Seizo Yamamoto

An aspartic acid repeat polymorphism in asporin inhibits chondrogenesis and increases susceptibility to osteoarthritis.

Osteoarthritis is the most common form of human arthritis. We investigated the potential role of asporin, an extracellular matrix component expressed abundantly in the articular cartilage of individuals with osteoarthritis, in the pathogenesis of osteoarthritis. Here we report a significant association between a polymorphism in the aspartic acid (D) repeat of the gene encoding asporin (ASPN) and osteoarthritis. In two independent populations of individuals with knee osteoarthritis, the D14 allele of ASPN is over-represented relative to the common D13 allele, and its frequency increases with disease severity. The D14 allele is also over-represented in individuals with hip osteoarthritis. Asporin suppresses TGF-β–mediated expression of the genes aggrecan (AGC1) and type II collagen (COL2A1) and reduced proteoglycan accumulation in an in vitro model of chondrogenesis. The effect on TGF-β activity is allele-specific, with the D14 allele resulting in greater inhibition than other alleles. In vitro binding assays showed a direct interaction between asporin and TGF-β. Taken together, these findings provide another functional link between extracellular matrix proteins, TGF-β activity and disease, suggesting new therapeutic strategies for osteoarthritis.

A functional polymorphism in the 5′ UTR of GDF5 is associated with susceptibility to osteoarthritis

Osteoarthritis (MIM 165720), characterized by degeneration of articular cartilage, is the most common form of human arthritis and a major concern for aging societies worldwide. Epidemiological and genetic studies have shown that osteoarthritis is a polygenic disease. Here, we report that the gene encoding growth differentiation factor 5 (GDF5) is associated with osteoarthritis in Asian populations. A SNP in the 5′ UTR of GDF5 (+104T/C; rs143383) showed significant association (P = 1.8 × 10−13) with hip osteoarthritis in two independent Japanese populations. This association was replicated for knee osteoarthritis in Japanese (P = 0.0021) and Han Chinese (P = 0.00028) populations. This SNP, located in the GDF5 core promoter, exerts allelic differences on transcriptional activity in chondrogenic cells, with the susceptibility allele showing reduced activity. Our findings implicate GDF5 as a susceptibility gene for osteoarthritis and suggest that decreased GDF5 expression is involved in the pathogenesis of osteoarthritis.

Prevalence of radiographic knee osteoarthritis and its association with knee pain in the elderly of Japanese population-based cohorts: The ROAD study

OBJECTIVE We investigated the prevalence of radiographic knee osteoarthritis (OA) and knee pain in the Japanese elderly using a large-scale population of a nationwide cohort study, Research on Osteoarthritis Against Disability (ROAD), and examined their association. METHODS From the baseline survey of the ROAD study, 2,282 participants > or =60 years (817 men and 1,465 women) living in urban, mountainous and seacoast communities were analyzed. The radiographic severity at both knees was determined by the Kellgren/Lawrence (KL) grading system. KL> or =2 and KL> or =3 knee OA were examined separately to assess osteophytosis and joint space narrowing (JSN). RESULTS The prevalence of KL> or =2 OA (47.0% and 70.2% in men and women, respectively) was much higher than that of previous studies in Caucasians, while that of KL> or =3 OA was not much different in men. Age, BMI, female sex and rural residency were risk factors for radiographic knee OA, knee pain and their combination. The prevalence of knee pain was age-dependent in women, but not in men. Knee pain was more strongly associated with KL> or =3 OA than with KL=2, and the association was higher in men than in women. Female sex was a strong risk factor even in the subgroup without radiographic knee OA (KL=0/1). CONCLUSION The present cross-sectional study revealed a high prevalence of radiographic knee OA in the Japanese elderly. Knee pain was strongly associated with JSN especially in men, while women tended to have knee pain even without radiographic OA.

Regulation by PGE2 of the production of interleukin‐6, macrophage colony stimulating factor, and vascular endothelial growth factor in human synovial fibroblasts

