In vitro evaluation of translating and rotating plates using a robot testing system under follower load.

BACKGROUND CONTEXT: Various modifications to standard "rigid" anterior cervical plate designs (constrained plate) have been developed that allow for some degree of axial translation and/or rotation of the plate (semi-constrained plate)-theoretically promoting proper load sharing with the graft and improved fusion rates. However, previous studies about rigid and dynamic plates have not examined the influence of simulated muscle loading. PURPOSE: The objective of this study was to compare rigid, translating, and rotating plates for single-level corpectomy procedures using a robot testing system with follower load. STUDY DESIGN: In-vitro biomechanical test. METHODS: N = 15 fresh-frozen human (C3-7) cervical specimens were biomechanically tested. The follower load was applied to the specimens at the neutral position from 0 to 100 N. Specimens were randomized into a rigid plate group, a translating plate group and a rotating plate group and then tested in flexion, extension, lateral bending and axial rotation to a pure moment target of 2.0 Nm under 100N of follower load. Range of motion, load sharing, and adjacent level effects were analyzed using a repeated measures analysis of variance (ANOVA). RESULTS: No significant differences were observed between the translating plate and the rigid plate on load sharing at neutral position and C4-6 ROM, but the translating plate was able to maintain load through the graft at a desired level during flexion. The rotating plate shared less load than rigid and translating plates in the neutral position, but cannot maintain the graft load during flexion. CONCLUSIONS: This study demonstrated that, in the presence of simulated muscle loading (follower load), the translating plate demonstrated superior performance for load sharing compared to the rigid and rotating plates.

Effects of secreted factors in culture medium of annulus fibrosus cells on microvascular endothelial cells: elucidating the possible pathomechanisms of matrix degradation and nerve in-growth in disc degeneration.

OBJECTIVE: To test whether the interaction between annulus fibrosus cells (AFCs) and endothelial cells (ECs) disrupts matrix homeostasis and stimulates production of innervation mediators. METHODS: Human microvascular ECs were cultured in the conditioned media of AF cell culture derived from degenerated human surgical specimen. Matrix-metalloproteinases (MMPs) and platelet-derived growth factor (PDGF) of ECs of this culture were analyzed by qRT-PCR, Western, and immunofluorescence. Vascular endothelial growth factor (VEGF), Interleukin-8 (IL-8), and nerve growth factor (NGF) in the media of this cell culture were assayed by ELISA. To determine the effects of ECs on AFCs, qRT-PCR was performed to determine mRNA levels of collagen I, II and aggrecan in AFCs cultured in EC conditioned media. RESULTS: Compared to ECs cultured in naïve media, ECs exposed to AFC conditioned media expressed higher mRNA and protein levels of key biomarkers of invasive EC phenotype, MMP-2 (2×), MMP-13 (4×), and PDGF-B (1.5-2×), and NGF (24.9 ± 15.2 pg/mL vs 0 in naïve media). Treatment of AF cells with EC culture conditioned media decreased collagen type II expression two fold. Considerable quantities of pro-angiogenic factors IL-8 (396.7 ± 302.0 pg/mL) and VEGF (756.2 ± 375.9 pg/mL) were also detected in the conditioned media of untreated AF cell culture. DISCUSSION: AFCs from degenerated discs secreted factors which stimulated EC production of factors known to induce matrix degradation, angiogenesis, and innervation. IL-8 and VEGF maybe the secreted factors from AFCs which mediate a pro-angiogenic stimulus often implicated in the development of disc degeneration.

Skeletal muscle-secreted myokine interleukin-6 induces white adipose tissue conversion into beige adipose tissue in myostatin gene knockout pigs.

