Growth Differentiation Factor 5-Induced Mesenchymal Stromal Cells Enhance Tendon Healing.

Mesenchymal stromal cells (MSCs) have immense potential for use in musculoskeletal tissue regeneration; however, there is still a paucity of evidence on the effect of tenogenic MSCs (TMSCs) in tendon healing in vivo. This study aimed to determine the effects of growth differentiation factor 5 (GDF5)-induced rabbit MSCs (rbMSCs) on infraspinatus tendon healing in a New Zealand white rabbit model. In this study, bone marrow-derived rbMSCs were isolated, and 100 ng/mL GDF5 was used to induce tenogenic differentiation in rbMSC. The effects of GDF5 on rbMSC in vitro were assessed by total collagen assay, gene expression analysis, and immunofluorescence staining of tenogenic markers; native tenocytes isolated from rabbit tendon were used as a positive control. In in vivo, a window defect was created on the infraspinatus tendons bilaterally. After 3 weeks, the rabbits (n = 18) were randomly divided into six groups and repaired with various interventions: (1) surgical suture; (2) fibrin glue (FG); (3) suture and FG; (4) suture, FG, and rabbit tenocytes (rbTenocyte); (5) suture, FG, and rbMSCs, and (6) suture, FG, and TMSC. All animals were euthanized at 6 weeks postoperatively. The in vitro GDF5-induced rbMSCs (or TMSC) showed increased total collagen expression, augmented scleraxis (SCX), and type-I collagen (COL1A1) mRNA gene expression levels. Immunofluorescence showed similar expression in GDF5-induced rbMSC to that of rbTenocyte. In vivo histological analysis showed progressive tendon healing in the TMSC-treated group; cells with elongated nuclei aligned parallel to the collagen fibers, and the collagen fibers were in a more organized orientation, along with macroscopic evidence of tendon callus formation. Significant differences were observed in the cell-treated groups compared with the non-cell-treated groups. Histological scoring showed a significantly enhanced tendon healing in the TMSC- and rbMSC-treated groups compared with the rbTenocyte group. The SCX mRNA expression levels, at 6 weeks following repair, were significantly upregulated in the TMSC group. Immunofluorescence showed COL-1 bundles aligned in parallel orientation; this was further confirmed in atomic force microscopy imaging. SCX, TNC, and TNMD were detected in the TMSC group. In conclusion, GDF5 induces tenogenic differentiation in rbMSCs, and TMSC enhances tendon healing in vivo compared with conventional suture repair.

Are Two- and Three-Dimensional Computed Tomographic Scan Measurements of Tibial Tubercle-Trochlear Groove Distance Equivalent? A Comparative Study.

Background: Accurate planning for patellar instability correction is important in obtaining good post-operative outcome. The main challenge in the current two-dimensional (2-D) computed tomographic (CT) scans method is the difficulty in choosing reliable bony landmarks. This study aimed to compare the reliabilities between the 2-D and three-dimensional (3-D) methods of measuring tibial tubercle-trochlear groove (TT-TG) distance. We hypothesize that the proposed 3-D method will result in measurements with narrower error margin, providing higher reliability and accuracy. Materials and Methods: We traced CT scans of 106 knees with no patellofemoral pathology from 59 subjects from the database system and converted all 2-D images into 3-D models to determine the values for each parameter. We compared the intra- and interobserver reliability of each method using intraclass correlation (ICC) and Bland-Altman method. Results: The values of TT-TG measured by 2-D and 3-D methods were 16.1 ± 4.6 mm and 16.2 ± 4.2 mm, respectively. The ICC values of both methods were comparable (95% limits of agreement between the same observer: - 3.3 to 3.8 mm versus - 2.4 to 2.7 mm and different observers: - 4.3 to 4.9 mm versus - 3.9 to 2.7 mm), with 3-D method results in narrower limits of agreement. Conclusion: TT-TG measurement is reliable using the 2-D method without using advanced radiographic software. The 3-D method of measuring TT-TG provides measurement with narrower variation when compared with the 2-D method. However, both TT-TG distances' measurement methods in the current study were comparable as the variations are not significant.

Variation in Kaplan fiber insertion to the distal femur and surgical implications: A cadaveric anatomical study comparing Asian and Caucasian knees.

