Human Lumbar Discs Research Articles

Rat disc torsional mechanics: effect of lumbar and caudal levels and axial compression load

Background context Rat models with altered loading are used to study disc degeneration and mechano-transduction. Given the prominent role of mechanics in disc function and degeneration, it is critical to measure mechanical behavior to evaluate changes after model interventions. Axial compression mechanics of the rat disc are representative of the human disc when normalized by geometry, and differences between the lumbar and caudal disc have been quantified in axial compression. No study has quantified rat disc torsional mechanics. Purpose Compare the torsional mechanical behavior of rat lumbar and caudal discs, determine the contribution of combined axial load on torsional mechanics, and compare the torsional properties of rat discs to human lumbar discs. Study design Cadaveric biomechanical study. Methods Cyclic torsion without compressive load followed by cyclic torsion with a fixed compressive load was applied to rat lumbar and caudal disc levels. Results The apparent torsional modulus was higher in the lumbar region than in the caudal region: 0.081±0.026 (MPa/°, mean±SD) for lumbar axially loaded; 0.066±0.028 for caudal axially loaded; 0.091±0.033 for lumbar in pure torsion; and 0.056±0.035 for caudal in pure torsion. These values were similar to human disc properties reported in the literature ranging from 0.024 to 0.21 MPa/°. Conclusions Use of the caudal disc as a model may be appropriate if the mechanical focus is within the linear region of the loading regime. These results provide support for use of this animal model in basic science studies with respect to torsional mechanics.

1,25(OH)2-Vitamin D3 Inhibits Proliferation and Decreases Production of Monocyte Chemoattractant Protein-1, Thrombopoietin, VEGF, and Angiogenin by Human Annulus Cells In Vitro

Human lumbar anulus tissue and cultured human lumbar anulus cells were used in retrospective studies of the immunocytochemical localization of the vitamin D receptor (VDR) in disc tissue, and of the in vitro effects of the active metabolite of vitamin D, 1,25(OH)2D3, on anulus cell proliferation, cytokine, and proteoglycan (PG) production. 24,25-D3 was also analyzed. Studies were approved by the authors' Human Subjects Institutional Review Board. Discs were obtained from surgical specimens and from control donors. To determine if human anulus cells express the VDR in vivo, and to test the effect of in vitro exposure to 1,25(OH)2D3 and 24,25-D3 on anulus cell proteoglycan and cytokine production in 3-dimensional culture. Intragenic polymorphisms in the VDR gene have been associated with disc degeneration. 1,25(OH)2D3 has well-recognized effects on calcium homeostasis and bone mineralization, and is a negative growth regulator of a variety of normal and tumor cells. Its effects on human disc cells, however, are unexplored. Immunocytochemistry was performed on human lumbar disc anulus tissue from 19 subjects; human disc cells were cultured to test the effect of 1,25(OH)2D3 on proliferation of anulus cells from 5 subjects. A paired experimental design was used to determine proteoglycan production in control or 1,25(OH)2D3-treated cells, or in control or 24,25-D3-treated cells using the dimethylmethylene blue assay. A paired experimental design was also used to identify differences in cytokine production in conditioned media from control or 1,25(OH)2D3-treated cells, or in control or 24,25-D3-treated cells using ELISA assays. Immunocytochemistry documented expression of the VDR in anulus cells. Young donor discs (aged newborn, 15 years) showed positive localization in all cells of the outer anulus, and some inner anulus cells. In adults (mean age, 38.9 years), some, but not all anulus cells, showed positive localization. Exposure to 10M 1,25(OH)2D3 in monolayer significantly reduced cell proliferation in vitro (P = 0.03). PG production in 3-dimensional was unchanged from control in both 1,25(OH)2D3- and 24,25-D3-treated cells. Cytokine production differed, however. 1,25(OH)2D3-treated cells showed significantly decreased production of vascular endothelial growth factor (VEGF) (P = 0.01), monocyte chemoattractant protein-1 (MCP-1) (P = 0.0006), angiogenin (P = 0.002), and thrombopoietin (P = 0.03) compared with controls. 24,25-D3-treated cells showed significantly elevated vascular endothelial growth factor-D (P = 0.01), beta-fibroblast growth factor (0.03), and significantly decreased interleukin-8, interferon-gamma, leptin, MCP-1, and TIMP-2 (tissue inhibitor of metalloproteinases-2) compared with controls (P <or= 0.01). Data suggest that 1,25(OH)2D3 and 24,25-D3 may play roles as regulators of cell proliferation and production of specific cytokines in the lumbar anulus.

