佐粧 孝久

Objective. To analyze the methylome of normal and osteoarthritic (OA) knee articular cartilage and to determine the role of DNA methylation in the regulation of gene expression in vitro. Methods. DNA was isolated from human normal (n = 11) and OA (n = 12) knee articular cartilage and analyzed using the Infinium HumanMethylation450 BeadChip array. To integrate methylation and transcription, RNA sequencing was performed on normal and OA cartilage and validated by quantitative polymerase chain reaction. Functional validation was performed in the human TC28 cell line and primary chondrocytes that were treated with the DNA methylation inhibitor 5-aza-2'deoxycytidine (5-aza-dC). Results. DNA methylation profiling revealed 929 differentially methylated sites between normal and OA cartilage, comprising a total of 500 individual genes. Among these, 45 transcription factors that harbored differentially methylated sites were identified. Integrative analysis and subsequent validation showed a subset of 6 transcription factors that were significantly hypermethylated and down-regulated in OA cartilage (ATOH8, MAFF, NCOR2, TBX4, ZBTB16, and ZHX2). Upon 5-aza-dC treatment, TC28 cells showed a significant increase in gene expression for all 6 transcription factors. In primary chondrocytes, ATOH8 and TBX4 were increased after 5aza-dC treatment. Conclusion. Our findings reveal that normal and OA knee articular cartilage have significantly different methylomes. The identification of a subset of epigenetically regulated transcription factors with reduced expression in OA may represent an important mechanism to explain changes in the chondrocyte transcriptome and function during OA pathogenesis.

症例は61歳女性で、10年前誘因なく右膝痛が出現し、変形性膝関節症と診断され、保存治療を受けた。初診8年後、X線で膝関節内の小腫瘤が指摘され、1年後、右膝の急激な疼痛、ロッキング症状が出現し、MRIにおいて多発する遊離体、腫瘤性病変を指摘された。初診10年後精査のため紹介受診となった。右膝に疼痛、腫脹がみられ、膝蓋跳動を認めた。可動域は伸展-10°、屈曲120°制限され、内側関節裂隙に圧痛を感じ、膝関節屈曲時のれき音が聴取された。初診時X線で膝関節内に多数の遊離体があり、MRIでも同様に多数の小腫瘤を認めた。滑膜性骨軟骨腫症を疑い関節鏡下に遊離体摘出、滑膜亜全切除を行った。seven-portal approachを用い、手術を施行した。HE染色で滑膜組織に連続し軟骨化成を認め滑膜性骨軟骨症と診断した。術後翌日から歩行可能で、歩行時、屈伸時の急激な疼痛は消失し、可動域は屈曲0°、伸展130°と改善し、特記すべき合併症はなく、術後7日目に退院した。術後2週目にCTを行い、2個遊離体残存を認めた。一つは半月板下、一つは膝蓋下脂肪体内に埋もれた遊離体と思われた。

Purpose: Our previous study showed that partial-thickness articular cartilage defects (PTCDs) created in immature rats spontaneously healed to resemble normal hyaline cartilage, but that of mature rats did not. To identify molecules involved in the spontaneous cartilage repair observed in this model, gene expression was compared between PTCD and sham-operated cartilage of immature and mature rats. Materials and Methods: Six sets of gene comparisons were made at 12, 24, and 48 hours after the creation of PTCDs in immature and mature rats using microarrays. All the genes upregulated in immature cartilage at 12 hours were selected for further analysis if their expression pattern was not irregular such that diminished at 24 hours and re-upregulated at 48 hours. Relationships among genes selected through the above steps were analyzed using Ingenuity Pathway Analysis (IPA) software. After deriving networks, important molecules were further narrowed down by location within a network. Genes were regarded as central if they had relationships with more than 10 molecules in a network. Protein localization in tissues was confirmed by immunohistochemistry. Results: Five networks were identified. Their functional annotations were gene expression, cell cycle, growth and proliferation, and cell signaling. Transforming growth factor-beta (TGF-beta) was centrally located in the network with the highest IPA score and mothers against decapentaplegic homolog-3 (Smad3) were centrally located in the second highest ranking network. Phosphorylated Smad3 was detected in the nuclei of chondrocytes in immature cartilage. Conclusions: Our data suggest the possible importance of Smad3 in the TGF-beta signaling in the spontaneous healing of PTCDs in immature rats.

Background Correction of pronation deformity and metatarsal primus varus is an important component of hallux valgus surgery, necessary to achieve a satisfactory correction and to prevent post-operative recurrence. Roundness of the lateral edge of the first metatarsal head (round sign) on the dorsoplantar radiograph of the foot has been empirically advocated as an indicator of first metatarsal pronation. The purpose of this study was to clarify the effect of rotation and inclination of the first metatarsal on the shape of the lateral edge of the first metatarsal head. Methods Computed tomographic images of feet in 30 subjects, without pathology of the first metatarsophalangeal joint, were included. Digitally reconstructed radiographs of the first metatarsal were created using the computed tomographic data. Thirty-nine images were created of each first metatarsal at different degrees of rotation (-10A degrees to 30A degrees of pronation) and inclination (-10A degrees to 20A degrees of plantarflexion). Then, the shape of the lateral edge of the first metatarsal head was classified into three types: angular, intermediate, and round. Generalized estimation equations were used to test if the shapes of the first metatarsal head were significantly different across the range of pronation and plantarflexion angles. Results The positive round sign changed to negative as the first metatarsal supinated. In most feet, these changes occurred as the pronation angle decreased from 10A degrees to 0A degrees. The positive round sign also changed to negative as the first metatarsal head plantarflexed. Conclusion Positive round sign of the first metatarsal head on the dorsoplantar radiograph of the foot was significantly associated with increased pronation as well as decreased inclination of the first metatarsal. Negative round sign may be used as an indicator of effective correction of first metatarsal pronation during hallux valgus surgery.

