Journal: Materials Today Bio
Article Title: Bi-phasic integrated silk fibroin/polycaprolactone scaffolds for osteochondral regeneration inspired by the native joint tissue and interface
doi: 10.1016/j.mtbio.2025.101737
Figure Lengend Snippet: Biocompatibility and osteogenic differentiation potential of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds. (a) The cell distribution inside 3DSP (1:3) /GelSP (1:4) . Dil-labeled ATDC5(red) were seeded on the SP (1:3) layer, and Dio-labeled MC3T3-E1(green) were seeded on the GelSP (1:4) layer of 3DSP (1:3) /GelSP (1:4) for 7days. (b) Cell proliferation of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds at days 1, 3, and 7, as measured by the CCK-8 assay. (c) The viability of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds by live/dead staining and its 3D fluorescent images for 3 days. (d) Cytoskeleton staining of MC3T3-E1 cultured on different scaffolds for 3 days. (e–g) The relative mRNA expression levels of OPN, Runx2, and OCN of MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds for 14 days. (h) Immunofluorescence images of osteocalcin (OCN) in MC3T3-E1 seeded on 2D/3D SP (1:3) /GelSP (1:4) scaffolds. (∗p < 0.05. ∗∗p < 0.01. ∗∗∗p < 0.001).
Article Snippet: After 14 days of in vitro culture, cells/scaffold complexes were fixed in 4 % paraformaldehyde, permeabilized with 0.1 % Triton X-100 for 10 min, and blocked with 3 % BSA for 1 h. The cells were then incubated with the primary antibody against OCN (Proteintech, China, 1:500) and collagen II (Proteintech, China, 1:500) overnight respectively, followed by 1 h co-incubation with Cy3-conjugated fluorescence secondary antibody (Abclonal, China, 1:200).
Techniques: Labeling, CCK-8 Assay, Staining, Cell Culture, Expressing, Immunofluorescence