Impact of radiocontrast agents and local anesthetic agents on the viability of human nucleus pulposus (NP) cells.

<p>NP cell–alginate 3D composites were exposed to saline, radiocontrast agent (iotrolan) or local anesthetic agents (lidocaine and bupivacaine) for 30, 60, or 120 min. After an additional 24 h, they were stained with calcein AM (live cells; green) and propidium iodide (PI; dead cells; red). (<b>A</b>) Confocal laser scanning micrographs of the NP cell–alginate 3D composites after the exposures. Scale bar: 100 μm. (<b>B</b>) Time-dependent effects on cell viability. (<b>C</b>) Dose-dependent effects on viability after 60 min. All data are from five independent experiments (mean ± SE; *, p<0.05). Bup, bupivacaine; Lid, lidocaine.</p

Impact of radiocontrast agents and local anesthetic agents on the viability of human nucleus pulposus (NP) cells.

<p>NP cell–alginate 3D composites were exposed to saline, radiocontrast agent (iotrolan) or local anesthetic agents (lidocaine and bupivacaine) for 30, 60, or 120 min. After an additional 24 h, they were stained with calcein AM (live cells; green) and propidium iodide (PI; dead cells; red). (<b>A</b>) Confocal laser scanning micrographs of the NP cell–alginate 3D composites after the exposures. Scale bar: 100 μm. (<b>B</b>) Time-dependent effects on cell viability. (<b>C</b>) Dose-dependent effects on viability after 60 min. All data are from five independent experiments (mean ± SE; *, p<0.05). Bup, bupivacaine; Lid, lidocaine.</p

Flow cytometric analysis of human nucleus pulposus (NP) cells.

<p>NP cell–alginate 3D composites were exposed to saline, radiocontrast agent (iotrolan), or local anesthetic agents (lidocaine and bupivacaine) for 30, 60, or 120 min, cultured 24 h, then the NP cells were isolated for analysis. (<b>A</b>) Flow cytometry scatter plots. (<b>B</b>) Time- and dose-dependent effects on cell viability (*, p<0.05 vs. untreated control and saline at the same exposure time). (<b>C</b>) Dose-dependent effects on cell apoptosis after 60 min (*, p<0.05). Data are from five independent experiments (mean ± SE). Bup, bupivacaine; Lid, lidocaine.</p

Implant rod angle of curvature at the concave and convex sides of the deformity.

<p>(<b>A</b>) θ1 and θ2 at the concave side of each patients. (<b>B</b>) θ1 and θ2 at the convex side of each patients. (<b>C</b>) Comparison between θ1 and θ2 at the concave side. (<b>D</b>) Comparison between θ1 and θ2 at the convex side. (<b>E</b>) Comparison between Δθ at the concave side and Δθ at the convex side.</p

Correlation analysis between rod deformation and variable in patients with rod deformation ≥14 ° at the concave side.

<p>Correlation analysis between rod deformation and variable in patients with rod deformation ≥14 ° at the concave side.</p

Rod angle before and after implantation.

<p>(<b>A</b>) Prior to implantation, the surgeon traced the rod shapes on paper. The angle between the proximal and distal tangential line was measured (θ1). (<b>B</b>) Postoperative implant rod geometry (θ2) was obtained after the surgical operation using computed tomography.</p

Western blot analysis of the apoptosis signaling pathways stimulated by lidocaine and bupivacaine.

<p>NP cell–alginate 3D composites were exposed 60 min to saline or a local anesthetic agent (lidocaine or bupivacaine), cultured 24 h, then the NP cell protein lysates were processed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting with primary antibodies against caspase-3, caspase-8, caspase-9, cleaved caspase-3, cleaved caspase-8, and cleaved caspase-9. β-actin was used as an internal control. (<b>A</b>) Representative Western blot. (<b>B</b>) Densitometry analysis performed via normalization to β-actin. Data are from five independent experiments (mean ± SE; *, p<0.05 vs. saline).</p

Associations between various factors and rod deformation at the concave side (°) using multiple stepwise linear regression analysis.

<p>Associations between various factors and rod deformation at the concave side (°) using multiple stepwise linear regression analysis.</p

Highly significant (<i>P</i><0.01) pathways based on KEGG database^*.

*<p>KEGG, Kyoto Encyclopedia of Genes and Genomes. “total gene count” means the number of genes in each pathway, which have already been registered in KEGG system, and “count” means the number of genes, which were expressed significantly in each pathway in this study.</p

Flow cytometric analysis of rat nucleus pulposus (NP) cells. Forty-eight hours after caspase 3 siRNA transfection, cells were serum-deprived and harvested after 48 h.

<p>(<b>A</b>) Representative graphs showing the cell cycle. (<b>B</b>) Comparison of the cell cycle in the G1 and S phase. (<b>C</b>) The dual parametric dot plots combining annexin V-FITC and PI fluorescence show the early apoptotic cells (FITC+/PI -), and the late apoptotic cells (FITC+/PI +). (<b>D</b>) Percentage of (early+late) apoptotic cells. Results are representative of three independent experiments. Values are expressed as the mean ± SD (* = P<0.05 versus all other groups, ** = P<0.05 versus serum-starved only and control siRNA).</p