Objective Biomechanical effects of laterally wedged insoles are assessed by reduction in the knee adduction moment. However, the degree of reduction may vary depending on the reference frame with which it is calculated. The purpose of this study was to clarify the effect of reference frame on the reduction in the knee adduction moment by laterally wedged insoles. Methods Twenty-nine healthy participants performed gait trials with a laterally wedged insole and with a flat insole as a control. The knee adduction moment, including the first and second peaks and the angular impulse, were calculated using four different reference frames: the femoral frame, tibial frame, laboratory frame and the Joint Coordinate System. Results There were significant effects of reference frame on the knee adduction moment first and second peaks (P < 0.001 for both variables), while the effect was not significant for the angular impulse (P = 0.84). No significant interaction between the gait condition and reference frame was found in either of the knee adduction moment variables (P = 0.99 for all variables), indicating that the effects of laterally wedged insole on the knee adduction moments were similar across the four reference frames. On the other hand, the average percent changes ranged from 9% to 16% for the first peak, from 16% to 18% for the second peak and from 17% to 21% for the angular impulse when using the different reference frames. Conclusion The effects of laterally wedged insole on the reduction in the knee adduction moment were similar across the reference frames. On the other hand, Researchers need to recognize that when the percent change was used as the parameter of the efficacy of laterally wedged insole, the choice of reference frame may influence the interpretation of how laterally wedged insoles affect the knee adduction moment.

背景:非ステロイド性抗炎症薬(NSAIDs)による消化管障害は広く知られているが,胃食道逆流症(gastroesophageal reflux disease,GERD)症状の頻度などについては不明である.目的:NSAIDs服用者のGERD症状の発生頻度を調べること.対象と方法:整形外科外来通院者の中でNSAIDsを4週間以上服用している患者53名を対象とし,Frequency Scale for the Symptoms of GERD(FSSG):Fスケール問診票を用いたアンケートを実施した.8点以上をGERDの疑いが強いと判断し,その他の因子との関係を調べた.結果:(1)85.0%に消化器用薬が処方されていた.(2)8点以上が17名(32.1%)であり,胃腸障害の既往が大きな因子であった.まとめ:4週以上のNSAIDs服用患者にGERD症状が高率に存在することがわかった.(著者抄録)

© 2014 Osteoarthritis Research Society International. Objective: To examine whether the detection of osteophytes anywhere in the knee could serve as a pre-radiographic biomarker for osteoarthritis (OA) development. Methods: Baseline magnetic resonance imaging (MRIs) of 132 participants in the Osteoarthritis Initiative (OAI) were studied. Based on radiographs, 66 knees were assessed as osteoarthritis-free (no-osteoarthritis [NOA], or Kellgren/Lawrence [K/L] severity grade 0/1 both at baseline and 48 months), and another 66 knees were assessed as having radiographic OA changes (pre-radiographic osteoarthritis [PROA], or with K/L grade 0/1at baseline and grade ≥2at 48 months). Using baseline MRI data, we examined eight sites of osteophyte formation: the medial and lateral femoral condyle (MFC and LFC, respectively); medial and lateral tibial plateau (MTP and LTP, respectively); medial and lateral facets of the patellofemoral joint (PM and PL, respectively); tibial spine (TS); and femoral intercondylar notch (IC). Knee joint osteophyte size was assessed via the 8-point marginal osteophytes item of the whole-organ magnetic resonance imaging score (WORMS). The frequencies and distribu

Partial thickness articular cartilage injuries (PTCIs) were not previously thought to heal spontaneously. Immature rats have the capacity for spontaneous repair of PTCIs, although it is a long-term process. Our aim has been to examine the spontaneous repair response mechanism in immature rats. Single linear PTCIs were created in 3-week-old and 12-week-old rats in the direction of joint motion. On day 1 and at 1, 2, and 4 weeks after PTCI, evaluations of histological changes and immunohistology at the injury site and in the surrounding cartilage were performed. Anti-CD105 and anti-CD166 antibodies (as stem cell markers to identify mesenchymal stem cells in reparative cartilage tissue) were used for immunohistological evaluations. To determine whether endogenous repair ability existed in articular cartilage, an ex vivo experiment was also carried out. Femoral condyles with PTCIs were incubated in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum for 1 day and for 1 and 2 weeks. Histological changes were subsequently examined. Immature cartilage showed a higher repair response than did mature cartilage, and the response occurred immediately after PTCI. In immature rats, CD105- and CD166-positive cells were found in the superficial and transitional zones of the articular cartilage. Few CD166-positive cells were identified in mature articular cartilage. No significant in vivo differences in the spontaneous repair responses to PTCIs were observed between mature and immature groups. Thus, the repair response to PTCIs seems to be associated not only with CD105- and CD166-positive cells, but also with other perichondral factors.

