Topical co-administration of zoledronate with recombinant human bone morphogenetic protein-2 can induce and maintain bone formation in the bone marrow environment.

BACKGROUND: Bone morphogenetic proteins (BMPs) induce osteogenesis in various environments. However, when BMPs are used alone in the bone marrow environment, the maintenance of new bone formation is difficult owing to vigorous bone resorption. This is because BMPs stimulate the differentiation of not only osteoblast precursor cells but also osteoclast precursor cells. The present study aimed to induce and maintain new bone formation using the topical co-administration of recombinant human BMP-2 (rh-BMP-2) and zoledronate (ZOL) on beta-tricalcium phosphate (ß-TCP) composite. METHODS: ß-TCP columns were impregnated with both rh-BMP-2 (30 µg) and ZOL (5 µg), rh-BMP-2 alone, or ZOL alone, and implanted into the left femur canal of New Zealand white rabbits (n = 56). The implanted ß-TCP columns were harvested and evaluated at 3 and 6 weeks after implantation. These harvested ß-TCP columns were evaluated radiologically using plane radiograph, and histologically using haematoxylin/eosin (H&E) and Masson's trichrome (MT) staining. In addition, micro-computed tomography (CT) was performed for qualitative analysis of bone formation in each group (n = 7). RESULTS: Tissue sections stained with H&E and MT dyes revealed that new bone formation inside the ß-TCP composite was significantly greater in those impregnated with both rh-BMP-2 and ZOL than in those from the other experimental groups at 3 and 6 weeks after implantations (p < 0.05). Micro-CT data also demonstrated that the bone volume and the bone mineral density inside the ß-TCP columns were significantly greater in those impregnated with both rh-BMP-2 and ZOL than in those from the other experimental groups at 3 and 6 weeks after implantations (p < 0.05). CONCLUSIONS: The topical co-administration of both rh-BMP-2 and ZOL on ß-TCP composite promoted and maintained newly formed bone structure in the bone marrow environment.

The posterior tibial slope does not influence the flexion angle in posterior-stabilized mobile-bearing total knee arthroplasty.

BACKGROUND: It remains uncertain whether an increase in the tibial slope leads to better flexion in posterior-stabilized (PS) total knee prostheses. PURPOSE: To compare the intra-operative flexion angle between standard and an additional 10° posterior slope inserts. PATIENTS AND METHODS: Between December 2014 and February 2015, 22 patients (25 knees) who underwent PS mobile-bearing primary total knee arthroplasty (TKA) were included. Flexion angles were measured using either standard or specially-made inserts. Differences in flexion angles between the two situations were analyzed to determine the relationship between changes in flexion angle and pre-operative flexion angle or body mass index (BMI), and between intra- and post-operative flexion angle. RESULTS: The difference between the average flexion angle of standard inserts and specially-made inserts was not statistically significant. Although the correlations between changes in flexion angle due to insert difference and flexion angle, pre-operative flexion angle or BMI were not significant, there was a positive correlation between intra-operative and post-operative flexion at 2 years. CONCLUSION: The results showed an additional posterior tibial slope by 10° did not affect the intra-operative flexion angle. Surgeons performing PS mobile-bearing TKA do not need to excessively slope the tibial bone cutting to improve the post-operative flexion angle. LEVEL OF EVIDENCE: I, Experimental study.

Medium-term clinical results of unicompartmental knee arthroplasty for the treatment for spontaneous osteonecrosis of the knee with four to 15 years of follow-up.

BACKGROUND: Unicompartmental knee arthroplasty (UKA) is an option for the treatment of spontaneous osteonecrosis of the knee (SONK). However, there are limited studies focusing on this area. This study presents medium-term clinical outcome data of UKA for SONK. METHODS: We reviewed 50 SONK knees in 48 patients that were treated by UKA. The mean age, height, and body weight were 73 years, 153 cm, and 57 kg, respectively. The mean follow-up was 8.4 years (range, four to 15 years). Preoperatively, we measured the size and the volume (estimated by width × length × depth) of the necrotic bone mass on T1-weighted magnetic resonance imaging. The clinical results were evaluated serially at follow-up visits radiographically and with the Knee Society Scoring (KSS) and Oxford Knee Scoring (OKS) systems. RESULTS: There were no revisions, re-operations, or major complications. The mean sizes of the necrotic lesions were 17.2 mm (14.7-22.3 mm) in width, 28.2 mm (6.2-34.7 mm) in length, and 11.3 mm (3.2-14.5 mm) in depth. The mean volume was approximately 5.4 cm3 (0.7-10.3 cm3). The mean flexion of the knee, KSS Knee Score, Function Score, and OKS increased from a preoperative 128.7-137.5°, 52.3-91.3, 39.7-90.2, and 21.6-40.2, respectively, at the latest follow-up. At the last follow-up, all patients had good or excellent OKS. CONCLUSIONS: This study demonstrates that UKA is a good option and is reliable for the treatment of SONK irrespective of necrotic bone mass size.

