Positive association between serum thymic stromal lymphopoietin and anti-citrullinated peptide antibodies in patients with rheumatoid arthritis.

Thymic stromal lymphopoietin (TSLP) has been suggested recently to play an important role in the pathophysiology of rheumatoid arthritis (RA). However, there is little information on serum TSLP concentrations in RA and its clinical significance. The present study investigated whether serum TSLP concentrations were affected in patients with RA. Using an enzyme-linked immunosorbent assay (ELISA), we measured TSLP concentrations in the serum obtained from 100 patients with RA, 60 patients with osteoarthritis (OA) and 34 healthy volunteers. We also investigated the correlation between serum TSLP concentrations and clinical parameters of disease activity in RA [disease activity score using 28 joint counts (DAS28)-C-reactive protein (CRP), DAS28-erythrocyte sedimentation rate (ESR), Clinical Disease Activity Index (CDAI]), patient's/-physician's Visual Analogue Scale (VAS), swollen joints count, tender joints count, CRP, ESR and matrix metalloproteinase-3 (MMP-3) concentrations]. In addition, we investigated the correlation between serum TSLP concentrations and anti-citrullinated peptide antibody (ACPA) and serum tumour necrosis factor (TNF)-α. Serum TSLP levels in patients with RA were significantly higher than those in patients with OA and in healthy volunteers. Interestingly, serum TSLP concentrations were correlated significantly with ACPA titres, but not with other clinical parameters. There was a significant increase in serum TSLP concentrations in patients with RA, which was correlated positively with serum ACPA titres. These findings suggest that in patients with RA, TSLP may play a role in ACPA production by B cells.

Do screws and screw holes affect osteolysis in cementless cups using highly crosslinked polyethylene? A 7 to 10-year follow-up case-control study.

BACKGROUND: The use of screws and the presence of screw holes may cause acetabular osteolysis and implant loosening in cementless total hip arthroplasty (THA) using conventional polyethylene. In contrast, this issue is not fully understood using highly crosslinked polyethylene (HXLPE), particularly in large comparative study. Therefore, we performed a case-control study to assess the influence of screw usage and screw holes on: (1) implant fixation and osteolysis and (2) polyethylene steady-state wear rate, using cases with HXLPE liners followed up for 7-10 years postoperatively. HYPOTHESIS: The screw usage and screw holes adversely affect the implant fixation and incidence of wear-related osteolysis in THA with HXLPE. PATIENTS AND METHODS: We reviewed 209 primary cementless THAs performed with 26-mm cobalt-chromium heads on HXLPE liners. To compare the effects of the use of screws and the presence of screw holes, the following groups were established: (1) with-screw (n=140); (2) without-screw (n=69); (3) no-hole (n=27) and (4) group in which a cup with screw holes, but no screw was used (n=42). Two adjunct groups (no-hole cups excluded) were established to compare the differences in the two types of HXLPE: (5) remelted group (n=100) and (6) annealed group (n=82). Implant stability and osteolysis were evaluated by plain radiography and computed tomography. The wear rate from 1 year to the final evaluation was measured using plain X-rays and PolyWare Digital software. RESULTS: All cups and stems achieved bony fixation. On CT-scan, no acetabular osteolysis was found, but there were 3 cases with a small area of femoral osteolysis. The mean steady-state wear rate of each group was (1) 0.031±0.022, (2) 0.033±0.035, (3) 0.031±0.024, (4) 0.029±0.018, (5) 0.030±0.018 and (6) 0.034±0.023mm/year, respectively. A comparison of the effects of screw usage or screw holes found no significant between-group differences in the implant stability, prevalence of osteolysis [no acetabular osteolysis and 3/209 at femoral side (1.4%)] and steady-state wear rate. DISCUSSION: This study suggests that there are no adverse effects on the results of THA with HXLPE from the use of cups with screw holes and the use of screws for cup fixation. LEVEL OF EVIDENCE: Level III retrospective case-control study.

Matrix metalloproteinase-3-dependent generation of a macrophage chemoattractant in a model of herniated disc resorption.

