Primer sequences used in quantitative real-time PCR (all primers were purchased from Invitrogen, Mississauga, Ontario, Canada).

<p>Primer sequences used in quantitative real-time PCR (all primers were purchased from Invitrogen, Mississauga, Ontario, Canada).</p

Ratios of mRNA expression levels of collagen type II (<i>Col2a1</i>) to collagen type I (<i>Col1a2</i>) and of aggrecan (AGG) to versican (VCN).

<p>Data represents mean ± SD; one-way ANOVA with Tukey's post hoc test: n.s. (not significant), * = <i>p</i><0.05, ** = <i>p</i><0.001, n = 10, N = 3.</p

Proliferation rate of human meniscus fibrochondroytes under normal and low oxygen tension in the presence of FGF-2.

<p>a) Mean cell doubling rate for P1 and P2 human meniscus fibrochondrocytes from 3 donors in the presence of FGF-2. b) Mean total population doubling of P2 human meniscus fibrochondrocytes. Data is expressed as mean ± SD of 3 donors (n = 3, N = 3). FGF-2, fibroblast growth factor 2; P1, passage 1; P2, passage 2. Student's t statistics; not significant (n.s.; <i>p</i>>0.05).</p

Gene expression profile of Duragen®-meniscus fibrochondrocytes constructs cultured for 21 days under normoxic (21%O₂) and hypoxic (3%O₂) conditions.

<p>The effect of oxygen tension on the matrix-forming capacity of differentially expanded MFCs prior to (monolayer cell culture; M) and after seeding and culture on Duragen® collagen scaffold (S) was investigated by real-time quantitative reverse-transcription-polymerase chain reaction (qRT-PCR). Total RNA isolated from monolayer cells culture (M) and cells-scaffold constructs (S) were analyzed for the mRNA expression of: the transcription factor, <i>Sox9</i>, and members of the collagen gene family and sulphated proteoglycans found in the extracellular matrix of meniscal fibrocartilage. All <i>y</i>-axes represent relative gene expression levels normalized to human RNA polymerase II (RPII). Data represents mean ± SD; one-way ANOVA with Tukey's post hoc test: n.s. (not significant), * = <i>p</i><0.05, ** = <i>p</i><0.001, n = 10, N = 3.</p

Histological and biochemical analysis of Duragen®-meniscus fibrochondrocytes (MFCs) constructs after culture for 21 days under normoxic (21% O₂) and hypoxic (3%O₂) conditions.

<p>a) Low (4×); c) medium (10×); e) high (40×) magnification photomicrographs of analysis of paraffin wax embedded sections (5 µm thickness) stained with alcian blue/neutral red stain (for sulphated GAG detection). Constructs were seeded with normoxia-expanded MFCs and were cultured under normoxia. b) Low (4×); d) medium (10×); f) high (40×) power magnification photomicrographs of alcian blue/neutral red (for sulphated GAG) stained constructs seeded with hypoxia-expanded MFCs and were cultured under hypoxia. Solid scale bar = 100 µm and dotted bar = 50 µm. (g) Biochemical analysis was used to evaluate the glycosaminoglycan (GAG) and DNA contents of constructs after 21 days culture in serum-free chondrogenic medium under normal (21%O₂) and low oxygen tension (3%O₂). Data is presented here as chondrogenic capacity (i.e. GAG levels normalized to DNA content) of constructs generated from 3 independent donors and it represents the mean ± SD (n = 10, N = 3). Student's t statistics; not significant (n.s.; <i>p</i>>0.05).</p