Efficacy of a Pelvic Lateral Positioner With a Mechanical Cup Navigator Based on the Anatomical Pelvic Plane in Total Hip Arthroplasty.

BACKGROUND: The acetabular component orientation in total hip arthroplasty (THA) is of critical importance to the good clinical results. However, traditional widely used cup alignment guides for cup placement are reported to be relatively unreliable. The present study aims at comparing a novel cup alignment guide, which can be attached to our anatomical pelvic plane (APP) pelvic lateral positioner for reducing discrepancies in sagittal pelvic tilt and indicate a targeted cup angle based on the APP, with a conventional cup alignment guide. METHODS: The subjects were 136 hips of 136 patients who underwent unilateral THA using the APP positioner. The procedure was performed with the conventional cup alignment guide (conventional group; 60 hips) and with the novel cup navigator (mechanical navigator group; 76 hips). Postoperative cup angles and discrepancies of postoperative cup angles (inclination and anteversion angles) from the targeted angles were compared between the 2 groups to evaluate the usefulness of these navigators. RESULTS: The mean cup angles in the conventional group were 39.0° ± 5.3° for the inclination angle and 21.7° ± 6.4° for the anteversion angle, whereas those in the mechanical navigator group were 40.6° ± 3.2° and 18.3° ± 4.6°, respectively (P = .018, P < .0001). The discrepancies from the targeted angles were 3.5° ± 3.1° for the inclination angle and 4.6° ± 3.4° for the anteversion angle in the conventional group and 2.3° ± 2.3° and 3.2° ± 2.7°, respectively, in the mechanical navigator group (P = .020, P = .012). CONCLUSION: The mechanical cup navigator easily attachable to the APP positioner is a tool that can improve the accuracy of cup placement in a simple, economical, and noninvasive manner in THA via the lateral position.

Efficacy of the Anatomical-Pelvic-Plane Positioner in Total Hip Arthroplasty in the Lateral Decubitus Position.

BACKGROUND: Total hip arthroplasty in the lateral position involves particularly large variance in the sagittal tilt of pelvis fixation, which affects the imprecision of the cup anteversion leading to poor outcomes. We have added an additional compression pad to an existing device, also to be used in the lateral position, but theoretically enabling fixation on the anatomical pelvic plane (APP) serving as the reference plane. The present study aims to evaluate the usefulness of this device in comparison with the conventional device. METHODS: We have studied 141 patients who underwent total hip arthroplasty at our hospital. Two frontal plain x-rays of the pelvis were obtained preoperatively for each patient after pelvis fixation; one with the conventional lateral fixation device and the other with an APP lateral fixation device. The sagittal tilt of the pelvis in each position was measured with 3D templating software, and variance in the sagittal tilt was compared between the 2 devices. RESULTS: The mean bias in sagittal tilt relative to the functional pelvic plane (FPP) in the conventional device was -5.0° ± 4.8° (minus mean backward tilt) and was within 5° relative to the functional pelvic plane in 43%. The mean bias in the sagittal tilt relative to the APP in the APP lateral position device was 1.7° ± 3.1° (forward tilt) and was within 5° relative to the APP in 89%. The APP lateral device significantly reduced the variance in the sagittal tilt. CONCLUSION: This device holds promise as a means of reducing the sagittal tilt in a simple, minimally invasive, and highly cost-effective manner.

Local co-application of zoledronate promotes long-term maintenance of newly formed bone induced by recombinant human bone morphogenetic protein 2.

Bone Morphogenetic Proteins (BMPs) strongly induce the recruitment and differentiation of mesenchymal progenitor cells into mature osteoblasts, but also directly and indirectly stimulate differentiation of osteoclast progenitor cells and acceleration of mature osteoclasts function leading excessive bone resorption. Bisphosphonates, such as zoledronate (ZOL), inhibit osteoclasts function and osteoclasts mediated bone resorption. The short or middle term effect of BMPs and bisphosphonates on bone formation were previously reported, but there was no study that argue about the long term effect of bisphosphonates on BMP-induced bone anabolism. The present study demonstrated that the local administration of ZOL with recombinant human BMP-2 (rh-BMP-2) using beta tricalcium phosphate (ß-TCP) as a carrier had superior efficacy not only to augment the BMP-induced new ectopic bone formation but to maintain the trabecular bone structure inside the new bone for long period. Histological analysis showed that rh-BMP-2/ß-TCP composite induced trabecular bone resorption especially inside the new bone nodules over time, whereas no trabecular bone resorption was seen in rh-BMP-2/ZOL/ß-TCP composite reducing the number of TRAP-positive cells. Thus, inhibition of bone resorption by bisphosphonate, such as ZOL, would be one of the advantageous ways to augment the new bone formation induced by rh-BMP-2, and moreover local co-application of ZOL using ß-TCP as a carrier can be a useful material for long term suppression of osteoclastic resorption and thereby maintain the structure of new bone formation induced by rh-BMP-2.

Fate, origin and roles of cells within free bone grafts.

