Fig. 6
α-SMA protein detection in the 3D matrices. alpha-smooth muscle actin (α-SMA) protein expression in the primary human normal lung fibroblasts was evaluated using western blot. Fibroblasts were cultured within 3D glycated DMEM matrices containing 10 % serum. a. Western Blot. α-SMA band, 42 kDa; α-tubulin band, 50 kDa. Albumin band (66 kDa, due to FBS and visible in cellular and acellular matrices blots’, probably because of the difficulty in rinsing all the media from the matrices prior to protein extraction). The experiments were repeated three times. a1. An insignificant amount of α-SMA was detected in all the cellular matrices. a2 and a3. Higher amounts of α-SMA were observed in all the cellular matrices on the 14th and 21st days of culture. a1, a2 and a3. No tubulin or α-SMA bands were detected in the non-cells matrices. b. Densitometric analysis of α-SMA levels (Ratio of α-SMA to α-tubulin). The results were obtained from three independent experiments. *p < 0.05, **p < 0.01. A gradual decrease of α-SMA was observed at higher ribose concentrations on the 7th day of culture (p > 0.05). Contractile phenotype was most strongly detected on the 14th and 21st days of culture. It was significantly higher at all ribose concentrations, also for controls without ribose, compared with the level on the 7th day. It suggests that the microambient ‘per se’ could induce a phenotype change and that cells contribute to the matrix contraction