Myostatin (MSTN) is primarily expressed in skeletal muscle and plays an important role in the regulation of muscle growth and development as well as fat deposition; however, little is known about the molecular mechanism through which MSTN regulates body fat deposition. Therefore, in this study, we sought to identify the signaling pathways through which MSTN regulates fat accumulation in pigs. MSTN knockout (MSTN-/-) pigs showed increased muscle mass, decreased fat mass, and a leaner body composition. In this study, we found that the adipose tissue of MSTN-/- pigs exhibits the characteristics of beige adipose tissue, and the mRNA expression levels of beige adipose marker genes, including UCP3, Cidea, and CD137, were significantly increased. Remarkably, the observed beige phenotype was not adipocyte autonomous but rather caused by muscle-secreted myokine interleukin (IL)-6. This occurrence results in increased AMP-activated protein kinase (AMPK) phosphorylation in adipose tissue, which subsequently activates peroxisome proliferator-activated receptor gamma coactivator 1α and the conversion of white adipocytes to beige in pigs. Therefore, we concluded that MSTN deficiency leads to increased IL-6 secretion in skeletal muscle and activates AMPK in adipocytes, thereby increasing the beige adipose tissue in MSTN-/- pigs.

p38 MAPK inhibition selectively mitigates inflammatory mediators and VEGF production in AF cells co-cultured with activated macrophage-like THP-1 cells.

OBJECTIVES: Recent data have suggested that macrophages are involved in the pathogenesis of discogenic back pain and enhance the secretion of inflammatory mediators in co-cultured annulus fibrosus (AF) cells. The purpose of these studies is to determine the role of p38 mitogen-activated protein kinase (p38 MAPK) signaling in the interactions between macrophage and AF cells. METHODS: Human AF cells were co-cultured with phorbol myristate acetate-stimulated macrophage-like THP-1 cells with and without p38 MAPK inhibition. Conditioned media from co-cultured cells were assayed for interleukin (IL)-6, IL-8, prostaglandin E2 (PGE2), PGF2alpha, and vascular endothelial growth factor (VEGF). Naïve and macrophage-exposed AF cell responses to 10ng/ml tumor necrosis factor-alpha (TNF-alpha) were compared using the same outcome measures. RESULTS: IL-6, IL-8, PGE2, PGF2alpha, and VEGF were secreted in greater quantities by cells maintained in co-culture compared to macrophages or AF cells cultured alone. SB202190 blunted IL-6, PGE2, and PGF2alpha production in a dose-dependent manner in co-culture. However, it did not suppress IL-8 and VEGF production. TNF-alpha-stimulated AF cell inflammatory mediators were up-regulated by macrophage exposure. SB202190 successfully suppressed IL-6, IL-8, PGE2, and PGF2alpha secretion in macrophage-exposed AF cells in response to TNF-alpha. CONCLUSIONS: Annular injury can result in macrophage infiltration, and this can cause enhanced inflammatory mediator and VEGF production by AF cells. The p38 MAPK pathway signals are responsible for much of IL-6 and PG secretion from AF cells with macrophage-like cells, suggesting that blockade of this signal may serve as a therapeutic approach to discogenic pain.

Histone deacetylase inhibitor M344 significantly improves nuclear reprogramming, blastocyst quality, and in vitro developmental capacity of cloned pig embryos.

M344 is a novel histone deacetylase inhibitor. There is no report on the effect of M344 treatment on the development of pig embryos after somatic cell nuclear transfer (SCNT). In the present study, we investigated the effect of M344 on the blastocyst formation rate in cloned embryos, acetylation level of histone H4 lysine 12 (AcH4K12), and the expression of pluripotency-related genes , , and . Our results indicated that treatment with 5 µ M344 for 6 h improved the development of porcine embryos, in comparison with the untreated group (25.1% ± 5.0 vs. 10.9% ± 2.4; < 0.05). Moreover, M344-treated embryos had increased average fluorescence intensity of AcH4K12 at the pseudo-pronuclear stage ( < 0.05). However, no differences exist in Oct4, NANOG, and SOX2 expression in M344-treated and untreated SCNT blastocysts. In evaluating the effect of M344 on in vivo development, 845 M344-treated embryos were transferred into 3 surrogates, 1 of whom became pregnant and developed 3 fetuses. These findings suggested that M344 elevated the level of histone acetylation, facilitated the nuclear programming, and subsequently improved the developmental competence of pig SCNT embryos.

Cervical intrathecal catheter placement for cerebrospinal fluid drainage: technical case report.

