Fig. 2

Histology, biomechanical properties and stability analyses of cryopreserved bioengineered tissues. Histological analysis: A fresh and thawed tissue sections for each condition were stained with hematoxylin/eosin (left) and their pictures were transformed into binary images (right) using ImageJ program for quantification of pore size (n = 6 random fields; scale bar: 50 µm); fresh fibrin-agarose mesh contrast with thawed tissue, which presents more and larger-sized pores and B bar graph representation of the percentage of interfibrillar spacing. Macroscopic images of NFAH: C Representative fresh (left) and thawed (right) NFAH show the tissue remains in a good macroscopic status after thawing. NFAH is sectioned in three parts, one halve is cryopreserved and two quarters are used for quality controls. Biomechanical properties analysis: D Young’s modulus, E stress at fracture break and F traction-deformation. Cell stability analysis: G LIVE/DEAD® Assay staining of the tissue H cell viability at different time points (scale bar: 100 µm). Live cells are shown in green and dead cells in red. Results are presented as mean ± SEM. One-way ANOVA differences *P < 0.05; ***P < 0.001; **** P < 0.0001