Herniated disc (HD) is a common health problem that is resolved by surgery unless spontaneous resorption occurs. HD tissue contains abundant macrophage infiltration and high levels of matrix metalloproteinases (MMPs) MMP-3 and MMP-7. We developed a model system in which disc tissue or isolated chondrocytes from wild-type or MMP-null mice were cocultured with peritoneal macrophages and used this system to investigate the role of MMPs and chondrocyte/macrophage interactions in disc resorption. We observed a marked enhancement of MMP-3 protein and mRNA in chondrocytes after exposure to macrophages. Chondrocytic MMP-3, but not MMP-7, was required for disc resorption, as determined by assaying for a reduction in wet weight and proteoglycan content after 3 days of coculture. Surprisingly, chondrocyte MMP-3 was required for the generation of a macrophage chemoattractant and the subsequent infiltration of the disc tissue by proteolytically active macrophages. We conclude that macrophage induction of chondrocyte MMP-3 plays a major role in disc resorption by mechanisms that include the generation of a bioactive macrophage chemoattractant.

Matrix metalloproteinase-7-dependent release of tumor necrosis factor-alpha in a model of herniated disc resorption.

Herniated disc (HD), one of the major causes of low back pain, is often resolved spontaneously without surgical intervention. Resorption is associated with a marked increase in infiltrating macrophages, and the matrix metalloproteinases (MMP) MMP-3 and MMP-7 have been implicated in this phenomenon. We developed a murine organ culture model in which intact intervertebral discs were cocultured with peritoneal macrophages to investigate the role of MMPs in HD resorption. Using macrophages isolated from MMP-null mice, we report that macrophage-produced MMP-7 was required for proteoglycan degradation, loss of wet weight, and macrophage infiltration of cocultured discs. The inability of MMP-7-deficient macrophages to infiltrate discs could not be attributed to a defect in macrophage migration. MMP-7 was required for the release of the cytokine TNF-alpha from peritoneal macrophages. The generation of soluble TNF-alpha was essential for the induction of MMP-3 in disc cocultures, which in turn is required for the generation of a macrophage chemoattractant and subsequent macrophage infiltration. TNF-alpha release from macrophages was necessary but insufficient for disc resorption, which required macrophage infiltration. We conclude that there is extensive communication between macrophages and chondrocytes in HD resorption and that an essential component of this communication is the requirement for MMPs to release soluble bioactive factors.

Hematometra-hematocolpos: a late complication of successful separation of conjoined twins.

A late complication of separation of conjoined twins, hematometra-hematocolpos, may appear with sexual maturation. An obstructed genital outflow tract can cause significant urologic and reproductive tract morbidity.

Sequential dynamics of monocyte chemotactic protein-1 expression in herniated nucleus pulposus resorption.

We previously demonstrated that the granulation tissues of herniated nucleus pulposus are composed of a marked infiltration of macrophages that strongly express monocyte chemotactic protein-1. Monocyte chemotactic protein-1 is a chemotactic cytokine that contributes to the activation and recruitment of macrophages. Relatively little is known about its role in the resorption process of herniated nucleus pulposus. To clarify the sequential dynamics of expression of monocyte chemotactic protein-1 in the granulation tissues of herniated nucleus pulposus, we introduced a rat autologous transplantation model of nuclear materials onto its lumbar dura mater and performed immunohistological analysis and competitive polymerase chain reaction assay using the grafted samples. Immunohistological analysis demonstrated that the majority of infiltrating mononuclear cells expressed monocyte chemotactic protein-1. Monocyte chemotactic protein-1 mRNA was expressed in the first 3 weeks after the procedure and was significantly and maximally upregulated at 1 week. To determine whether human recombinant monocyte chemotactic protein-1 facilitates the resorption process of herniated nucleus pulposus, we introduced another model of autologous transplantation, wherein the nuclear materials were grafted to the abdominal subcutaneous tissues and recombinant monocyte chemotactic protein-1 was subsequently applied to these materials. When monocyte chemotactic protein-1 was injected into the murine nucleus pulposus tissues, they reduced in size more rapidly than in the control group. These findings suggest that monocyte chemotactic protein-1 plays an important role in the recruitment of macrophages in the early phase of the resorption process of herniated nucleus pulposus and that its application may physiologically facilitate the resorption process of the nucleus pulposus.

Chemonucleolysis with human stromelysin-1.

