Improvement of lower-extremity stability by rotational leg press training.

BACKGROUNDS: Previous reports revealed that balance and stability are essential for human physical activity. The present study aimed to evaluate the comparative outcome of a novel beginning movement load training (BMLT) versus conventional leg press training in terms of improvement of lower-extremity stability. RESEARCH QUESTION: Does the leg press training improve the stability of lower extremities? METHODS: Twenty adult, asymptomatic and active amateur basketball players were enrolled in this study and randomized in equal numbers into a BMLT group (study group) and a conventional leg press training group (control group). RESULTS: The results after eight-week course of training showed that conventional leg press training had an opposite effect on dynamic stability in the left leg (p = 0.015) and backward movement of the right leg (p = 0.038). The athletes in the study group revealed superior dynamic stability maintenance in both legs backwards (p = 0.001), the left leg (p = 0.013), the right leg backwards (p = 0.022) and overall stability (p = 0.002). SIGNIFICANCE: The study group showed better postural stability in terms of the medial/lateral index (p = 0.017), and the athletes in the control group exhibited a significant inability to maintain postural stability in the medial/lateral aspect after the training (p = 0.016). Rotational BMLT lower-extremity press training yielded superior enhancement of dynamic stability and maintenance of center of gravity and postural stability.

Clinical and radiologic outcomes of the modified phemister procedure with coracoclavicular ligament augmentation using mersilene tape versus hook plate fixation for acute acromioclavicular joint dislocation.

BACKGROUND: The clinical superiority of surgical treatment for acromioclavicular (AC) joint dislocation remains controversial. The aim of this study was to compare the clinical and radiological outcomes of the modified Phemister procedure with CC ligament augmentation using Mersilene tape to those of hook plate fixation for acute AC joint dislocation. METHODS: In this study, patients who received modified Phemister surgery with CC ligament augmentation using Mersilene tape (PM group) or hook plate fixation (HK group) for acute unstable AC joint dislocation with a minimum 5-year follow-up period were retrospectively reviewed. The clinical outcomes were evaluated according to blood loss during surgery, surgical duration, visual analogue scale (VAS), Constant-Murley score (CMS), University of California at Los Angeles (UCLA) shoulder rating scale, and the occurrence of complications. Radiological outcomes were assessed from radiographs according to multiple parameters, including CC distance maintenance, acromion osteolysis, and the presence of distal clavicle osteolysis. RESULTS: A total of 35 patients completed follow-up for more than 5 years and were analyzed in this study (mean = 74.08 months). There were 18 patients in the PM group and 17 in the HK group. The PM group exhibited similar improvement in functional outcome to the HK group. Regarding radiological outcomes, the HK group had a superior performance in terms of CC distance maintenance, of statistical significance (CCDR: 94.29 ± 7.01% versus 111.00 ± 7.69%, p < 0.001) after a one-year follow-up period. However, there were 4 cases of acromion osteolysis and 2 cases of distal clavicle osteolysis in the HK group. CONCLUSION: Hook plate fixation was found to be superior to the modified Phemister technique with CC ligament augmentation using Mersilene tape in terms of CC distance maintenance, but there was no significant difference in the functional outcome after 5 years of follow-up. Both surgical methods are reliable options for the treatment of acute AC joint dislocation. Modified Phemister surgery with CC ligament augmentation using Mersilene tape is a relatively lower-cost option for acute AC joint dislocation without the need of a second surgery for implant removal.

Inferior outcome of rotator cuff repair in chronic hemodialytic patients.