OBJECTIVE AND IMPORTANCE: The purpose of this report is to describe the placement and use of a cervical subarachnoid catheter for cerebrospinal fluid diversion. This technique provides an important alternative drainage system for patients whose clinical situations preclude lumbar spinal fluid diversion. CLINICAL PRESENTATION: Two patients were involved in accidents that resulted in traumatic dural tears. Both patients required thoracolumbar spinal reconstruction with instrumentation. Cerebrospinal fistulae developed, which were refractory to surgical reexploration. TECHNIQUE: A commercially available catheter was successfully placed in the cervical subarachnoid space under fluoroscopic guidance using a C1-C2 approach in both patients. Spinal fluid drainage was maintained for 5 days via this route, and this proved effective in resolving the cerebrospinal fluid fistula. No complications were observed with the placement or maintenance of the catheter. CONCLUSION: Cervical spinal fluid drainage is a feasible alternative for patients in whom lumbar access cannot be obtained or is contraindicated.

Sagittal measurements of the cervical spine in subaxial fractures and dislocations. An analysis of two hundred and eighty-eight patients with and without neurological deficits.

We analyzed three factors involved in fractures and dislocations of the cervical spine and their relation to the degree of injury of the spinal cord. The three factors were the space available for the spinal cord at the level of the injury, the sagittal diameter of the spinal canal at the uninjured levels, and the Pavlov ratio at the uninjured levels. Of the 288 patients analyzed, eighty-three had a complete injury of the spinal cord, ninety-two had an incomplete injury of the spinal cord, thirty had an isolated nerve-root injury, and eighty-three had no neurological deficit. The mean space available for the spinal cord at the level of the injury was 10.5 millimeters for the patients who had a complete injury of the spinal cord, 13.1 millimeters for those who had an incomplete injury of the spinal cord, 15.9 millimeters for those who had an isolated nerve-root injury, and 16.7 millimeters for those who had no neurological deficit. The difference between the groups was significant (p < 0.001) except for the difference between the patients who had an isolated nerve-root injury and those who had no neurological deficit. The mean sagittal diameter of the canal at the uninjured levels was 16.1 millimeters for the patients who had a complete injury of the spinal cord, 16.1 millimeters for those who had an incomplete injury of the spinal cord, 17.9 millimeters for those who had an isolated nerve-root injury, and 18.1 millimeters for those who had no neurological deficit. The difference between the patients who had a complete injury of the spinal cord and those who had an incomplete injury of the spinal cord and that between the patients who had an isolated nerve-root injury and those who had no neurological deficit were not significant (p > 0.05). However, the patients who had a complete injury of the spinal cord and those who had an incomplete injury of the spinal cord were significantly different from the patients who had an isolated nerve-root injury and those who had no neurological deficit (p < 0.001). The mean Pavlov ratio at the uninjured levels was 0.82 for the patients who had a complete injury of the spinal cord, 0.84 for those who had an incomplete injury of the spinal cord, 0.96 for those who had an isolated nerve-root injury, and 0.96 for those who had no neurological deficit.(ABSTRACT TRUNCATED AT 400 WORDS)

Ingrowth and formation of bone in defects in an uncemented fiber-metal total hip-replacement model in dogs.

An osseous defect was created in the acetabulum and femur of twenty dogs, and then a fiber-metal total hip prosthesis was inserted. A comparison was made between the effects of leaving the defects unfilled, filling the defects with an autogenous bone graft, and filling them with a 50:50 mixture of autogenous bone graft and a biphasic ceramic composed of hydroxyapatite and tricalcium phosphate. The characteristics of formation of bone within the sites of the defects and the extent of the ingrowth of bone into the underlying porous surface were analyzed. At six and twelve weeks after implantation, the dogs in the control group (unfilled defects) had the least amount of bone in the sites of the defects. In the dogs that had had an autogenous bone graft and those in which the defects had been filled with a mixture of autogenous bone graft and biphasic ceramic, there were excellent osteoconductive properties in the filling of the sites of the defects with new bone. A comparison was made between the amount of bone that formed in the osseous defects and the amount that formed in the spaces of the porous-surfaced fiber-metal components of the prostheses, directly underlying and adjacent to the defects. At six weeks, the greatest amount of ingrowth of bone into the spaces of the underlying titanium fiber-metal acetabular components was seen in the control group (22 per cent of the porous surface), followed by the group in which the defects had been filled with a mixture of autogenous bone graft and biphasic ceramic (19 per cent). The defects that had been filled only with autogenous bone graft had poor ingrowth of bone into the porous surface (4 per cent) and predominantly fibrous ingrowth. At twelve weeks, these defects again showed the least amount of ingrowth of bone (15 per cent) compared with the defects in the control group (24 per cent) and those that had been filled with a mixture of autogenous bone graft and biphasic ceramic (24 per cent). No statistically significant differences were found in the amount of ingrowth of bone into the defects that had been created in the femur in the three groups of dogs at either six or twelve weeks. However, the trends appeared to correlate with the data for the acetabular defects.

