Nonlinear dynamics of cell migration in anisotropic microenvironment

Cell migration in anisotropic microenvironment plays an important role in the development of normal tissues and organs as well as neoplasm progression, e.g., osteogenic differentiation of embryonic stem cells was facilitated on stiffer substrates, indicating that the mechanical signals greatly affect both early and terminal differentiation of embryonic stem cells. However, the effect of anisotropy on cell migration dynamics, in particular, in terms of acceleration profiles which is important for recognizing dynamics modes of cell migration and analyzing the regulation mechanisms of microenvironment in mechanical signal transmission, has not been systematically investigated. In this work, we firstly rigorously investigate and quantify the differences between persistent random walk and anisotropic persistent random walk models based on the analysis of cell migration trajectories and velocity auto-covariance function, both qualitatively and quantitatively. Secondly, we introduce the concepts of positive and negative anisotropy based on the motility parameters to study the effect of anisotropy on acceleration profiles, especially the nonlinear decrease and non-monotonic behaviors. We particularly elaborate and discuss the mechanisms, and physical insights of non-monotonic behaviors in the case of positive anisotropy,focusing on the force exerted on migrating cells. Finally, we analyze two types of in vitro cell migration experiments and verify the universality of nonlinear decrease and the consistence of non-monotonic behaviors with numerical results.We conclude that the anisotropy of microenvironment is the cause of the non-monotonic and nonlinear dynamics, and the anisotropic persistent random walk can be as a suitable tool to analyze in vitro cell migration with different combinations of motility parameters. Our analysis provides new insights into the dynamics of cell migration in complex microenvironment,which also has implications in tissue engineering and cancer research.     

Migration and shape of cells on different interfaces

Impacts of microenvironments on cell migration have been reported in various interaction modes. A rapid tumor metastasis occurs along topological interfaces in vivo, such as the interface between the blood vessels and nerves. In this work, we culture MDA-MB231 cells at dish-liquid, dish-hydrogel, and hydrogel-liquid interfaces, respectively, to study how these different interfaces influence cell dynamics and morphology. Our results show that the migration mode of cells changes from an amoeboid motion to a mesenchymal motion but their speed do not change obviously if the interface changes from hydrogel-liquid to dish-liquid. In contrast, the migration mode of cells at a dish-hydrogel interface maintains as a mesenchymal motion, whereas their speed increases significantly.     

Cancer cell recognition--mechanical phenotype

The major characteristics of cancer metastasis is the ability of the primary tumor cells to migrate by way of the blood or lymph vessels and to form tumors at multiple, distant sites. There are evidences that cancer progression is characterized by disruption and/or reorganization of cytoskeleton (i.e. cellular scaffold). This is accompanied by various molecular alterations influencing the overall mechanical resistance of cells. Current approach in diagnosis focuses mainly on microbiological, immunological, and pathological aspects rather than on the biomechanics of diseases. The determination of mechanical properties of an individual living cell has became possible with the development of local measurement techniques, such as atomic force microscopy, magnetic or optical tweezers. The advantage of them lies in the capability to measure living cells at a single cell level and in liquid conditions, close to natural environment. Here, we present the studies on mechanical properties of single cells originating from various cancers. The results show that, independently of the cancer type (bladder, melanoma, prostate, breast and colon), single cells are characterized by the lower Young's modulus, denoting higher deformability of cancerous cells. However, the obtained Young's modulus values were dependent on various factors, like the properties of substrates used for cell growth, force loading rate, or indentation depth. Their influence on elastic properties of cells was considered. Based on these findings, the identification of cancerous cells based on their elastic properties was performed. These results proved the AFM capability in recognition of a single, mechanically altered cell, also in cases when morphological changes are not visible. The quantitative analysis of cell deformability carried out using normal (reference) and cancerous cells and, more precisely, their characterization (qualitative and quantitative) can have a significant impact on the development of methodological approaches toward precise identification of pathological cells and would allow for more effective detection of cancer-related changes.     

Migration and proliferation dichotomy in tumor-cell invasion

We propose a two-component reaction-transport model for the migration-proliferation dichotomy in the spreading of tumor cells. By using a continuous time random walk (CTRW), we formulate a system of the balance equations for the cancer cells of two phenotypes with random switching between cell proliferation and migration. The transport process is formulated in terms of the CTRW with an arbitrary waiting-time distribution law. Proliferation is modeled by a standard logistic growth. We apply hyperbolic scaling and Hamilton-Jacobi formalism to determine the overall rate of tumor cell invasion. In particular, we take into account both normal diffusion and anomalous transport (subdiffusion) in order to show that the standard diffusion approximation for migration leads to overestimation of the overall cancer spreading rate.     

