The purpose of this study was to measure chondrocytes detachment from cellularized constructs cultured in a perfusion bioreactor, and to evaluate the effect of different scaffold coatings on cell adhesion under a fixed flow rate. The scaffolds were polyurethane foams, treated to promote cell attachment and seeded with human chondrocytes. In a preliminary static culture experiment, the scaffolds were imbibed with fetal bovine serum (FBS) and then cultured for 4 weeks. To quantify cell detachment, the number of detached cells from the scaffold treated with FBS was estimated under different interstitial perfusion flow rates and shear stress levels (0.005 mL/min equivalent to 0.05 mPa, 0.023 mL/min equivalent to 0.23 mPa, and 0.045 mL/min equivalent to 0.45 mPa). Finally, groups of scaffolds differently treated (FBS, plasma plus FBS, plasma plus collagen type I) were cultured under a fixed perfusion rate of 0.009 mL/min, equivalent to a shear stress of 0.09 mPa, and the detached cells were counted. Static cultivation showed that cell proliferation increased with time and matrix biosynthesis decreased after the first week of culture. Perfused culture showed that the number of detached cells increased with the perfusion rate on FBS-treated constructs. The plasma-treated/collagen-coated scaffolds showed the highest resistance to cell detachment. To minimize cell detachment, the perfusion rate must be maintained in the order of 0.02 mL/min, giving a shear stress of 0.2 mPa. Our set-up allowed estimating the resistance to cell detachment under interstitial perfusion in a repeatable manner, to test other scaffold coatings and cell types.
Bioreactors ; Cell Adhesion ; Cell Proliferation ; Chondrocytes* ; Collagen ; Humans ; Perfusion* ; Plasma ; Polyurethanes* ; Tissue Engineering