Cervical disc herniation in a patient with congenital insensitivity to pain: a case report.

STUDY DESIGN: A case report of a patient with a known diagnosis of congenital insensitivity to pain who developed a herniated cervical disc. OBJECTIVES: To study the clinical manifestations of cervical radiculopathy in a patient with congenital insensitivity to pain and the long-term outcome after surgical treatment. SUMMARY OF BACKGROUND DATA: There have been no reports in the English literature documenting such a patient. METHODS: Retrospective case report and long-term clinical and radiographic follow-up. RESULTS: This patient with a known diagnosis of congenital insensitivity to pain had neurologic motor weakness with "neck and shoulder pain." Clear radicular pattern could not be elicited. The patient underwent a successful anterior discectomy and fusion with long-term clinical and radiographic results. CONCLUSION: Patients with congenital insensitivity to pain who develop a cervical disc herniation may present with atypical symptoms not manifesting in the classic radicular pattern. Higher index of suspicion by the clinician must be practiced to make the appropriate diagnosis. Successful surgical outcome may be achieved in these patients.

Toward a biochemical understanding of human intervertebral disc degeneration and herniation. Contributions of nitric oxide, interleukins, prostaglandin E2, and matrix metalloproteinases.

STUDY DESIGN: Normal and herniated human intervertebral disc specimens were cultured to study the effects of interleukin-1 beta on the production of nitric oxide, interleukin-6, prostaglandin E2, and matrix metalloproteinases. The effects of endogenously produced nitric oxide on the synthesis of other mediators also were studied. OBJECTIVES: To test the hypothesis that the cells of the intervertebral disc are metabolically active and are capable of responding to biochemical stimuli such as interleukin-1 beta in a manner that could engender degenerative changes. As part of this study, the authors also investigated some of the possible autocrine regulatory mechanisms that may operate during the biochemical responses of disc cells. SUMMARY OF BACKGROUND DATA: The authors previously showed, for the first time, that herniated cervical and lumbar disc specimens spontaneously produce increased amounts of nitric oxide, interleukin-6, prostaglandin E2, and certain matrix metalloproteinases. These results suggest that these biochemical agents are in some manner involved with degenerative processes in the intervertebral disc. This novel hypothesis merits further evaluation; the current communication reports the results of experiments designed to do so. METHODS: Fourteen normal, nondegenerated discs (control group) were obtained from seven patients undergoing anterior spinal surgery for trauma or lumbar scoliosis. Thirty-six herniated discs (18 lumbar and 18 cervical) were obtained from 30 patients undergoing surgery for persistent radiculopathy. The specimens were placed into tissue culture and incubated for 72 hours in the presence or absence of interleukin-1 beta and NG-monomethyl-L-arginine, and inhibitor of nitric oxide synthases, and the media were subsequently collected for biochemical analysis. Biochemical assays for matrix metalloproteinases, nitric oxide, interleukin-6, and prostaglandin E2 were performed. RESULTS: Normal, control disc specimens significantly increased their production of matrix metalloproteinases, nitric oxide, interleukin-6, and prostaglandin E2 in response to interleukin-1 beta. Herniated lumbar and cervical discs, which were spontaneously releasing increased levels of these biochemical agents, further increased their production of nitric oxide, interleukin-6, and prostaglandin E2 in response to interleukin-1 beta. Blocking the biosynthesis of nitric oxide in interleukin-1 beta-stimulated disc cells provoked a large increase in the production of interleukin-6. CONCLUSIONS: Cells of the intervertebral discs are biologically responsive and increase their production of matrix metalloproteinases, nitric oxide, interleukin-6, and prostaglandin E2 when stimulated by interleukin-1 beta. The effect is more dramatic in normal, nondegenerated discs where spontaneous synthesis of these mediators is low. Nevertheless, cells of the herniated degenerated discs where spontaneous production was high were still capable of further increasing their synthesis of several of these biochemical agents in response to interleukin-1 beta. Endogenously produced nitric oxide appears to have a strong inhibitory effect on the production of interleukin-6, which suggests that autocrine mechanisms play an important role in the regulation of disc cell metabolism.

