A comparison of the torsional stiffness of the lumbar spine in flexion and extension.

OBJECTIVE: The main mechanism of injury to the spine is torsion especially when coupled with compression. In this study, the in vitro torsional stiffness of the lumbar spine segments is compared in flexion and extension positions by cyclic and failure testing. METHODS: Fifteen lumbar spines were sectioned from fresh cadavers into 15 L2/3 and 15 L45 motion segments. Each vertebral segment was then potted superiorly and inferiorly in polymethylmethacrylate, effectively creating a bone-disk-bone construct. The potted spinal segments were mounted in a mechanical testing system, preloaded in compression to 300 N, and axially rotated to 3 degrees in both directions at a load rate of 1 degrees /s. This was done over 3 cycles for each motion segment in the flexion and extension positions. Each specimen was then tested to torsional failure in either flexion or extension. Stiffness, torque, and energy were determined from cyclic and failure testing. RESULTS: The results showed that in all cases of cyclic testing, the higher segment extension resulted in higher torsional stiffness. In relative extension, the lumbar specimens were stiffer, generated higher torque values, and generally absorbed more energy than the relative flexion condition. There were no differences found in loading direction or failure testing. CONCLUSIONS: Increasing the effective torsional stiffness of the lumbar spine in extension could provide a protective mechanism against interverbral disk injury. Restoration of segmental extension through increasing the lumbar lordosis may decrease the strain and reinjury of the joints, which can help reduce the extent of pain in the lumbar spine.

In vitro evaluation of the effects of candidate immunosuppressive drugs: flow cytometry and quantitative real-time PCR as two independent and correlated read-outs.

BACKGROUND: Immune monitoring may use flow cytometry or molecular biology techniques. Flow cytometry assays cells that are phenotypically characterized, whereas TaqMan RT-PCR starts with RNA extraction from unfractionated heterogeneous cell populations. We therefore wondered how the effects of immunosuppressive drugs on cytokine production in stimulated whole blood, as determined by flow cytometry, would correlate with those obtained with quantitative real-time PCR (TaqMan RT-PCR). METHODS: Blood drawn from naive cynomolgus monkeys was exposed to incremental amounts of cyclosporine (CsA; 300, 600, 900 and 1200 ng/ml) or tacrolimus (TRL; 8, 20, 40 and 80 ng/ml) before lectin stimulation in vitro. Blood was then either stained for CD3, IFN-gamma, IL-2, IL-4, and TNF-alpha and analyzed on a flow cytometer with various gating strategies, or submitted to RNA extraction for analysis of the above mentioned cytokines mRNA transcripts using TaqMan RT-PCR. RESULTS: Both methods revealed a parallel dose-dependent inhibition of cytokine production in stimulated blood. The 50% inhibitory concentrations (IC(50)'s) ranged from 511-771 ng/ml (CsA) and 15-29 ng/ml (TRL) with flow cytometry, and from 275-529 ng/ml (CsA) and 11-48 ng/ml (TRL) with TaqMan RT-PCR for T-helper 1 cytokines. Both assays correlated well with a Pearson product moment correlation of 0.76. Extending gating from a CD3(+) gate to a lymphocyte gate improved correlation (r = 0.85) for all cytokines investigated (except IL-2; unchanged) whereas further extending gating resulted, to the contrary, in lower correlations. Independent of gating strategy a high correlation (r = 0.97) was observed when drug IC(50)'s were considered. CONCLUSIONS: Flow cytometry and TaqMan RT-PCR may be used interchangeably to monitor the effects of candidate immunosuppressive drugs on cytokine mRNA production in lectin-stimulated whole blood.