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Abstract Background The evaluation of lateral ankle laxity remains challenging when diagnosing chronic lateral ankle instability (CLAI). Several studies have reported that internal rotation of the talus as an indicator of rotational lateral ankle laxity (RLAL) increases in patients with CLAI. However, there is no established method for detecting and evaluating the RLAL. This study aimed to report a novel method for evaluating the RLAL in the gravity stress position by measuring the talofibular distance (TFD) using ultrasonography (US) and show the normative value of the TFD. Methods The TFDs in the subjects with healthy ankles were prospectively measured 10 mm distal to the ankle joint in the neutral ankle position and gravity stress position using US. The differences in the TFD between the two ankle positions were evaluated. The differences in the TFD by gender and ankle laterality were also evaluated. Results A total of 52 healthy ankles of 28 subjects (mean age, 24.0 ± 1.6; male/female, 12/16) were finally included. There was a significant difference in the TFD between the neutral ankle position (6.9 ± 0.9 mm) and gravity stress position (9.0 ± 0.9 mm) (p

Abstract Background Alkaptonuria (AKU) is a rare hereditary disease. Ochronotic arthropathy, tendinopathy, and osteopenia/osteoporosis are generally musculoskeletal manifestations in patients with AKU. Because of the ochronotic tendinopathy and osteopenia, the surgical strategy for chronic ochronotic Achilles tendon rupture may be challenging. No studies have reported the surgical treatment of chronic Achilles tendon rupture in patients with AKU. Case presentation We report a case of AKU that required surgical treatment for chronic Achilles tendon rupture. A 60-year-old woman was referred to our department for the assessment of left hindfoot pain that persisted for more than nine months after an ankle sprain. Three years prior to the first presentation to our hospital, she was diagnosed with AKU due to pigmented hip cartilage at the time of total hip arthroplasty. The patient was diagnosed as chronic Achilles tendon rupture based on the results of physical examination and magnetic resonance imaging (MRI). The MRI showed enlarged scar tissue of the Achilles tendon with an intrasubstance high signal intensity. We performed resection of the central part of the scar tissue (total length, 24 mm) followed by direct repair using the proximal and distal stumps of the scar tissue. The MRI at 12months after surgery showed continuity of the repaired Achilles tendon. At 18 months after surgery, the Achilles Tendon Total Rupture Score improved from 22 points preoperatively to 84 points postoperatively. The foot and ankle outcome score also improved. Conclusions We reported a case of AKU with chronic Achilles tendon rupture in which direct repair using scar tissue between the tendon stumps was effective. Careful selection of the surgical procedure for the treatment of chronic ochronotic Achilles tendon rupture is recommended because of concomitant ochronotic tendinopathy and osteopenia/osteoporosis.

Abstract Background Clavicle fractures are common injuries, especially in young, active individuals. Operative treatment is recommended for completely displaced clavicle shaft fractures, and plate fixation is stronger than the use of intramedullary nails. Few studies have reported on iatrogenic injuries to the muscle attached to the clavicle during fracture surgery. The aim of this study was to clarify the area of the insertion sites of muscles attached to the clavicle in Japanese cadavers using gross anatomy and three-dimensional (3D) analysis. We also aimed to compare the effects of anterior plate templating and superior plate templating on clavicle shaft fractures using 3D images. Methods Thirty-eight clavicles from Japanese cadavers were analyzed. We removed all clavicles to identify the insertion sites and measured the size of the insertion area of each muscle. Three-dimensional templating was performed on both the superior and anterior plates of the clavicle using data obtained from computed tomography. The areas covered by these plates on the muscles attached to the clavicle were compared. Histological examination was performed on four randomly selected specimens. Results The sternocleidomastoid muscle was attached proximally and superiorly; the trapezius muscle was attached posteriorly and partly superiorly; and the pectoralis major muscle and deltoid muscles were attached anteriorly and partially superiorly. The non-attachment area was located mainly in the posterosuperior part of the clavicle. It was difficult to distinguish the borders of the periosteum and pectoralis major muscles. The anterior plate covered a significantly broader area (mean 6.94 ± 1.36 cm2) of the muscles attached to the clavicle than did the superior plate (mean 4.11 ± 1.52 cm2) (p

