Fig. 5.

Surface coating by films formation is feasible with Trypsin-treated DCC and it promotes in vitro cartilage differentiation. A) Schematic of surface coating procedure and further testing. B) Relationship between used DCC amount and formed film thickness (n = 3), C) Quantification of film thickness after 28 days incubation at 37 °C and 5%CO₂ (data normalized to film thickness at day 0) (n = 5), and inserted images of films stained with Alcian Blue as representative of film stability or dilution. D) Representative AFM image showing the surface roughness of T films and their average roughness measurement (n = 6). E-H) cell culture studies on Trypsin-obtained DCC films (T). E-F) Time-course studies of, E) Cell morphology and viability observed by CalceinAM, a viable cell-permeant green-fluorescent dye. Dead cells are observed by EtBr red-fluorescent nuclear staining. Surface roughness is provided by superposition of the transmitted-light image. F) Quantification of cell metabolic activity at different time point by (Resazurin-Resorufin fluorescence assay). G-H) Gene expression study of selected cartilage markers at cells cultured 21 days on control plastic surface or Trypsin-obtained DCC films (Data normalized to plastic grown cells) (n = 3), E) Cells cultured at control media, F) Cells cultured at cartilage differentiation media. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)