Modeling of micropipette aspiration and optical tweezers stretching of erythrocytes with or without Malaria parasite

The erythrocytes play an important role in the human body. The healthy erythrocytes can undergo extremely large deformation while passing through small capillaries. Their infection by Malaria Plasmodium falcipurum （P.f.） will lead to capillary blockage and blood flow obstruction. Many experimental and computational methods have been applied to study the increase in stickiness and decrease in deformability of the Malaria （P.f.） infected erythrocytes. The novelty of this paper lies in the establishment of an multi-component model for investigating mechanical properties of Malaria （P.f.） infected erythrocytes, especially of their enclosed parasites. Finite element method was applied to simulate the erythrocytes＇ deformation in micropipette aspiration and optical tweezers stretching using the computational software ABAQUS. The comparisons between simulations and experiments were able to quantitatively conclude the effects of stiffness and stickiness of the parasitophorous vacuole membrane on the cells＇ deformation, which could not be obtained from experiments directly.

Integrin betal mediates hepatocellular carcinoma cells chemotaxis to laminin

BACKGROUND: Chemotaxis is an important step durin the invasion of carcinoma cells. And integrins are most im portant receptors mediating interaction between cells an extracellular matrix ( ECM). This study was designed t study integrin betal mediating chemotaxis of hepatocellula carcinoma (HCC) cells to laminin (LN). METHODS: A micropipette technique was adopted to in vestigate the effect of blockade of integrin betal on pseudo pod protrusion of HCC cells in response to LN stimulation Chemotactic pseudopod protrusion of a HCC cell was eva luated using a dual-pipette set-up, in which two pipette filled with LN solution were positioned in close contact wit the same cell, and pseudopod protrusion into each pipett was viewed dynamically and recorded with a tape recorder The lengths of pseudopods were measured and plotte against time to obtain a pseudopod growth curve. The in tegrin betal subunit on the surfaces of HCC cells were ana lyzed by flow cytometry. RESULTS: In dual pipette chemotaxis experiment, whe the two pipettes were filled with LN(50 μg/ml, 200 μg/ml) pseudopods extended from the HCC cell into each of th pipettes nearly symmetrically, ie, with nearly identica maximum pseudopod length and similar pseudopod growth curves. Upon addition of anti-CD29 (20 μg/ml) to one o the pipettes, pseudopod protrusion was blocked nearly completely while protrusion into the opposite pipette be came more evidently, with a larger maximum length. Ex pression of integrin betal was up to 95.78% to cells chosen in the experiment. CONCLUSION: Integrin betal subunit was an importan constituent receptor subunit for mediating chemotactic pseudopod protrusion of HCC cell to LN.

Adhesive properties of hepatoma cells to collagen Ⅳ coated surfaces

Objectives: To quantitatively study the adhesive pro- perties of hepatoma cells to collagen Ⅳ coated artifi- cial basement membrane and to investigate the rele- vance of cell adhesive forces to the concentration of collagen Ⅳ. Methods: Synchronous G1 and S phase cells were a- chieved using thymine-2-desoxyriboside and cochicine sequential blockage method and double thymine-2- desoxyriboside blockage method respectively. The adhesive forces of hepatoma cells were investigated by micropipette aspiration technique. Results: The adhesive forces of hepatoma cells to ar- tificial basement membrane were (107.78±65.44) ×10^(-10)N, (182.60±107.88)×10^(-10)N, (298.91± 144.13)×10^(-10)N when the concentration of the membrane coated by 1, 2, 5μg/ml collagen Ⅳ re- spectively (P<0.001). The adhesive forces of G1 and S phases hepatoma cells to artificial basement membrane were (275.86±232.80)×10^(-10)N and (161.16±120.40)×10^(-10)N respectively when the concentration of the membrane coated by 5μg/ml collagen Ⅳ (P<0.001). Conclusions: The adhesive forces of hepatoma cells to artifical basement membrane in direct proportion to the concentration of collagen Ⅳ suggests that the in- crease of basement membrane might be conducive to the chemotactic motion and adhesiveness of tumor cells. G1 phase cells are more capable of adhering to basement membrane than S phase cells. Hepatoma cells, especially G1 phase cells, may survive in blood circulation, and sequest and adhere in microcircula- tion, and get through basement membrane for re- mote metastasis.

