Wirat Kongcharoensombat

Silencing microRNA-34a inhibits chondrocyte apoptosis in a rat osteoarthritis model in vitro

OBJECTIVE miRNAs, which are non-coding RNAs, play a role in the pathogenesis of disease including OA. miRNA (miR)-34a is induced by p53, subsequently leading to cell apoptosis, which is one of the major factors in the pathogenesis of OA. The purpose of this study is to investigate the effect of silencing miR-34a on IL-1β-induced chondrocyte apoptosis in a rat OA model in vitro. METHODS Locked nucleotide analogue (LNA)-modified miR-34a-specific anti-sense was transfected into rat chondrocyte monolayer culture. After that, IL-1β was added to the chondrocytes to create an OA model in vitro. The effect of silencing miR-34a on the prevention of chondrocyte apoptosis was analysed by assessment of the expression levels of Col2a1 and iNOS, also through assessment of cell viability and TUNEL staining. RESULTS The expression of miR-34a was significantly up-regulated by IL-1β. Silencing of miR-34a significantly prevented IL-1β-induced down-regulation of Col2a1, as well as IL-1β-induced up-regulation of iNOS. Finally, MiR-34a inhibitor could also reduce TUNEL-positive cells. CONCLUSION Silencing of miR-34a by LNA-modified anti-sense could effectively reduce rat chondrocyte apoptosis induced by IL-1β. This present study revealed that silencing of miR-34a might develop a novel intervention for OA treatment through the prevention of cartilage degradation.

Articular Cartilage Repair With Magnetic Mesenchymal Stem Cells

Background: Cell therapies are hampered by the difficulty of delivering cells to and retaining them in target tissues long enough to repair or regenerate local tissues. Hypothesis: Magnetic-assisted delivery of magnetically labeled mesenchymal stem cells (m-MSCs) would be rapid, allowing for chondrogenic differentiation and functional joint repair without replacement. Study Design: Controlled laboratory study. Methods: Sixteen mini-pigs aged 6 to 7 months were used. A full-thickness cartilage defect was created in the center of the patella with a cylindrical punch (diameter, 6 mm). At 4 weeks after creation of the cartilage defects, the animals were divided into 3 treatment groups: In the M group, m-MSCs (5 × 106 cells) were injected and accumulated to the cartilage defect using an external magnetic force (1.5 T) for 10 minutes; in the G group, the patella was faced upward, filled with MSCs (5 × 106 cells), and held for 10 minutes; and in the C group, only phosphate-buffered saline was injected. The regenerated cartilage was evaluated in 5 knees in each of the 3 groups by arthroscopic surgery at 6 and 12 weeks and histological and ultrasound evaluation at 12 and 24 weeks. Results: The mean arthroscopic scores at 6 weeks were 10.4 ± 1.10 in the M group, 8.8 ± 0.84 in the G group, and 7.4 ± 0.89 in the C group. There was a statistically significant difference between the M group and the other 2 groups. The mean arthroscopic scores at 12 weeks were 12.8 ± 1.30 (M group), 10.5 ± 1.30 (G group), and 9.5 ± 0.58 (C group), with a statistically significant difference between the M and C groups. The mean histological scores using the Wakitani scoring system at 12 weeks were 2.8 ± 0.96 (M group), 5.4 ± 0.55 (G group), and 6.0 ± 2.20 (C group), and the mean histological scores at 24 weeks were 2.4 ± 1.50 (M group), 3.5 ± 0.56 (G group), and 5.3 ± 1.50 (C group). The mean histological scores at 12 weeks were significantly better in the M group than in the other groups, and the M group maintained a significantly better histological score than did the C group at 24 weeks. Conclusion: The m-MSCs had no adverse effect on chondrogenic differentiation, and m-MSCs delivered by magnetic field application repaired cartilage defects. Clinical Relevance: The clinical application of this novel stem cell delivery system is a potential therapeutic option for treating cartilage defects and may be more applicable throughout the body than traditional methods.

Double bundle arthroscopic Anterior Cruciate Ligament reconstruction with remnant preserving technique using a hamstring autograft

BackgroundPreservation of the Anterior Cruciate Ligament (ACL) remnant is important from the biological point of view as it enhances revascularization, and preserves the proprioceptive function of the graft construct. Additionally, it may have a useful biomechanical function. Double bundle ACL reconstruction has been shown to better replicate the native ACL anatomy and results in better restoration of the rotational stability than single bundle reconstruction.MethodsWe used the far anteromedial (FAM) portal for creation of the femoral tunnels, with a special technique for its preoperative localization using three dimensional (3D) CT. The central anteromedial (AM) portal was used to make a longitudinal slit in the ACL remnant to allow visualization of the tips of the guide pins during anatomical creation of the tibial tunnels within the native ACL tibial foot print. The use of curved hemostat allow retrieval of the wire loop from the apertures of the femoral tunnels through the longitudinal slit in the ACL remnant thereby, guarding against impingement of the reconstruction graft against the ACL remnant as well as the roof of the intercondylar notch.ConclusionOur technique allows for anatomical double bundle reconstruction of the ACL while maximally preserving the ACL remnant without the use of intra-operative image intensifier.

