Figure legends
Figure 1: Schematic illustration of the designs of the hybrid
scaffold combining 3D printed polycaprolatone coated with Silk Fibroin
Methacryloyl, epithelium, BMSCs and Kartogenin for
reconstruction
of rabbit tracheal window-shape defect.
Figure 2: The macroscopic appearance of SilMA in different
status (A: curing ring; B: pure 10% SilMA; C, D: 15%, 20% SilMA after
culturing in DMEM-F12 medium; E: freeze-dried status of Fig. D).
Figure 3: The swelling ratio and remaining mass of SilMA in
different concentration and different status (A, D: The swelling ratio
and remaining mass of pure SilMA; C: The degradation process of pure
SilMA; B, E: The swelling ratio and remaining mass of freeze-dried
SilMA). Data represent mean ± SD; **P < 0.01.
Figure 4: A1: At day 4, almost no RBCs left and BMSCs grew well
and fast; A2: The 2nd passage BMSCs; A3: Flow
cytometry analysis revealed BMSCs matched the characteristics of MSCs
that CD29, CD44, and CD90 were highly expressed and few cells were
positive in CD34. The epithelial cells were cultured by the tissue
extracted from autologous trachea mucosa. B1, B2: At day 5,
the cells gathered into clusters;
B3, B4: Cells reached 90% confluence at day 13. (Magnification: A1, A2:
×40; B1, B3: ×100; B2, B4: ×200).
Figure 5: A: CCK-8 test of SilMA in different concentration. B
represented the H&E staining of 15% SilMA and C represented 20%, red
arrows indicate the alive cells (Magnification: B, C: ×100; b, c: ×200).
D represented the quantified column chart. Data represent mean ± SD;
*P < 0.05, **P < 0.01.
Figure 6: The appearance
and
Giemsa staining of BMSCs around the scaffold of each group after
inoculation for 48 h. In vitro cytotoxicity test shows 20% SilMA has
good cytocompatibility (Magnification: a-l: ×40).
Figure 7: Scanning electron microscopy (SEM) observation of
PCL, 20% SilMA, PCL+20% SilMA and SilMA+BMSCs (Magnification: a: ×50,
d and g: ×500, b, c, e, f, h and i: ×1000, j: ×3000).
Figure 8: Mechanical test shows hybrid scaffold has favorable
biomechanical properties. In the upper left corner of the figure, from
left to right are native trachea, PCL scaffold and hybrid scaffold
(PCL+20% SilMA).
Figure 9: a-c: Intraoperative images of tracheal partial
window-shape defect and repair. Black arrow indicates the SilMA hydrogel
droplet containing BMSCs and KGN; d, e: X-ray images of 1, 2 months
after operation; f: Bronchoscopic images of the luminal area of the
graft at 2 months,
white
arrow indicates the transplantation site; g-i: Gross view of native
trachea and the patch after tracheal reconstruction for 2 months. Red
arrow and white dotted boxes indicate the transplantation patch.
Figure 10: a-c: H&E staining (S: SilMA hydrogel; P: PCL; C:
cartilage rings); d-f: Immunohistochemistry staining of CK-18; g-i:
Immunofluorescence staining of CK-18. Red arrow indicates the
vascular-like structures, white dotted lines mark the anastomosis, green
arrows indicate the native tracheal epithelium and yellow arrows show
the nascent epithelial cell layer on the inner surface of the repair
site (Magnification: a, d: ×40, e: ×100, b, c, f: ×200, g, h, i: ×200).