Herniated cervical intervertebral discs spontaneously produce matrix metalloproteinases, nitric oxide, interleukin-6, and prostaglandin E2.

STUDY DESIGN: Herniated cervical disc specimens were obtained from patients undergoing surgical discectomy for persistent radiculopathy and cultured in vitro to determine whether various biochemical agents were being produced. OBJECTIVES: Our hypothesis is that biochemical mediators of inflammation and tissue degradation play a role in cervical intervertebral disc degeneration and in the pathophysiology of cervical radiculopathy. SUMMARY OF BACKGROUND DATA: Neck pain with or without radiculopathy is a common clinical problem, but the etiology of neck pain and the exact pathophysiology of radiculopathy remain uncertain. We have previously reported the production of various biochemical agents by herniated lumbar disc specimens in vitro. Because of a lack of such studies in the literature with respect to the cervical spine, the purpose of this study was to determine whether similar biochemical agents of inflammation and tissue degradation were being produced by herniated cervical disc specimens. METHODS: Eighteen herniated cervical discs were obtained from 15 patients undergoing anterior disc surgery. The specimens were cultured and incubated for 72 hours, and the media were subsequently collected for biochemical analysis. Biochemical assays for matrix metalloproteinases, nitric oxide, prostaglandin E2, and a variety of cytokines were performed. As a control group, six cervical discs specimens were obtained from three patients undergoing anterior surgery for traumatic burst fractures, and similar biochemical analyses were performed. RESULTS: The culture media from the herniated cervical disc specimens showed increased levels of matrix metalloproteinase activity compared with the control discs. Similarly, the levels of nitric oxide, prostaglandin E2, and interleukin-6 were significantly higher in the herniated disc specimens compared with the control discs. Interleukin-1 alpha, interleukin-1 beta, tumor necrosis factor-alpha, interleukin-1 receptor antagonist protein, and substance P were not detected in the culture media of the herniated or control discs. CONCLUSIONS: Herniated cervical disc specimens were making spontaneously increased amounts of matrix metalloproteinases, nitric oxide, prostaglandin E2, and interleukin-6. These results were similar to those obtained in herniated lumbar disc specimens that we have previously reported. These products may be intimately involved in the biochemistry of disc degeneration and the pathophysiology of radiculopathy.

Human intervertebral disc cells are genetically modifiable by adenovirus-mediated gene transfer: implications for the clinical management of intervertebral disc disorders.

STUDY DESIGN: Human intervertebral disc cells were cultured in monolayer and treated with adenovirus-containing marker genes to determine the susceptibility of the cells to adenovirus-mediated gene transfer. OBJECTIVES: To test the efficacy of the adenovirus-mediated gene transfer technique for transferring exogenous genes to human intervertebral disc cells in vitro. SUMMARY OF BACKGROUND DATA: Upregulated proteoglycan synthesis after direct in vivo adenovirus-mediated transfer of growth factor genes to the rabbit intervertebral disc has previously been reported. Before contemplating extending this approach to the treatment of human disc disease, it is necessary to demonstrate that human intervertebral disc cells are indeed susceptible to adenovirus-mediated gene transduction. METHODS: Human intervertebral disc cells were isolated from disc tissue obtained from 15 patients during surgical disc procedures. The cells were cultured in monolayer and treated with saline containing five different doses of adenovirus carrying the lacZ gene (Ad/CMV-lacZ), saline containing adenovirus carrying the luciferase gene (Ad/CMV-luciferase), or saline alone. Transgene expression was analyzed by 5-bromo-4-chloro-3-indolyl-beta-galactosidase (X-Gal) staining and luciferase assay. RESULTS: Adenovirus efficiently transferred lacZ and luciferase marker genes to cells from degenerated discs as well as to cells from nondegenerated discs. A minimum dose of 150 MOI Ad/CMV-lacZ was found to be sufficient to achieve transduction of approximately 100% of disc cells-regardless of patient age, sex, surgical indication, disc level, and degeneration grade. No statistically significant difference in the luciferase activities could be detected in disc cell cultures from degenerated and nondegenerated discs treated with Ad/CMV-luciferase. CONCLUSIONS: In vitro transducibility of human intervertebral disc cells by adenovirus is relatively insensitive to disc degeneration grade. Because the rate-limiting step for successful gene therapy is the ability to transfer genes efficiently to the target tissue, the achievement of efficient gene transfer to human intervertebral disc cells(using a direct, adenovirus-mediated approach) is an important and necessary step in the development of gene therapy strategies for the management of human intervertebral disc disorders.

