Fig 1.
Grain sizes of nanoBHA and nanoHA based on transmission electron microscopy (a). Hydrodynamic particle size and polydispersity index of nanoBHA after fabrication using the wet ball milling method (b). Comparison of nanoBHA and nanoHA on deionized water and growth medium (c).
Fig 2.
Scanning electron microscopy images (a). XRD (b) and FTIR spectra (c) of BHA, HA, nanoBHA, and nanoHA.
Table 1.
Elemental weight of nanoBHA, nanoHA, BHA, and HA (%).
Table 2.
Zeta potential value of nanoBHA, nanoHA, BHA, and HA (mV).
Fig 3.
LDH release (a), cell viability (b), live/dead cell viability (c), and proliferation of preosteoblasts (d) and induced osteoblasts (e) from nanoBHA, nanoHA, BHA, and HA groups. Scale bar: 200 μm.
Fig 4.
Morphology of preosteoblasts after 14 days of differentiation induction.
Green, F-actin; blue, nuclei. Material concentration = 50 μg/mL. Scale bar: 50 μm.
Fig 5.
ALP staining results (a) and the corresponding quantification (b) and calcium deposition of osteoblasts after 14 days of differentiation inducement (c). Scale bar: 10 μm.
Fig 6.
Gene expression of osteogenic differentiation markers: Runx2 (a), ALP (b), OPN (c), and OCN (d), as well as OPN protein level (e).
Fig 7.
Results of Western blot analysis of p-ERK1/2 and ERK1/2 (a) and the corresponding quantification of p-ERK/ERK (b). Immunofluorescence imaging of p-ERK1/2 (c) and the corresponding quantification of p-ERK1/2 intensity (d). OPN protein level, with and without pretreatment of ERK1/2 signaling pathway inhibitor (e). The quantifications were based on fold change from the negative control. Scale bar: 50 μm. Blots from b were observed for each antibody as separated in white space, the original blots are presented in S1 Raw image.
Fig 8.
Representative micro-CT images of the augmented region after 6 weeks of treatment.
Scale bar: 100 μm.
Fig 9.
Representative histological images of the augmented region after 6 weeks of treatment.
H&E, hematoxylin–eosin; MT, Masson’s trichrome. Scale bar: 100 μm.