Masataka Deie

Expression of MicroRNA-146a in osteoarthritis cartilage.

OBJECTIVE A role of microRNA, which are approximately 22-nucleotide noncoding RNAs, has recently been recognized in human diseases. The objective of this study was to identify the expression pattern of microRNA-146a (miR-146a) in cartilage from patients with osteoarthritis (OA). METHODS The expression of miR-146a in cartilage from 15 patients with OA was analyzed by quantitative reverse transcription-polymerase chain reaction (RT-PCR) and by in situ hybridization. Induction of the expression of miR-146a by cultures of normal human articular chondrocytes following stimulation with interleukin-1beta (IL-1beta) was examined by quantitative RT-PCR. RESULTS All cartilage samples were divided into 3 groups according to a modification of the Mankin score (grade I = mild OA scored 0-5, grade II = moderate OA scored 6-10, and grade III = severe OA scored 11-14). In grade I OA cartilage samples, the expression of miR-146a and COL2A1 was significantly higher than that in the other groups (P < 0.05). In grades II and III OA cartilage, the expression of miR-146a and COL2A1 was decreased, whereas the expression of matrix metalloproteinase 13 (MMP-13) was elevated in grade II OA cartilage. These data showed that miR-146a is expressed intensely in cartilage with a low Mankin grade and that miR-146a expression decreases in parallel with the level of MMP-13 expression. Tissue section in situ hybridization of primary miR-146a (pri-miR-146a) revealed that pri-miR-146a was expressed in chondrocytes residing in all tissue layers, especially in the superficial layer, where it was intensely expressed. The expression of miR-146 was markedly elevated by IL-1beta stimulation in human chondrocytes in vitro. CONCLUSION This study shows that miR-146 is intensely expressed in low-grade OA cartilage and that its expression is induced by stimulation of IL-1beta. Thus, miR-146 might play a role in OA cartilage pathogenesis.

Reconstruction of the medial patellofemoral ligament for the treatment of habitual or recurrent dislocation of the patella in children

We investigated the clinical outcome of a reconstructive procedure of the medial patellofemoral ligament for the treatment of habitual or recurrent dislocation of the patella in four children (6 knees), with a minimum follow-up of four years. The technique involves transfer of the tendon of semitendinosus to the patella using the posterior one-third of the femoral insertion of the medial collateral ligament as a pulley. There was no recurrence of dislocation after surgery. The mean Kujala score at follow-up was 96.3 points. Radiological assessment showed that the congruence angle, the tilt angle and the lateral shift radio were restored to normal. The lateral and medial stress shift ratios and the Insall-Salvati ratio remained abnormal. We conclude that this technique can be recommended for the treatment of habitual or recurrent patellar dislocation in children, although hypermobility and patella alta are not fully corrected.

MicroRNA-146a expresses in interleukin-17 producing T cells in rheumatoid arthritis patients

BackgroundInterleukin (IL)-17 is an important factor in rheumatoid arthritis (RA) pathogenesis. MicroRNA (miRNA)s are a family of non coding RNAs and associated with human diseases including RA. The purpose of this study is to identify the miRNAs in the differentiation of IL-17 producing cells, and analyze their expression pattern in the peripheral blood mononuclear cells (PBMC) and synovium from RA patients.MethodsIL-17 producing cells were expanded from CD4+T cell. MiRNA microarray was performed to identify the miRNAs in the differentiation of IL-17 producing cells. Quantitative polymerase chain reaction was performed to examine the expression patterns of the identified miRNAs in the PBMC and synovium from RA and osteoarthritis (OA) patients. Double staining combining in situ hybridization and immunohistochemistry of IL-17 was performed to analyze the expression pattern of identified miRNA in the synovium.ResultsSix miRNAs, let-7a, miR-26, miR-146a/b, miR-150, and miR-155 were significantly up regulated in the IL-17 producing T cells. The expression of miR-146a and IL-17 was higher than in PBMC in the patients with low score of Larsen grade and short disease duration. MiR-146a intensely expressed in RA synovium in comparison to OA. MiR-146a expressed intensely in the synovium with hyperplasia and high expression of IL-17 from the patients with high disease activity. Double staining revealed that miR-146a expressed in IL-17 expressing cells.ConclusionThese results indicated that miR-146a was associated with IL-17 expression in the PBMC and synovium in RA patients. There is the possibility that miR-146a participates in the IL-17 expression.