BACKGROUND: Repair of rotator cuff tears has yielded excellent functional outcomes in recent decades; however, poor outcomes and dissatisfaction have been noted in specific groups. Spontaneous tendon rupture has been reported in patients receiving long-term hemodialysis owing to alteration of tendon structure, which might impede functional recovery after rotator cuff repair. The purpose of the present study was to compare the clinical outcomes between hemodialysis and non-hemodialysis patients after rotator cuff repair. METHODS: We retrospectively reviewed patients who underwent mini-open rotator cuff repair from Jan 2013 to Jan 2017. A total of 14 patients under chronic hemodialysis (HD) were matched to non-hemodialysis (NHD) patients at a 1:2 ratio according to age, gender, tear size, severity of fatty infiltration and history of diabetes. Pre- and post-operative functional outcome was assessed using the simple shoulder test (SST), American Shoulder and Elbow Surgeons (ASES), Shoulder Rating Scale of the University of California at Los Angeles (UCLA) and visual analog scale (VAS) scores. Clinical functional outcome at the last follow-up was adopted for comparison of the HD and NHD groups. RESULTS: A total of 42 patients were enrolled in this comparative study, with a mean age of 66.64 ± 1.68 years in the HD group and 65.71 ± 5.40 years in the NHD group. At the final clinical assessment, the post-operative functional outcome was significantly improved in both groups (p < 0.001). However, the functional outcome of the HD group was significantly inferior to that of the NHD group in terms of the SST score (6.50 ± 2.24 vs 9.39 ± 1.87, p < 0.001), ASES score (63.17 ± 15.93 vs 86.96 ± 11.43, p < 0.001), UCLA score (20.14 ± 7.71 vs 29.82 ± 5.08, p < 0.001) and VAS score (3.00 ± 0.96 vs 1.21 ± 1.03, p < 0.001). CONCLUSION: The improvement of pain and functional improvement of long-term hemodialysis patients were inferior to those of patients without hemodialysis after mini-open rotator cuff repair.

Efficacy of Extracorporeal Shockwave Therapy on Calcified and Noncalcified Shoulder Tendinosis: A Propensity Score Matched Analysis.

BACKGROUND: Extracorporeal shock wave therapy (ESWT) had been proved to be beneficial in calcific tendinosis; however, the treatment efficacy in noncalcific tendinosis of rotator cuff still remains controversial. The present study was to compare the outcomes ESWT among the noncalcific rotator cuff tendinosis and different types of calcific tendinosis on the basis of similar shoulder functional status. METHODS: A retrospective, comparative study was conducted with the enrollment of 20 patients in each group through propensity score matching in a 1:1:1 ratio from 291 patients who underwent single ESWT for painful shoulder tendinosis. The patients were divided into three groups which included noncalcified tendinosis (NCTS), type I dense calcified tendinosis of shoulder (DCTS), and type II and type III translucent calcified tendinosis of shoulder (TCTS) according to Gartner and Heyer classification. The clinical evaluation included the subjective pain score with visual analog scale (VAS) and functional outcome with Constant and Murley score (CMS). RESULTS: Twelve months after ESWT, the VAS in TCTS (1.5 ± 2.48) was statistically significant lower than NCTS (2.9 ± 2.86) and DCTS (3.8 ± 2.46) (p=.011). For the functional outcome, the overall CMS was superior in TCTS than the NCTS and DCTS (86.9 ± 19.7 versus 78.7 ± 18.3 and 71.1 ± 17.8, p=.014). Besides, the subscales of pain score, strength, and range of motion in TCTS improved significantly better than NCTS and DCTS. 70% of patients were complaint-free in TCTS group, which was higher than the NCTS group (15%) and DCTS group (25%) (p<0.05). CONCLUSION: The present study indicated that the high-dose ESWT posed superior clinical efficacy in type II/III calcification tendinosis rather than type I calcification and noncalcific shoulder tendinosis.

Outcomes and factors of elbow arthroscopy upon returning to sports for throwing athletes with osteoarthritis.