BACKGROUND: The iliotibial band together with its attachment to the distal femur, known eponymously as the Kaplan fibers, has been shown to contribute to anterolateral rotatory stability of the knee. However, there remains paucity of data regarding the detailed anatomy of the Kaplan fibers to aid us in anatomical-based surgical reconstruction of these structures. The aim of our study was to identify and compare the presence and quantitative anatomy of the Kaplan fibers in Caucasian and Asian cadavers. METHODS: Twenty paired fresh-frozen Asian and 20 unpaired embalmed Caucasian cadaveric knees were dissected using a standard outside-in technique. The presence, qualitative and quantitative measurements of any distal femoral attachments of the iliotibial band were recorded. Mean values of its dimensions were calculated and compared between the Caucasian and Asian groups. RESULTS: A consistent supracondylar attachment of the iliotibial band was found in all Caucasian knees and in 19 of 20 Asian knees. The fibers were noted to have two variations with regard to their insertion to the distal femur, either as a single limb or as a double limb attachment, averaging between 30 mm and 40 mm from the lateral femoral epicondyle. No statistically significant difference was noted in dimensions of the fibers between the Caucasian and Asian groups. CONCLUSIONS: The Kaplan fibers are a consistent attachment of the iliotibial band at the distal femur in both Caucasian and Asian knees with two different patterns of insertion. Surgical reconstructions should aim to recreate these attachments to be as anatomically based as possible.

Biomechanical comparison of new Achilles tendon rupture repair technique the "Giftbox" versus the Krackow technique in New Zealand white rabbits: An experimental animal study.

INTRODUCTION: To compare the strength between the Achilles tendons repaired with the "Giftbox" and the Krackow techniques in New Zealand white rabbits post six weeks of tendon healing. MATERIALS AND METHODS: Eight rabbits were randomized into Giftbox and Krackow groups. Tenotomy was performed on the Achilles tendon of one side of the lower limb and repaired with the respective techniques. The contralateral limb served as control. Subjects were euthanized six weeks post-operative, and both repaired and control Achilles tendons were harvested for biomechanical tensile test. RESULTS: The means of maximum load to rupture and tenacity in the Giftbox group (156.89 ± 38.49 N and 159.98 ± 39.25 gf/tex) were significantly different than Krackow's (103.55 ± 27.48 N and 104.91 ± 26.96 gf/tex, both p = 0.043). CONCLUSION: The tendons repaired with Giftbox technique were biomechanically stronger than those repaired with Krackow technique after six weeks of tendon healing.

A Biomechanical and Ease of Learning Comparison Study of Arthroscopic Sliding Knots.

INTRODUCTION: This study aims to compare the biomechanical properties and ease of learning and tying of our novel knot (UM Knot) with other commonly used arthroscopic sliding knots. MATERIALS AND METHODS: The Duncan, HU, SMC, Pretzel, Nicky's and square knots were selected for comparisons with UM knot. All knots were prepared with size 2 HiFi® suture by a single experienced surgeon and tested with cyclic loading and load to failure tests. The ease of learning was assessed objectively by recording the time to learn the first correct knot and the total number of knots completed in 5 min by surgeons and trainees. RESULTS: The UM knot average failure load is significantly superior to the HU knot (p < 0.05) and comparable to Duncan, SMC, Pretzel and Nicky's knots. According to the ease of learning assessment, UM, Duncan, SMC, Pretzel and Nicky's knots took statistically less time to learn than the HU knot. Although not significant, the failure count due to slippage is fewer in UM knot compared with other knots. CONCLUSIONS: This study showed that UM knot is among the easiest knot to learn and tie, along with Duncan, SMC, Pretzel and Nicky's knots. Their biomechanical properties are comparable and their loads to failure were superior to the HU knot.

Improving knowledge of doctors and paramedics through effective bone procurement workshop: A cognitive approach.