A Minimum 2-Year Follow-up After Selective Anteromedial or Posterolateral Bundle Anterior Cruciate Ligament Reconstruction

PURPOSE The purpose of this study was to investigate the 2-year follow-up clinical results of 45 patients who had undergone our anterior cruciate ligament (ACL) augmentation procedure using an autogenous semitendinosus tendon and the EndoButton CL (Smith & Nephew, Andover, MA). METHODS We followed up 45 patients for more than 2 years after their ACL augmentations. Of these, 37 were anteromedial (AM) bundle reconstructions and 8 were posterolateral (PL) bundle reconstructions. They were assessed using a KT-2000 knee arthrometer (MEDmetric, San Diego, CA) at 30 lb by joint position sense, Lysholm knee score, and magnetic resonance imaging (MRI). RESULTS The mean side-to-side difference of anterior displacement measured by the KT-2000 knee arthrometer at 30 degrees of knee flexion preoperatively was 3.3 +/- 2.4 mm, significantly improving to a mean of 0.5 +/- 2.7 mm 2 years after surgery. The preoperative joint position sense inaccuracy was 1.6 degrees +/- 1.8 degrees , which improved significantly to 0.3 degrees +/- 2.0 degrees after surgery. The median Lysholm knee score significantly improved from 74 (range, 44 to 95) to 100 points (range, 81 to 100) after surgery. In 20 of 29 patients who had the postoperative MRI examination, the augmented ACL resembled 1 bundle on the sagittal planes of the postoperative MRI. CONCLUSIONS Selective AM or PL bundle reconstruction showed improved joint stability, joint position sense, and Lysholm scores postoperatively. This procedure can be a treatment option for patients whose ACL remnants are left in certain conditions.

A Novel Cell Delivery System Using Magnetically Labeled Mesenchymal Stem Cells and an External Magnetic Device for Clinical Cartilage Repair

PURPOSE The purpose of this study was to investigate whether it is possible to successfully accumulate magnetically labeled mesenchymal stem cells (MSCs), under the direction of an external magnetic force, to the desired portion of osteochondral defects of the patellae after intra-articular injection of the MSCs. METHODS MSCs were cultured from bone marrow and were labeled magnetically. Osteochondral defects were made in the center of rabbit and swine patellae, and magnetically labeled MSCs were injected into the knee joints either under the direction of an external magnetic force or with no magnetic force applied. In the rabbit model we evaluated the patellae macroscopically and histologically, and in the swine model we observed the patellae arthroscopically. RESULTS Accumulation of magnetically labeled MSCs to the osteochondral defect was shown macroscopically and histologically in the rabbit model and was shown by arthroscopic observation to be attached to the chondral defect in the swine model. CONCLUSIONS We showed the ability to deliver magnetically labeled MSCs to a desired place in the knee joint. CLINICAL RELEVANCE Our novel approach is applicable for human cartilage defects and may open a new era of repairing cartilage defects caused by osteoarthritis or trauma by use of a less invasive technique.

Effects of transforming growth factor-β 1 on the early stages of healing of the Achilles tendon in a rat model

We studied the effects of transforming growth factor-β 1 (TGF-β 1) on the genetic expression of procollagen type I and III and its effects on structural properties in the early stages of healing in rat Achilles tendon. The Achilles tendons in 90 rats were transsected and repaired immediately. TGF-β 1 dissolved in phosphate-buffered saline was injected locally at the repair site using two different doses, and outcomes in both groups were compared to that in the control group given phosphate-buffered saline only. Five animals in each group were killed at one, two, and four weeks postoperatively, and the healing tendon was evaluated. A dose-dependent increase in the expression of procollagen type I and III mRNA was found one week postoperatively. The failure load and stiffness of the healing tendon were increased by TGF-β 1 at two and four weeks.