BACKGROUND: Elbow arthroscopy had good functional outcome for throwing athletes. Returning to sports is a major concern for all athletes, but only a few reports have investigated the clinical factors related to the duration of returning to sports. The present study evaluates the efficacy of elbow arthroscopic surgery on throwing elbows with osteoarthritis and defines the clinical factors related to the duration of the returning to sports. METHODS: This was a retrospective study with fifteen active baseball throwing athletes with elbow osteoarthritis who were treated with elbow arthroscopy. Perioperative clinical factors were analyzed for functional outcomes. A multiple linear regression analysis was used to analyze the clinical factors associated with the duration of returning to training and sports. RESULTS: The 15 patients' mean age was 27 years. The mean follow-up time was 2.6 years. The mean procedural complexity was 3.1 ± 1.6 (range 1-6). The elbow total range of motion (ROM) improved significantly from 100.7 ± 28.7° to 125.7 ± 18.5° (p = 0.001). The terminal flexion range of the elbow increased significantly from 116.0 ± 22.6° to 130.0 ± 13.2° (p = 0.001), and the terminal extension range improved from 15.3 ± 11.1° to 4.3 ± 5.9° (p = 0.001). Before the operation, the average subjective patient outcome for return to sports (SPORTS) score was 3.4 ± 1.5, which increased significantly to 9.67 ± 0.45 (p = 0.003) at the last follow-up. The multiple linear regression analysis revealed that higher procedural complexity hinders the athletes from returning to competition. CONCLUSIONS: Elbow arthroscopy offered highly satisfactory results in the throwing elbows of elite athletes and significantly improved the range of motion and SPORTS score. The procedural complexity was significantly related to the duration of returning to competition. Early and aggressive arthroscopic intervention is recommended for elite throwing athletes with elbow osteoarthritis who fail to respond to conservative treatment.

Comparative outcomes of extracorporeal shockwave therapy for shoulder tendinitis or partial tears of the rotator cuff in athletes and non-athletes: Retrospective study.

BACKGROUND: Refractory shoulder tendinitis or partial thickness rotator cuff tears (PTRCTs) are common findings in overhead athletes. Previous studies have examined the effectiveness of extracorporeal shockwave therapy (ESWT) for shoulder tendinitis. MATERIALS AND METHODS: In the current study, we recruited 36 shoulders and performed a comparison between the professional athletes (13 shoulders, athletic group; AG) and the non-athletic population (23 shoulders, non-athletic group, NAG) with PTRCTs or shoulder tendinitis of the shoulder after ESWT. Patients with symptomatic tendinitis of the shoulder with or without a partial tear of the rotator cuff tendon and failed oral medication and physical therapy for more than 3 months were treated with electrohydraulic mode of ESWT. All patients that met the inclusion criteria were categorized into two groups according to their pre-treatment activity level. RESULTS: We found that NAG exhibited significant aging and degenerative change around the glenohumeral joint and subacromial space. After ESWT treatment, the patients in AG were with 53.8% high satisfaction rating and patients in NAG were 52.1% by one-year followed up. CONCLUSION: The results showed ESWT was equally effective treatment in both AG and NAG. In light of its efficacy and less-invasive nature, we suggest ESWT can be used to treat athletes with refractory tendinitis or PTRCTs before proceeding to arthroscopic intervention.

Dosage effects of extracorporeal shockwave therapy in early hip necrosis.

BACKGROUND: This study investigated the effects of different dosages of extracorporeal shockwave therapy (ESWT) in early osteonecrosis of the femoral head (ONFH). MATERIALS AND METHODS: Thirty-three patients (42 hips) were randomly divided into three groups. Group A (10 patients with 16 hips) received 2000 impulses of ESWT at 24 Kv to the affected hip. Group B (11 patients with 14 hips) and Group C (12 patients with 12 hips) received 4000 and 6000 impulses of ESWT respectively. The evaluations included clinical assessment, radiographs, dynamic contrast-enhanced MRI for microcirculation (Ktrans) and plasma volume (Vp), and blood tests for biomarker analysis (NO3, VEGF, BMP-2, osteocalcin, TNF-α, IL-6, substance P, CGRP, DKK-1 and IGF). RESULTS: Significant differences of pain and Harris hip scores were noticed between Group A and C in 6 months after ESWT (all P < 0.05). The pain score decreased, but not Harris hip score improved over the observation time period from 6 to 24 months. Total hip arthroplasty was performed in 3 patients (4 hips) in Group A, but none in Groups B and C. Group C showed significant changes in serum biomarkers for angiogenesis, osteogenesis, anti-inflammation, pain threshold and tissue regeneration between one week and one month after treatment (all P < 0.05). However, no significant changes in the infarction volume in image studies were noted in all groups (all P > 0.05). The post-treatment Ktrans and Vp in the peri-necrotic areas of Group B and C were significantly greater than pre-treatment data (both P < 0.05). CONCLUSIONS: High dosage ESWT is more effective in early stage ONFH. The systemic beneficial effects of ESWT may ultimately enhance angiogenesis with improvement of microcirculation of the peri-necrotic areas, that in turn, can improve subchondral bone remodeling and prevent femoral head collapse.