STUDY DESIGN: Immunohistologic analysis was performed on surgically removed samples of herniated nucleus pulposus to examine the expression of stromelysin-1. We performed in vitro and in vivo experiments to determine whether recombinant human (rh) stromelysin-1 is capable of degrading nucleus pulposus. OBJECTIVE: To analyze the production of stromelysin-1 in various types of herniated nucleus pulposus, and to examine the effects of this recombinant protein on nucleus pulposus tissues. SUMMARY OF BACKGROUND DATA: The authors previously demonstrated a progressive decrease in herniated nucleus pulposus size in some of the transligamentous and sequestration types of herniated nucleus pulposus using magnetic resonance imaging. An increased production of stromelysin-1, a cartilage proteoglycan degrading enzyme, in herniated nucleus pulposus was reported recently. The authors speculated that if stromelysin-1 is involved in the degradation of herniated nucleus pulposus, stromelysin-1 itself may be used as a chemonucleolytic agent. METHODS: Immunohistologic analysis using streptoavidin-biotin method was performed on 20 herniated nucleus pulposus samples to investigate the expression of stromelysin-1. Five herniated nucleus pulposus samples were incubated in a tissue culture medium in the presence or absence of rh stromelysin-1. After 24 hours of incubation, their weight changes were measured, and the loss of proteoglycan was assessed by Safranin O staining. Rat nucleus pulposus tissues were obtained from coccygeal intervertebral discs, and autologous subcutaneous transplantation was performed. Rh stromelysin-1 was injected into the grafted materials, and the reduction in size was followed by two-dimensional measurements from the skin surface, using engineer's calipers. RESULTS: Immunohistologic analysis demonstrated the production of stromelysin-1 in the granulation tissues of herniated nucleus pulposus. When stromelysin-1 was injected into the murine nucleus pulposus tissues, they reduced in size more rapidly than the control group. In addition, human herniated nucleus pulposus materials obtained at surgery showed significant weight loss when treated with stromelysin-1 in an organ culture system. Safranin O staining revealed extensive depletion of proteoglycan in these herniated nucleus pulposus samples. CONCLUSIONS: Stromelysin-1 is a possible key enzyme in herniated nucleus pulposus resorption, and stromelysin-1 may be a good candidate for use in chemonucleolysis. Administration of human stromelysin-1 may physiologically facilitate the resorption process of herniated nucleus pulposus, increase the healing rate and decrease complications after chemonucleolysis.

Contrast-enhanced magnetic resonance imaging in conservative management of lumbar disc herniation.

STUDY DESIGN: This study was designed to investigate the morphologic changes in contrast-enhanced magnetic resonance imaging that occur during conservative treatment of patients with unilateral leg pain resulting from herniated nucleus pulposus without significant lumbar canal stenosis. OBJECTIVES: To compare the morphologic results with clinical outcomes to ascertain whether enhanced magnetic resonance imaging contributes to the management of lumbar disc herniation. SUMMARY OF BACKGROUND DATA: Contrast-enhanced magnetic resonance imaging has already been reported to be useful in the postoperative examination of the lumbar spine and in visualization of symptomatic nerve roots. However, there have been few reports about its usefulness in the conservative management of herniated nucleus pulposus or about the correlation between herniated nucleus pulposus regression and enhanced effect. The study population consisted of 48 patients with radiculopathy. All patients primarily reported unilateral leg pain, and 94% had positive tension signs. Additionally, 38% exhibited muscle weakness corresponding to the symptomatic nerve root. METHODS: All patients were studied twice or more using gadolinium-magnetic resonance imaging during conservative therapy, at a mean interval of 191 days. Changes in the size of the herniated nucleus pulposus on precontrast images fell into four categories, with changes in enhancement on postcontrast images classified into two categories: "enlargement" and "no change." RESULTS: In all cases of migrating type herniated nucleus pulposus, circular enhancement was seen on postcontrast images. In 17 of 22 cases, the enhanced area gradually thickened and intruded into the migrated disc materials as the size of the herniated nucleus pulposus decreased; the herniated nucleus pulposus disappeared in nine cases and showed a marked decrease in seven cases. These cases showed good clinical courses of sciatica. In the other five patients, in whom there were no changes in the enhanced area, there was less of a tendency for the herniated nucleus pulposus to decrease in size, and there were poorer clinical results. In six cases of extruding-type herniated nucleus pulposus, no enhanced effects were observed throughout the follow-up period. The other 20 cases showed enhancement that was relatively weaker than that of migrating disc herniation. Extension or expansion of the enhanced area was observed in the follow-up images of 15 cases, though only four showed obvious changes in the size of the herniated nucleus pulposus. These 15 cases had better clinical results than the other cases, in which enhanced effects did not change or were not observed. CONCLUSION: Contrast-enhanced magnetic resonance imaging is a useful prognostic parameter, and multiple use contributes to the proper management of lumbar disc herniation.