Herniated lumbar intervertebral discs spontaneously produce matrix metalloproteinases, nitric oxide, interleukin-6, and prostaglandin E2.

STUDY DESIGN: Herniated lumbar disc specimens were obtained from patients undergoing surgical discectomy for persistent radiculopathy and cultured in vitro to determine whether various biochemical agents were being produced. OBJECTIVES: Our hypothesis is that biochemical mediators of inflammation and tissue degradation play a role in intervertebral disc degeneration and in the pathophysiology of radiculopathy. SUMMARY OF BACKGROUND DATA: Low back pain with or without radiculopathy is a significant clinical problem, but the etiology of low back pain and the exact pathophysiology of radiculopathy remain elusive. The biochemical events that occur with intervertebral disc degeneration and, in particular, the role of biochemical mediators of inflammation and tissue degradation have received sparse attention in the literature. There is some preliminary evidence that inflammatory mediators may have an important role in the pathophysiology of radiculopathy. METHODS: Eighteen herniated lumbar discs were obtained from 15 patients undergoing disc surgery. The specimens were cultured and incubated for 72 hours, and the media were collected subsequently for biochemical analysis. Biochemical assays for matrix metalloproteinases, nitric oxide, prostaglandin E2, and a variety of cytokines were performed. As a control group, eight lumbar disc specimens were obtained from four patients undergoing anterior surgery for scoliosis and traumatic burst fractures, and similar biochemical analyses were performed. RESULTS: The culture media from the herniated lumbar discs showed increased levels of matrix metalloproteinase activity compared with the control discs. Similarly, the levels of nitric oxide, prostaglandin E2, and interleukin-6 were significantly higher in the herniated discs compared with the control discs. Interleukin 1 alpha, interleukin-1 beta, tumor necrosis factor-alpha, interleukin-1 receptor antagonist protein, and substance P were not detected in the culture media of either the herniated or control discs. CONCLUSIONS: Herniated lumbar discs were making spontaneously increased amounts of matrix metalloproteinases, nitric oxide, prostaglandin E2, and interleukin-6. These products may be involved intimately in the biochemistry of disc degeneration and the pathophysiology of radiculopathy. Their exact roles certainly need further investigation, but their mere presence implicates biochemical processes in intervertebral disc degeneration.

Modulation of the biologic activity of the rabbit intervertebral disc by gene therapy: an in vivo study of adenovirus-mediated transfer of the human transforming growth factor beta 1 encoding gene.

