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tail and abdominal region at 1, 24, and 48 h, and at 1 week after injection. We confirmed that
there were no signals at the defect area in the cranium at any time point. In the PBS group,
fluorescent signals were observed in the tail and in the abdominal area at 24 and 48h after
injection, and very low signals were observed in the breast and cranial area after 1 week. In
the MSC-CM group, a moderate increase in signal intensity was observed in the area of the
tail and the abdominal region during the first 24h after injection. At 48h after injection, signal
intensity in the MSC-CM-implanted area of the parietal bone started to increase. The maximum
fluorescent signal in the implanted area was observed 1 week after injection (Fig. 2).
In vivo imaging analysis shows that DiR-labeled rMSCs that were injected into the caudal vein
just after implantation of the materials into the calvarial bone defects started to mi-grate
immediately after injection. At 24 h, 48 h, 72h and 1 week after injection, the signal of the
fluorescent-labeled rMSCs was only detected in the tail and abdominal region in the control
PBS group. In the MSC-CM group, a moderate increase in signal intensity was observed in the
abdominal region during the first 24 h after injection. Forty-eight hours after injection, the
MSC-CM-implanted area of calvarial bone started to increase in signal intensity, and, 1 week
after injection, the MSC-CM implanted area, as well as the implanted bone cavity, showed the
highest fluorescent signal of the experimental groups.
Figure 2.
In vivo imaging of injected rMSC migration to implants.
2.5. MSC-CM enhanced osteogenic and angiogenic marker gene expression
The levels of expression of the
ALP
,
OCN
,
Runx2
,
VEGF-A
,
ANG-1,
and
ANG-2
genes were
significantly upregulated in rMSCs cultured with MSC-CM compared with rMSCs cultured
in EM (Fig. 3).
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