We examined the effects of endogenous prostaglandin E2 (PGE2) on the production of interleukin‐6 (IL‐6), macrophage colony stimulating factor (M‐CSF), and vascular endothelial growth factor (VEGF) by interleukin‐1β (IL‐1β)‐stimulated human synovial fibroblasts. NS‐398 (1 μM), a cyclo‐oxygenase‐2 (COX‐2) inhibitor, inhibited IL‐6 and VEGF production (35±4% and 26±2%, respectively) but enhanced M‐CSF production (38±4%) by IL‐1β (1 ng ml−1) in synovial fibroblasts isolated from patients with osteoarthritis (OA) and rheumatoid arthritis (RA). Exogenous PGE2 completely abolished the effects of NS‐398 on the production of each mediator by OA fibroblasts stimulated with IL‐1β. 8‐Bromo cyclic AMP and dibutyryl cyclic AMP, cyclic AMP analogues, mimicked the effects of PGE2 on IL‐6, M‐CSF, and VEGF production by OA fibroblasts. The EP2 selective receptor agonist ONO‐AE1‐259 (2 nM) and the EP4 selective receptor agonist ONO‐AE1‐329 (2 or 20 nM), but not the EP1 selective receptor agonist ONO‐DI‐004 (1 μM) and the EP3 selective receptor agonist ONO‐AE‐248 (1 μM), replaced the effects of PGE2 on IL‐6, M‐CSF, and VEGF production by OA and RA fibroblasts stimulated with IL‐1β in the presence of NS‐398. Both OA and RA fibroblasts expressed mRNA encoding EP2 and EP4 but not EP1 receptors. In addition, up‐regulation of EP2 and EP4 receptor mRNAs was observed at 3 h after IL‐1β treatment. These results suggest that endogenous PGE2 regulates the production of IL‐6, M‐CSF, and VEGF by IL‐1β‐stimulated human synovial fibroblasts through the activation of EP2 and EP4 receptors with increase in cyclic AMP.

Effects of β-adrenergic agonists on bone-resorbing activity in human osteoclast-like cells

In the present study, we demonstrate for the first time that beta-adrenergic agonists stimulate bone-resorbing activity in human osteoclast-like multinucleated cells (MNCs). Osteoclast-like MNCs constitutively expressed mRNA for alpha1B-, alpha2B- and beta2-adrenergic receptor (AR) in addition to characteristic markers of mature osteoclast, such as calcitonin receptor (CT-R), tartrate-resistant acid phosphatase (TRAP), alphaV-chain of integrin (Int alphaV), carbonic anhydrase II (CA-II) and cathepsin K (Cathe K). Epinephrine (1 microM; alpha,beta-adrenergic agonist) up-regulated expression of Int alphaV, CA-II and Cathe K in the osteoclast-like MNCs. Osteoclastic resorbing activity was markedly increased by isoprenaline (1 microM; beta-adrenergic agonist), moderately by epinephrine, but poorly by phenylephrine (1 microM; alpha1-adrenergic agonist). The actin ring, which was suggested to be correlated with bone-resorbing activity, was clearly observed in osteoclast-like MNCs treated with isoprenaline and epinephrine, but faintly in those treated with phenylephrine. These findings suggest that beta-adrenergic agonists directly stimulate bone-resorbing activity in matured osteoclasts.

Prevalence of radiographic lumbar spondylosis and its association with low back pain in elderly subjects of population-based cohorts: the ROAD study

Objectives: Although lumbar spondylosis is a major cause of low back pain and disability in elderly people, few epidemiological studies have been performed. The prevalence of radiographic lumbar spondylosis was investigated in a large-scale population study and the association with low back pain was examined. Methods: From a nationwide cohort study (Research on Osteoarthritis Against Disability; ROAD), 2288 participants aged ⩾60 years (818 men and 1470 women) living in urban, mountainous and coastal communities were analysed. The radiographic severity at lumbar intervertebral levels from L1/2 to L5/S was determined by Kellgren/Lawrence (KL) grading. Results: In the overall population the prevalence of radiographic spondylosis with KL⩾2 and ⩾3 at the severest intervertebral level was 75.8% and 50.4%, respectively, and that of low back pain was 28.8%. Although KL⩾2 spondylosis was more prevalent in men, KL⩾3 spondylosis and low back pain were more prevalent in women. Age and body mass index were risk factors for both KL ⩾2 and KL⩾3 spondylosis. Although KL = 2 spondylosis was not significantly associated with low back pain compared with KL = 0 or 1, KL⩾3 spondylosis was related to the pain only in women. Conclusions: This cross-sectional study in a large population revealed a high prevalence of radiographic lumbar spondylosis in elderly subjects. Gender seems to be distinctly associated with KL⩾2 and KL⩾3 lumbar spondylosis, and disc space narrowing with or without osteophytosis in women may be a risk factor for low back pain.

