Bone structure and mechanical competence for the 4 groups of samples.

<p>Morphometric parameters (BV/TV  =  bone volume fraction, BS/TV  =  bone surface density, Tb.Th.  =  trabecular thickness, Tb.N.  =  trabecular number, Tb.Sp.  =  trabecular separation, Conn. D.  =  connectivity density, SMI  =  structure model index) and calculated apparent Young's modulus (E app) for the 4 groups of samples are indicated. No statistical differences were found. Values are expressed as mean ± standard deviation (n = 6 samples/group).</p

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<p>A) Light microscopic image showing an osteoid seam; B) Fluorescence micrograph showing the same area of cell activity with the corresponding alizarin bone surface labelling. C) Close-up of osteoclast as visualized by light microscopy; D) Close-up of the same area of cell activity as visualized by fluorescence labelling. Close-up scale bar represents 25 µm.</p

Outcomes of the correlation analyses between the bone turnover markers (ProCI and NTx) released by the samples with the volume (expressed in percentage) strained above different strain effective levels.

<p>Reported are the strain thresholds for which the coefficient of determination R² was maximum (only when the <i>p</i>-value associated with the Spearman correlation analyses was significant). ‘Direction’ indicates whether the bone markers were increasing (↑) or decreasing (↓) with increasing the strained bone volume.</p>a<p>, <i>p</i><0.05.</p

ProCI (µM) released by each bone sample.

<p>ProCI (µM) released by each sample in the culture medium during the first week (medium analyzed at the 6^th day) and the second week (medium analyzed at the 12^th day) of the experiment for the four groups of samples. A) Data relative to the samples dynamically loaded at microgravity; B) Data relative to the samples statically loaded at microgravity; C) Data relative to the samples dynamically loaded at normal gravity; D) Data relative to the samples statically loaded at normal gravity. ^a<i>p</i><0.05, n = 6. ^b<i>p</i><0.05, n = 5 (sample ‘10G’ is an outlier and was excluded for statistical analyses).</p

Label penetration depth at two penetration sites (from lower and upper surfaces, and from circumference edge).

<p>Calcein green penetration, which can be defined as time point zero, was significantly increased (*<i>p</i><0.01) compared to the alizarin complexon penetration (end time point). Box plots show the median line, the 25% and the 75% quartiles which define the box, the 1.5x interquartile-range whiskers, as well as outliers (○).</p

Light microscopic micrographs of Giemsa and eosin stained sections of the bone explants.

<p>A) Micrograph shows bone matrix labelled (red), haematopoietic bone marrow (blue), and the presence of a multinucleated osteoclast (blue) lying in its resorption lacunae; B) Micrograph shows bone matrix (red), haematopoietic bone marrow (blue), and the presence of an osteoid seam (blue); C) Close-up of osteoclast; D) Close-up of osteoid. Close-up scale bar represents 25 µm.</p

Schematic illustration of the FreqBone bioreactor and culture system.

<p>A) The front side shows the culture chambers that are arranged in a circle with a rotating loading stack in the centre; B) The medium reservoirs and the pump system are visible on the back side.</p