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A B

C D

E F G

I
H J

Figure 7. Effects of the trabeculae-like biomimetic bone-filling material (TBM) on in situ bone regeneration in calvarial defect mice. (A) Schematic
diagram of the TBM treatment on calvarial defect mice. (B) Representative hematoxylin and eosin staining images of mice calvarial defect region
(scale bar: 200 μm; magnification: ×32.5). (C) Microcomputing tomography reconstructed images of calvaria defect after 3 weeks (scale bar:
5 mm; magnification: ×2.16). Red arrows indicate a bone defect area. (D-F) Quantitative analyzes of bone mineral density (BMD; D), bone volume
to tissue volume (BV/TV; E), and bone mineral content (BMC; F) in (C). (G and H) Representative images show the mineral apposition rate of
mice’s calvarial defection region (G; scale bar: 10 μm; magnification: ×700) and quantitative analysis of the mineral apposition rate (H). Red
arrows indicate bone formation line distance labeled with calcein. (I and J) Osteocalcin (OCN) staining images after 3 weeks as detected by
immunohistochemical staining (I; scale bar: 30 μm; magnification: ×350) and quantification of related integrated optical density (IOD) values (J;
data represented as mean ± SD, n = 3). Red arrows indicate OCN-positive cells.
Notes: All data in bar graphs are represented as mean ± SD, n = 3; Blank: No treatment for the bone defect region; TBM: TBM filling the bone defect
region; Ber: TBM loaded with bergamottin filling the bone defect region; Polyvinylamine (PVAm): TBM loaded with PVAm filling the bone defect
region; MSA: TBM loaded with empty recombinant tRNA (loaded by PVAm) filling the bone defect region; anti138: TBM loaded with recombinant
miR-138-5p antagonist (loaded by PVAm) filling the bone defect region; *p<0.05; **p<0.01; ***p<0.001.

while osteogenic drug-loaded TBM markedly promoted and 7D-H). Moreover, immunohistochemical staining
bone repair (Figures 6B and C, 7B and C). The values of bone for OCN in osteoblast demonstrated significantly higher
apposition rates, BMD, BV/TV, BMC, and tibial Tb.N at bone intensity in osteoblast within the defected regions treated
defect regions were significantly increased (Figures 6D-I with either bergamottin or miR-138-5p antagonist-loaded

Volume 1 Issue 2 (2025) 13 doi: 10.36922/OR025040003