| Objective:to apply the micropipette aspiration technique for sodium alginate cultured rabbit knee articular cartilage cell mechanical properties of quantitative analysis for cartilage tissue engineering, mechanical factors to provide the corresponding technical parameters and for the ultimate use of tissue engineering in the treatment of cartilage injury related theoretical basis.Methods:Eight two-month-old New Zealand white rabbits, male and female, They were disposed with the requirement of animal ethics. Under sterilized conditions, separation of its double knee, to culture medium prepared0.4%Protease enzyme,0.025%type Ⅱ collagenase digestion of rabbit cartilage cell separation sequence, with a4x106/mL concentration and Sodium Alginate Blend, by silica gel plate molded into a circular cylindrical cell plate (25L/a), CaC12solution gel5minutes, DMEM/F12serum-free medium plus20%FBS (fatal blood solution) in a culture plate with6holes in culture.In1,2,4weeks (1W,2W,,4W) take out the cell plate, using micropipette aspiration technique combined with semi infinite body model (Half-space model) analysis of cartilage cell mechanical properties (instantaneous modulus E0, equilibrium modulus E infinity, apparent viscosity mu,).The use of flow cytometry determination of1W,2W and4W cartilage cell apoptosis.Results:1W,2W,4W chondrocytes compared with pairwise cartilage cells of early apoptosis and living cells were not statistically significant (P>0.05).4W cultured cartilage cell viscoelasticity was significantly lower than that in1W and2W (P<0.005),1W,2W between comparison was not statistically significant (P>0.005);1W,2W,4W cartilage cells are typical viscoelastic solid creep characteristics.Conclusion:by micropipette aspiration of mechanical measuring sodium alginate three-dimensional culture of chondrocytes in the mechanical characteristics of relatively stable, suitable for construction of tissue engineered cartilage seed cell. |
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