BACKGROUND: Procurement of bone allograft must be performed by trained personnel. Improper handling and lack of knowledge during bone procurement will lead to contamination hence jeopardizing quality of the procured bones and expose bone recipients to risks of infection in post-operative phase. Bone procurement workshop is the fundamental training programme to enhance skill among personnel who has been or will be involved in bone procurement. This study evaluated the effectiveness of the workshop contents including teaching materials by assessing the knowledge on bone procurement among the participants before and after the workshop. METHODS: Bone procurement workshop was held for 2 days for doctors and paramedics. The knowledge on bone procurement was evaluated in pre- and post-assessments by answering self administration questionnaire before and after the workshop, respectively. RESULTS: A total of 50 participants comprised of doctors and paramedics attended the workshop however only 15 (55.6%) doctors and 12 (44.4%) paramedics completed the assessments. Overall, the mean total score for the post-assessment (61.4%) was significantly higher (p < 0.05) than that of the pre-assessment score (32.2%). The mean values of correct responses for the post-assessment was significantly higher (p < 0.05) than that of the pre-assessment in all five topics given during the workshop. The correct responses for the pre- and the post- assessments in the respective group of the doctors and paramedics were also statistically significant (p < 0.05). In the pre-assessment, the doctors had the highest score in Surgical Approach & Reconstruction (50%) while the paramedics had the highest score in Donor Screening & Selection Criteria (33.3%). In the post-assessment, the doctors had the highest score in Donor Screening & Selection Criteria (70%) while the paramedics in Packaging & Transportation (65.8%). CONCLUSIONS: The assessment managed to show that the workshop contents and teaching materials were effective in improving the cognitive knowledge of the personnel who would get involved in bone procurement under the National Donation Programme.

Low-Cost Self-Made Arthroscopic Training Camera Is Equally as Effective as Commercial Camera: A Comparison Study.

PURPOSE: To assess the effectiveness of a low-cost self-made arthroscopic camera (LAC) in basic arthroscopic skills training compared with a commercial arthroscopic camera (CAC). METHODS: One hundred fifty-three orthopaedic residents were recruited and randomly assigned to either the LAC or CAC. They were allocated 2 practice sessions, with 20 minutes each, to practice 4 given arthroscopic tasks: task 1, transferring objects; task 2, stacking objects; task 3, probing numbers; and task 4, stretching rubber bands. The time taken for participants to complete the given tasks was recorded in 3 separate tests; before practice, immediately after practice, and after a period of 3 months. A comparison of the time taken between both groups to complete the given tasks in each test was measured as the primary outcome. RESULTS: Significant improvements in time completion were seen in the post-practice test for both groups in all given arthroscopic tasks, each with P < .001. However, there was no significant difference between the groups for task 1 (P = .743), task 2 (P = .940), task 3 (P = .932), task 4 (P = .929), and total (P = .944). The outcomes of the tests (before practice, after practice, and at 3 months) according to repeated measures analysis of variance did not differ significantly between the groups in task 1 (P = .475), task 2 (P = .558), task 3 (P = .850), task 4 (P = .965), and total (P = .865). CONCLUSIONS: The LAC is equally as effective as the CAC in basic arthroscopic skills training with the advantage of being cost-effective. CLINICAL RELEVANCE: In view of the scarcity in commercial arthroscopic devices for trainees, this low-cost device, which trainees can personally own and use, may provide a less expensive and easily available way for trainees to improve their arthroscopic skills. This might also cultivate more interest in arthroscopic surgery among junior surgeons.

Crossing Suture Technique for the Osteochondral Fractures Repair of Patella.

Osteochondral fracture of the patella is a common concomitant injury of the knee, especially in lateral patellar instability, and the importance of early stable fixation with minimal complication and early mobilization should be emphasized. Screws and Kirschner wires both absorbable and nonabsorbable have been the common mode of fixation of these fractures. Nevertheless, these fixation techniques require larger osteochondral fragments and are associated with cartilage abrasion, hardware prominence, synovitis, and foreign body reaction. In contrast, suture fixation can adequately stabilize smaller osteochondral fragments without comminution and prevent some of the possible complications of other techniques of fixation. We created 4 holes in a rectangular pattern on the patella oriented perpendicular to its anteroposterior surface. We used readily available, and affordable, no. 2 Ultrabraid sutures inserted into the holes and looped around the osteochondral fragment, compressing it to the patella. The technique is very simple and is relatively easy to learn. It provides secure fixation and allows early mobilization. And it spares the knee from subsequent surgical procedure for removal of metallic implants.

Radiological analysis of the medial epicondyle in the adolescent throwing athlete.