Influence of matrix-metalloproteinase inhibitor on the interaction between cancer cells and matrigel

Various behaviors of cancer cells are strongly influenced by their interaction with extracellular matrices(ECM). We investigate how this interaction may be influenced if the cancer cells' ability of secreting matrix metalloproteinases(MMPs)to degrade ECM is inhibited by adding the MMP inhibitor. We use MDA-MB-231-GFP cells as model cells and use matrigel to mimic ECM. It is found that the added MMP inhibitor significantly reduces the migration speed of cancer cells covered by matrigel but has little influence on the migration persistence and shape factor of the cells and that with the MMP inhibitor added the presence of matrigel on the top has no influence on the migration speed of the cells but increases the cells' shape factor and migration persistence.     

物理学在肿瘤细胞的极性及迁移研究中的应用

肿瘤细胞和所处微环境的物理性质, 以及它们之间的相互物理作用对于肿瘤的产生、发展与转移都有极大的影响, 这使得从物理学角度探索肿瘤研究成为了必然趋势. 肿瘤转移是癌症致死的最大因素, 而肿瘤细胞迁移中的极化是肿瘤转移的重要一步. 本文总结了物理学实验和模型在揭示细胞迁移和极化机理方面的贡献. 实验上应用最新的微流控芯片技术与表面微模型化技术等手段, 研究空间维度、黏附行为、机械力等物理信号对于细胞极性的建立与保持以及细胞迁移行为的影响后, 发现物理信号与生化反应之间的相互耦合对于细胞迁移有着至关重要的作用; 理论上基于扩散反应方程, 已经建立了一系列表征细胞极化的模型. 今后的研究将结合物理实验建立肿瘤细胞迁移中的极化模型, 进而发展针对肿瘤细胞感知物理信号的新的治疗肿瘤转移方法.     

Influence of matrigel on the shape and dynamics of cancer cells

The interaction between extracellular matrices and cancer cells plays an important role in regulating cancer cell behaviors. In this article, we use matrigel to mimic extracellular matrices and investigate experimentally how matrigel influences the shape and dynamics of breast cancer cells(MDA-MB-231-GFP cells). We find that matrigel facilitates cancer cells' migration and shape deformation. The influences of the matrigel concentration are also reported.     

癌细胞对血清光谱的影响

研究了在血清中加入不同剂量的癌细胞对血清光谱的影响。比较了两种癌细胞的光谱 ,并与正常血清的谱线做了比较。实验结果表明 :随着癌细胞个数的降低 ,吸收谱和荧光谱强度均呈增加趋势 ,但增加到一定程度时又开始下降 ;正常血清的谱线强度明显较强。结论 :荧光光谱表明癌细胞抑制了正常血清中某种基团的振动 ,吸收光谱则表明癌细胞抑制了酪氨酸对光的吸收。     

Cell-cycle-dependent variations in the FTIR spectroscopy of HeLa cells treated with trichostatin A

It is quite complex to evaluate the mechanism of action for antitumor drugs on cancer cells.Studies have pointed out that there is an unique advantage of Fourier transform infrared spectrum to obtain a fingerprint of all molecules present in the cells when cancer cells were exposed to anti-cancer drugs.Trichostatin A (TSA) is a most potent reversible inhibitor of mammalian histone deacetylases.It can inhibit cancer cell growth in vitro and in vivo.In the present study,HeLa cells were exposed to 0,50,100,200,300 and 400 nmol · L-1 TSA,and FTIR spectra were applied to evaluate the effect of TSA on cancer cells.Results show that there is some significant relationship between the changes in FTIR absorption and cell cycle arresting.On the other hand,this investigation shows that the concentration of TSA had to be more than 200 nmol · L-1 in order to ensure A1080 cm-1/A1540cm-1 ≥1 for inhibiting cell proliferation.     

High frequency ultrasound imaging of whole blood gelation and retraction during in vitro coagulation

Blood coagulation is a series of biochemical reactions resulting in the mechanical transformation of liquid blood into a gel. As a consequence, ultrasound, being mechanical waves, can provide specific details on the dynamics of coagulation. In fact, previous high-frequency ultrasound monitoring studies have shown drastic changes in ultrasound velocity and attenuation during whole blood coagulation and a model discussing the observed mechanical transformations was proposed. In this paper, a technique of visualization of the clotting mechanism is introduced, which complements and revises the previous hypotheses. This method is based on the monitoring of scatterers (red blood cells) movement through a time correlation of 20 MHZ rf signals. It allows the computing of both a displacement map revealing local details and disparities and a parameter quantifying the global structural behavior. Qualitative results for two typical samples show that the technique provides new insights on the gelation dynamics. A quantitative analysis computed from 12 healthy subjects found that the changes in the structural parameters are significantly correlated to the changes in velocity and attenuation, both dependent on the mechanical transformations in the sample. The previous model is therefore revised and a new way to measure gel and retraction times is proposed.     