In vivo bioluminescence imaging of transplanted bone marrow mesenchymal stromal cells using a magnetic delivery system in a rat fracture model

For the treatment of ununited fractures, we developed a system of delivering magnetic labelled mesenchymal stromal cells (MSCs) using an extracorporeal magnetic device. In this study, we transplanted ferucarbotran-labelled and luciferase-positive bone marrow-derived MSCs into a non-healing femoral fracture rat model in the presence of a magnetic field. The biological fate of the transplanted MSCs was observed using luciferase-based bioluminescence imaging and we found that the number of MSC derived photons increased from day one to day three and thereafter decreased over time. The magnetic cell delivery system induced the accumulation of photons at the fracture site, while also retaining higher photon intensity from day three to week four. Furthermore, radiological and histological findings suggested improved callus formation and endochondral ossification. We therefore believe that this delivery system may be a promising option for bone regeneration.

The Transverse Ligament as a Landmark for Tibial Sagittal Insertions of the Anterior Cruciate Ligament: A Cadaveric Study

PURPOSE The purpose of this study was to determine the relation between the position of the transverse ligament, the anterior edge of the anterior cruciate ligament (ACL) tibial footprint, and the center of the ACL tibial insertion. We used arthroscopy for localization of the anatomic landmarks, followed by insertions of guide pins under direct visualization, and then the position of these guide pins was checked on plain lateral radiographs. METHODS The transverse ligament and the anterior aspect of the ACL tibial footprint were identified by arthroscopy in 20 unpaired cadaveric knees (10 left and 10 right). Guide pins were inserted with tibial ACL adapter drill guides under direct observation at the transverse ligament, the anterior aspect of the tibial footprint, and the center of tibial insertion of the ACL. Then, plain lateral radiographs of specimens were taken. The Amis and Jakob line was used to define the attachment of the ACL tibial insertion and the transverse ligament. A sagittal percentage of the location of the insertion point was determined and calculated from the anterior margin of the tibia in the anteroposterior direction. RESULTS The transverse ligament averaged 21.20% ± 4.1%, the anterior edge of the ACL tibial insertion averaged 21.60% ± 4.0%, and the center of the ACL tibial insertion averaged 40.30% ± 4.8%. There were similar percent variations between the transverse ligament and the anterior edge of the ACL tibial insertion, with no significant difference between them (P = .38). Intraobserver and interobserver reliability was high, with small standard errors of measurement. CONCLUSIONS This study shows that the transverse ligament coincides with the anterior edge of the ACL tibial footprint in the sagittal plane. CLINICAL RELEVANCE The transverse ligament can be considered as a new landmark for tibial tunnel positioning during anatomic ACL reconstruction.

Repair of a large osteochondral defect in the knee joint using autologous and artificial bone graft combined with motion preserving distraction arthroplasty: a case report

The biological reconstruction of a large osteochondral defect in the weight-bearing area of the knee joint has long been a challenge to orthopedic surgeons. We present a case of a large posttraumatic defect in the weight-bearing area of knee joint treated with a novel distraction arthroplasty device after reconstruction of the joint surface using combined autologous and artificial bone graft.

Knee Articulated Distraction Arthroplasty for the Middle-aged Osteoarthritic Knee Joint

Objective We developed a knee distraction arthroplasty device that allows continuous joint movement. The objective of this article is to show the surgical procedure of knee distraction arthroplasty with a bone marrow-stimulating technique, for treatment of osteoarthritis of the knee and to evaluate the clinical results. Methods As we showed in Arthroscopy in 2007, we performed this distraction arthroplasty to 6 knees (in 6 patients, aged 42 to 63 y). Then we compared preoperative findings with postoperative ones. The fixation period for the distraction device ranged from 7 to 12 weeks and the follow-up period ranged from 24 months to 53 months (average 36 mo). Results The Japan Orthopaedic Association knee score, range of motion, and the values of the joint spaces were significantly improved in all cases at the latest follow-up (P<0.05). Visual analog pain scales were also significantly improved (P<0.05). Conclusions We conclude that treatment using this arthroplasty device in combination with a bone marrow stimulating method is effective for osteoarthritic knees in middle-aged patients.

Augmentation Procedure for Partial Rupture of the Anterior Cruciate Ligament

Arthroscopic examination before anterior cruciate ligament (ACL) reconstruction shows the presence of several types of ACL remnants within the intercondylar notch. It is thought that 10% to 20% of the ACL injury cases represent a partial rupture of the ACL. In these cases, although rupture of the anteromedial or posterolateral bundle can be seen, the other bundle is preserved with an attachment of the anatomical femoral origin. We considered it beneficial to perform the ACL augmentation procedure without sacrificing the remaining remnant in terms of proprioception, biomechanical functions, and vascularization of the graft. This study will present the ACL augmentation procedure for patients with partial ACL rupture. We believe that the described technique can be a treatment option for patients whose ACL remnants are left in certain conditions.