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Tobacco smoking increases the risk of intervertebral disc degeneration (IDD) and back pain, but the mechanisms underlying the adverse effects of smoking are largely unknown. Current hypotheses predict that smoking contributes to IDD indirectly through nicotine-mediated vasoconstriction which limits the exchange of nutrients between the discs and their surroundings. We alternatively hypothesize that direct contact of disc cells, that is, cells in the outermost annulus and those present along fissures in degenerating discs, with the vascular system containing soluble tobacco smoking constituents could perturb normal metabolic activities resulting in IDD. In this study, we tested our hypothesis by comparing the effects of direct exposure of human disc cells to tobacco smoke condensate and nicotine on cell viability and metabolic activity. We showed that smoke condensate, which contains all of the water-soluble compounds inhaled by smokers, exerts greater detrimental effects on human disc cell viability and metabolism than nicotine. Smoke condensate greatly induced an inflammatory response and gene expression of metalloproteinases while reduced active matrix synthesis and expression of matrix structural genes. Therefore, we have demonstrated that disc cell exposure to the constituents of tobacco smoke has negative consequences which have the potential to alter disc matrix homeostasis., (Copyright © 2011 Orthopaedic Research Society.)

As clinical evidence suggests that mechanical forces can have both reparative and traumatic effects on the spine, we investigated responses to different magnitudes, frequencies, and durations of applied tensile strain in an in vitro system. We examined the interactions of inflammatory and mechanical stimuli on cells isolated from the annulus fibrosus. Rabbit annulus fibrosus fibrochondrocytes were cultured in the presence or absence of an inflammatory stimulus. Cells were exposed to various magnitudes and frequencies of tensile strain for 4 or 24 h, and mRNA expression of catabolic mediators of inflammation and matrix degradation was measured by quantitative real time PCR and compared to control cells. Conditioned media were analyzed for matrix metalloprotease activity and production of prostaglandin E(2) . Application of low magnitudes and frequencies of tensile strain resulted in down-regulation of catabolic mediators, particularly under inflammatory stress. However, loss of this protective effect was observed at higher frequency and magnitude, and after prolonged duration. These in vitro data confirm the existence of magnitude, frequency, and duration based effects, which determine biochemical response of disc tissue resulting in either anti- or pro-catabolic outcomes. This may help to explain the beneficial effects of motion-based therapies as well as the destructive effect of traumatic levels of applied strain., (Copyright © 2011 Orthopaedic Research Society.)

Intervertebral disc degeneration (IDD) is a common and debilitating disorder that results in reduced flexibility of the spine, pain, and reduced mobility. Risk factors for IDD include age, genetic predisposition, injury, and other environmental factors such as smoking. Loss of proteoglycans (PGs) contributes to IDD with advancing age. Currently there is a lack of a model for rapid investigation of disc aging and evaluation of therapeutic interventions. Here we examined progression of disc aging in a murine model of a human progeroid syndrome caused by deficiency of the DNA repair endonuclease, ERCC1-XPF (Ercc1(-/Δ) mice). The ERCC1-deficient mice showed loss of disc height and degenerative structural changes in their vertebral bodies similar to those reported for old rodents. Compared to their wild-type littermates, Ercc1(-/Δ) mice also exhibit other age-related IDD characteristics, including premature loss of disc PG, reduced matrix PG synthesis, and enhanced apoptosis and cell senescence. Finally, the onset of age-associated disc pathologies was further accelerated in Ercc1(-/Δ) mice following chronic treatment with the chemotherapeutic agent mechlorethamine. These results demonstrate that Ercc1(-/Δ) mice represent an accurate and rapid model of disc aging and provide novel evidence that DNA damage negatively impacts PG synthesis., (© 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.)

Objective: To examine serum markers of matrix turnover in an animal model of disk degeneration., Design: Randomized prospective in vivo study., Setting: Laboratory for Orthopaedic and Spine Research and Department of Large Animal Research., Participants: Twenty-one New Zealand White rabbits., Intervention: Rabbits were randomly grouped into control (n = 8), sham surgery (n = 5), or stab surgery (n = 8). The stab surgical group underwent annulotomy of L2-3, L3-4, and L4-5 to induce intervertebral disk degeneration. The sham surgical group underwent surgical exposure without annulotomy, and the control group received no intervention., Outcome Measurements: Lumbar spine magnetic resonance imaging (MRI) and serum samples were obtained before intervention and at 0, 3, 6, and 12 weeks thereafter. MRIs were analyzed for evidence of intervertebral disk degeneration via measurement of MRI index. The serum was assayed at 0, 3, 6, and 12 weeks for the aggrecan biosynthesis marker CS846 and the C-telopeptide of collagen II (CTX-II)., Results: The stabbed disks demonstrate degeneration apparent by MRI criteria. CTX-II increased with time in the stabbed group compared to the control and sham surgery groups regardless of baseline levels. Aggrecan showed no statistically significant difference among groups., Conclusions: CTX-II shows promise as a useful serum biomarker for intervertebral disk degeneration.

Monolayer cultures of lapine articular chondrocytes were transduced with first-generation adenoviral vectors carrying lacZ or transforming growth factor beta1 genes under the transcriptional control of the human cytomegalovirus early promoter. High concentrations of transforming growth factor beta1 were produced by chondrocytes following transfer of the transforming growth factor beta1 gene but not the lacZ gene. Transduced chondrocytes responded to the elevated endogenous production of transforming growth factor beta1 by increasing their synthesis of proteoglycan, collagen, and noncollagenous proteins in a dose-dependent fashion. The increases in collagen synthesis were not accompanied by alterations in the collagen phenotype; type-II collagen remained the predominant collagen. Transforming growth factor beta1 could not, however, rescue the collagen phenotype of cells that had undergone phenotypic modulation as a result of serial passaging. These data demonstrate that chondrocytes can be genetically manipulated to produce and respond to the potentially therapeutic cytokine transforming growth factor beta1. This technology has a number of experimental and therapeutic applications, including those related to the study and treatment of arthritis and cartilage repair.