Radiography shows bone formation in DBM/AF constructs that were implanted into the dorsal skin pocket of athymic mice.

<p>Rabbit annulus tissue was dissected and fabricated into a DBM cylinder and the constructs were implanted into the dorsal skin pocket of mice. Specimens were harvested at different time points. X-ray images A) in <i>vitro</i>, and, B) <i>in vivo</i>. Scale bar = 1 mm.</p

Primer Sequences for real-time PCR.

<p>Primer Sequences for real-time PCR.</p

Immunohistochemitry detection of subcutaneously implanted DBM/AF (A) or AF alone constructs (B).

<p>Separate sections were treated with collagen I, collagen II, collagen X, and osteocalcin primary antibodies and corresponding secondary biotinylated IgG. The signals were developed with 3,3′-diaminobenzidine and photographed on an Axioskop 2 Zeiss microscope. Five sections from four discs were used at each time point. Arrows showing hypertrophy chondrocytes.</p

Hypertrophic chondrocytes and osteoblasts in punctured intervertebral disc specimens with Safranin-O staining.

<p>The rat intervertebral discs including endplates, NP, and AF tissues were punctured with a needle and implanted into the dorsal skin pocket of athymic mice. The specimens were harvested at 1, 2, 4, and 6 months postoperatively. Tissue sections stained with Safranin-O for disc structure and constituent proteoglycan staining shown in red (A, 4 x magnification; B,10 x magnification). Arrow: hypertrophy chondrocytes. Five sections from four discs were used at each time point.</p

Chondrogenesis of rabbit annulus fibrosus cells <i>in vitro</i>.

<p>Rabbit annulus fibrosus cells were cultured in a pellet culture system, for three weeks, with chondrogenic medium or control medium (DMEM+1%FBS+1% ITS). A) Gross morphology of cell-pellet. B) GAG in the cell-pellet was measured with the dimethymethylene blue colorimetric assay using chondroitin sulfate as a standard; values are normalized to cell DNA content. C) Expression of collagen II and aggrecan genes are significantly increased in the chondrogenic culture medium compared to control medium. D) Representative images of Safranin-O staining for proteoglycan and immunostaining for type II collagen in the cell-pellet. Scale bar = 500 µm. * p<0.05; ** p<0.01.</p