissues are composed of cells, extracellular matrix, and interstitial fluid, freezing of biological tissues induces complex cell-fluid-matrix interaction. Quantitative understanding of this cell-fluid-matrix interaction is crucial to the design and optimization of a wide variety of cryomedicine applications. However, quantitative measurement of the interaction is extremely challenging due to the lack of reliable non-invasive measurement techniques during freezing and thawing. In the present study, a new measurement technique was developed to dynamically measure microscale tissue deformation during freezing/thawing and its feasibility was demonstrated. In this method, which is named “Cell Image Deformetry” (CID), engineered tissues with pre-labeled cells with quantum dots are imaged under a fluorescence microscope. Then, the tissue deformation is evaluated by cross-correlating cell locations between sequential microscopic images with known time intervals based on the particle image velocimetry (PIV) data processing technique.


Author(s):  
Ka Yaw Teo ◽  
J. Craig Dutton ◽  
Frederick Grinnell ◽  
Bumsoo Han

Long-term cryopreservation of functional engineered tissues (ETs) is a key enabling technology for tissue engineering and regenerative medicine. However, a limited understanding of tissue-level biophysical phenomena during freeze/thaw (F/T) and their effects on cells and ECM microstructure poses significant challenges for i) preserving tissue functionality, and ii) controlling highly tissue-type dependent cryopreservation outcomes.


2018 ◽  
Author(s):  
R. J. Murphy ◽  
P. R. Buenzli ◽  
R. E. Baker ◽  
M. J. Simpson

AbstractMechanical heterogeneity in biological tissues, in particular stiffness, can be used to distinguish between healthy and diseased states. However, it is often difficult to explore relationships between cellular-level properties and tissue-level outcomes when biological experiments are performed at a single scale only. To overcome this difficulty we develop a multi-scale mathematical model which provides a clear framework to explore these connections across biological scales. Starting with an individual-based mechanical model of cell movement, we subsequently derive a novel coarse-grained system of partial differential equations governing the evolution of the cell density due to heterogeneous cellular properties. We demonstrate that solutions of the individual-based model converge to numerical solutions of the coarse-grained model, for both slowly-varying-in-space and rapidly-varying-in-space cellular properties. Applications of the model are discussed, including determining relative cellular-level properties and an interpretation of data from a breast cancer detection experiment.


Author(s):  
R. J. Murphy ◽  
P. R. Buenzli ◽  
R. E. Baker ◽  
M. J. Simpson

Mechanical heterogeneity in biological tissues, in particular stiffness, can be used to distinguish between healthy and diseased states. However, it is often difficult to explore relationships between cellular-level properties and tissue-level outcomes when biological experiments are performed at a single scale only. To overcome this difficulty, we develop a multi-scale mathematical model which provides a clear framework to explore these connections across biological scales. Starting with an individual-based mechanical model of cell movement, we subsequently derive a novel coarse-grained system of partial differential equations governing the evolution of the cell density due to heterogeneous cellular properties. We demonstrate that solutions of the individual-based model converge to numerical solutions of the coarse-grained model, for both slowly-varying-in-space and rapidly-varying-in-space cellular properties. We discuss applications of the model, such as determining relative cellular-level properties and an interpretation of data from a breast cancer detection experiment.


Author(s):  
Zhengbin Xu ◽  
Altug Ozcelikkale ◽  
Young L. Kim ◽  
Bumsoo Han

Quality and functionality of engineered tissues are closely related to the microstructures and integrity of their extracellular matrix (ECM). However, currently available methods for characterizing ECM structures are often labor-intensive, destructive, and limited to a small fraction of the total area. These methods are also inappropriate for assessing temporal variations in ECM structures. In this study, to overcome these limitations and challenges, we propose an elastic light scattering approach to spatiotemporally assess ECM microstructures in a relatively large area in a nondestructive manner. To demonstrate its feasibility, we analyze spectroscopic imaging data obtained from acellular collagen scaffolds and dermal equivalents as model ECM structures. For spatial characterization, acellular scaffolds are examined after a freeze/thaw process mimicking a cryopreservation procedure to quantify freezing-induced structural changes in the collagen matrix. We further analyze spatial and temporal changes in ECM structures during cell-driven compaction in dermal equivalents. The results show that spectral dependence of light elastically backscattered from engineered tissue is sensitively associated with alterations in ECM microstructures. In particular, a spectral decay rate over the wavelength can serve as an indicator for the pore size changes in ECM structures, which are at nanometer scale. A decrease in the spectral decay rate suggests enlarged pore sizes of ECM structures. The combination of this approach with a whole-field imaging platform further allows visualization of spatial heterogeneity of EMC microstructures in engineered tissues. This demonstrates the feasibility of the proposed method that nano- and micrometer scale alteration of the ECM structure can be detected and visualized at a whole-field level. Thus, we envision that this spectroscopic imaging approach could potentially serve as an effective characterization tool to nondestructively, accurately, and rapidly quantify ECM microstructures in engineered tissue in a large area.


1997 ◽  
Vol 78 (02) ◽  
pp. 934-938 ◽  
Author(s):  
Hsiun-ing Chen ◽  
Yueh-I Wu ◽  
Yu-Lun Hsieh ◽  
Guey-Yueh Shi ◽  
Meei-Jyh Jiang ◽  
...  

