Masataka Fujii

Tensile strain increases expression of CCN2 and COL2A1 by activating TGF-β-Smad2/3 pathway in chondrocytic cells

Physiologic mechanical stress stimulates expression of chondrogenic genes, such as multifunctional growth factor CYR61/CTGF/NOV (CCN) 2 and α1(II) collagen (COL2A1), and maintains cartilage homeostasis. In our previous studies, cyclic tensile strain (CTS) induces nuclear translocation of transforming growth factor (TGF)-β receptor-regulated Smad2/3 and the master chondrogenic transcription factor Sry-type HMG box (SOX) 9. However, the precise mechanism of stretch-mediated Smad activation remains unclear in transcriptional regulation of CCN2 and COL2A1. Here we hypothesized that CTS may induce TGF-β1 release and stimulate Smad-dependent chondrogenic gene expression in human chondrocytic SW1353 cells. Uni-axial CTS (0.5Hz, 5% strain) stimulated gene expression of CCN2 and COL2A1 in SW1353 cells, and induced TGF-β1 secretion. CCN2 synthesis and nuclear translocalization of Smad2/3 and SOX9 were stimulated by CTS. In addition, CTS increased the complex formation between phosphorylated Smad2/3 and SOX9. The CCN2 promoter activity was cooperatively enhanced by CTS and Smad3 in luciferase reporter assay. Chromatin immunoprecipitation revealed that CTS increased Smad2/3 interaction with the CCN2 promoter and the COL2A1 enhancer. Our results suggest that CTS epigenetically stimulates CCN2 transcription via TGF-β1 release associated with Smad2/3 activation and enhances COL2A1 expression through the complex formation between SOX9 and Smad2/3.

Postoperative change in medial meniscal length in concurrent all-inside meniscus repair with anterior cruciate ligament reconstruction

PurposeMeniscus repair can restore meniscal function that transfers the axial compressive force to circumferential tensile strain. However, few reports have investigated the relationship between concurrent meniscus repair with acute anterior cruciate ligament (ACL) reconstruction and postoperative meniscal position. This study aimed to evaluate medial meniscal size and clinical results in patients who underwent ACL reconstruction and concomitant all-inside medial meniscus repair.MethodsTwenty patients underwent ACL reconstruction and concurrent medial meniscus repair of a peripheral longitudinal tear using the FasT-Fix meniscal repair device. Medial tibial plateau length (MTPL) and width (MTPW) were determined by radiographic images. We evaluated the Lysholm score, anteroposterior instability, meniscal healing and magnetic resonance imaging (MRI)-based medial meniscal length (MML) and width (MMW). Correlations between MRI-based meniscal size, radiographic measurement and height were investigated.ResultsAll patients showed complete healing of the repaired meniscus in arthroscopic evaluation. However, one patient needed a subsequent meniscus repair during the follow-up period. Lysholm score and anteroposterior instability improved significantly. A better correlation was observed between MMW and MTPW than between MML and MTPL. Concurrent all-inside medial meniscus repair with ACL reconstruction significantly increased MML percentage (%MML) (100 MML/MTPL) but did not affect MMW percentage (%MMW) (100 MMW/MTPW).ConclusionsConcurrent all-inside medial meniscus repair with ACL reconstruction had satisfactory clinical results. %MML was increased by concurrent medial meniscus repair without affecting %MMW. Our results suggest that medial meniscus repair associated with ACL reconstruction may restore meniscal function by adjusting the anteroposterior length of the torn medial meniscus.

Chondromodulin-I derived from the inner meniscus prevents endothelial cell proliferation†

The meniscus is a fibrocartilaginous tissue that plays an important role in controlling complex biomechanics of the knee. A perimeniscal capillary plexus supplies the outer meniscus, whereas the inner meniscus is composed of avascular tissue. Anti‐angiogenic molecules, such as chondromodulin‐I (ChM‐I) and endostatin, have pivotal roles in preserving the avascularity of cartilage. However, the anti‐angiogenic role of ChM‐I is unclear in the meniscus. We hypothesized that the inner meniscus might maintain its avascular feature by expressing ChM‐I. Immunohistochemical analyses revealed that ChM‐I was mainly detected in the inner and superficial zones of the meniscus. On the other hand, endostatin distribution was similar between the inner and outer meniscus. In Western blot, ChM‐I was detected only in the inner meniscus, whereas endostatin was equally observed in both inner and outer menisci. In addition, ChM‐I concentration of the inner meniscus‐derived conditioned medium was higher than that of the outer meniscus‐derived medium. ChM‐I removal from the inner meniscus‐derived medium and functional blocking of ChM‐I significantly increased endothelial cell proliferation. In this study, we demonstrated that the inner meniscus contained larger amounts of ChM‐I, and that the inner meniscus‐derived ChM‐I inhibited endothelial cell proliferation. Our results suggest that ChM‐I may be a key anti‐angiogenic factor for maintaining the avascularity of the inner meniscus.