Morphology of C5 ventral nerve rootlets as part of dissociated motor loss of deltoid muscle.

STUDY DESIGN: This study analyzed anatomic characteristics of cervical ventral rootlets. After total vertebrectomy, detailed morphology of the ventral rootlets was studied from the anterior. SUMMARY OF BACKGROUND DATA: The clinical study showed the predominance of ventral root lesion. There are few studies concerning the morphologic pathogenesis of cervical amyotrophy and detailed cervical ventral rootlet anatomy. METHODS: Thirty-six embalmed adult human cadavers were studied. The measurements for the ventral rootlets of C5 to C8 were made as follows: 1) angle between the rootlet and spinal cord, 2) longitudinal width of the ventral rootlet origin, and 3) length of the ventral rootlets. RESULTS: The C5 ventral rootlets were shorter and issued more obtusely from the cervical spinal cord than lower rootlets. The spinal cord segment of the deltoid muscle, indicated by the longitudinal widths of the C5 and C6 ventral rootlet exits from the spinal cord, were wider than the C7 and C8 segments. Preforaminal anterior compression at the C4-C5 disc level might affect the lower part of the C5 ventral rootlets and upper part of the C6 ventral rootlets. CONCLUSION: Short C5 ventral rootlets appeared to become taut and easily injured by hemilateral anterior compression. Spinal cord lesion resulting from localized anterior compression at the single disc level might not play as important a role in the pathogenesis of dissociated motor loss of the deltoid muscle because of the wider spinal segments of C5 and C6.

Regression of cervical disc herniation observed on magnetic resonance images.

STUDY DESIGN: A retrospective study of cervical disc herniation using results of repeated magnetic resonance imaging examinations. OBJECTIVES: To clarify the cervical disc herniation morphological changes over time in order to establish a strategy for treatment. SUMMARY OF BACKGROUND DATA: In the authors' previous magnetic resonance imaging follow-up study of patients with lumbar disc herniation, spontaneous regression was observed in the sequestration-type lesions, and it was found that the tendency toward regression differed based on the anatomic position of extruded disc material. METHODS: Thirty-eight patients with cervical disc herniation who underwent repeated magnetic resonance imaging examinations were studied. The changes over time in herniated disc size were evaluated using this imaging technique. Evaluation showed the characteristics of those in whom spontaneous regression was found, such as extrusion pattern, and the clinical outcome was evaluated by symptoms. RESULTS: In 15 patients (40%), the volume of herniated material was decreased. The interval from onset of symptoms to the initial examination was significantly shorter in the regression group than in the group that showed no change in disc herniation. By extrusion pattern, cervical disc herniation, which was divided into migration type on sagittal view and lateral type on axial view, most frequently exhibited spontaneous regression. All of the patients with radicular pain and upper limb amyotrophy were treated successfully with conservative therapy. CONCLUSION: Although the possibility of the combination of hemorrhage and disc material could not be denied, active resorption of herniated material probably occurred during the acute phase. Extruded material exposed to the epidural space may be resorbed more quickly than that beneath the ligament. Vascular supply probably plays a role in the mechanism of resorption. The phase and position of extrusion were the significant factors affecting cervical disc herniation resorption. It was demonstrated that examination performed during the acute phase using magnetic resonance imaging is necessary for elucidation of the pathogenesis of cervical disc herniation, and that migrating, lateral-type herniations regress so frequently that conservative treatment should be chosen not only for patients with radicular pain, but also for those with upper limb amyotrophy.

Upregulated expression of chemokines in herniated nucleus pulposus resorption.

STUDY DESIGN: Immunohistologic examination was performed on surgically removed samples of herniated nucleus pulposus. OBJECTIVES: To determine what cell types predominate in the granulation tissues of herniated nucleus pulposus, and to elucidate whether chemokines are involved in the resorption process of herniated nucleus pulposus. SUMMARY OF BACKGROUND DATA. The study population consisted of 30 patients suffering from herniated nucleus pulposus. Five macroscopically normal discs were obtained from spinal cord tumor and spinal cord injury managed with anterior discectomy (age range, 27-63 years) as a healthy control group. METHODS: Immunohistochemical analysis was used to analyze the expression of chemokines. RESULTS: A marked infiltration of macrophage and vascular proliferation was identified with a T lymphocyte infiltration of mild degree in the granulation tissues. This tendency was more prominent in the exposed group compared with the nonexposed group. Infiltrating macrophages, fibroblasts, and endothelial cells in the granulation tissues strongly expressed monocyte chemotactic protein-1 and macrophage inflammatory protein-1 alpha. Statistical analysis demonstrated that the exposed group was more abundant in Factor VIII, monocyte chemotactic protein-1, and macrophage inflammatory protein-1 alpha positive cells than the unexposed group. CONCLUSIONS: Inflammatory cells and their positivity for chemokines, such as monocyte chemotactic protein-1 and macrophage inflammatory protein-1 alpha, are associated with blood vessels. Chemokines, such as monocyte chemotactic protein-1 and macrophage inflammatory protein-1 alpha, were overexpressed in macrophages, fibroblasts, and endothelial cells, suggesting that these chemokines contribute to activation and recruitment of macrophages in a paracrine or autocrine fashion.