STUDY DESIGN: In vivo studies using a rabbit model to determine the biologic effects of direct, adenovirus-mediated transfer of a therapeutic gene to the intervertebral disc. OBJECTIVES: 1) To deliver an exogenous therapeutic gene to rabbit lumbar intervertebral discs in vivo, 2) to quantify the resulting amount of gene expression, and 3) to determine the effect on the biologic activity of the discs. SUMMARY OF BACKGROUND DATA: Although growth factors such as transforming growth factor beta 1 appear to have promising therapeutic properties, there currently is no practical method for sustained delivery of exogenous growth factors to the disc for the management of certain chronic types of disease (e.g., disc degeneration). A possible solution is to modify the disc cells genetically through gene transfer such that the cells manufacture the desired growth factors endogenously on a continuous basis. METHODS: Saline, with or without virus, was injected directly into lumbar discs of 22 skeletally mature female New Zealand white rabbits. Group 1 (n = 11) received the adenovirus construct Ad/CMV-hTGF beta 1 containing the therapeutic human transforming growth factor beta 1-encoding gene. Group 2 (n = 6) received adenovirus containing the luciferase marker gene. Group 3 (n = 5) received saline only. The rabbits were killed 1 week after injection. Immunohistochemical staining for human transforming growth factor beta 1 was performed on the disc tissues of one rabbit from Group 1. Nucleus pulposus tissues from the remaining rabbits were cultured in serumless medium. Bioassays were performed to determine human transforming growth factor beta 1 production and proteoglycan synthesis. RESULTS: Discs injected with Ad/CMV-hTGF beta 1 exhibited extensive and intense positive immunostaining for transforming growth factor beta 1. The nucleus pulposus tissues from the discs injected with Ad/CMV-hTGF beta 1 exhibited a 30-fold increase in active transforming growth factor beta 1 production, and a 5-fold increase in total (active + latent) transforming growth factor beta 1 production over that from intact control discs (P < 0.05). Furthermore, these tissues exhibited a 100% increase in proteoglycan synthesis compared with intact control tissue, which was statistically significant (P < 0.05). CONCLUSIONS: The results of this study suggest that the intervertebral disc is an appropriate site for adenovirus-mediated transfer of exogenous genes and subsequent production of therapeutic growth factors. Gene therapy therefore may have useful applications for study of the basic science of the intervertebral disc and for clinical management of degenerative disc disease.

Degenerative lumbar listhesis and bone mineral density in elderly women. The study of osteoporotic fractures.

STUDY DESIGN: A cross-sectional and prospective study. OBJECTIVES: To investigate the association between lumbar listhesis in elderly white women and bone mineral density at the spine, hip, radius, and calcaneus. SUMMARY OF BACKGROUND DATA: Several types of degenerative spinal changes have been found to be associated with high bone mineral density at the spine and other body sites. METHODS: Lateral radiographs of the lumbar spine for 1400 elderly women enrolled in the Study of Osteoporotic Fractures were digitized. Listhesis (antero and retro) was assessed at L3-L4, L4-L5, and L5-S1. Bone mineral density was measured at the spine, hip, calcaneus, and the distal and proximal radius. RESULTS: After adjusting the data for age and body mass index, retrolisthesis at L3-L4, L4-L5, and L5-S1 was associated with mean spinal bone mineral density levels that were 9% to 13% higher compared with those levels in women with no listhesis (P < 0.0001). In addition, bone mineral density at the hip and appendicular sites increased from 4% to 9%. The mean lumbar spinal bone mineral density of women with anterolisthesis at L3-L4 was 12% higher (P < 0.05) than that of women with no listhesis; it was the same for both groups at L4-L5 and was 7% lower (P < 0.005) at L5-S1. At L5-S1 the bone mineral density level at the hip and appendicular sites was also lower among the women with anterolisthesis at that level. CONCLUSIONS: This study suggests that retrolisthesis, like other spinal degenerative diseases, is associated with increased spinal bone mineral density. Anterolisthesis, however, may involve a different etiology, because its association with bone mineral density varies by spinal level.

Adenovirus-mediated gene transfer to nucleus pulposus cells. Implications for the treatment of intervertebral disc degeneration.

STUDY DESIGN: In vitro and in vivo studies using a rabbit model were performed to determine the feasibility of adenovirus-mediated gene transfer to the intervertebral disc. OBJECTIVES: This study was conducted to determine whether it is possible to transfer genes to cells within the intervertebral disc by direct injection of an adenovirus and to determine the duration of gene expression obtained by this method. SUMMARY OF BACKGROUND DATA: Although growth factors have the potential to stimulate the regeneration of nucleus pulposus, sustained delivery of growth factors to a degenerated disc is clinically unfeasible with present technology. Novel approaches such as gene transfer should be investigated as possible solutions to this problem. METHODS: The lacZ marker gene was used to evaluate gene delivery to cells within intervertebral discs. For the in vitro study, cell cultures were established from the nucleus pulposus tissue of New Zealand white rabbits and infected with an adenovirus encoding the lacZ gene (Ad-lacZ). For the in vivo study, the anterior aspects of lumbar intervertebral discs were surgically exposed, and Ad-lacZ in saline solution was directly injected into the nucleus pulposus. An equal volume of saline only was injected into control discs. Expression of the transferred gene was detected by staining with 5-bromo-4-chloro-3-indolyl-beta-galactosidase (X-Gal). RESULTS: The in vitro experiments confirmed that nucleus pulposus cells were efficiently transduced by an adenoviral vector carrying the lacZ gene. In vivo injection of Ad-lacZ into the nucleus pulposus resulted in the transduction of a considerable number of cells. Marker gene expression in vivo persisted at an apparently undiminished level for at least 12 weeks. No staining was noted in control discs. CONCLUSIONS: The results show the feasibility of adenovirus-mediated gene transfer to the intervertebral disc. Expression of the marker gene persisted at least 12 weeks in vivo. This successful demonstration of exogenous gene transfer to the disc and sustained, long-term expression suggests that the adenoviral vector may be suitable for delivery of appropriate genes to the disc for the treatment of spinal disorders.

