The detailed osteogenic medium component list.

<p>The detailed osteogenic medium component list.</p

Fluorescent image of DAPI-labeled cells on pore surfaces of scaffolds.

<p>BMSCs adhered in the vicinity of the pores and its edges of a nHA-CS scaffold. Red arrows indicate labeled cells (figure magnification 100×).</p

Micrographs of nHA-CS.

<p>The nHA-CS scaffold exhibited a three-dimensional porous structure in which the nHA particles were evenly distributed throughout the pore surface (figure magnification 500×).</p

BMSCs and identification of osteogenic differentiation at passage 3.

<p><b>(A)</b> BMSCs cultured in basic medium exhibited typical elongated fibroblast-like morphology. <b>(B)</b> ALP expression was identified by calcium-cobalt staining of black particle deposition in cells after one week of osteogenic induction. <b>(C)</b> Calcified nodules that formed in the extracellular matrix were detected by Alizarin red staining after three weeks of osteogenic induction. The red arrow indicates a calcified nodule (figure magnification 100×).</p

CT evaluation of the implants.

<p><b>(A)</b> Representative CT images of the implants scanned at 2, 4, 6, 8, and 12 weeks post-implantation. Red arrows indicate the implants. <b>(B)</b> The CT values of the implants in the nHA-CS+cells group were significantly greater than those of the nHA-CS group (*<i>P</i> < 0.05), indicating that <i>de novo</i> bone formation was greater in the nHA-CS+cells group.</p

Histological examination of HE staining.

<p><b>(A)</b> Representative HE images of implants from the two groups harvested at different time points (figure magnification 100×). <b>(B)</b> The analysis of the proportions of newly formed bone area indicated that the nHA-CS+cells group gained greater amounts of <i>de novo</i> bone compared with the nHA-CS group (*<i>P</i> < 0.05). <b>(C)</b> The analysis of the proportions of the scaffold area revealed that the nHA-CS+cells group exhibited significant scaffold degradation than the nHA-CS group (*<i>P</i> < 0.05).</p

Histological examination of Masson's trichrome staining.

<p><b>(A)</b> Representative Masson's trichrome staining images showing the expression of collagen in the two groups at different time points (figure magnification 100×). <b>(B)</b> The analysis of the proportions of the collagen area indicate that significantly more collagen was present in the nHA-CS+cells group than the nHA-CS group (*<i>P</i> < 0.05), indicating that the nHA-CS+cells group formed more <i>de novo</i> bone than the nHA-CS+cells group.</p