Figure 4.

Content of proteins released into conditioned media. Conditioned media (CM) from vital (VC) and high hydrostatic pressure treated cartilage (HHPTC) were analyzed in supernatants generated during days 0 to 2 (2 d), 4 to 7 (7 d), 11 to 14 (14 d) and 32 to 35 (35 d) of cultivation as indicated in the separate graphs. Concentrations of procollagen type II (A), free active transforming growth factor (TGF-)β1 (C) and insulin-like growth factor-1 binding protein (IGFBP)3 (D) were quantified by enzyme-linked immunosorbent assays (ELISA; all n ≥ 3). Glycosaminoglycans (GAGs) (B) were quantified by Blyscan™ assay after enzymatic digestion with papain (n = 4). All concentrations are depicted as weight per volume. Data are presented as boxplots with minimum, 25th percentile, median, 75th percentile and maximum. As data were normally distributed according to Shapiro–Wilk test statistical analysis was performed with unpaired t test or unpaired t test with Welch’s correction depending on variances being significantly different. * p < 0.05, ** p < 0.01 and *** p < 0.001: significance between vital and high hydrostatic pressure treated cartilage. ^§ p < 0.05: significant difference between 2 and 35 days, ^$ p < 0.05: significant difference between 7 and 35 days and ^# p < 0.05: significant difference between 14 and 35 days for the same treatment.