Activation of microbubbles by low-intensity pulsed ultrasound enhances the cytotoxicity of curcumin involving apoptosis induction and cell motility inhibition in human breast cancer MDA-MB-231 cells

Ultrasound and microbubbles-mediated drug delivery has become a promising strategy to promote drug delivery and its therapeutic efficacy. The aim of this research was to assess the effects of microbubbles (MBs)-combined low-intensity pulsed ultrasound (LPUS) on the delivery and cytotoxicity of curcumin (Cur) to human breast cancer MDA-MB-231 cells. Under the experimental condition, MBs raised the level of acoustic cavitation and enhanced plasma membrane permeability; and cellular uptake of Cur was notably improved by LPUS–MBs treatment, aggravating Cur-induced MDA-MB-231 cells death. The combined treatment markedly caused more obvious changes of cell morphology, F-actin cytoskeleton damage and cell migration inhibition. Our results demonstrated that combination of MBs and LPUS may be an efficient strategy for improving anti-tumor effect of Cur, suggesting a potential effective method for antineoplastic therapy.     

拉曼光谱研究复方鹿仙草颗粒对SMMC-7721肝癌细胞的作用

应用拉曼光谱研究了不同浓度的鹿仙草溶液与肝癌细胞SMMC-7721的相互作用,通过对药物作用前后细胞的光谱变化进行分析,从而为阐明鹿仙草与肝癌细胞的作用方式提供重要的依据。拉曼光谱显示,加入鹿仙草后,细胞的许多峰都发生了变化。归属于磷酸骨架振动的785和1 092 cm-1的两个峰强度下降,对应碱基A和G的峰1 312和1 585 cm-1 等也有不同程度的降低,表明鹿仙草可能插入DNA碱基对之间,使DNA复制受到抑制,导致细胞DNA含量下降,而且会引起DNA单、双链的断裂。同时,研究还发现属于蛋白质的振动峰(1 005, 1 360, 1 656 cm-1)强度也有不同程度下降,说明蛋白质二级结构以及侧链氨基酸的环境均发生了改变。此外,鹿仙草对细胞的作用效果随浓度增加逐步增强。     

The impact of environmental changes upon the microrheological response of adherent cells

The mechanical behaviour of adherent cells cultured in vitro is known to be dependent on the mechanical properties of the substrate. We show that this mechanical behaviour is also strongly affected by the cells’ environment. We focus here on the impact of temperature and pH . Experiments carried out on individual cells in a tuneable environment reveal that the intra-cellular mechanical behaviour exhibits large and fast changes when the external cell environment is changed. Fast passive microrheometry measurements allow for the precise characterisation of the transient regime observed during a temperature drop. When maintained at a non-physiological temperature, the cells reach a stabilised state distinct from the state observed in physiological conditions. The perturbation can be reversed but exhibits hysteretic behaviour when physiological conditions are restored. The transient regime observed during the recovery process is found to be different from the transient regime observed when leaving physiological conditions. A modified generalized Stokes-Einstein equation taking into account the cell activity through an effective temperature is proposed here to fit the experimental results. Excellent agreement between the model and the measurements is obtained for time lags from 10^-3 to 1s considered in this study.     

Realistic mechanical tuning in a micromechanical cochlear model

Two assumptions were made in the formulation of a recent cochlear model [P.J. Kolston, J. Acoust. Soc. Am. 83, 1481-1487 (1988)]: (1) The basilar membrane has two radial modes of vibration, corresponding to division into its arcuate and pectinate zones; and (2) the impedance of the outer hair cells (OHCs) greatly modifies the mechanics of the arcuate zone. Both of these assumptions are strongly supported by cochlear anatomy. This paper presents a revised version of the outer hair cell, arcuate-pectinate (OHCAP) model, which is an improvement over the original model in two important ways: First, a model for the OHCs is included so that the OHC impedance is no longer prescribed functionally; and, second, the presence of the OHCs enhances the basilar membrane motion, so that the model is now consistent with observed response changes resulting from trauma. The OHCAP model utilizes the unusual spatial arrangement of the OHCs, the Deiters cells, their phalangeal processes, and the pillars of Corti. The OHCs do not add energy to the cochlear partition and hence the OHCAP model is passive. In spite of the absence of active processes, the model exhibits mechanical tuning very similar to those measured by Sellick et al. [Hear. Res. 10, 93-100 (1983)] in the guinea pig cochlea and by Robles et al. [J. Acoust. Soc. Am. 80, 1364-1374 (1986)] in the chinchilla cochlea. Therefore, it appears that mechanical response tuning and response changes resulting from trauma should not be used as justifications for the hypothesis of active processes in the real cochlea.     

