Figure 1.
Build-up monitoring of the CHT/CS polyelectrolyte multi-layered using QCM for film constructed.
A) Normalized frequency (?f7/7) and dissipation changes (?D7) obtain at 35 MHz, 1) deposition of CHT, 2) washing step and 3) deposition of CS; B) Estimated thickness (th) evolution and SEM micrographs of the multilayer surface with 10 double layers (inset image).
Figure 2.
Live/dead assay and SEM micrographs of BCH seeded on glass coverslips coated with chitosan and chondroitin sulphate at day 1 (A, B, C), 3 (D, E, F), 7 (G, H, I), 14 (J, K, L) and 21 (M, N) of culture in proliferation medium.
Figure 3.
A) Production steps of scaffolds: LbL and leaching of free-packet paraffin spheres, B)Digital photograph of the scaffold after all the steps C) Optical Microscopy image of the scaffolds after the leaching of the core material, D, E) SEM micrographs of cross-sections (two different magnifications) and Histological cross-sections of the scaffolds after staining with alcian blue (F) and eosin (G).
Figure 4.
Physicochemical characterization of scaffolds.
A) FTIR measurements of CHT/CS scaffolds and pure polysaccharides (CHT and CS), B) Swelling test up to 3 days (The inset graphic expands the water uptake for the first 5 hours), C) Weight loss of the scaffolds in PBS (▴) and in an enzymatic solution at 37°C (▪).
Figure 5.
Variations of (A) Storage modulus (E') and (b) loss factor (tanδ) of the CHT/CS scaffolds obtained by LbL methodology.
Experiments are reported for dry samples (▪) and hydrated samples in PBS at 37°C (•).
Figure 6.
Live/dead assay, MTT assay and cross-section SEM micrographs of BCH seeded on scaffold at day 1(A, B, C), 3(D, E, F), 14 (G, H, I) and 21(J, K, L) of culture in proliferation medium.
Figure 7.
Live/dead assay, MTT assay and cross-section SEM micrographs of hMSCs seeded on scaffold at day 1(A, B, C), 3(D, E, F), 14 (G, H, I) and 21(J, K, L) of culture in proliferation medium.
Figure 8.
DNA assay on the scaffolds seeded with BCH and hMSCs in differentiation medium.
Significant differences between each cell type at different time points were found for p<0.05(*) and p<0.01(**).
Figure 9.
Histological cross-sections of scaffolds seeded with BCH and hMSCs stained by H&E and Alcian blue at different days of culture in differentiation medium.