Postoperative change in the length and extrusion of the medial meniscus after anterior cruciate ligament reconstruction

PurposeThe medial meniscus is a secondary stabilizer of anterior tibial translation in anterior cruciate ligament (ACL)-deficient knees. ACL reconstruction effectively restores an increased anterior tibial translation in the ACL-deficient knee. However, knee osteoarthritis sometimes develops in ACL-reconstructed patients during a long-term follow-up period. We hypothesized that the medial meniscal position would be different between the ACL-deficient and reconstructed knees. The aim of this study was to investigate pre-operative and postoperative location of the medial meniscus in patients who underwent ACL reconstruction.MethodsACL-reconstructed knees (28 knees) and normal knees (27 knees) were investigated. Medial tibial plateau length (MTPL) and medial tibial plateau width (MTPW) were determined using radiographic images. Magnetic resonance imaging (MRI)-based medial meniscal length (MML), medial meniscal width (MMW), and medial meniscal extrusion (MME) were measured. Postoperative change in the MML, MMW, and MME were evaluated and compared with those in normal knees.ResultsNo significant differences between the ACL-deficient (pre-operative) and normal groups were noted. The ACL-reconstructed (postoperative) group showed an increase in the MML, in the percentage of the MML (%MML = 100 MML/MTPL), and in the MME. Significant differences between postoperative and normal groups were observed in the MML, %MML, and MME. MMW and MMW percentage (100 MMW/MTPW) were similar in all groups.ConclusionsThe anteroposterior length and radial extrusion of the medial meniscus increased after ACL reconstruction. Transposition of the medial meniscus may be a possible cause of developing further degenerative knee joint disorders after ACL reconstruction.

Pullout repair of a medial meniscus posterior root tear using a FasT-Fix ® all-inside suture technique

A medial meniscus posterior root tear (MMPRT) may increase the tibiofemoral contact pressure by decreasing the tibiofemoral contact area. Meniscal dysfunction induced by posterior root injury may lead to the development of osteoarthritic knees. Repair of a MMPRT can restore medial meniscus (MM) function and prevent knee osteoarthritis progression. Several surgical procedures have been reported for treating a MMPRT. However, these procedures are associated with several technical difficulties. Here, we describe a technique to stabilize a torn MM posterior root using the FasT-Fix® all-inside meniscal suture device and a new aiming device. The uncut free-end of the FasT-Fix® suture can be used as a thread for transtibial pullout repair. Our procedure might help overcome the technical difficulties in arthroscopic treatment of a MMPRT.

Comparison between normal and loose fragment chondrocytes in proliferation and redifferentiation potential

PurposeLoose fragments in osteochondritis dissecans (OCD) of the knee require internal fixation. On the other hand, loose fragments derived from spontaneous osteonecrosis of the knee (SONK) are usually removed. However, the difference in healing potential between OCD- and SONK-related loose fragments has not been elucidated. In this study, we investigated proliferative activity and redifferentiation potential of normal cartilage-derived and loose fragment-derived chondrocytes.MethodsCells were prepared from normal articular cartilages and loose fragment cartilages derived from knee OCD and SONK. Cellular proliferation was compared. Redifferentiation ability of pellet-cultured chondrocytes was assessed by real-time PCR analyses. Mesenchymal differentiation potential was investigated by histological analyses. Positive ratio of a stem cell marker CD166 was evaluated in each cartilaginous tissue.ResultsNormal and OCD chondrocytes showed a higher proliferative activity than SONK chondrocytes. Chondrogenic pellets derived from normal and OCD chondrocytes produced a larger amount of safranin O-stained proteoglycans compared with SONK-derived pellets. Expression of chondrogenic marker genes was inferior in SONK pellets. The CD166-positive ratio was higher in normal cartilages and OCD loose fragments than in SONK loose fragments.ConclusionsThe OCD chondrocytes maintained higher proliferative activity and redifferentiation potential compared with SONK chondrocytes. Our results suggest that chondrogenic properties of loose fragment-derived cells and the amount of CD166-positive cells may affect the repair process of osteochondral defects.

