Multinucleated TRAP⁺ OCLs.

<p>The images show sRANKL-treated RAW264.7 cells cultured in direct contact with the samples: (A) HA-LF, (B) HA, (C) LF and (D) cells only, respectively. White arrows indicate large and multinucleated cells positive for tartrate-resistant acid phosphatase (mature OCLs). Scale bar 100 μm.</p

OBs viability and apoptosis.

<p>(A) shows the percentage of live OBs respect to the total cells counted and (B) shows the percentage of apoptotic OBs respect to the total cells counted. Mean and standard error (n = 3) represented as the percentage of the total counted cells, after 7 and 14 days of culture in direct contact with all the tested samples. Statistical significant differences among the samples are indicated in both graphs: *p≤0.05. (C) Different examples of nuclear fragmentation in OBs stained with DAPI are indicated with red arrows. Scale bar 50 μm.</p

MSCs viability.

<p>Percentage of MSCs viability grown in Sr2% and 5%-BCs compared to the cells grown in BC. Statistical analysis showed no differences among the samples tested and over the experimental time points.</p

OBs gene expression analysis.

<p>Relative quantification of gene expression after 7 and 14 days of OBs grown on Sr-BCs. The graph showed the fold change expression of Osterix, Bglap and IBSP, relative to the expression of the OBs grown on BCs, used as a control.</p

Samples identification.

<p>Labelling of the samples, weight percentage (wt%) of LF loaded onto HA and LF and HA concentration (μg/ml) of the samples tested in the study.</p><p>Samples identification.</p

OCLs gene expression analysis.

<p>Relative quantification of gene expression after 7 and 14 days of OCLs grown on Sr-BCs. The graph showed the fold change expression of Oscar, CtsK and Itgβ3 relative to the expression of the OCLs grown on BCs, used as a control. Statistical differences exist for Oscar and CtsK expression between day 7 and day 14 of culture (*p≤0.05; ** p≤ 0.01).</p

Cells morphology.

<p>The upper level on the panel showed phalloidin staining: in green the cytoplasm of the cells and in blue the nuclei. At 3d, MSCs (A) and OBs (B) were well spread on BC surface exhibiting their characteristic morphology without any difference among each sample; scale bars 200 μm. Image (C) showed a big multinucleate OCLs (*) and groups of undifferentiated monocytes (white arrows); scale bar 50 μm. On the lower level of the panel SEM images are showed: D and E showed the cytoplasmic extension (white arrows) of MSCs and OBs, respectively; scale bars 5 μm. (F) One OCLs grown on BCs surface, exhibiting the typical apical-basal polarised resorbing morphology (yellow arrow); scale bar 10 μm.</p

Ion concentration.

<p>Cumulative Ca²⁺ (A) and Sr²⁺ (B) ion release concentrations from the cements in Dulbecco's modified Eagle's medium and resulting ion concentration (in weight %) in respect to the initial amount of Sr²⁺ in the precursor powders (C). (*p≤0.05; **p≤0.01, ***p≤0.001).</p

OBs gene expression analysis.

<p>(a) and (c) Relative quantification (2^-ΔΔCt) of gene expression after 7 and (b) and (d) 14 days of OBs cultured in direct contact with all the tested samples. The graph shows the average and standard error of the technical triplicate of Osterix and IBSP, respect to the expression of the cells only, used as a control. Statistical significant differences among the samples are indicated in the graphs: *p≤0.05 and ***p≤0.001.</p

MSCs gene expression analysis.

<p>Relative quantification of gene expression after 7 and 14 days of MSCs grown on Sr-BCs. The graph showed the fold change expression of RUNX2 and ALP, relative to the expression of the MSCs grown on BCs, used as a control (*p≤0.05; **p≤0.01).</p