Cancer stem cell marker CD90 inhibits ovarian cancer formation via ß3 integrin.

Cancer stem cell (CSC) markers have been identified for CSC isolation and proposed as therapeutic targets in various types of cancers. CD90, one of the characterized markers in liver and gastric cancer, is shown to promote cancer formation. However, the underexpression level of CD90 in ovarian cancer cells and the evidence supporting the cellular mechanism have not been investigated. In the present study, we found that the DNA copy number of CD90 is correlated with mRNA expression in ovarian cancer tissue and the ovarian cancer patients with higher CD90 have good prognosis compared to the patients with lower CD90. Although the expression of CD90 in human ovarian cancer SKOV3 cells enhances the cell proliferation by MTT and anchorage-dependent growth assay, CD90 inhibits the anchorage-independent growth ability in vitro and tumor formation in vivo. CD90 overexpression suppresses the sphere-forming ability and ALDH activity and enhances the cell apoptosis, indicating that CD90 may reduce the cell growth by the properties of CSC and anoikis. Furthermore, CD90 reduces the expression of other CSC markers, including CD133 and CD24. The inhibition of CD133 is attenuated by the mutant CD90, which is replaced with RLE domain into RLD domain. Importantly, the CD90-regulated inhibition of CD133 expression, anchorage-independent growth and signal transduction of mTOR and AMPK are restored by the ß3 integrin shRNA. Our results provide evidence that CD90 mediates the antitumor formation by interacting with ß3 integrin, which provides new insight that can potentially be applied in the development of therapeutic strategies in ovarian cancer.

Long-term outcomes of extracorporeal shockwave therapy for chronic foot ulcers.

BACKGROUND: Recent studies showed that extracorporeal shockwave therapy (ESWT) is effective in the treatment of chronic foot ulcers in short term. However, the long-term effects of ESWT in chronic foot ulcers are unknown. The purpose of this study was to evaluate the long-term outcomes of ESWT in chronic foot ulcers with 5-y follow-up. METHODS: The study cohort consisted of 67 patients with 72 ulcers including 38 patients with 40 ulcers in the diabetes mellitus (DM) group and 29 patients with 32 ulcers in the non-diabetes mellitus (non-DM) group. Each patient received ESWT to the affected foot twice per week for 3 wk for a total of six treatments. The evaluations included clinical assessment for the ulcer status, local blood flow perfusion, and analysis of mortality and morbidity. RESULTS: The results showed completely healed ulcers in 55.6% and 57.4% of total series, 48% and 43% of DM group, and 66% and 71% of non-DM group at 1 and 5 y (P = 0.022 and P = 0.027), respectively. The mortality rate was 15% in total series, 24% in DM group, and 3% in non-DM group (P = 0.035). The rate of amputation was 11% in total series, 17% in DM group, and 3.6% in non-DM group (P = 0.194). The blood flow perfusion rate significantly increased after ESWT for up to 1 yr but decreased from 1-5 y in both groups. However, the non-DM group showed significantly better blood flow perfusion than the DM group at 5 y (P = 0.04). CONCLUSIONS: ESWT appears effective in chronic diabetic and nondiabetic foot ulcers. However, the effects decreased from 1-5 y after treatment.

Long-term results of extracorporeal shockwave therapy and core decompression in osteonecrosis of the femoral head with eight- to nine-year follow-up.