Pharmacological enhancement of disc diffusion and differentiation of healthy, ageing and degenerated discs

Pharmacological enhancement of disc diffusion and differentiation of healthy, ageing and degenerated discs

Novel Synthetic Total Disk Model for Mechanical Testing of Nucleus Replacement Devices

We are presenting a novel, non-biologic model of the healthy human annulus. This lumbar Total Disc Model (TDM) ideally would be both biofidelic and sufficiently robust to withstand long-term fatigue testing without breaking down or tearing apart. Mechanical validation testing was performed to confirm that the compressive and torsional properties were similar to literature values of denucleated human lumbar discs in two-body constructs. Long-term fatigue tests were performed to establish the durability of the model. We have reported data for both our empty TDM and the TDM filled with a representative nucleus replacement device (NRD). The silicone model is geometrically equivalent to the healthy human lumbar disc, including the discoid cavity present following a total nuclectomy, the annulus fibrosis with micro-annulotomy, and the cartilaginous endplates. The pressure transmitted through the center of the disc went from negligible when the TDM was empty to over 40% when the TDM was filled. The compression stiffness was 992±15 N/mm for the empty TDM and 1583±136 N/mm for the filled TDM. The torsional stiffness was 0.505±0.024 Nm/° for the empty and 0.550±0.056 Nm/° for the filled TDM. Lastly, the only mechanical damage suffered by either empty or filled TDMs during dynamic testing came from debonding from the endplates at higher torque levels. No damage was seen during dynamic compression testing. After determining the appropriate geometry for the TDM, validation testing was performed to ensure that the load sharing, compressive, and torsional properties were similar to the native human disc. The silicone model was durable enough to avoid tearing or mechanical failure at physiologic loads. This study demonstrated that a silicone total disc model was developed with appropriate properties necessary for determining the mechanical degradation properties of a NRD and will not mechanical fail at physiologic loads.

Comparison of Animal Discs Used in Disc Research to Human Lumbar Disc

Experimental measurement and normalization of in vitro disc axial compression mechanics and glycosaminoglycan and water content for several animal species used in intervertebral disc research. To compare normalized axial mechanical properties and glycosaminoglycan and water content from other species to those of the human disc to aid in selection and interpretation of results in animal disc studies. There is a lack of mechanical and biochemical comparative data from animal intervertebral discs with respect to the human disc. Intervertebral disc axial mechanical properties, glycosaminoglycan, and water content were evaluated for 9 disc types in 7 mammalian species: the calf, pig, baboon, sheep, rabbit, rat and mouse lumbar, and the cow and rat tail. Disc area and height were used for calculation of the normalized mechanical parameters. Glycosaminoglycan content was normalized by dry weight. Many directly measured mechanical parameters varied by orders of magnitude. However, these parameters became comparable and often did not show significant differences after geometric normalization. Both glycosaminoglycan and water content revealed similarity across species. Disc axial mechanics are very similar across animal species when normalizing by the geometric parameters of disc height and area. This suggests that the disc tissue material properties are largely conserved across animal species. These results provide a reference to compare disc axial mechanics and glycosaminoglycan and water composition of experimental animal models to the human lumbar disc, to aid in both selection and interpretation of experimental disc research.

Relationship between disc injury and manual lifting: A poroelastic finite element model study

Understanding how failure originates in a lumbar motion segment subjected to loading conditions that are representative of manual lifting is important because it will pave the way for a better formulation of the exposure-injury relationship. The aim of the current investigation was to use a poroelastic finite element model of a human lumbar disc to determine its biomechanical characteristics under loading conditions that corresponded to three different, commonly occurring lifting activities and to identify the most hazardous type of loading with regard to damage to the disc. The current study showed that asymmetric lifting may increase the risk of back injury and pain. Lifting that involved lateral bending (asymmetric lifting) of the trunk was found to produce stresses at a localized area in the annulus, annuluar fibres, end plates, and facet joints that were higher than their respective tissue failure strength. Thus asymmetric lifting, if performed over a large number of cycles, might help to propagate this localized failure of the disc tissue to a larger area, owing to fatigue. The analyses also showed that largest fluid exchange between the nucleus and the end plates occurred during asymmetric lifting. If the fluid exchange is restricted owing to end plate calcification or sclerosis of the subchondral bone, high intradiscal pressure might develop, leading to higher disc bulge causing back pain.