Abstract Background Recently, cryotherapy has become a common practice for postoperative pain management. The current accepted practice in Japan is the use of cryotherapy at 5 °C after arthroscopic shoulder surgery. However, this therapy has been reported to be highly intense because the sustained low temperature causes discomfort for patients. The optimum temperature and duration of cooling required for comfortable and effective cryotherapy after arthroscopic shoulder surgery were investigated. Methods Because pain levels might differ depending on the condition, we selected 52 patients with rotator cuff injuries, which were the most common disorders indicated for arthroscopic shoulder surgery. Patients were treated with cryotherapy at 5 °C or 10 °C for 16 h or 24 h. The pain level was determined using the visual analogue scale, and deep shoulder joint temperatures were recorded at different time points for analysis. Results Pain after arthroscopic shoulder surgery was found to be related to the presence of a brachial plexus block using the interscalene approach during surgical anesthesia. To obtain effective analgesia with cryotherapy, the cooling temperature and duration of cryotherapy had to be changed based on the presence or absence of the brachial plexus block. Patients who received brachial plexus blocks had the lowest recorded pain scores after receiving cryotherapy at 5 °C for 24 h after surgery. Patients who did not receive the block had the lowest recorded pain scores when receiving cryotherapy at either 5 °C for 16 h or 10 °C for 24 h. Conclusions Using universal cryotherapy intensity and duration settings regardless of the use of other interventions is likely to unintentionally increase postoperative pain levels. This study revealed that cryotherapy at 5 °C for 24 h was optimal for patients who received an anesthesia block and at 5 °C for 16 h or at 10 °C for 24 h for those who did not receive the anesthesia block. These results can be used as a reference for setting the temperature and duration of cryotherapy after arthroscopic shoulder surgery.

Abstract Background High-Mobility Group Box 1 (HMGB1) and HMGB2 are chromatin-associated proteins that belong to the HMG protein family, and are involved in the regulation of DNA transcription during cell differentiation, proliferation and regeneration in various tissues. However, the role of HMGB2 in ovarian folliculogenesis is largely unknown. Methods We investigated the functional role of HMGB1 and HMGB2 in ovarian folliculogenesis and fertilization using C57BL/6 wild type (WT) and HMGB2-knockout (KO) mice. Ovarian tissues were obtained from WT and HMGB2-KO mice at postnatal days 0, 3, 7, and 2, 6 months of age, then performed immunohistochemistry, qPCR and Western blotting analyses. Oocyte fertilization capability was examined by natural breeding and in vitro fertilization experiments. Results In HMGB2-KO mice, ovary weight was decreased due to reduced numbers of oocytes and follicles. Natural breeding and in vitro fertilization results indicated that HMGB2-KO mice are subfertile, but not sterile. Immunohistochemistry showed that oocytes expressed HMGB2, but not HMGB1, in neonatal and adult WT ovaries. Interestingly, in HMGB2-KO ovaries, a compensatory increase in HMGB1 was found in oocyte nuclei of neonatal and 2-month-old mice; however, this was lost at 6 months of age. Conclusions The depletion of HMGB2 led to alterations in ovarian morphology and function, suggesting that HMGB2 plays an essential role in ovarian development, folliculogenesis and fertilization.

Abstract Background A number of studies have evaluated risk factors for lateral ankle sprain (LAS) or chronic lateral ankle instability (CLAI). However, the definitive risk factors for LAS or CLAI remain controversial. The purpose of this study was to evaluate whether the contralateral healthy ankles of subjects with ipsilateral mechanical lateral ankle laxity (group I) show greater lateral ankle laxity in comparison to the healthy ankles of bilateral healthy controls (group B). Methods From March 2020, anterior talofibular ligament (ATFL) lengths of young adult volunteers were cross-sectionally measured in non-stress and stress positions using a previously reported stress ultrasonography (US) procedure. The ATFL ratio (the ratio of stress ATFL/non-stress ATFL length) was calculated as an indicator of lateral ankle laxity. The manual anterior drawer test (ADT) was also performed. The US findings of healthy ankles from groups I and B were compared. Results A total of 154 subjects in group B (mean age, 24.5 ± 2.8 years; male/female, 84/70) and 40 subjects in group I (mean age, 24.4 ± 2.3 years; male/female, 26/14) were included in the study. There was no significant difference in the ADT between the groups. There were no significant differences in the non-stress ATFL length (19.4 ± 1.8 vs. 19.3 ± 1.9, p = 0.84), stress ATFL length (20.8 ± 1.8 vs. 20.9 ± 1.9, p = 0.66), length change (1.5 ± 0.6 vs. 1.6 ± 0.6, p = 0.12) and ATFL ratio (1.08 ± 0.03 vs. 1.08 ± 0.03, p = 0.13) between the groups. Conclusion No significant difference was detected between the contralateral healthy ankles of subjects with ipsilateral mechanical lateral ankle laxity and those of bilateral healthy controls.