BACKGROUND: This study analyzed the long-term outcomes of extracorporeal shockwave therapy (ESWT) and core decompression for early osteonecrosis of the femoral head (ONFH) with 8- to 9-year follow-up. METHODS: The study cohort consisted of 48 patients with 57 hips including 23 patients with 29 hips in the ESWT group and 25 patients with 28 hips in the surgical group. Patients in ESWT group received shockwave therapy to the affected hip. Patients in surgical group underwent core decompression and autogenous cancellous bone and allogenous fibular graft. The average length of follow-up was 103.5 ± 3.4 (ranged 93-106) months and 104.5 ± 4.3 (ranged 95-108) months for the ESWT and the surgical group, respectively. The evaluations included clinical assessment for pain and function, X-ray and MRI of the affected hips. RESULTS: The overall clinical results were 76% good or fair and 24% poor for the ESWT group; and 21% good or fair and 79% poor for the surgical group. THA was performed in 3% and 21% at one year, 10% and 32% at 2 years and 24% and 64% at 8-9 years for ESWT and the surgical group respectively. Significant differences in pain and Harris hip scores were observed at different time intervals favoring the ESWT group. There was a trend of decrease in the size of the lesion in the ESWT group when compared with the surgical group. CONCLUSION: ESWT appears to be more effective than core decompression and bone grafting for early ONFH with 8- to 9-year long-term follow-up.

Molecular changes in diabetic foot ulcers.

AIM: This study investigated the molecular changes of extracorporeal shockwave therapy (ESWT) and hyperbaric oxygen therapy (HBOT) in chronic diabetic foot ulcers. METHODS: A cohort study consisted of 39 patients (44 ulcers) in the ESWT group and 38 patients (40 ulcers) in the HBOT group with similar demographic characteristics. The ESWT group received shockwave therapy twice per week for total six treatments. The HBOT group received hyperbaric oxygen therapy daily for total 20 treatments. Biopsy was performed from the periphery of the ulcer before and after treatment. The specimens were immuno-stained, and the positive immuno-activities of vWF, VEGF, eNOS, PCNA, EGF and TUNEL expressions were examined and quantified microscopically. RESULTS: Significant increases in vWF, VEGF, eNOS, PCNA and EGF expressions and a decrease in TUNEL expression were noted after ESWT (P<0.05), whereas the changes after HBOT were statistically not significant (P>0.05). The differences of vWF, VEGF, eNOS, PCNA, EGF and TUNEL expressions between the two groups were comparable before treatment (P>0.05), however, the differences became statistically significant after treatment (P<0.05) favoring the ESWT group. CONCLUSION: ESWT showed significant increases in angiogenesis and tissue regeneration over HBOT in diabetic foot ulcers.

Extracorporeal shockwave therapy shows chondroprotective effects in osteoarthritic rat knee.

PURPOSE: This study investigated the effects of extracorporeal shockwave therapy (ESWT) on the subchondral bone and articular cartilage in the initiation of osteoarthritis of the knee in rats. METHODS: Anterior cruciate ligament transected (ACLT) osteoarthritis (OA) rat knee model was used in this study. Twenty-seven male Sprague-Dawley rats were divided into three groups. The control group underwent sham surgery without ACLT and received no ESWT. The ACLT group underwent ACLT, but received no ESWT. The ACLT plus ESWT group underwent ACLT and received ESWT immediately after surgery. The evaluation parameters included radiograph, bone mineral density, serum levels of cartilage oligometric protein and osteocalcin, and urinary concentration of C-telopeptide of type II collagen (CTX-II), and histomorphological examination. RESULTS: At 12 weeks, OA of the knee was radiographically verified in the ACLT group, but very subtle changes were noticed in the control and the ACLT plus ESWT groups. On articular cartilage, the ACLT group showed significant increases in cartilage degradation and chondrocyte apoptosis compared to the control and ACLT plus ESWT groups. The ACLT plus ESWT group demonstrated significant decrease in the cartilage degradation and an increase in chondrocyte activity comparable to the control. In subchondral bone, the ACLT group showed a significant decrease in bone remodeling as compared to the control and ACLT plus ESWT groups. The ACLT plus ESWT group showed significant improvement in bone remodeling comparable to the control. CONCLUSION: Extracorporeal shockwave therapy shows chondroprotective effect associated with improvement in subchondral bone remodeling in the initiation of ACLT OA knee model in rats.

Treatment of diabetic foot ulcers: a comparative study of extracorporeal shockwave therapy and hyperbaric oxygen therapy.