Histological Changes and Temperature Distribution Studies of a Novel Bipolar Radiofrequency Heating System in Degenerated and Nondegenerated Human Cadaver Lumbar Discs

The purpose of this experimentation was to investigate the safety of a novel cooled bipolar radiofrequency system by examining histology and monitoring temperature distribution in the disc, epidural space, and adjacent to the nerve roots. In our study we used two human cadaver lumbar spines, one moderately to severely degenerated and the other mildly degenerated. Radiofrequency ablation of the disc posterior annulus is a theoretically plausible technique to ablate the nociceptors and to modify collagen of the annulus fibrosus. A novel cooled bipolar radiofrequency system is used to perform a procedure called intervertebral disc biacuplasty to heat the posterior annulus for the treatment of discogenic pain. Four lumbar intervertebral discs were treated in each spine sample using the bipolar system while two lumbar discs of each spine were used as controls. Temperatures developed in the posterior annulus of the disc were on average 52.35 +/- 5.07 degrees C, while in the intervertebral foramen and in the spinal canal were 38.84 +/- 1.7 degrees C and 38.29 +/- 2.04 degrees C, respectively. There was no histological evidence of damage to any other structures including vertebral end plates, epidural space, or nerve roots. Additionally, there were no histological changes in the posterior annulus that were consistent with heat-induced changes to collagen structure. Temperatures reached in the posterior annulus during transdiscal biacuplasty were greater than required (45 degrees C) for neuroablation. Temperatures reached at the neural foramina and epidural were low enough to avoid neural damage.

Characterization of an in vitro intervertebral disc organ culture system

Intervertebral disc organ culture has the capacity to control mechanical and chemical boundary conditions while keeping the tissue largely intact, and allowing interventions that would be impossible or unethical on animal studies. Recent studies on ex vivo organ culture has mostly involved small animals, or been limited to development and validation studies. In this study, bovine caudal discs were used. The large animal model design ensures that sufficient tissue is available for measurement of multiple dependent variables on the same disc, and a similar aspect ratio, diffusion distance, composition and rate of proteoglycan synthesis to human lumbar discs. The first goal of this study was to refine a set of dependent variables capable of characterizing the response of the intervertebral disc to culturing and to develop a technique to measure cell viability in all three regions of the disc. The second goal was to use these variables to compare static and diurnal loading as a method of maintaining intervertebral disc structure, composition, and cell metabolism similar to the in vivo state. Static (0.2 MPa) and diurnal loading (0.1 and 0.3 MPa alternating at 12 h intervals) were applied and intervertebral discs were examined after 4 or 8 days with dependent variables including changes in geometry (disc height and diameter), composition (tissue water content, tissue proteoglycan content and proteoglycan content lost to the culture media), cell viability and metabolism (proteoglycan synthesis). Results indicate that there was a decrease in disc height and water content after culture regardless of culture duration or loading condition. Cell viability significantly decreased with culture duration in the inner annulus and nucleus; however, a significant reduction in cell viability for the diurnal versus static loading condition was only observed after 8 days in the nucleus region. No significant differences were seen in viability of the outer annulus region with time, or in any loading groups. We conclude that our system is capable of keeping bovine caudal discs alive for at least 8 days without significant changes in GAG content, or cell metabolism, and that static loading was slightly better able to maintain cell viability than diurnal loading. This system offers promise for the future studies on large intervertebral discs requiring measurements of multiple mechanical and biological dependent variables on the same tissue.

Comparison of Animals Used in Disc Research to Human Lumbar Disc Geometry

Measurement and normalization of disc geometry parameters for several animal models used in disc research. To compare normalized values of disc geometry to the human disc geometry to aid in the selection and interpretation of animal model studies. Animal models are widely used to study intervertebral disc degeneration and to evaluate disc treatment methods because of the availability of the tissue, the decreased variability between subjects compared with humans, and the feasibility to perform in vivo experiments. There is a general lack of comparative data with respect to the human disc analog for animal models. The disc height, lateral width, AP width, area, and the nucleus pulposus lateral width, AP width, area, and centroid offset were all measured and normalized by 2 scaling factors, lateral width and disc area, for comparison to human. The species studied were ranked according to the average percent deviation of the normalized disc height, AP width and nucleus pulposus area from human geometry as: mouse lumbar (12%), rat lumbar (15%), mouse tail (18%), baboon (19%), bovine tail (22%), rabbit (26%), sheep (31%), and rat tail (46%). This paper provides a reference to compare disc geometries of experimental animal models to the human lumbar disc, to aid both in interpretation of and in planning for experimental disc research, and to provide normalized disc geometry parameters for computational models.