INTRODUCTION: Injuries to the medial structures of the elbow due to overhead throwing games are well documented. However, variations of medial epicondyles are not well described, especially in athletes with fused medial epicondyles. In this study, we evaluated variations in the medial epicondyle of baseball players who were aged 15-17 years and had fused epicondyles. METHODS: In this cross-sectional observational study, 155 skeletally mature baseball players with unilateral medial elbow pain and 310 elbow radiographs were reviewed by two independent reviewers. The medial epicondyles were categorised into three groups: normal, elongated or separated. RESULTS: Among the 155 patients, 65 (41.9%) had normal epicondyles, 41 (26.5%) had elongated epicondyles and 49 (31.6%) had separated epicondyles. The medial epicondyle was larger on the dominant arm for 125 (80.6%) patients; the mean surface area on the dominant arm was 222.50 ± 45.77 mm2, while that of the non-dominant arm was 189.14 ± 39.56 mm2 (p < 0.01). Among the three categories of medial epicondyles, separated epicondyles had the largest surface area, followed by elongated and normal epicondyles. CONCLUSION: Medial epicondyles in adolescent throwing athletes can be categorised into three different groups according to their shape (normal, elongated and separated). We observed a correlation between the shape and the surface area of the medial epicondyle in adolescent throwing athletes, with separated medial epicondyles having the largest surface area. Further studies and follow-up are needed to determine the prognostic value and clinical significance of these morphological variations.

Soft-Tissue Loop for Medial Patellofemoral Ligament Reconstruction.

A patient with patellar instability frequently presents with anterior knee pain, patellar subluxation, or dislocation. Medial patellofemoral ligament (MPFL) has a key role for normal patella tracking and stability. Reconstruction of the MPFL using a hamstring graft is a commonly used procedure for the treatment of chronic lateral subluxation of patella. Anchor sutures and bony tunnels are used for the patellar attachment of the graft. This can be associated with complications such as patella fracture; besides, it does not produce an anatomical reconstruction for the native MPFL that can alter the direction of tension applied on the patella. To overcome these problems, a soft-tissue loop technique is used for MPFL reconstruction. During this procedure, a semitendinosus graft was passed through the prepatellar extensor retinaculum and secured with sutures. The free ends of the graft were then passed between the second and third layers of the medial patellofemoral retinaculum and fixed to a femoral tunnel on the medial femoral condyle with an interference screw. The desired amount of tension on the graft is achieved under direct vision of patella tracking arthroscopically. We found this method to be relatively safe and fast. It is more anatomical and can avoid the complications during the conventional bony procedures.

Correlation between Focal Nodular Low Signal Changes in Hoffa's Fat Pad Adjacent to Anterior Femoral Cartilage and Focal Cartilage Defect Underlying This Region and Its Possible Implication.

Purpose. This study investigates the association between focal nodular mass with low signal in Hoffa's fat pad adjacent to anterior femoral cartilage of the knee (FNMHF) and focal cartilage abnormality in this region. Method. The magnetic resonance fast imaging employing steady-state acquisition sequence (MR FIESTA) sagittal and axial images of the B1 and C1 region (described later) of 148 patients were independently evaluated by two reviewers and categorized into four categories: normal, FNMHF with underlying focal cartilage abnormality, FNMHF with normal cartilage, and cartilage abnormality with no FNMHF. Results. There was a significant association (p = 0.00) between FNMHF and immediate adjacent focal cartilage abnormality with high interobserver agreement. The absence of focal nodular lesions next to the anterior femoral cartilage has a very high negative predictive value for chondral injury (97.8%). Synovial biopsy of focal nodular lesion done during arthroscopy revealed some fibrocollagenous tissue and no inflammatory cells. Conclusion. We postulate that the FNMHF adjacent to the cartilage defects is a form of normal healing response to the cartilage damage. One patient with FHMHF and underlying cartilage abnormality was rescanned six months later. In this patient, the FNMHF disappeared and normal cartilage was observed in the adjacent region which may support this theory.

Identification of Pathways Mediating Growth Differentiation Factor5-Induced Tenogenic Differentiation in Human Bone Marrow Stromal Cells.