OBJECTIVE: This study compared the effectiveness of extracorporeal shockwave therapy (ESWT) and hyperbaric oxygen therapy (HBOT) in chronic diabetic foot ulcers. PATIENTS AND METHODS: The ESWT group (39 patients/44 feet) received shockwave therapy twice per week for total six treatments. The HBOT group (38 patients/40 feet) received hyperbaric oxygen therapy daily for total 20 treatments. Evaluations included clinical assessment, blood flow perfusion scan and histopathological examination. RESULTS: The overall clinical results showed completely healed ulcers in 57% and 25% (P = 0.003); ≥ 50% improved ulcers in 32% and 15% (P = 0.071); unchanged ulcers in 11% and 60% (P < 0.001) and none worsened for the ESWT and the HBOT group respectively. The blood flow perfusion rates were comparable between the two groups before treatment (P = 0.245), however, significant differences were noted after treatment favoring the ESWT group (P = 0.002). Histopathological examination revealed considerable increases in cell proliferation and decreases in cell apoptosis in the ESWT group as compared to the HBOT group. CONCLUSION: ESWT is more effective than HBOT in chronic diabetic foot ulcers. ESWT-treated ulcers showed significant improvement in blood flow perfusion rate and cell activity leading to better healing of the ulcers relative to HBOT in chronic diabetic foot ulcers.

Extracorporeal shockwave shows regression of osteoarthritis of the knee in rats.

BACKGROUND: This study investigated the effects of extracorporeal shockwave technology (ESWT) in osteoarthritis of the knee in rats. MATERIALS AND METHODS: Thirty-six Sprague-Dawley rats were randomly divided into three groups with 12 rats in each group. Group I was the control group and received neither anterior cruciate ligament transection (ACLT) nor ESWT. In groups II and III, ACLT was performed in left knee and osteoarthritis (OA) was verified at 12 wk. Group II received no ESWT, and group III received ESWT at 12 wk after ACLT. Radiographs and bone mineral density (BMD) were obtained at 0, 12, and 24 wk. The animals were sacrificed at 24 wk. One half of the animals were subjected to bone strength test, and the other half for histomorphologic examination and immunohistochemical analysis. RESULTS: Radiographs of the left knee showed progressive OA changes at 12 and 24 wk in group II, whereas, very subtle OA changes were noted in group I and group III. BMD and bone strength were significantly lower in group II compared with groups I and III, but no difference was noted between group I and group III. The cartilage degradations were significantly higher in group II compared with groups I and III, but no difference was noted between group I and group III. The subchondral bone remodeling was significantly less pronounced in group II compared with groups I and III, but no difference was noted between group I and group III. CONCLUSIONS: Application of ESWT to the subchondral bone of the medial tibia condyle showed regression of osteoarthritis of the knees in rats.

The effects of shockwave on systemic concentrations of nitric oxide level, angiogenesis and osteogenesis factors in hip necrosis.

This study investigated the effects of shockwave on systemic concentrations of nitric oxide (NO) level, angiogenic and osteogenic and anti-inflammatory factors in hips with osteonecrosis of the femoral head (ONFH). Thirty-five patients (47 hips) with ONFH were enrolled in this study. Each hip was treated with 6,000 impulses of shockwave at 28 kV in a single session. Ten milliliters of peripheral blood was obtained for the measurements of serum NO level, angiogenic factors (VEGF, vWF, FGF basic and TGF-ß1); osteogenic factors (BMP-2, osteocalcin, alkaline phosphatase, DKK-1 and IGF); and anti-inflammation markers (sICAM and sVCAM) before treatment and at 1, 3, 6 and 12 months after treatment. The hips were evaluated with clinical assessment, serial radiograph and MRI. At 12 months, the overall results showed 83% improved and 17% un-improved. Total hip was performed in 4 cases (8.5%). Serum NO3 level showed significant elevation at 1 month after treatment, but the changes at 3, 6 and 12 months were not significant. For angiogenesis, significant elevations of VEGF, vWF and FGF basic and a decrease in TGF-ß1 were observed at 1 month, but the changes at 3, 6 and 12 months were non-significant. For osteogenesis, BMP-2, osteocalcin, alkaline phosphatase and IGF were significantly elevated, while DKK-1 was decreased at 1 month, but the changes at 3, 6 and 12 months were not significant. For anti-inflammation markers, significant decreases in sICAM and sVCAM were noted at 1 month after treatment, but the changes at 3, 6 and 12 months were non-significant. Local ESWT application results in significant elevations of serum NO level, angiogenic and osteogenic and anti-inflammatory factors in ONFH.