LATERAL BENDING AND FLEXION PLACE INTERVERTEBRAL DISC TISSUE AT GREATEST RISK OF INJURY

During certain motions, the disc is at risk of annular injury. Axial compression coupled with various combinations of excessive flexion, lateral bending or axial rotation has been shown to lead to disc injury. However, similar injuries have also been caused by repetitive activity at lower, more physiological ranges of motion. The primary objectives of this study were to determine the regions of largest shear strain experienced by disc tissues in six degrees of freedom (DOF), since shear is considered a likely tissue failure criterion, and to identify the physiological motions that may place the disc at greatest risk of injury. A grid of wires was inserted into the mid-transverse plane of nine human lumbar discs that were subjected to each of six principal displacements and rotations. Stereo-radiographs were taken in each position and digitised for reconstruction of the 3D position of each grid intersection. Maximum shear strains (MSS) were calculated from relative grid-intersection displacements and normalised by the input displacement or rotation. Physiological MSS were calculated using the maximum reported physiological lumbar segmental motion for each DOF. The largest MSS were found in the posterior, posterolateral and lateral regions of the disc. For the translation motions, lateral shear and compression produced the largest MSS (approx. 9%/mm). For the rotation motions, lateral bending had significantly larger MSS than all other tests (5.8±1.6 %/°, P The physiological MSS was greatest for lateral bending, being significantly larger than all other motions (57.8±16.2%, P This study has identified lateral bending and flexion as the lumbar segmental motions that may place the disc at greatest risk of injury. The exact failure criterion for intervertebral disc tissue is not known, and MSS was used because it is related to maximum and minimum principal strains, and it was shown that disc tears may be initiated by large interlamellar shear strains that dominate over radial and circumferential annular fibre strains. These results provide improved understanding of disc behaviours under loading and may also be of value validating finite element models.

Direct measurement of intervertebral disc maximum shear strain in six degrees of freedom: Motions that place disc tissue at risk of injury

Human intervertebral disc specimens were tested to determine the regions of largest maximum shear strain (MSS) experienced by disc tissues in each of three principal displacements and three rotations, and to identify the physiological rotations and displacements that may place the disc at greatest risk for large tissue strains and injury. Tearing of disc annulus may be initiated by large interlamellar shear strains. Nine human lumbar discs were tagged with radiographic markers on the endplates, disc periphery and with a grid of wires in the mid-transverse plane and subjected to each of the six principal displacements and rotations. Stereo-radiographs were taken in each position and digitized for reconstruction of the three-dimensional position of each marker. Maximum tissue shear strains were calculated from relative marker displacements and normalized by the input displacement or rotation. Lateral shear, compression, and lateral bending were the motions that produced the mean (95% confidence interval) largest mean MSS of 9.6 (0.7)%/mm, 9.0 (0.5)%/mm, and 5.8 (1.6)%/°, respectively, and which occurred in the posterior, posterolateral and lateral peripheral regions of the disc. After taking into account the reported maximum physiological range of motion for each degree of freedom, motions producing the highest physiological MSS were lateral bending (57.8 (16.2)%) and flexion (38.3 (3.3)%), followed by lateral shear (14.4 (1.1)%) and compression (12.6 (0.7)%).

Analysis of Tissue Distribution of TNF‐α, TNF‐α‐Receptors, and the Activating TNF‐α–Converting Enzyme Suggests Activation of the TNF‐α System in the Aging Intervertebral Disc

We immunohistochemically analyzed the expression and localization of TNF-alpha, its receptors TNF-RI and -RII, and the TNF-alpha-activating enzyme TACE in human autopsy (n=63) and surgical (n=35) lumbar intervertebral disc samples. In parallel, the TNF-alpha-mRNA was quantified by reverse transcriptase-polymerase chain reaction (RT-PCR). All samples were morphometrically evaluated for the proportion of positively labeled cells in the different anatomical regions of the disc. We detected a significant and comparable expression of all four parameters beginning in young adult age (c. 18 years) and being most extensive in the nucleus pulposus. This level was slightly reduced in older age discs. The annulus fibrosus contained significantly less labeled cells. In accord, the number of TNF-alpha-transcripts was elevated in most cases with immunohistochemical TNF-alpha expression. We provide clear evidence that TNF-alpha is expressed in discs of increasing age, which correlates with histomorphological signs of disc degeneration. In consequence, TNF-alpha seems to be activated (by the converting enzyme TACE) and biologically active through its receptors in human lumbar disc tissue.