Abstract Background Different methods of acetabular reconstruction with total hip arthroplasty (THA) for Crowe II and III of adult developmental dysplasia of the hip (DDH) acetabular bone defect have been implemented clinically. However, the biomechanical effect of different augmented materials for acetabular reconstruction in THA on shell stability has never been discussed. Methods In the present study, autologous bone graft (BG)and metal (Ti6Al4V) augment (MA) were simulated with several acetabular bone defect models of DDH in THA. The contact pressure and micromotion between the shell and host bone were measured for evaluating the shell stability using a finite element method. Results The peak contact stress between shell and host bone was higher in the MA situation (12.45 vs 8.71 MPa). And the load transfer path was different, for BG models, the high local contact stresses were found at the junction of bone graft and host bone while for MA models the concentrated contact stresses were at the surface of MA. The peak relative micromotion between shell and host bone was higher in the MA situation (12.61 vs 11.13 µm). However, the peak micromotion decreased in the contact interface of MA and cup compared to the BG models. Conclusions The higher micromotion was found in MA models, however, enough for bone ingrowth, and direct stronger fixation was achieved in the MA-cup interface. Thus, we recommended the MA can be used as an option, even for Crowe III, however, the decision should be made from clinical follow-up results.

Abstract Background Lateral ankle sprain (LAS) is one of the most common musculoskeletal injuries. Numerous studies regarding LAS have been performed. However, there are few studies evaluating the current clinical practice of orthopaedic surgeons regarding LAS. The purpose of this study was to evaluate the current clinical practice of general orthopaedic surgeons in the treatment of LAS. Methods A questionnaire survey was conducted from September 2020 to December 2020 in Miyazaki, Japan, to evaluate the clinical practice of general orthopaedic surgeons in the treatment of LAS. The survey was composed of 12 questions that were developed with consideration of the recommendations in the current clinical practice guidelines (CPGs) published by the Dutch orthopaedic society. The questions in this study were focused on the diagnosis, conservative treatment, rehabilitation, and the criteria for return to sports (RTS). Results The survey response rate was 82.7% (129/156). Among the respondents, 95.3% did not consider the Ottawa Ankle Rules in the decision to perform plain radiography for patients. Rehabilitation following LAS was performed in 58.9% of patients. Eighty-five (65.9%) of the surgeons used only one factor as the criterion for RTS. The absence of pain was the most frequently used criterion (45.7%). No objective criteria were used for the RTS decision in athletes with LAS. Conclusions The present study suggested that most general orthopaedic surgeons do not provide the care for patients with LAS recommended by the current CPGs. No objective criteria for the RTS decision are used for athletes with LAS.

Abstract Background Intertrochanteric curved varus osteotomy (CVO) has been widely used to remove the necrotic bone away from the weight-bearing portion in the treatment of osteonecrosis of the femoral head (ONFH). However, whether all types of necrosis will benefit from CVO, in terms of the stress level, the effect of different center-edge (CE) angles of acetabulum on stress distribution of necrosis after CVO, and the relationship between the intact ratio and the stress of necrosis, has never been addressed. The purpose of the study was to evaluate the influence of CVO on the stress reduction in necrotic bone using a finite element analysis (FEA) with different CE angles. Methods CVO finite element models of the hip joint were simulated with a lesion of 60°. The osteotomy angles were divided into four configurations (15°, 20°, 25°, and 30°), and three types (A, B, and C1) of lesions were established based on the Japanese Investigation Committee (JIC) classification. In addition, two CE angles (18° and 33°) of acetabulum were considered. The maximum and mean von Mises stress were analyzed in terms of the necrotic bone by a physiological loading condition. Moreover, the correlation of the intact ratio measured in 3D and the stress distribution after CVO was analyzed. Results Stress reduction was obtained after CVO. For type B, the CVO angle was 20° (0.61 MPa), and for type C1, the CVO angle was 30° (0.77 MPa), if the mean stress level was close to type A (0.61 MPa), as a standard. The maximum and mean von Mises stress were higher in the CE angle of 18°models, respectively. The intact ratio measured in 3D had a good negative correlation with stress after CVO and had more influence on stress distribution in comparison to other geometric parameters. Conclusions For making decisions about the biomechanics of CVO, a CVO angle of > 20° was recommended for type B and > 30° was safe for type C1. The risk of progressive collapse was increased in the insufficient situation of the weight-bearing portion after CVO. The intact ratio could provide information about clinical outcomes and stress distribution after CVO.