DNA Binding Property and Antitumor Evaluation of Xanthone with Dimethylamine Side Chain

In this work, a xanthone derivative was obtained by cationic modification of the free hydroxyl group of xanthone with dimethylamine group of high pKa value. The interactions of xanthones with DNA were investigated by spectroscopic methods, electrophoretic migration assay and polymerase chain reaction test. Results indicate that xanthones can intercalate into the DNA base pairs by the hydrophobic plane and the xanthone with dimethylamine side chain may also bind the DNA phosphate framework by the basic amine alkyl chain, thus showing a better DNA binding ability than the xanthone. Furthermore, inhibition on tumor cells (ECA109, SGC7901, GLC-82) proliferation of xanthones were evaluated by MTT method. Analysis results show that the xanthone with dimethylamine side chain exhibits more effective inhibition activity against three cancer cells than the xanthone. The effects on the inhibition of tumor cells in vitro agree with the studies of DNA binding. It means that the amine alkyl chain would play an important role in its antitumor activity and DNA binding property.     

Evaluation of Stability and Sensitivity of Cell Fluorescent Labels When Used for Cell Migration

The directed migration of mammalian cells is a foundation of development and growth. A variety of processes such as tissue development, wound healing, pathogen recognition/destruction as well as cancer metastasis are the result of regulated or dysregulated cell migration. While the ability to measure a cell’s propensity to migrate has clinical relevance in several settings, no universal protocol has been established to measure cell migration. A variety of techniques are currently used to measure migration including manual counting, flow cytometry or Coulter counting, microfluidic devices, computerized spectroscopic methods, or the use of various tracking dyes interfaced with fluorescent or non-fluorescent plate readers. In order to expedite the measurement of migration, we compared several common cytoplasmic and lipophilic cell tracking dyes to determine the best dye for determining migration of rare population of cells. CellVue® Burgundy was found to be superior over calcein AM, Cell Tracker Green CMFDA (chloromethyl fluorescein diacetate), Vybrant CFDA (carboxy fluorescein diacetate succinimidyl ester) in its retention within cells, superior to CellVue® NIR 815, PKH67, and CM DiI with regard to signal to noise ratio, and superior to PKH26 with regard to instrument versatility.     

Silicon nanocrystals as photo- and sono-sensitizers for biomedical applications

Silicon nanocrystals (nc-Si) with sizes of about 2÷5 nm prepared from porous Si act as photosensitizers of the singlet oxygen generation revealed by direct detection of the luminescence at 0.98 eV (1270 nm). In vitro experiments show a suppression of the cancer cell proliferation because of the effect of photo-excited nc-Si. Aqueous suspensions of nc-Si formed by mechanical grinding (milling) of crystalline and porous Si exhibit properties of efficient sonosensitizers. A destruction of cancer cells under ultrasound irradiation in the presence of nc-Si is observed in vitro. Possible applications of nc-Si as photo- and sono-sensitizers for cancer therapy are discussed.     

Synergistic effects of negative-charged nanoparticles assisted by ultrasound on the reversal multidrug resistance phenotype in breast cancer cells

We have fabricated a negative-charged nanoparticle (Heparin-Folate-Tat-Taxol NP, H-F-Tat-T NP) with dual ligands, tumor targeting ligand folate and cell-penetrating peptide Tat, to deliver taxol presenting great anticancer activity for sensitive cancer cells, while it fails to overcome multidrug resistance (MDR) in MCF-7/T cells (taxol-resistant breast cancer cells). Ultrasound (US) can increase the sensitivity of positive-charged NPs thereby making it possible to reverse MDR through inducing NPs’ drug release. However, compared with the negative-charged NPs, positive-charged NPs may cause higher toxic effect. Hence, the combination of negative-charged NPs and US may be an efficient strategy for overcoming MDR. The conventional procedure to treat with NPs followed by US exposure possibly destruct multifunctional NPs resulting in its bioactivity inhibition. Herein, we have further improved the operating approach to eliminate US mechanical damage and keep the integrity of negative-charged NPs: cells are exposed to US with microbubbles (MBs) prior to the treatment of H-F-Tat-T NPs. Superior to the conventional method, US sonoporation affects the physiological property of cancer cells while preventing direct promotion of drug release from NPs. The results of the present study displayed that US in condition (1 MHz, 10% duty cycle, duration of 80 s, US intensity of 0.6 W/cm² and volume ratio of medium to MBs 20:1) combined with H-F-T-Tat-T NPs can achieve optimal reversal MDR effect in MCF-7/T cells. Mechanism study further disclosed that the individual effect of US was responsible for the enhancement of cell membrane permeability, inhibition of cell proliferation rate and down-regulation of MDR-related genes and proteins. Simultaneously, US sonoporation on resistant cancer cells indirectly increased the accumulation of NPs by inducing endosomal escape of negative-charged NPs. Taken together, the overcoming MDR ability for the combined strategy was achieved by the synergistic effect from individual function of NPs, physiological changes of resistant cancer cells and behavior changes of NPs caused by US.