Chitosan flakes (molecular weight 50,000–190,000 Da based on viscosity, 75–85% deacetylated), silver nitrate and acetic acid were purchased from Sigma-Aldrich Chemie GmbH, Steinheim, Germany. Solutions were prepared with MilliQ water. Poly-L-lactic acid films of 50 µm thick were purchased from Goodfellow Cambridge Ltd, Huntingdon, UK and were cut in square shapes of 5 × 5 cm size.
Culture media and solutions: Alpha Minimum Essential Medium (α MEM, with ribonucleosides, deoxyribonucleosides, 2 mMl -glutamine and 1 mM sodium pyruvate, without ascorbic acid GIBCO, Custom Product, Catalog No. A1049001); Bovine fetal serum (BFS); Penicillin/Streptomycin/Neomycin solution (P/S/N) for cell culture; Phosphate Buffered saline (PBS) for cell culture; 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), solution in PBS (5 mg/mL) were purchased from Sigma-Aldrich Chemie GmbH, Steinheim, Germany.
Cells: Preosteoblasts of MC3T3-E1 line, subclone 4 (passage 21) (purchased from Sigma-Aldrich Chemie GmbH, Steinheim, Germany) were thawed and multiplied in culture flasks with a surface of 75 cm2, in culture media α MEM, without ascorbic acid, supplemented with 10% BFS and 1% mixture of antibiotic. The initial density of cells culture was 2000 cells/cm2 culture surface.
Culture media and solutions: Alpha Minimum Essential Medium (α MEM, with ribonucleosides, deoxyribonucleosides, 2 mM
Cells: Preosteoblasts of MC3T3-E1 line, subclone 4 (passage 21) (purchased from Sigma-Aldrich Chemie GmbH, Steinheim, Germany) were thawed and multiplied in culture flasks with a surface of 75 cm2, in culture media α MEM, without ascorbic acid, supplemented with 10% BFS and 1% mixture of antibiotic. The initial density of cells culture was 2000 cells/cm2 culture surface.