CHITIN AND CHITOSAN FOR TISSUE ENGINEERING APPLICATION - Intelligent Scaffolds for Tissue Engineering and Regenerative Medicine - page 446

Figure 23.6. SEM microographs of chitosan bead scaffolds prepared in
various chitosan concentrations. (a, e) 0.5%, (b, f) 1.0%, (c, g) 1.5%, and
(d,h)2.0%chitosanin1%aceticacidsolution.(a-d)Surfaceand(e-h)cross
sectionofchitosanbeadscaffolds(magnification 1.2mm). 7 × 25,scalebar =

architecture. Figure 23.6 shows morphologies of the surface and
cross section of the chitosan bead scaffolds prepared in various chi-
tosan concentrations. The microstructure of the chitosan bead scaf-
folds became looserwitha decrease in the chitosan concentration.

23.4.2 Cell Culture on Chitosan Bead Scaffolds
Chondrocytes from rabbit costal cartilage at passage 3 were used
and inoculated into the chitosan bead scaffolds at a concentra-
tion of 2.4 × 10 4 cells/100 μ L/well (7 beads/well). The inoculated
chondrocytes were cultured in a medium (MSCGM BulletKit, Lonza,
Walkersville, MD, USA), including basic fibroblast growth factor
(b-FGF) (R&D Systems, Inc.) (1 ng b-FGF in 1 mL media) in an incu-
bator under 95% humidity at 37 ◦ Cwith5% CO 2 .
Figure23.7showsthemorphologiesofchondrocytesculturedon
the surface ofthe chitosan bead scaffolds after 14 days of culture.
The cultured chondrocytes and ECM were shown on the surface
of the bead scaffolds.
In the results of cellular viability and proliferation by the MTT
assay, initial cell proliferation in the bead scaffolds with microp-
oreswashigherthanthatinthebeadscaffoldswithoutmicropores. 7
The safrain-O staining images of the bead scaffolds showed that the