Finite element analysis of micropipette aspiration considering finite size and compressibility of cells

Micropipette aspiration(MA) is widely applied in cell mechanics, however, at small deformations a common model corresponding to the MA is the half-space model wherein the finite cell size and cell compressibility are neglected. This study extends the half-space model by accounting for the influence of cell geometry and compressibility(sphere model). Using a finite element analysis of cell aspiration into a micropipette, an elastic approximation formula of the aspirated length was derived for the sphere model. The approximation formula includes the geometry parameter of the sphere model(ζ = R/a, R is the radius of the cell, and a is the inner radius of the micropipette) and the Poisson's ratio v of the cell. The results indicate that the parameter and Poisson's ratio v markedly affect the aspirated length, particularly for small and v. When ζ→∞ and v→0.5,the approximation formula tends to the analytical solution for the half-space model. In the incompressible case(v = 0.5), within the general experimental range(ζ varying from 2 to 4), the difference between the analytical solution and the approximate one is significant, and is up to 29% of the approximation solution when ζ= 2. Additionally, parametere was introduced to evaluate the error of elastic moduli between the half-space model and sphere model. Based on the approximation formula, the ζ thresholds, beyond which e becomes larger than 10% and 20%, were derived.

Normal age-related viscoelastic properties of chondrons and chondrocytes isolated from rabbit knee

Background The mechanical microenvironment of the chondrocytes plays an important role in cartilage homeostasis and in the health of the joint. The pericellular matrix, cellular membrane of the chondrocytes, and their cytoskeletal structures are key elements in the mechanical environment. The aims of this study are to measure the viscoelastic properties of isolated chondrons and chondrocytes from rabbit knee cartilage using micropipette aspiration and to determine the effect of aging on these properties. Methods Three age groups of rabbit knees were evaluated： （1） young （2 months, n=10）; （2） adult （8 months, n=10）; and （3） old （31 months, n=10）. Chondrocytes were isolated from the right knee cartilage and chondrons were isolated from left knees using enzymatic methods. Micropipette aspiration combined with a standard linear viscoelastic solid model was used to quantify changes in the viscoelastic properties of chondrons and chondrocytes within 2 hours of isolation. The morphology and structure of isolated chondrons were evaluated by optical microscope using hematoxylin and eosin staining and collagen-6 immunofluorescence staining. Results In response to an applied constant 0.3-0.4 kPa of negative pressure, all chondrocytes exhibited standard linear viscoelastic solid properties. Model predictions of the creep data showed that the average equilibrium modulus （E~）, instantaneous modulus （E0）, and apparent viscosity （~） of old chondrocytes was significantly lower than the young and adult chondrocytes （P 〈0.001）; however, no difference was found between young and adult chondrocytes （P 〉0.05）. The adult and old chondrons generally possessed a thicker pericellular matrix （PCM） with more enclosed cells. The young and adult chondrons exhibited the same viscoelastic creep behavior under a greater applied pressure （1.0-1.1 kPa） without the deformation seen in the old chondrons. The viscoelastic properties （E,, E0, and/~） of young and adult chondrons were significantly greater than that observed in young and adult cells, respectively （P 〈0.001）. The adult chondrons were stiffer than the young chondrons under micropipette aspiration （P 〈0.001）. Conclusions Our findings provide a theoretical model to measure the viscoelastic properties of the chondrons as a whole unit by micropipette aspiration, and further suggest that the properties of the chondrocytes and PCM have an important influence on the biomechanical microenvironment of the knee joint cartilage degeneration that occurs with aging.

ADHESION STRENGTH AND MORPHOLOGIES OF r BMSCs DURING INITIAL ADHESION AND SPREADING

Cell adhesion plays an important role in cell physiology. A better understanding of this process could facilitate many clinical therapies. In this study, Rat bone marrow-derived mesenchymal stem cells （rBMSCs） were cultured on glass substrate, and the morphology and adhesion strength were characterized. The cell morphology was defined as spherical, adhesive, and spreading. The adhesion strengths of the different morphologies exhibited different distribu- tion patterns. The spherical cells exhibited low adhesion strength; the adhesive cells exhibited rapidly increasing adhesion strength while their diameters remained relatively constant. The ad- hesion strength increased with the cell diameter in the spreading cells. These findings suggest that adhesion strength can be quickly assessed by examining the cell morphology.