The cell compatibility was detected according to previously described procedures [37 (link)]. The test samples (films Col/Chi, Col/Chi-Ta and DC-Col/Chi) were sterilized by UV radiation prior to testing. Mouse embryonic fibroblast cell line (ATCC CRL-1658 NIH/3T3; Marlboro, MA, USA) was used to test the adhesion and proliferation of cells on the surfaces. ATCC-formulated Dulbecco’s modified Eagle’s medium (Biosera, Nuaille, France) containing 10% calf serum (Biosera) and 100 U·mL−1 penicillin/streptomycin (PAA, Trasadingen, Switzerland) was used as the culture medium.
In the case of adhesion, the cells were seeded on the samples in the concentration of 1.106 cells mL−1. After one hour, the cells were stained with Hoechst 33258 (Molecular Probes, Carlsbad, CA, USA). To determine the ability of cells to proliferate on the surfaces, the cells were seeded at an initial concentration of 1.105 cells mL−1 and cultivated for 48 h. After 48 h the cells were fixed and stained with Hoechst 33258 and ActinRed 555 (Thermo Fisher Scientific, Waltham, MA, USA). Micrographs were taken using the fluorescence microscope Olympus IX 81 (Olympus, Tokyo, Japan).
In the case of adhesion, the cells were seeded on the samples in the concentration of 1.106 cells mL−1. After one hour, the cells were stained with Hoechst 33258 (Molecular Probes, Carlsbad, CA, USA). To determine the ability of cells to proliferate on the surfaces, the cells were seeded at an initial concentration of 1.105 cells mL−1 and cultivated for 48 h. After 48 h the cells were fixed and stained with Hoechst 33258 and ActinRed 555 (Thermo Fisher Scientific, Waltham, MA, USA). Micrographs were taken using the fluorescence microscope Olympus IX 81 (Olympus, Tokyo, Japan).
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