Regional variations in the density and arrangement of elastic fibres in the anulus fibrosus of the human lumbar disc

Elastic fibres are critical components of the extracellular matrix in dynamic biological structures that undergo extension and recoil. Their presence has been demonstrated in the anulus fibrosus of the human lumbar intervertebral disc; however, a detailed regional analysis of their density and arrangement has not been undertaken, limiting our understanding of their structural and functional roles. In this investigation we have quantitatively described regional variations in elastic fibre density in the anulus fibrosus of the human L3-L4 intervertebral disc using histochemistry and light microscopy. Additionally, a multiplanar comparison of patterns of elastic fibre distribution in the intralamellar and interlamellar zones was undertaken. Novel imaging techniques were developed to facilitate the visualization of elastic fibres otherwise masked by dense surrounding matrix. Elastic fibre density was found to be significantly higher in the lamellae of the posterolateral region of the anulus than the anterolateral, and significantly higher in the outer regions than the inner, suggesting that elastic fibre density in each region of the anulus is commensurate with the magnitude of the tensile deformations experienced in bending and torsion. Elastic fibre arrangments in intralamellar and interlamellar zones were shown to be architecturally distinct, suggesting that they perform multiple functional roles within the anulus matrix structural hierarchy.

Penetration of Cephazolin in Human Lumbar Intervertebral Disc

Levels of the antibiotic cephazolin were measured in disc and blood from a consecutive series of patients undergoing lumbar spinal fusion. To determine if levels of cephazolin in the human disc reach the stated minimum inhibitory concentration against Staphylococcus aureus (1 mg/L) following intravenous administration, and to determine if there is a therapeutic relationship between serum concentration and disc concentration. Disc space infection is a potential complication of surgical procedures that violates the intervertebral disc. Bacteria, often from normal skin flora, can be introduced into the disc space causing inflammation with destruction of the adjacent vertebral endplate. Although there is substantial evidence from animal studies that prophylactic antibiotics reduce the incidence of discitis, similar evidence in human beings is not strong. A total of 30 patients, including 15 females and 15 males, with a mean age 42 years (range 21-63), received 1-g cephazolin (intravenous) during 1 or 2-level lumbar spinal fusion surgery. Venous blood was collected before administration of cephazolin and again at disc removal. Disc cephazolin concentrations were measured at the same time serum levels were measured for each patient. The interval between cephazolin administration and tissue sampling ranged from 7 to 137 minutes. Cephazolin concentration in the serum (range 31.1-148 mg/L) was higher than in the disc (range 0-9.5 mg/L). The concentration of cephazolin peaked in the serum at 7 minutes, and in the disc between 37 and 53 minutes. More than 70% of the disc samples had detectable levels of cephazolin at the time the disc was removed, although only half of these samples had cephazolin levels higher than 1 mg/L. All serum cephazolin concentrations were higher than 1 mg/L. Serum cephazolin concentration did not relate to disc concentration at a given time. The time when antibiotic concentration is highest in the disc varies among patients. More than 70% of disc samples had detectable levels of cephazolin, although only half of the discs contained cephazolin higher than the stated minimum inhibitory concentration for S. aureus(>1 mg/L). Factors such as molecular charge, degree of disc degeneration, and disc size may influence antibiotic penetration into the disc.