The anterior cruciate ligament-lateral meniscus complex: A histological study

ABSTRACT The anterior root of the lateral meniscus (LM) dives underneath the tibial attachment of the anterior cruciate ligament (ACL). Although the distinct role of meniscal attachments has been investigated, the relationship between the LM anterior insertion (LMAI) and ACL tibial insertion (ACLTI) remains unclear. This study histologically analyzed the LMAI and ACLTI. Samples were divided into four regions in an anterior-to-posterior direction. Histological measurements of these insertion sites were performed using safranin O-stained coronal sections. Distribution and signal densities of type I and II collagen were quantified. The ACLTI and LMAI formed the ACL–LM complex via fiber connections. The anterior part of the ACLTI had a widespread attachment composed of dense fibers. Attachment fibers of the LMAI became dense and wide gradually at the middle-to-posterior region. The ACL–LM transition zone (ALTZ) was observed between the LMAI and the lateral border of the ACLTI at the middle part of the ACL tibial footprint. Type II collagen density of the LMAI was higher than that of the ACLTI and ALTZ. Our results can help create an accurate tibial bone tunnel within the dense ACL attachment during ACL reconstruction surgery.

Location of the tibial tunnel aperture affects extrusion of the lateral meniscus following reconstruction of the anterior cruciate ligament

The anterior root of the lateral meniscus provides functional stability to the meniscus. In this study, we evaluated the relationship between the position of the tibial tunnel and extrusion of the lateral meniscus after anterior cruciate ligament reconstruction, where extrusion provides a proxy measure of injury to the anterior root. The relationship between extrusion and tibial tunnel location was retrospectively evaluated from computed tomography and magnetic resonance images of 26 reconstructed knees, contributed by 25 patients aged 17–31 years. A measurement grid was used to localize the position of the tibial tunnel based on anatomical landmarks identified from the three‐dimensional reconstruction of axial computed tomography images of the tibial plateaus. The reference point‐to‐tibial tunnel distance (mm) was defined as the distance from the midpoint of the lateral edge of the grid to the posterolateral aspect of the tunnel aperture. The optimal cutoff of this distance to minimize post‐operative extrusion was identified using receiver operating curve analysis. Extrusion of the lateral meniscus was positively correlated to the reference point‐to‐tibial tunnel distance (r 2 = 0.64; p < 0.001), with a cutoff distance of 5 mm having a sensitivity to extrusion of 83% and specificity of 93%. The mean extrusion for a distance >5 mm was 0.40 ± 0.43 mm, compared to 1.40 ± 0.51 mm for a distance ≤5 mm (p < 0.001). Therefore, a posterolateral location of the tibial tunnel aperture within the footprint of the anterior cruciate ligament decreases the reference point‐to‐tibial tunnel distance and increases extrusion of the lateral meniscus post‐reconstruction.

A new aiming guide can create the tibial tunnel at favorable position in transtibial pullout repair for the medial meniscus posterior root tear

INTRODUCTION Injuries to the medial meniscus (MM) posterior root lead to accelerated cartilage degeneration of the knee. An anatomic placement of the MM posterior root attachment is considered to be critical in transtibial pullout repair of the medial meniscus posterior root tear (MMPRT). However, tibial tunnel creation at the anatomic attachment of the MM posterior root is technically difficult using a conventional aiming device. The aim of this study was to compare two aiming guides. We hypothesized that a newly-developed guide, specifically designed, creates the tibial tunnel at an adequate position rather than a conventional device. MATERIALS AND METHODS Twenty-six patients underwent transtibial pullout repairs. Tibial tunnel creation was performed using the Multi-use guide (8 cases) or the PRT guide that had a narrow twisting/curving shape (18 cases). Three-dimensional computed tomography images of the tibial surface were evaluated using the Tsukadas measurement method postoperatively. Expected anatomic center of the MM posterior root attachment and tibial tunnel center were evaluated using the percentage-based posterolateral location on the tibial surface. Percentage distance between anatomic center and tunnel center was calculated. RESULTS Anatomic center of the MM posterior root footprint located at a position of 78.5% posterior and 39.4% lateral. Both tunnels were anteromedial but tibial tunnel center located at a more favorable position in the PRT group: percentage distance was significantly smaller in the PRT guide group (8.7%) than in the Multi-use guide group (13.1%). DISCUSSION The PRT guide may have great advantage to achieve a more anatomic location of the tibial tunnel in MMPRT pullout repair. LEVEL OF EVIDENCE III.

The figure-of-nine leg position for anatomic anterior cruciate ligament reconstruction.

Anatomic double-bundle anterior cruciate ligament (ACL) reconstruction can restore the function and kinematics of the knee in ACL-deficient patients. Several outside-in drilling systems for accurate femoral tunnel creations have been developed. However, the femoral tunnel creation at the lower position of the intercondylar notch can be difficult in a usual leg position with the knee flexed at 90° without varus stress. This technical note describes that the figure-of-nine leg position provides a better arthroscopic view to safely clean up the ACL femoral footprint located at the lower area of the lateral intercondylar wall. This position is useful to create the optimal femoral tunnels using the outside-in drilling technique, without damaging the lateral meniscus posterior root, lateral tibial eminence, and supplemental fibers that bridge the gap between the lateral meniscus and the ACL tibial insertion.