Comparison of statin users and a comparison group of statin non-users.

<p>Comparison of statin users and a comparison group of statin non-users.</p

Body mass index and collagen organization.

<p>The entire cohort (n = 66) was categorized as having a body composition that was normal (18.5 to 24.9, n = 24), overweight (25 to 29.9, n = 21) or obese (30+, n = 11).[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0199645#pone.0199645.ref021" target="_blank">21</a>] Statin users are shown as open circles, controls are yellow. Obese individuals (comprising 5 male statin-users, 1 female statin user, and 5 male comparison participants,) demonstrated a reduced collagen organization (Kruskal-Wallis H = 7.4, df = 2, p = 0.02) compared to those of normal body weight.</p

Primary antibodies used for analyses.

<p>Primary antibodies used for analyses.</p

Aggregation and Chondrogenesis of sfMPCs.

<p>Normal and OA CD90+ sfMPCs were induced to differentiate without prior micro-mass aggregation. By day 14 normal sfMPCs expressed Sox9, Collagen2 and Aggrecan mRNA (A,D) while OA sfMPCs expressed severely reduced levels (A,D). Furthermore, normal sfMPCs generated a tissue-like structure that stained positive for Alcian Blue (B,G), whereas OA sfMPCs did not (C,F) and remained as a monolayer. Scale bars represent 200 µM.</p

Staining and TEM of Derived Tissue from Normal sfMPCs.

<p>The resultant tissue-like masses were stained with antibodies to Collagen 2 (A) and Aggrecan (C), nuclei were stained with TOTO3 (B, D). Cells were found in closer association to each other (E), with higher magnification showing production of collagen fibres (F). Cells were also found in proximity with the produced collagen ECM (E, F). Scale bars represent 200 µM.</p

qRT-PCR probes used for analysis.

<p>qRT-PCR probes used for analysis.</p

Multi-potent differentiation of CD90+/− sfMPCs.

<p>Normal and OA CD90+ or CD90− sfMPCs were induced to differentiate into osteoblasts or adipocytes and stained with Akaline phosphatase (A) to indentify osteoblasts, or Oil Red-O (B) to identify lipid-containing cells (adipocytes). qRT-PCR was used to determine the relative level of adipocyte specific genes (Adiponectin, PPAR-gamma, [C]) and osteoblastic specific genes (Osteonectin, Sp7/Osterix [D]). * = p>0.05.</p

FACS characterization of human sfMPCs.

<p>Normal and OA CD90+ and CD90− subpopulations were assayed for expression of CD105, CD90, CD73, CD45 and CD11b prior to chondrogenic differentiation.</p

Micro-mass differentiation of sfMPCs.

<p>Normal (human N = 5 A–D & canine N = 2 H–K) and OA (human N = 5 E–G & canine N = 2 L–N) derived sfMPCs where aggregated using the micro-mass technique and induced to differentiate over a 14 day period with media supplements. qRT-PCR results demonstrated that Sox9, Collagen 2 and Aggrecan are significantly elevated at days 3, 5, 8 and 14 of differentiation (A,E,H,L), furthermore, at day 14 Collagen 2 protein is expressed in the cell cultures (B,F,I,M). Secondary controls (C,J) demonstrate minimal non-specific staining. Scale bars represent 50 µM. * = p>0.05.</p

NK1 and Trk expression in Achilles tendon.

<p>Expression of NK1 and Trk in intact and ruptured tendon of Wistar and diabetic GK rats at day 14 post-rupture. Original magnification is 20x; bars = 100 μm. (I; Intact part of the tendon and R; Ruptured area).</p