VEGF modulates angiogenesis and osteogenesis in shockwave-promoted fracture healing in rabbits.

OBJECTIVE: Strong vascular endothelial growth factor (VEGF) expression of osteoprogenitors was found in callus site during fracture healing. The aim of this study was to investigate whether VEGF modulates the angiogenesis and osteogenesis in shockwave-promoted fracture healing in rabbits. MATERIALS AND METHODS: Twenty-seven Japanese rabbits were used in the study. A fracture of left tibia with 5 mm gap was created, and the fracture was stabilized with an external fixator. The rabbits were randomly divided into three groups. Group I was the control group and received no shockwave therapy. Group II received shockwave therapy, and group III was pretreated with bevacizumab, a monoclonal antibody against VEGF, before receiving shockwave. Radiographs of the tibia were obtained at 1, 4, and 8 wk. Bone mineral density was performed at 8 wk. The rabbits were euthanized at 8 wk, and the bone specimens were subjected to histomorphological examination and immunohistochemical analysis. RESULTS: At 8 wk, radiographs showed considerably better bone healing and remodeling of the fracture in group II compared with groups I and III, whereas no discernable difference was noted between group I and group III. The BMD values were significantly higher in group II than groups I and III, but no difference noted between group I and group III. In histomorphological examination, significant increases in bone tissue was were noted in group II compared with groups I and III, but no difference was noted between group I and group III. In immunohistochemical analysis, significant increases in VEGF, vWF, PCNA, BMP-2 and osteocalcin, and a decrease in TUNEL expression were observed in group II compared with groups I and III, but no statistical difference was noted between group I and group III. CONCLUSION: Significant increases in VEGF and angiogenic and osteogenic growth factors were noted in shockwave-promoted bone healing. Pre-treatment with bevacizumab inhibited VEGF and in turn, attenuated the effect of shockwave. It appears that VEGF modulates angiogenesis and osteogenesis in shockwave-promoted bone healing in rabbits.

pCMV-BMP-2-transfected cell-mediated gene therapy in anterior cruciate ligament reconstruction in rabbits.

PURPOSE: This study investigated the effect of plasmid cytomegalovirus (pCMV)-bone morphogenetic protein 2 (BMP-2) gene therapy on the healing of the tendon-bone interface after anterior cruciate ligament (ACL) reconstruction in rabbits. METHODS: The pCMV-BMP-2 was synthesized from full-length human BMP-2 complementary deoxyribonucleic acid, followed by cloning into pCMV Script vector (Clontech Laboratories, Inc., San Jose, CA), and was delivered by a xenogeneic (rat kidney) cell line. The ACL was reconstructed by the transfer of extensor digital tendon in the proximal tibia. In the study group the pCMV-BMP-2 gene-transfected normal rat kidney cells mixed with calcium alginate gel were placed at the tendon-bone interface, whereas no pCMV-BMP-2 was used in the control group. The evaluations included radiography, bone mineral density, magnetic resonance imaging, biomechanical study, histologic examination, and immunohistochemical analysis. RESULTS: Bone mineral density showed no significant difference between the groups (P > .05). Magnetic resonance imaging showed significantly better contact between tendon and bone in the study group compared with the control group (P < .0001). In the biomechanical study, significantly higher failure load and maximal graft tension were noted in the study group compared with the control group (P = .034). The modes of graft failure were rupture of the tendon proper in 78% and graft pullout from the bone tunnel in 22% of specimens in the study group versus graft rupture in 22% and graft pullout in 78% in the control group (P = .018). On histologic examination, the study group showed significantly better integration between tendon and bone, as well as more bone tissue around the tendon graft, than the control group (P = .0004). On immunohistochemical analysis, the study group showed significantly higher expressions of von Willebrand factor, vascular endothelial growth factor, proliferation cell nuclear antigen, and BMP-2 than the control group (P < .05). CONCLUSIONS: The pCMV-BMP-2 gene therapy significantly improved the healing of tendon to bone and promoted angiogenesis and osteogenesis at the tendon-bone interface after ACL reconstruction in the rabbit model. CLINICAL RELEVANCE: Application of pCMV-BMP-2 gene therapy may be an effective adjunct therapy in ACL reconstruction.