Tendon-Bone Insertion Repair and Regeneration Using Polyglycolic Acid Sheet in the Rabbit Rotator Cuff Injury Model

Background The quality of tendons has considerable limitations regarding torn rotator cuff tendons. Tissue-engineering techniques using a biodegradable scaffold offer potential alternatives for recreating a valid tendon-to-bone interface. Hypothesis A polyglycolic acid (PGA) sheet could facilitate the regeneration of the rotator cuff tendon insertion in vivo. Study Design Controlled laboratory study. Methods An implant consisting of a PGA sheet, a rapidly absorbable material, was used to replace a completely resected infraspinatus tendon insertion in 33 adult Japanese white rabbits. The contralateral infraspinatus tendon was replaced by poly-L-lactate-epsilon-caprolactone (PLC), a slowly absorbable material, by the same methods based on the results of the pilot study. Histological comparisons were made at 4, 8, and 16 weeks, and mechanical evaluations were performed at 4 and 16 weeks in both groups. Unrepaired defects were created in a control group. Results: In the control group, the rotator cuff defects were covered with thin fibrous membranes with many fibroblasts arranged in an irregular pattern. In the PLC group, some chondrocytes were seen in the tendon insertion; however, these were not arranged along the long axis for a 16-week period. In the PGA group, a well-arranged fibrocartilage layer could be found in the regenerated tendon insertions; however, these tendon insertions were mainly regenerated by type III collagen. In mechanical examinations, the PGA group had significantly higher values in the maximum failure load, tensile strength, and Youngs modulus for the 4-week and 16-week periods. These 3 categories statistically improved from 4 to 16 weeks postoperatively in both groups except for the Youngs modulus in the PGA group (E = 5.66 at 4 weeks to 5.53 at 16 weeks). Conclusion The PGA sheet scaffold material allows for tendon insertion regeneration with a fibrocartilage layer but displays mechanical properties inferior to those of the normal tendon in an animal model. Clinical Relevance The PGA sheet represent a possible alternative scaffold material for tendon regeneration in rotator cuff repair.

Potential Risks of Femoral Tunnel Drilling Through the Far Anteromedial Portal: A Cadaveric Study

PURPOSE The purpose of this study was to estimate the potential risks when drilling femoral tunnels through the far anteromedial portal in double-bundle anterior cruciate ligament reconstruction in cadaveric knees. METHODS Ten cadaveric knees were used. We drilled the anteromedial bundle (AMB) and posterolateral bundle (PLB) through the far anteromedial portal at 3 different knee flexion angles: 70 degrees, 90 degrees, and 110 degrees. We measured the shortest distance to the common peroneal nerve and the posterior articular cartilage of the lateral femoral condyle and the femoral tunnel length. RESULTS At 70 degrees, the distance to the nerve was less than 10 mm in 7 AMB cases and in 9 PLB cases, and the distance to the cartilage was less than 10 mm in all the AMB and PLB cases. At 90 degrees, the distance to the nerve was less than 10 mm in 1 AMB and 5 PLBs, and the distance to the cartilage was less than 10 mm in 2 AMBs and all the PLBs. On the other hand, at 110 degrees , the distance to the nerve was greater than 10 mm in all the AMBs and PLBs, and the distance to the cartilage did not exceed 10 mm in just 2 of the PLBs. CONCLUSIONS In our cadaveric study we found that the low knee flexion angles when drilling femoral tunnels through the far anteromedial portal might have the potential risks of damage to the common peroneal nerve and the posterior articular cartilage, and the risks would be decreased at higher degrees of knee flexion. However, we found there was a 20% risk of damage to the cartilage while drilling the PLB at 110 degrees. CLINICAL RELEVANCE High knee flexion angles are recommended to avoid damage to the nerve and the cartilage when drilling femoral tunnels through the far anteromedial portal in double-bundle anterior cruciate ligament reconstruction.