SummaryTo investigate whether the endothelium-platelet interactions may be altered by plasminogen activation, cultured human umbilical vein endothelial cells (ECs) were treated with tissue-type plasminogen activator (t-PA) in the presence of plasminogen, and platelet adhesion to ECs was subsequently measured by using a tapered flow chamber. Our results demonstrated that platelets adhered more readily to t-PA treated EC monolayer than to the control monolayer at all shear stress levels tested. This phenomenon was treatment time-dependent and dose-dependent, and it could be blocked by adding plasmin inhibitors, such as e-amino caproic acid and aprotinin. Adherent platelets on t-PA treated EC monolayer underwent more severe shape change than those on the control monolayer. While the extracellular matrix directly treated with t-PA attracted less platelets than the control matrix did, platelet adhesion to the matrix that was produced by t-PA-treated ECs was unaltered. These data suggest that t-PA treatment on ECs compromised antiplatelet-adhesion capability on their apical surface without altering the reactivity of their extracellular matrix towards platelets.


2008 ◽  
Vol 56 (3) ◽  
pp. 341-348
Author(s):  
P. Pepó ◽  
A. Kovács

Cryopreservation appears to be a suitable solution for the maintenance of potato germplasms. The protocol described in this paper can be applied for the vitrification and preservation of meristems. During histo-cytological studies it is possible to observe modifications at the cellular level and to understand the adaptive mechanism to low temperatures. Control potato meristem tissue contained a number of meristematic cells with a gradient of differentiation. After freezing there were a large number of vacuolated cells, some of which exhibited broken cell walls and plasmolysis. The thickening of the cell wall, giving them a sinuous appearance, was observed after freezing and thawing the meristems, with ruptures of the cuticle and epidermal layer.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yoshifumi Asakura ◽  
Yohei Kondo ◽  
Kazuhiro Aoki ◽  
Honda Naoki

AbstractCollective cell migration is a fundamental process in embryonic development and tissue homeostasis. This is a macroscopic population-level phenomenon that emerges across hierarchy from microscopic cell-cell interactions; however, the underlying mechanism remains unclear. Here, we addressed this issue by focusing on epithelial collective cell migration, driven by the mechanical force regulated by chemical signals of traveling ERK activation waves, observed in wound healing. We propose a hierarchical mathematical framework for understanding how cells are orchestrated through mechanochemical cell-cell interaction. In this framework, we mathematically transformed a particle-based model at the cellular level into a continuum model at the tissue level. The continuum model described relationships between cell migration and mechanochemical variables, namely, ERK activity gradients, cell density, and velocity field, which could be compared with live-cell imaging data. Through numerical simulations, the continuum model recapitulated the ERK wave-induced collective cell migration in wound healing. We also numerically confirmed a consistency between these two models. Thus, our hierarchical approach offers a new theoretical platform to reveal a causality between macroscopic tissue-level and microscopic cellular-level phenomena. Furthermore, our model is also capable of deriving a theoretical insight on both of mechanical and chemical signals, in the causality of tissue and cellular dynamics.


2014 ◽  
Vol 11 (97) ◽  
pp. 20140352 ◽  
Author(s):  
Alfonso Bueno-Orovio ◽  
David Kay ◽  
Vicente Grau ◽  
Blanca Rodriguez ◽  
Kevin Burrage

Impulse propagation in biological tissues is known to be modulated by structural heterogeneity. In cardiac muscle, improved understanding on how this heterogeneity influences electrical spread is key to advancing our interpretation of dispersion of repolarization. We propose fractional diffusion models as a novel mathematical description of structurally heterogeneous excitable media, as a means of representing the modulation of the total electric field by the secondary electrical sources associated with tissue inhomogeneities. Our results, analysed against in vivo human recordings and experimental data of different animal species, indicate that structural heterogeneity underlies relevant characteristics of cardiac electrical propagation at tissue level. These include conduction effects on action potential (AP) morphology, the shortening of AP duration along the activation pathway and the progressive modulation by premature beats of spatial patterns of dispersion of repolarization. The proposed approach may also have important implications in other research fields involving excitable complex media.


Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1760
Author(s):  
Joshua J. A. Poole ◽  
Leila B. Mostaço-Guidolin

Biological tissues are not uniquely composed of cells. A substantial part of their volume is extracellular space, which is primarily filled by an intricate network of macromolecules constituting the extracellular matrix (ECM). The ECM serves as the scaffolding for tissues and organs throughout the body, playing an essential role in their structural and functional integrity. Understanding the intimate interaction between the cells and their structural microenvironment is central to our understanding of the factors driving the formation of normal versus remodelled tissue, including the processes involved in chronic fibrotic diseases. The visualization of the ECM is a key factor to track such changes successfully. This review is focused on presenting several optical imaging microscopy modalities used to characterize different ECM components. In this review, we describe and provide examples of applications of a vast gamut of microscopy techniques, such as widefield fluorescence, total internal reflection fluorescence, laser scanning confocal microscopy, multipoint/slit confocal microscopy, two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG, THG), coherent anti-Stokes Raman scattering (CARS), fluorescence lifetime imaging microscopy (FLIM), structured illumination microscopy (SIM), stimulated emission depletion microscopy (STED), ground-state depletion microscopy (GSD), and photoactivated localization microscopy (PALM/fPALM), as well as their main advantages, limitations.


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