Sequences of rabbit primer pairs, gene bank accession numbers used for real-time PCR analysis and size of the PCR products.

<p>Sequences of rabbit primer pairs, gene bank accession numbers used for real-time PCR analysis and size of the PCR products.</p

Schematic overview of <i>in vivo</i> experimental design.

<p>A) Schematic overview of the chondrogenic potential of differentially preconditioned rabbit adipose stromal cells (rASC). rASC were isolated and cultured under normoxic conditions (21% O₂) in control medium or chondrogenic medium or under hypoxic conditions (5% O₂) in chondrogenic medium. As a positive control, rabbit nasal chondrocytes (RNC) were used. Preconditioned rASC and RNC were finally associated with Si-HPMC hydrogel and implanted in rabbit articular cartilage defects for 18 weeks. B) Schematic overview of the chondrogenic potential of differentially preconditioned human adipose stromal cells (hASC). hASC were isolated and cultured under normoxic conditions (21% O₂) in control medium or chondrogenic medium or under hypoxic conditions (5% O₂) in chondrogenic medium. As a positive control, horse nasal chondrocytes (HoNC) were used. Preconditioned hASC and HoNC were finally associated with Si-HPMC hydrogel and implanted in nude mice subcutis for 5 weeks.</p

Sequences of human primer pairs, gene bank accession numbers used for real-time PCR analysis and size of the PCR products.

<p>Sequences of human primer pairs, gene bank accession numbers used for real-time PCR analysis and size of the PCR products.</p

Chondrogenic potential of differentially preconditioned rabbit ASC (rASC).

<p><b>A)</b> rASC were cultured under normoxic conditions (21% O₂) in control medium (NCT) and chondrogenic medium (NCH) or under hypoxic conditions (5% O₂) in chondrogenic medium (HCH). The expression of transcripts encoding type II collagen (<i>col2a1</i>) and aggrecan (<i>acan</i>) was measured by real-time PCR. The results are expressed as relative expression levels. ND: not detected. # <i>p</i><0.05 compared with NCT; * <i>p</i><0.05 compared with NCH. <b>B)</b> rASC were cultured in NCT (a, b, c, d), NCH (e, f, g, h), or HCH (i, j, k, l) and implanted with the Si-HPMC hydrogel in rabbit osteochondral defects. Rabbit nasal chondrocytes (RNCs) incorporated into the Si-HPMC hydrogel were used as a control (m, n, o, p). After 18 weeks of implantation, the defects were macroscopically observed [gross appearance (a, e, i, m)], histologically stained using Movat's pentachrome (b, f, j, n) and alcian blue (c, g, k, o) and immunostained for type II collagen (d, h, l, p). a, e, i, m: bar indicates 1 mm. b–d; f–h, j–l, n–p: bar indicates 100 µm. <b>C)</b> A semi-quantitative analysis of the regenerated tissue was performed using O′Driscoll's repair score as described in the “Materials and Methods” section. The results are expressed as a mean O′Driscoll score.</p

Chondrogenic potential of differentially preconditioned human ASC (hASC).

<p><b>A)</b> hASC were cultured under normoxic conditions (21% O₂) in control medium (NCT) and chondrogenic medium (NCH) or under hypoxic conditions (5% O₂) in chondrogenic medium (HCH). The expression of transcripts encoding type II collagen (<i>COL2A1</i>) and aggrecan (<i>ACAN</i>) was measured using real-time PCR. The results are expressed as relative expression levels. ND: not detected * <i>p</i><0.05 compared with NCH. <b>B)</b> hASC were cultured in NCT (a, b), NCH (c, d) or HCH (e, f) and implanted with the Si-HPMC hydrogel into subcutaneous pockets of nude mice. Horse nasal chondrocytes (HoNCs) incorporated into the Si-HPMC hydrogel were used as a control (g, h). After five weeks, the samples were harvested, histologically stained using alcian blue (a, c, e, g) and immunostained for type II collagen (b, d, f, h). Bar indicates 20 µm.</p