Acceleration of skeletal muscle regeneration in a rat skeletal muscle injury model by local injection of human peripheral blood-derived CD133-positive cells.

Muscle injuries in sport activities can pose challenging problems in traumatology and sports medicine. The best treatment for muscle injury has not been clearly established except for the conservative treatment that is routinely performed. We investigated the potential of human adult CD133+ cells to contribute to skeletal muscle regeneration in an athymic rat model. We tested whether CD133+ cells locally transplanted to the skeletal muscle lacerated models could (a) induce vasculogenesis/angiogenesis, (b) differentiate into endothelial and myogenic lineages, and (c) finally promote histological and functional skeletal myogenesis. Granulocyte colony stimulating factor‐mobilized peripheral blood (PB) CD133+ cells, PB mononuclear cells, or phosphate‐buffered saline was locally injected after creating a muscle laceration in the tibialis anterior muscle in athymic rats. After treatment, histological and functional skeletal myogenesis was observed significantly in the CD133+ group. The injected CD133+ cells differentiated into endothelial and myogenic lineages. Using real‐time polymerase chain reaction analysis, we found that the gene expressions related to microenvironment conduction for host angiogenesis, fibrosis, and myogenesis were ideally up/downregulated. Our results show that CD133+ cells have the potential to enhance the histological and functional recovery from skeletal muscle injury rather via indirect contribution to environment conduction for muscular regeneration. It would be relatively easy to purify this cell fraction from PB, which could be a feasible and attractive autologous candidate for skeletal muscle injuries in a clinical setting. These advantages could accelerate the progression of cell‐based therapies for skeletal muscle injuries from laboratory to clinical implementation. STEM CELLS 2009;27:949–960

Medial Patellofemoral Ligament Reconstruction Fixed With a Cylindrical Bone Plug and a Grafted Semitendinosus Tendon at the Original Femoral Site for Recurrent Patellar Dislocation

Background: The medial patellofemoral ligament (MPFL) is the most important factor for stabilizing the patella and preventing lateral patellar dislocation. Medial patellofemoral ligament reconstruction is an accepted surgical technique to restore patellofemoral stability after lateral patellar dislocation. The authors recently developed a new anatomical MPFL reconstruction method using a cylindrical bone plug and grafted semitendinosus tendon at the anatomical femoral attachment site to mimic the native MPFL. This study evaluated the new technique for stabilizing recurrent patellar dislocation. Hypothesis: This new MPFL reconstruction technique will improve knee symptoms and function with excellent clinical results. Study Design: Case series; Level of evidence, 4. Method: Thirty-one knees were evaluated from 29 cases of recurrent patellar dislocation that were surgically treated using the anatomical MPFL reconstruction technique. The average patient age was 22.2 years (range, 12-34 years); postsurgery follow-up was 2 to 5 years (average, 3.2 years). The patients were clinically evaluated based on the Kujala score, range of motion, and signs of apprehension. The Merchant view was used to measure congruence and tilting angles. Results: Of the 31 knees, 30 showed good clinical results after surgery, while 1 patient showed remaining signs of apprehension. The Kujala score improved from an average of 64 points (range, 35-70) initially to an average of 94.5 points (range, 79-100) at the final follow-up. Range of motion improved for all patients, with an average knee extension of 0° ± 2° and knee flexion of 145° ± 3° at final follow-up. No patellar redislocation was reported. Radiological assessment indicated significant improvement to the congruence angle from 13° ± 4° before surgery to −5° ± 5° at the final follow-up, while the tilting angle went from 8° ± 7° before surgery to 7° ± 4° at the final follow-up. Conclusion: This study demonstrated excellent results using the new procedure for recurrent dislocation of the patella, with instability in only 1 of 31 knees (3.2%).