Abstract Background Turner’s syndrome, discoid meniscus, and Blount’s disease have all been studied in isolation, but, to the best of our knowledge, there have been no studies reporting a patient with all three. Thus, the first case of Turner’s syndrome with discoid meniscus and Blount’s disease is presented. Case presentation A 5-year-old Japanese girl with a history of Turner’s syndrome and Blount’s disease complained of pain in her left knee. Magnetic resonance imaging showed a discoid lateral meniscus tear, and arthroscopic partial meniscectomy was performed, providing a good outcome. Conclusions In this report, some possible explanations regarding the concomitant presence of these three diseases are discussed. A possible explanation in this case is that the patient with Turner’s syndrome had a discoid lateral meniscus that might have been induced by some genetic factors associated with Turner’s syndrome, and then the discoid lateral meniscus might have been the mechanical stress that caused Blount’s disease.

Abstract Background Although shadow pitching, commonly called “towel drill,” is recommended to improve the throwing motion for the rehabilitation of pitching disorders before the initiation of a throwing program aimed at returning to throwing using a ball, the motion differs from that of normal throwing. Learning improper motion during ball release (BR) may increase shoulder joint forces. Abnormal throwing biomechanics leads to injures. However, there has been no study of shadow pitching focusing on the BR position. The purpose of the present study was to evaluate the BR position and kinematic differences between shadow pitching and normal throwing. In addition, the effect of setting a target guide for BR position on throwing motion was examined in shadow pitching. Methods The participants included in this study were 20 healthy male students who were overhand right-handed pitchers with no pain induced by a throwing motion. Participants performed normal throwing (task 1), shadow pitching using a hand towel (task 2), and shadow pitching by setting a target of the BR position (task 3). A motion capture system was used to evaluate kinematic differences in throwing motions, respectively. Examination items comprised joint angles and the differences in BR position. Results BR position of task 2 shifted significantly toward the anterior, leftward, and downward directions compared with task 1. The distance of BR position between tasks 1 and 2 was 24 ± 10%. However, task 3 had decreased BR deviation compared with task 2 (the distance between 3 and 1 was 14 ± 7%). Kinematic differences were observed among groups at BR. For shoulder joint, task 2 showed the highest value in abduction and horizontal adduction among groups. In spine flexion, left rotation and thorax flexion, task 2 was significantly higher than task 1. Task 3 showed small differences compared with task 1. Conclusions The BR position of shadow pitching deviated significantly in the anterior, leftward, and downward directions compared with normal throwing. Furthermore, we demonstrated that the setting of BR target reduces this deviation. Thus, the target of BR position should be set accurately during shadow pitching exercises in the process of rehabilitation.

Abstract Background Spondylolysis is the main cause of low back pain (LBP) in young athletes. There are few studies analyzing the difference of spondylolysis among young athletes with different sports activities. The purpose of this study was to compare the clinical factors and distribution of the lesions of spondylolysis on magnetic resonance imaging (MRI) scans in young soccer and baseball players with symptomatic spondylolysis. Methods The medical records of 267 young athletes aged 7 to 18 years old who underwent MRI to evaluate the cause of LBP between 2017 and 2020 were retrospectively reviewed to identify patients with spondylolysis. Of the young athletes with symptomatic spondylolysis, clinical factors and MRI findings in soccer and baseball players were retrospectively evaluated. The clinical factors were age, sex, interval from onset of LBP to MRI, and side of the dominant leg in the sports field. MRI findings included number, lumbar level, and side of the lesions. Results A total of 33 soccer players (mean age, 15.4 ± 1.4 years) and 49 baseball players (mean age, 15.4 ± 1.6 years) with symptomatic spondylolysis were enrolled. All patients were male. No significant differences were noted in age and the interval from onset of LBP to MRI between the groups. Soccer players had greater numbers of multiple (p < 0.001) and bilateral (p < 0.001) lesions than baseball players. The dominant side of the hand for pitching or batting was correlated with the contralateral-side lesions in baseball players (p = 0.001). Conclusions The distribution of the lesions of spondylolysis differed in young soccer and baseball players. Pitching or batting with the dominant-side hand would be associated with contralateral-side lesions in baseball players. Sports-specific movements and the side of the dominant leg should be considered when treating young athletes with symptomatic spondylolysis.