In Vitro Organ Culture of the Bovine Intervertebral Disc

Whole bovine coccygeal discs were cultured under static load, with or without vertebral endplates (VEPs), and assessed for cell viability, biochemical stability, biosynthetic activity, and biosynthetic responsiveness to changes in mechanical load. To assess the effects of VEPs on biochemical and cellular stability of disc cells during in vitro culture of large disc explants. To determine whether cultured discs could respond to mechanical perturbation. Previous methods for culturing the intervertebral disc have focused on rabbit and rat discs, but the small size of these discs limits the relevance of these culture systems to the human condition. Bovine coccygeal discs have similar dimensions to the human lumbar disc (i.e., similar size and nominal stresses), but long-term culture of these discs has not been reported. Bovine coccygeal discs were harvested with or without VEPs, cultured under static load (5 kg, approximately 0.25 MPa, in situ swelling pressure) for up to 1 week, and evaluated for changes in hydration, glycosaminoglycan content, cell viability, and biosynthetic activity. Additionally, the biochemical and biosynthetic response of discs cultured without VEP to increasing the load to a 20-kg (approximately 1 MPa, the estimated stress in human lumbar disc during heavy lifting) static load for 6 hours was assessed. During the first 24 hours, culturing discs with endplates was moderately better with regards to maintaining in situ anulus hydration and nucleus glycosaminoglycan levels. The endplates, however, obstructed media flow to the disc, resulting in a marked decrease in cell viability after 1 week of culture. Nucleus pulposus cell viability was maintained in discs cultured without endplates, but there was a significant drop in biosynthetic activity within 2 days of culture. Despite this drop, the disc cells in the discs without VEP remained biosynthetically responsive to changes in mechanical loading. It is possible to maintain cell viability and the biosynthetic responsiveness of large discs for up to 1 week in vitro when the discs are cultured under static load and without VEP.

Correlation of HR-MAS Spectroscopy Derived Metabolite Concentrations With Collagen and Proteoglycan Levels and Thompson Grade in the Degenerative Disc

A quantitative high-resolution magic angle spinning (HR-MAS) NMR study of human lumbar discs was conducted to determine biomarkers of disc degeneration. To correlate HR-MAS quantification of compounds relevant to human lumbar disc degeneration to conventional methods of disc grading such as Thompson grading and biochemical analysis. It has been shown that there is poor correlation between MRI and CT morphologic findings, spinal biomechanics, and patient symptoms in degenerative disc disease and low back pain. There is a need for an objective, quantitative measurement of biochemical status, morphology, and function. A total of 17 cadaveric human lumbar intervertebral discs were harvested from patients ranging from 20 to 85 years of age. Quantitative HR-MAS data were acquired, and proteoglycan and collagen biochemical analyses were conducted on 3-mm biopsy punches taken from the anulus fibrosus and nucleus pulposus of each sample. HR-MAS data were fitted and analyzed for hydroxyproline (3.42 ppm), glycine (3.56 ppm), and the N-acetyl peak (2.04 ppm) associated with proteoglycans in comparison with an internal standard. These concentrations were then compared directly to biochemical analyses and Thompson grade. HR-MAS data correlated well with Thompson grade (P < 0.001). An increase was seen in the levels of unbound hydroxyproline and glycine in annular tissue, which is directly associated to collagen breakdown. This trend also correlates with the changes of total collagen measured by a collagen biochemical assay. HR-MAS also detected a decrease in concentration of nucleus pulposus proteoglycans with degeneration. This proteoglycan decrease was verified by a standard proteoglycan biochemical assay. Changes in disc chemical composition can be accurately quantified using quantitative HR-MAS NMR spectroscopy ex vivo. This noninvasive method of qualitatively and quantitatively assessing disc degeneration supports the utility of these biomarkers and underlines the need for developing in vivo magnetic resonance spectroscopic imaging (MRSI) for characterizing intervertebral disc degeneration.

5:52 137. Mitochondrial involvement in Fas-mediated apoptosis of human lumbar disc cells

5:52 137. Mitochondrial involvement in Fas-mediated apoptosis of human lumbar disc cells