The biochemical and histological effects of artificial ligament wear particles: in vitro and in vivo studies.

Growing evidence suggests that biochemical mechanisms play a role in the pathogenesis of arthritis. Cartilaginous wear particles have been shown to induce destructive enzymes and cytokines. To assess the biocompatibility of artificial ACL replacements, the effects of wear particles from the following ligaments were analyzed biochemically and histologically: GORETEX, Stryker Dacron Ligament Prosthesis, Versigraft carbon, Kennedy LAD, Xenograft, Leeds-Keio, and human patellar tendon allograft. Ligaments were frozen and ground to produce wear particles similar to those seen clinically and were added to lapine synovial cell cultures. The resulting conditioned medium was analyzed for collagenase, gelatinase, and chondrocyte activating factor (CAF) production. All of the ligaments induced significantly elevated enzyme and CAF production by the synoviocytes, with Xenograft and carbon inducing significantly higher enzyme levels than those of the other five ligaments. Five milligrams of wear particles were injected into the knees of 4 kg to 5 kg rabbits that were analyzed histologically after 14 weeks. Wear particles accumulated in the periarticular synovial folds and induced modest to severe macrophage infiltration in the synovium. A hypothetical model explaining the role of artificial ligament wear particles in the pathogenesis of arthritis is presented.

Biochemistry of intervertebral disc degeneration and the potential for gene therapy applications.

BACKGROUND CONTEXT: Low back pain continues to be a major cause of morbidity in the United States and the world. Although the exact cause has yet to be defined, the intervertebral disk and its age-related changes have been most frequently implicated. PURPOSE: This article represents a brief summary of intervertebral disk structure and function, both in the "normal" and degenerative states. STUDY DESIGN/SETTING: Review article. A Medline search from 1966 to present was performed to identify pertinent articles related to the topic of the intervertebral disc and degeneration. METHODS: This review article describes the pertinent anatomy, as well as the biochemical and biomechanical changes that occur in the intervertebral disc over time. It presents many of the current theories implicated as causing these changes. RESULTS: Recent studies have shown that gene therapy (the transfer of therapeutic gene[s] into a cell), may have promise as a method of slowing down, or preventing some of the changes seen in the intervertebral disc. CONCLUSION: Intervertebral disc degeneration is a complex phenomenon, likely the result of a combination of biochemical and biomechanical factors that are known to occur in the disk. Ongoing research efforts in the area of gene therapy show promise as a way to prevent, or even reverse, some of these changes.

Thoracic and lumbar fusions for degenerative disorders: rationale for selecting the appropriate fusion techniques.

This article defines the indications for spinal fusion surgery based on the current literature as well as a rationale for selecting the appropriate spinal fusion techniques for the more common degenerative lumbar and thoracic conditions.

Analysis of power law models for the creep of nucleus pulposus tissue.

Tissue engineering approaches are now being investigated for altering the course of intervertebral disc degeneration (IDD). Because the disease changes the mechanical properties of the load bearing tissues of the disc, viscoelastic tissue behavior is a key measure for comparing the efficacy of treatments. To investigate the basic viscoelastic behavior of nucleus pulposus tissue, tissue from the rabbit disc was tested in torsional creep. Both the Andrade and Nutting creep models had a good fit to the data, however, the Andrade creep model gave a much better prediction of the longer term creep. This is the first application of Andrade creep to biological tissue and results indicate that this model may be particularly well suited for characterizing the viscoelastic behavior of very soft biological tissues.