An amendment to this paper has been published and can be accessed via the original article.

Abstract Background Bone tunnel enlargement following primary anterior cruciate ligament (ACL) reconstruction with soft tissue graft might be a severe disadvantage for revision surgery. The postoperative rehabilitation protocol including the non-weight-bearing periods were different depending on the surgeon or institute. To determine the relationship between femoral bone tunnel enlargement and the postoperative non-weight-bearing period after double-bundle ACL reconstruction with hamstring grafts. Methods Forty-two patients who underwent primary double-bundle ACL reconstruction with hamstring grafts were divided into two postoperative non-weight-bearing protocol groups: 1-week non-weight-bearing postoperatively (group A, n = 19); and 2-week non-weight-bearing (group B, n = 18). Five cases were excluded due to additional knee injury, pregnancy, and lost to follow-up. Bone tunnel enlargement was evaluated by computed digital radiographs (anteroposterior (A-P) and lateral views) taken on the first postoperative day and at 12 months. Each tunnel diameter was shown as a percentage to the maximum joint width of the proximal tibia in the A-P view, or a percentage of the maximum diameter of the patella in the lateral view. To determine the incidence of tunnel enlargement, percentage diameter changes of more than 10% were defined as an enlarged tunnel. The magnitude of tunnel enlargement and the standard clinical evaluation were also evaluated. Results There were no significant differences between groups in the incidences of anteromedial and posterolateral bone tunnel enlargement, both in the A-P and lateral views (2 × 2 Chi-squared test). The magnitude of femoral posterolateral bone tunnel enlargement was significantly greater in group A in the A-P view (p = 0.01) and lateral view (p = 0.03) (Mann Whitney U-test). Twelve months after surgery, the Lysholm score and Tegner activity level scale were not significantly different between the groups. Conclusions This prospective, clinical and radiographical study showed that early weight-bearing protocol after double-bundle ACL reconstruction with hamstring grafts might have the potential risk of significant postoperative femoral bone tunnel enlargement of the posterolateral bundle. There was no significant difference in clinical outcomes by postoperative non-weight-bearing period. To reduce and prevent the femoral bone tunnel enlargement, the comprehensive management could be considered and required to establish the suitable early stage rehabilitation protocol after surgery. Trial registration Trial registration number; UMIN000036212. Scientific title: Prospective comparisons of femoral tunnel enlargement with two different postoperative non weight bearing periods after double-bundle anterior cruciate ligament reconstruction with hamstring grafts. Registered date: 15 Mar 2019 (retrospectively registered).

Abstract Background Patients undergoing total or partial arthroscopic meniscectomy for treating traumatic meniscal tears are at greater risk of developing knee osteoarthritis (OA) due to increased mechanical load. The purpose of this study was to evaluate the effects of a valgus unloader brace in the medial meniscectomized knee joint during the gait cycle. Methods A three-dimensional finite element model of the knee joint was developed using the substructures segmented from magnetic resonance images. Experimentally measured forces and moments for one complete gait cycle, without brace and with brace at three different alignment angles (0°, 4°, and 8°), were applied to the finite element model, and the changes in the tibiofemoral contact mechanics were estimated. Results The brace in 0°/4°/8° valgus alignment modes reduced the total contact force in the medial compartment by 16%/46%/82% at opposite toe off and 18%/17%/29% at opposite initial contact events, while it increased the total contact force in the lateral compartment by 31%/81%/110% at opposite toe off and 30%/38%/45% at opposite initial contact events, respectively, when compared to the unbraced meniscectomized knee. Conclusions Increasing the valgus alignment from 0° to 4° and 8° resulted in a greater reduction of contact conditions (total contact force, total contact area, peak contact pressure) in the medial compartment and vice versa in the lateral compartment. This decrease in contact conditions in the medial compartment infers enhanced knee joint function due to a valgus unloader brace, which translates to increased knee-related confidence. Results suggest choosing a higher valgus alignment angle could potentially increase the risk for the onset of osteoarthritis in the lateral compartment, and this computational model could be used in validating the effectiveness of braces on joint health.

