GAG/DNA ratio of hACPCs, after 4 week of culture.

<p>The total amount of GAG and the amount of DNA was quantified and GAG production was normalised to DNA content. Results are displayed as averages ± standard deviation (n = 12). * = significantly different from the control loaded group; $ = significantly different from the BMP-2 loaded group (<i>p ≤ 0</i>.<i>05</i>).</p

Relative gene expression of hACPCs.

<p>Gene expression analyses (a. Runx2, b. SOX9. c. Aggrecan, d. collagen I, e. collagen X, f. ALP) were conducted using the comparative ΔΔC_T method and 18s RNA as internal control. * = significant difference between control and BMP-2 (comparing unloaded to unloaded and loaded to loaded); # = significant difference between unloaded and loaded; $ = significant difference between control unloaded and BMP-2 loaded, (<i>p ≤ 0</i>.<i>05</i>).</p

Self-designed forward primers, reverse primers and probes used for real-time PCR.

<p>The primers and probes were designed and validated within our group using the primer express design software V.1.5 and were synthesized by Microsynth.</p

Overview of the eight different experimental groups, which were tested in the study.

<p>Overview of the eight different experimental groups, which were tested in the study.</p

Calcium incorporation in alginate beads with MSCs.

<p>For MSC beads, we found an increase in the uptake of AR from day -21 (A) before differentiation and day 0 (B). The Dex-treated beads then showed a much more intense staining, and at higher magnification, the Dex-treated cells (D) showed a pericellular matrix and dark red calcium depositions around the cells, which was not seen at this intensity in the other, T3-treated samples (C).</p

Proteoglycan visualization in alcian blue staining of alginate beads.

<p>Freshly prepared beads (A) only have slight background staining of alcian blue, almost all proteoglycans are removed from the cells during the digestion process. In the overview after serum preculture and five weeks of culture (B), the cells impose as round, singular cells with surrounding deep-blue pericellular matrix and further removed matrix in the gel. No differences can be visualized in the GAG content between T3-treated (C) and Dex-treated (D) chondrocytes.</p

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