Association of radiographic and symptomatic knee osteoarthritis with health-related quality of life in a population-based cohort study in Japan: the ROAD study

OBJECTIVE Knee osteoarthritis (OA) is a major public health issue causing chronic pain and disability. However, there is little information on the impact of this disease on quality of life (QOL) in Japanese men and women. The objective of the present study was to clarify the impact of radiographic and symptomatic knee OA on QOL in Japan. METHODS This study examined the association of radiographic and symptomatic knee OA with QOL parameters such as the Medical Outcomes Study Short Form-8 (SF-8), EuroQOL (EQ-5D) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Radiographic knee OA was defined according to Kellgren/Lawrence (KL) grades, and symptomatic knee OA was defined as KL=3 or 4 with knee pain. We also examined the independent association of symptomatic knee OA and grip strength with QOL. RESULTS From the 3040 participants in the Research on Osteoarthritis Against Disability (ROAD) study, the present study analyzed 2126 subjects older than 40 years who completed the questionnaires (767 men and 1359 women; mean age, 68.9+/-10.9 years). Subjects with KL=3 or 4 had significantly lower physical QOL as measured by the physical component summary (PCS) score of the SF-8 and pain domains of the WOMAC, whereas mental QOL, as measured by the mental component summary (MCS) score of the SF-8, was higher in subjects with KL=3 or 4 than KL=0 or 1. Symptomatic knee OA was significantly more likely than radiographic knee OA without pain to be associated with physical QOL loss as measured by the PCS score and physical domains of the WOMAC. Symptomatic knee OA and grip strength were independently associated with physical QOL. CONCLUSION This cross-sectional study revealed that subjects with symptomatic knee OA had significantly lower physical QOL than subjects without it.

Spontaneous anterior interosseous nerve palsy with hourglass-like fascicular constriction within the main trunk of the median nerve

Interfascicular neurolysis was performed in nine patients with spontaneous anterior interosseous nerve palsy. In eight of these patients, an hourglass-like constriction in the fascicles forming the anterior interosseous nerve was found within the main trunk of the median nerve at 2-7.5 cm above the medial epicondyle. The clinical signs and symptoms of these eight patients were similar to those that have been described to isolated neuritis. While the etiology remains unknown, when spontaneous anterior interosseous nerve palsy is suspected to be caused by isolated neuritis, interfascicular neurolysis should be performed to confirm the lesion and to discover whether fascicular constriction is present.

Regional differences in chondrocyte metabolism in osteoarthritis: A detailed analysis by laser capture microdissection

OBJECTIVE To determine the change in metabolic activity of chondrocytes in osteoarthritic (OA) cartilage, considering regional difference and degree of cartilage degeneration. METHODS OA cartilage was obtained from knee joints with end-stage OA, at both macroscopically intact areas and areas with various degrees of cartilage degeneration. Control cartilage was obtained from age-matched donors. Using laser capture microdissection, cartilage samples were separated into superficial, middle, and deep zones, and gene expression was compared quantitatively in the respective zones between OA and control cartilage. RESULTS In OA cartilage, gene expression changed markedly with the site. The expression of cartilage matrix genes was highly enhanced in macroscopically intact areas, but the enhancement was less obvious in the degenerated areas, especially in the upper regions. In contrast, in those regions, the expression of type III collagen and fibronectin was most enhanced, suggesting that chondrocytes underwent a phenotypic change there. Within OA cartilage, the expression of cartilage matrix genes was significantly correlated with SOX9 expression, but not with SOX5 or SOX6 expression. In OA cartilage, the strongest correlation was observed between the expression of type III collagen and fibronectin, suggesting the presence of a certain link(s) between their expression. CONCLUSION The results of this study revealed a comprehensive view of the metabolic change of the chondrocytes in OA cartilage. The change of gene expression profile was most obvious in the upper region of the degenerated cartilage. The altered gene expression at that region may be responsible for the loss of cartilage matrix associated with OA.

Fully automatic quantification of knee osteoarthritis severity on plain radiographs

OBJECTIVE Although knee osteoarthritis (OA) is a major public health issue causing chronic disability, there is no objective or accurate method for measurement of the structural severity in general clinical practice. Here we have established a fully automatic program KOACAD (knee OA computer-aided diagnosis) to quantify the major OA parameters on plain knee radiographs, validated the reproducibility and reliability, and investigated the association of the parameters with knee pain. METHODS KOACAD was programmed to measure joint space narrowing at medial and lateral sides, osteophyte formation, and joint angulation. Anteroposterior radiographs of 1979 knees of a large-scale cohort population were analyzed by KOACAD and conventional categorical grading systems. RESULTS KOACAD automatically measured all parameters in less than 1s without intra- or interobserver variability. All parameters, especially medial joint space narrowing, were significantly correlated with the conventional gradings. In the parameters, osteophyte formation was associated with none of the joint space parameters, suggesting different etiologic mechanisms between them. Multivariate logistic regression analysis after adjustment for age and confounding factors revealed that medial joint space narrowing and varus angulation of knee joints were risk factors for the presence of pain (594/1979 knees), while neither lateral joint space nor osteophyte area was. CONCLUSION KOACAD was shown to be useful for objective, accurate, simple and easy evaluation of the radiographic knee OA severity in daily clinical practice. This system may also serve as a surrogate measure for the development of disease-modifying drugs for OA, just as bone mineral density does in osteoporosis.