To date, the molecular signalling mechanisms which regulate growth factors-induced MSCs tenogenic differentiation remain largely unknown. Therefore, a study to determine the global gene expression profile of tenogenic differentiation in human bone marrow stromal cells (hMSCs) using growth differentiation factor 5 (GDF5) was conducted. Microarray analyses were conducted on hMSCs cultures supplemented with 100 ng/ml of GDF5 and compared to undifferentiated hMSCs and adult tenocytes. Results of QuantiGene® Plex assay support the use and interpretation of the inferred gene expression profiles and pathways information. From the 27,216 genes assessed, 873 genes (3.21% of the overall human transcriptome) were significantly altered during the tenogenic differentiation process (corrected p<0.05). The genes identified as potentially associated with tenogenic differentiation were ARHGAP29, CCL2, integrin alpha 8 and neurofilament medium polypeptides. These genes, were mainly associated with cytoskeleton reorganization (stress fibers formation) signaling. Pathway analysis demonstrated the potential molecular pathways involved in tenogenic differentiation were: cytoskeleton reorganization related i.e. keratin filament signaling and activin A signaling; cell adhesion related i.e. chemokine and adhesion signaling; and extracellular matrix related i.e. arachidonic acid production signaling. Further investigation using atomic force microscopy and confocal laser scanning microscopy demonstrated apparent cytoskeleton reorganization in GDF5-induced hMSCs suggesting that cytoskeleton reorganization signaling is an important event involved in tenogenic differentiation. Besides, a reduced nucleostemin expression observed suggested a lower cell proliferation rate in hMSCs undergoing tenogenic differentiation. Understanding and elucidating the tenogenic differentiation signalling pathways are important for future optimization of tenogenic hMSCs for functional tendon cell-based therapy and tissue engineering.

Femoral head diameter in the Malaysian population.

INTRODUCTION: Hip arthroplasty is commonly performed worldwide. The objective of this study was to determine the diameter of the femoral head in the Malaysian population in relation to gender and race (i.e. among Malay, Chinese and Indian patients). METHODS: This was a retrospective cross-sectional study performed between January 1995 and December 2006, evaluating the femoral head diameters of all patients aged 50 years and above who underwent hemiarthroplasty at two major hospitals in Malaysia. RESULTS: A total of 945 femoral heads (663 women, 282 men) were evaluated. The mean age of the patients in our cohort was 75.2 ± 9.4 (range 50-101) years. The mean femoral head diameter (with intact articular cartilage) was 44.9 ± 3.2 (range 38-54) mm. In our study, men had a significantly larger mean femoral head diameter than women (47.7 ± 2.8 mm vs. 43.7 ± 2.4 mm; p < 0.05). Patients of Chinese ethnicity were also found to have significantly larger femoral head diameters, when compared among the three races studied (p < 0.05). CONCLUSION: Malaysians have a mean femoral head diameter of 44.9 ± 3.2 mm. Among our patients, Chinese patients had a significantly larger femoral head size than Malay and Indian patients. We also found that, in our cohort, men had significantly larger femoral head diameters than women.

Effect of growth differentiation factor 5 on the proliferation and tenogenic differentiation potential of human mesenchymal stem cells in vitro.

The use of growth differentiation factor 5 (GDF-5) in damaged tendons has been shown to improve tendon repair. It has been hypothesized that further improvements may be achieved when GDF-5 is used to promote cell proliferation and induce tenogenic differentiation in human bone marrow-derived mesenchymal stem cells (hMSCs). However, the optimal conditions required to produce these effects on hMSCs have not been demonstrated in previous studies. A study to determine cell proliferation and tenogenic differentiation in hMSCs exposed to different concentrations of GDF-5 (0, 5, 25, 50, 100 and 500 ng/ml) was thus conducted. No significant changes were observed in the cell proliferation rate in hMSCs treated at different concentrations of GDF-5. GDF-5 appeared to induce tenogenic differentiation at 100 ng/ml, as reflected by (1) a significant increase in total collagen expression, similar to that of the primary native human tenocyte culture; (2) a significant upregulation in candidate tenogenic marker gene expression, i.e. scleraxis, tenascin-C and type-I collagen; (3) the ratio of type-I collagen to type-III collagen expression was elevated to levels similar to that of human tenocyte cultures, and (4) a significant downregulation of the non-tenogenic marker genes runt-related transcription factor 2 and sex determining region Y (SRY)-box 9 at day 7 of GDF-5 induction, further excluding hMSC differentiation into other lineages. In conclusion, GDF-5 does not alter the proliferation rates of hMSCs, but, instead, induces an optimal tenogenic differentiation response at 100 ng/ml.