A preclinical model of double- versus single-unit unrelated cord blood transplantation.

Cord blood transplantation (CBT) with units containing total nucleated cell (TNC) dose >2.5 x 10(7)/kg is associated with improved engraftment and decreased transplant-related mortality. For many adults no single cord blood units are available that meet the cell dose requirements. We developed a dog model of CBT to evaluate approaches to overcome the problem of low cell dose cord blood units. This study primarily compared double- versus single-unit CBT. Unrelated dogs were bred and cord blood units were harvested. We identified unrelated recipients that were dog leukocyte antigen (DLA)-88 (class I) and DLA-DRB1 (class II) allele-matched with cryopreserved units. Each unit contained

The effect of shock wave treatment at the tendon-bone interface-an histomorphological and biomechanical study in rabbits.

PURPOSE: This study was performed to investigate the effect of shock wave treatment on the healing at tendon-bone interface in rabbits. MATERIALS AND METHODS: Thirty-six New Zealand White rabbits were used in this study. The anterior cruciate ligament was excised and replaced with the long digital extensor. The right knees (study group) were treated with 500 impulses of shock waves at 14 kV, while the left knees (control group) received no shock waves. Histomorphological studies were performed in 24 rabbits at 1, 2, 4, 8, 12 and 24 weeks. Biomechanical studies were performed in 12 rabbits at 12 and 24 weeks. RESULTS: There was significantly more trabecular bone around the tendons noted in the study group compared with the control group at different time intervals after 4 weeks (P<0.05). The contacting between bone and tendon was significantly better in the study group than the control group after 8 weeks (P<0.05). The tensile strength of the tendon-bone interface was significantly higher in the study group than the control group at 24 weeks (P=0.018), whereas similar modes of graft failure were noted between the two groups. CONCLUSION: Shock wave treatment significantly improves the healing rate of the tendon-bone interface in a bone tunnel in rabbits. The effect of shock waves appears to be time-dependent.

Nitric oxide mediates ultrasound-induced hypoxia-inducible factor-1alpha activation and vascular endothelial growth factor-A expression in human osteoblasts.

Vascular endothelial growth factor (VEGF) is an important regulator for angiogenesis and endochondral bone formation. Although low-intensity pulsed ultrasound (US) has been recently used for accelerating fracture healing, the effect of US stimulation on angiogenic factor production by osteoblasts remains undetermined. Here, we found that US elevation of VEGF-A expression in human osteoblasts to be mediated by nitric oxide (NO) and hypoxia-inducible factor-1alpha (HIF-1alpha). Human osteoblasts were treated with or without US stimulation (200 micros pulse, 1 kHz at 30 mW/cm2) for 20 min. Cells were subjected to assessment of VEGF-A expression, NO production, nitric oxide synthase (NOS) catalytic activities, and HIF-1alpha transactivation. Results showed that US significantly increased VEGF-A mRNA and protein levels in 6 h. US augmentation of VEGF level was transcriptionally mediated. Early inhibition of NO production, but not calcium or prostaglandin E2, significantly reduced US-enhanced VEGF-A levels. Osteoblasts responded to US treatment by increasing NO production, NOS catalytic activities, iNOS immunoexpression, nuclear HIF-1alpha activation, and binding to the VEGF-A promoter. Inhibition of NOS activity by N-nitro-L-arginine methyl ester (L-NAME) or blockade of guanylate cyclase activity by ODQ reduced US-augmented HIF-1alpha transactivation and VEGF-A levels. Conditioned medium harvested from US-treated osteoblasts promoted tube formation of human umbilical vein endothelial cells (HUVEC). Monoclonal VEGF-A antibody neutralization or L-NAME pretreatment reduced the promoting effect of conditioned medium on angiogenesis of HUVEC. Together, these findings show that NO plays an important role in mediating extracellular stimuli released by US and triggering intracellular response of osteoblasts to produce angiogenic factor after US treatment.