Abstract Background The stability of acetabulum reconstructions using reinforcement rings and hooks is important for successful replacement surgery. The objective of this study was to biomechanically determine the effects of the hook on stress and the related micromotions of the acetabular reinforcement ring during the immediate postoperative period. Methods Acetabular reinforcement ring models were developed using a nonlinear, three-dimensional, finite element method. Using a pre-prepared template, we constructed without-hook and bone graft models of varying volumes and material properties. Results The stress on the inferior margin of the acetabulum was higher in the with-hook model than in the without-hook model, especially with increased bone graft volumes, and the stiffness of the bone graft material was decreased. Relative micromotions in the without-hook model were higher than in the with-hook models. The highest relative micromotion was observed in the model with increased bone graft volume and lower stiffness of bone graft material. Conclusions In biomechanical analyses, the hook effectively dispersed stress and improved the initial fixation strength of the acetabular reinforcement ring.

Abstract Background Achieving stability of the tibial implant is essential following cementless total knee arthroplasty with bone grafting. We investigated the effects of bone grafting on the relative micromotion of the tibial implant and stress between the tibial implant and adjacent bone in the immediate postoperative period. Methods Tibial implant models were developed using a nonlinear, three-dimensional, finite element method. On the basis of a preprepared template, several bone graft models of varying sizes and material properties were prepared. Results Micromotion was larger in the bone graft models than in the intact model. Maximum micromotion and excessive stress in the area adjacent to the bone graft were observed for the soft and large graft models. With hard bone grafting, increased load transfer and decreased micromotion were observed. Conclusions Avoidance of large soft bone grafts and use of hard bone grafting effectively reduced micromotion and undue stress in the adjacent area.

Abstract Background Although a loss of rotator cuff integrity leads to the superior migration of the humeral head, the parameters that characterize the anterolateral migration of the humeral head have not been established. The purpose of this study was to investigate the correlation between the translation of the humeral head scale (T-scale) and clinical outcomes of rotator cuff repair, as well as the correlation between the T-scale and radiologic parameters. Methods One hundred thirty-five consecutive patients with full-thickness rotator cuff tears underwent primary rotator cuff repair. The T-scale, which indicates the distance from the center of the humeral head to the lateral coracoacromial arch, was measured on axial computed tomography scans, and the acromiohumeral interval (AHI) was measured radiographically. The correlation of the two parameters with the clinical scores of the Japanese Orthopaedic Association and University of California–Los Angeles scores and active forward elevation (FE) were evaluated at the preoperative and postoperative stages, respectively. Results The postoperative T-scale and AHI correlated well with the postoperative FE and clinical scores in the patients with large-massive tears but not in those patients with small-medium tears and preoperative large-massive tears. A significant correlation was observed between the postoperative T-scale and AHI. The T-scale was subject to cuff repair integrity. Conclusions We demonstrated that the postoperative T-scale was well correlated with the clinical results and postoperative AHI after rotator cuff repair for large-massive tears, indicating that poor outcomes are associated with combined superior and anterolateral migration of the humeral head following retears.

Abstract Background ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) proteins play an important pathological role in matrix degeneration. Aggrecan degradation is a significant and critical event in early-stage osteoarthritis. To determine the effect of hemoglobin (Hb) on the ability of synovial tissues to produce ADAMTS family members, we examined the influence of Hb by synovial cells in an in vitro experimental system. Methods Synovial tissues were obtained from five young patients with meniscal injury under arthroscopic surgery. Primary cultures of human knee synovial cells were treated with different doses of human Hb (0, 25, 50, 100 μg/ml). The culture media were collected 24 h after Hb-treatment. In the time-course studies, cells were treated with and without 100 μg/ml Hb, and culture media were taken at 6, 12, and 24 h. To identify the proteins responsible for aggrecanase activity, Western blot analysis using antibodies against human ADAMTS-5, −8, −9, and −10; enzyme-linked immunosorbent assay (ELISA); and gene expression for ADAMTS-5 and -9 were examined. Statistical comparisons between each group were performed using paired t-tests. Results Western blot analysis revealed that Hb-treatment resulted in the expression of ADAMTS-5 and -9. Neither control group nor Hb-treated medium showed immunoreactivity against ADAMTS-8 or −10. In a dose-dependency study, the Hb-treated group showed significantly higher levels of ADAMTS-5 and -9 compared with the control (p