Objective: To investigate the effectiveness of quantitative Magnetic resonance imaging (MRI) for evaluating the quality of cartilage repair over time following allograft chondrocyte implantation using a threedimensional scaffold for osteochondral lesions. Design: Thirty knees from 15 rabbits were analyzed. An osteochondral defect (diameter, 4 mm; depth, 1 mm) was created on the patellar groove of the femur in both legs. The defects were filled with a chondrocyte-seeded scaffold in the right knee and an empty scaffold in the left knee. Five rabbits each were euthanized at 4, 8, and 12 weeks and their knees were examined via macroscopic inspection, histological and biochemical analysis, and quantitative MRI (T2 mapping and dGEMRIC) to assess the state of tissue repair following allograft chondrocyte implantation with a three-dimensional scaffold for osteochondral lesions. Results: Comparatively good regenerative cartilage was observed both macroscopically and histologically. In both chondrocyte-seeded and control knees, the T2 values of repair tissues were highest at 4 weeks and showed a tendency to decrease with time. Delta R1 values of dGEMRIC also tended to decrease with time in both groups, and the mean Delta R1 was significantly lower in the CS-scaffold group than in the control group at all time points. Delta R1 - 1/r (R1post -R1pre), where r is the relaxivity of Gd-DTPA(2-), R1 -1/T1 (longitudinal relaxation time). Conclusion: T2 mapping and dGEMRIC were both effective for evaluating tissue repair after allograft chondrocyte implantation. DR1 values of dGEMRIC represented good correlation with histologically and biochemically even at early stages after the implantation. (C) 2014 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Objectives. To document the reliability of Abe's classification and to clarify the predictive factors for acetabular labral lesions in osteoarthritis of the hip with radial magnetic resonance (MR) imaging. Methods. Reliability trial for the classification of acetabular labral lesion was performed by six orthopedic surgeons, grading 20 radial MR images in a blinded fashion at an interval of 4-5 weeks. Radial MR images of 275 hips in 263 patients were prospectively analyzed to determine the relationship between acetabular labral lesions, their distribution, age, and the acetabular coverage. Results. Cohen's quadratic weighted kappa of inter-observer reliability was 0.784 for the grade and 0.812 for the shape category. The weighted kappa of intra-observer reliability was 0.852 for the grade and 0.90 for the shape category. Multiple regression analysis revealed that both the grade and the shape were associated with age, acetabular coverage, and location of the labrum. Conclusions. Abe's classification of labral lesions was reliable for both the grade and shape categories. Aging, acetabular dysplasia, and the anterosuperior portion would be predictive factors for degeneration of the acetabular labrum using radial MR imaging.

人工膝関節手術の情報管理としてRadio frequency identification技術の応用を提案し、そのRFタグをUltra-high molecular weight polyethylene(UHMPE)インサートに埋め込んだ際の読み取りの可否を献体膝で検証した。RFタグはμ-ChipとFerVID familyを使用し、GUR1050を最高温度220℃、最高圧力245kgf/cm2でインサートの側面に埋め込まれるように成型した。リーダライタにより通信感度と通信可能距離を測定したところ、献体膝内に挿入することでμ-Chipの通信感度は-60.0dBmから-71.6dBmに、FerVID familyは-37.9dBmから-55.2dBmに低下し、大腿骨コンポーネントではCo-Cr-MoよりもTi-6Al-4Vの方が低かった。通信可能距離は、μ-Chipが200mmから50mmに、FerVID familyは310mmから100mmに低下した。いずれも挿入後は低下したが、実際の読み取りには十分な通信性能を有していると考えられた。

This study investigated the effect of MMP-13 gene knock down on cartilage degradation by injecting small interfering RNA (siRNA) into knee joints in a mouse model of osteoarthritis (OA). OA was induced in male C57BL/6 mice by destabilization of medial meniscus (DMM) surgery. Change of Mmp13 expression over time was determined by qPCR analysis from 3 days to 6 weeks after surgery. Mmp13 and control chemically modified siRNA were injected into the knee joint 1 week after surgery and expression levels were assessed in synovium by qPCR 48 h later. Cartilage degradation was histologically assessed 8 weeks after DMM surgery according to OARSI recommendations. Mmp13 expression levels were elevated 1 week after surgery and peaked at 77 fold at 2 weeks compared to expression at 3 days. A 55% decrease of Mmp13 levels in cartilage was observed 48 h after injection of Mmp13 siRNA (p = 0.05). Significant reduction in the histological score at 8 weeks after surgery was observed in the Mmp13 siRNA-treated group compared to the control siRNA group (p < 0.001). Intra-articular injection of Mmp13 siRNA at the early phase of OA development resulted in effective knock down of Mmp13 expression and delay in cartilage degradation in vivo. (C) 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.