The association of degeneration of the intervertebral disc with 5a/6a polymorphism in the promoter of the human matrix metalloproteinase-3 gene.

It has been suggested that matrix metalloproteinase-3 (MMP-3, stromelysin-1) has an important role in the degeneration of intervertebral discs (IVDs). A human MMP-3 promoter 5A/6A polymorphism was reported to be involved in the regulation of MMP-3 gene expression. We suggest that IVD degeneration is associated with 5A/6A polymorphism. We studied 54 young and 49 elderly Japanese subjects. Degeneration of the lumbar discs was graded using MRI in the younger group and by radiography in the elderly. 5A/6A polymorphism was determined by polymerase-chain reaction-based assays. We found that the 5A5A and 5A6A genotype in the elderly was associated with a significantly larger number of degenerative IVDs than the 6A6A (p < 0.05), but there was no significant difference in the young. In the elderly, the IVD degenerative scores were also distributed more highly in the 5A5A and 5A6A genotypes (p = 0.0029). Our findings indicate that the 5A allele is a possible risk factor for the acceleration of degenerative changes in the lumbar disc in the elderly.

Up-regulated expression of matrilysin and neutrophil collagenase in human herniated discs.

Spontaneous resorption of herniated nucleus pulposus (HNP) is commonly observed when there is substantial contact of the disc with the spinal canal. We already demonstrated the expression of matrix metalloproteinase (MMP)-3 (stromelysin-1) in the granulation tissues of HNP, suggesting its role in the resorption process of HNP. Recent studies of osteoarthritic cartilages reported an up-regulated expression of metalloproteinases including MMP-7 (matrilysin) and MMP-8 (neutrophil collagenase), suggesting their roles in the matrix degradation. To clarify the expression of MMP-7 and MMP-8 in HNP, immunohistological analysis of various types of HNP was performed. We found MMP-7 was expressed in infiltrated mononuclear cells and chondrocytes, whereas MMP-8 was specifically expressed in chondrocytes. The positive rate for both MMP-7 and MMP-8 significantly increased when HNP was exposed to the epidural space (p < 0.01). Our data suggest that not only MMP-3 but also MMP-7 and MMP-8 may play a role in the resorption process of HNP.

Osteopontin, a novel substrate for matrix metalloproteinase-3 (stromelysin-1) and matrix metalloproteinase-7 (matrilysin).

Osteopontin (OPN) is a secreted phosphoprotein shown to function in wound healing, inflammation, and tumor progression. Expression of OPN is often co-localized with members of the matrix metalloproteinase (MMP) family. We report that OPN is a novel substrate for two MMPs, MMP-3 (stromelysin-1) and MMP-7 (matrilysin). Three cleavage sites were identified for MMP-3 in human OPN, and two of those sites were also cleaved by MMP-7. These include hydrolysis of the human Gly166-Leu167, Ala201-Tyr202 (MMP-3 only), and Asp210-Leu211 peptide bonds. Only the N-terminal Gly-Leu cleavage site is conserved in rat OPN (Gly151-Leu152). These sites are distinct from previously reported cleavage sites in OPN for the proteases thrombin or enterokinase. We found evidence for the predicted MMP cleavage fragments of OPN in vitro in tumor cell lines, and in vivo in remodeling tissues such as the postpartum uterus, where OPN and MMPs are co-expressed. Furthermore, cleavage of OPN by MMP-3 or MMP-7 potentiated the function of OPN as an adhesive and migratory stimulus in vitro through cell surface integrins. We predict that interaction of MMPs with OPN at tumor and wound healing sites in vivo may be a mechanism of regulation of OPN bioactivity.