Mitochondrial Involvement in Fas-Mediated Apoptosis of Human Lumbar Disc Cells

Two main pathways of Fas-mediated apoptosis have been identified: the Type-I (death-inducing signaling complex) pathway and the Type-II (mitochondrial) pathway. While apoptotic cell death has been implicated in lumbar degenerative disc disease, we are not aware of any studies in which surgically removed discs from live humans have been examined to determine which of the two pathways is involved in the apoptosis of disc cells. As an initial step in the development of therapies to inhibit inappropriate or premature apoptosis of disc cells, our objective was to determine which pathway is involved. We examined thirty-two samples of herniated lumbar disc tissue with use of immunohistochemical staining and Western blot analysis to determine the presence of several proteins, including caspase-8 (associated with the Type-I pathway); BID (BH3 interacting domain death agonist), cytochrome-c, and caspase-9 (associated with the Type-II pathway); and caspase-3 (an executioner of apoptosis). The TUNEL (terminal deoxynucleotidyl transferase [TDT]-mediated dUTP nick end labeling) assay was performed to confirm the occurrence of apoptosis of the disc cells. The proteins associated with the Type-II pathway (BID, cytochrome-c, and caspase-9) stained positively in all samples. Although the protein associated with the Type-I pathway (caspase-8) was not detected on immunohistochemical analysis, a small amount of caspase-8 was detected on Western blot analysis. However, the expression of Type-II proteins was still higher than the expression of caspase-8 on Western blot analysis. The expression of caspase-3 was identified in all samples with immunohistochemical and Western blot analysis. TUNEL-positive disc cells were identified in all samples. The results of the present study suggest that human disc cells are Type-II cells, which undergo apoptotic cell death through mitochondrial involvement.

In vivo age- and sex-related creep of human lumbar motion segments and discs in pure centric tension

In vivo creep of human lumbar motion segments and discs subject to pure centric tension is presented, in terms of aging, sex and disc level. Time-related elongations of segments L3-4, L4-5 and L5-S1 were measured during the usual 20 min long traction hydrotherapy of patients, by using a computerized subaqual ultrasound measuring method [Kurutz et al., 2002a. Orvosi Hetilap 143 (13), 673–684; Kurutz et al., 2003. Journal of Bioengineering and Biomechanics 5 (1), 67–92]. Elongation of segments was considered as a change of the distance between two adjacent spinous processes. Based on these experiments, in vivo creep of human lumbar FSUs was investigated in centric tension, in terms of sex, age and disc level. Three-parameter rheological models were used to determine viscoelastic tensile moduli of human lumbar FSUs and discs. From three time-related measured elongation values, in vivo damping constants with creep functions were calculated for each segment, in terms of sex, aging and disc level. It has been demonstrated that initial elastic elongations decrease, concerning stiffness increase with aging. Similarly, tensile creep elongations decrease, damping properties increase with aging. Former observations concerning the difference in deformation propagation of men and women in time, have been verified by means of creep analysis: although males have higher initial elastic deformability, due to a smaller damping of females, the deformation propagation of women overtakes men in creep process. This tendency is more significant with aging. Increasing damping was observed in distal direction, both for males and females.

Feasibility of using interstitial ultrasound for intradiscal thermal therapy: a study in human cadaver lumbar discs

Application of heat in the spine using resistive wire heating devices is currently being used clinically for minimally invasive treatment of discogenic low back pain. In this study, interstitial ultrasound was evaluated for the potential to heat intradiscal tissue more precisely by directing energy towards the posterior annular wall while avoiding vertebral bodies. Two single-element directional applicator design configurations were tested: a 1.5 mm OD direct-coupled (DC) applicator which can be implanted directly within the disc, and a catheter-cooled (CC) applicator which is inserted in a 2.4 mm OD catheter with integrated water cooling and implanted within the disc. The transducers were sectored to produce 90° spatial heating patterns for directional control. Both applicator configurations were evaluated in four human cadaver lumbar disc motion segments. Two heating protocols were employed in this study in which the temperature measured 5 mm away from the applicator was controlled to either T = 52 °C, or T > 70 °C for the treatment period. These temperatures (thermal doses) are representative of those required for thermal necrosis of in-growing nociceptor nerve fibres and disc cellularity alone, or with coagulation and restructuring of annular collagen in the high-temperature case. Steady-state temperature maps, and thermal doses (t43) were used to assess the thermal treatments. Results from these studies demonstrated the capability of controlling temperature distributions within selected regions of the disc and annular wall using interstitial ultrasound, with minimal vertebral end-plate heating. While directional heating was demonstrated with both applicator designs, the CC configuration had greater directional heating capabilities and offered better temperature control than the DC configuration, particularly during the high-temperature protocol. Further, ultrasound energy was capable of penetrating within the highly attenuating disc tissue to produce more extensive radial thermal penetration, lower maximum intradiscal temperature, and shorter treatment times than can be achieved with current clinical intradiscal heating technology. Thus, interstitial ultrasound offers potential as a more precise and faster heating modality for the clinical management of low back pain.