Objective: To explore the effect of three-dimensional (3D) and two-dimensional (2D) acellular fibrotic lung matrices on fibroblast-to-myofibroblast differentiation.
Methods: Twenty adult SD rats were randomly divided into control group and idiopathic pulmonary fibrosis (IPF) group. The pulmonary fibrosis was induced by Bleomycin. Normal and fibrotic decellularized lungs were made, then sections with 500 µm (3D) and 10 µm (2D) thick were cut by a standard Vibratome. The lung fibroblasts were isolated from the lungs of SD rats at postnatal day 1, which were seeded dropwise into different slices (2D/3D normal and fibrotic scaffolds), then cultured for 72 hours in vitro. The protein expression of alpha smooth muscle actin (α-SMA) was measured by immunofluorescence staining and western blot.
Results: There was no spatial structure both in 2D normal scaffold and fibrotic scaffold. The spatial structure was different between 3D normal scaffold and fibrotic scaffold. The relative expression amount of α-SMA in 2D normal and fibrotic scaffold group and none scaffold group were (4.64 ± 0.36, 4.86 ± 0.37, 4.67 ± 0.35, respectively), there were no difference among them (P>0.05). In 3D fibrotic scaffold group, the relative expression amount of α-SMA was significantly increased compared with those in 3D normal scaffold group and 2D fibrotic scaffold group (6.29 ± 0.85 vs 4.85 ± 0.56, 4.86 ± 0.37, both P<0.05).
Conclusion: 3D acellular fibrotic lung scaffold can promote fibroblast-to-myofibroblast differentiation.