BACKGROUND: The efficacy of autologous bone grafting in repairing nonunion fractures, large bone defects and spinal instability is widely accepted. However, the cellular and molecular mechanisms underlying new bone formation in bone grafting have yet to be fully elucidated. The purpose of this study was to clarify the fate, origin and the contribution of the cells within the grafted bone. METHODS: This study was designed to investigate the role and fate of cells contained in the grafted bone and their contribution to new bone formation in the graft in an animal model. Middiaphyseal cylindrical bone samples obtained from green fluorescent protein (GFP) transgenic and wild-type rats were transplanted into the back muscle of wild-type and GFP rats, respectively. The transplanted bones were evaluated by immunohistochemistry, in situ hybridization and quantitative reverse transcription polymerase chain reaction. RESULTS: Immunohistochemical analyses showed that all the cells in the newly formed bone originated from the grafted bone, and osteoblasts were gradually replaced by host cells. Conversely, osteoclasts were immediately replaced by host cells 2 weeks after the bone graft. In addition, expression of bone morphogenetic protein (Bmp)-4, Bmp receptors and Noggin in the grafted bone was significantly upregulated before new bone formation occurred, indicating that the grafted cells might contribute to the recruitment of mesenchymal cells into the graft bed. CONCLUSION: This study revealed the possible molecular mechanisms of the contribution of cells contained in grafted bone to facilitate new bone formation.

Number and severity of prevalent vertebral fractures and the risk of subsequent vertebral fractures in Japanese women with osteoporosis: results from the minodronate trial.

The aim was to evaluate the risk of new vertebral fractures with the increasing number and severity of prevalent vertebral fractures in women who received placebo or minodronate in a post hoc analysis of a 2-year randomized, double-blind, placebo-controlled study. The subjects were women aged 55-80 years old with 1-5 fragility fractures between the T4 and L4 vertebrae and bone mineral density <80 % of the young adult mean. A total of 704 subjects were randomized to take minodronate 1 mg (n = 359) or placebo (n = 345) once a day for 24 months. In the placebo group, the risk of incident vertebral fractures during the 2-year observational period was significantly related to the number and severity of prevalent vertebral fractures at baseline. The number of prevalent vertebral fractures was an independent risk factor for incident vertebral fracture in multivariate analysis. The relative risk reductions of vertebral fractures by minodronate treatment were 45.2, 61.1, and 64.2 % for patients with 1, 2, and ≥3 prevalent vertebral fractures, respectively, and 87.8, 64.6, and 50.1 % for patients with mild, moderate, and severe prevalent vertebral fractures, respectively. In conclusion, the number of prevalent vertebral fractures is an independent risk factor for incident vertebral fracture and minodronate reduces the fracture risk even in patients at a higher risk for fracture.

Impact of initial conservative treatment interventions on the outcomes of patients with osteoporotic vertebral fractures.

STUDY DESIGN: Prospective multicenter study. OBJECTIVE: To examine whether initial conservative treatment interventions for osteoporotic vertebral fractures (OVF) influence patient outcomes. SUMMARY OF BACKGROUND DATA: OVFs have been described as stable spinal injuries and, in most cases, are managed well with conservative treatment. However, systematic treatments for OVF have not been clearly established. METHODS: A total of 362 patients with OVF (59 males and 303 females; mean age, 76.3 yr) from 25 institutes were enrolled in this clinical study. All the patients were treated conservatively without any surgical interventions. The patient outcomes were evaluated 6 months after the fractures on the basis of Short Form-36 (SF-36) physical component summary (PCS) and mental component summary (MCS), activities of daily living (the Japanese long-term care insurance system), back pain (visual analogue scale), cognitive status (mini-mental state examination), and vertebral collapse, which were used as response variables. Furthermore, brace type, hospitalization, bisphosphonates after injury, and painkillers after injury were explanatory variables for the treatment interventions. To evaluate the independent effects of treatment interventions on patient outcomes, we performed multivariate logistic regression analyses and obtained odds ratios that were adjusted for the potential confounding effects of age, sex, level of fracture, presence of middle-column injury, pain visual analogue scale at enrollment, mini-mental state examination score at enrollment, and previous use of steroids. RESULTS: There was no significant difference for treatment intervention factors including brace type, hospitalization, bisphosphonates after injury, and painkillers after injury. For adjusting factors, the presence of middle-column injury was significantly associated with SF-36 PCS ≤ 40, reduced activities of daily living, prolonged back pain, and vertebral collapse. Female sex and advanced age were associated with SF-36 PCS ≤ 40. Low mini-mental state examination scores at enrollment were associated with SF-36 PCS ≤ 40 and reduced activities of daily living. The previous use of steroids was associated with SF-36 MCS ≤ 40, prolonged back pain, and vertebral collapse. No other examined variables were significant risk factors for patient outcomes. CONCLUSION: These results showed that treatment intervention factors did not affect patient outcomes 6 months after OVF. Middle-column injury was a significant risk factor for both clinical and radiological outcomes. In the future, establishing systematic treatments for cases with middle-column injuries is needed. LEVEL OF EVIDENCE: 2.

Regenerative repair of bone defects with osteoinductive hydroxyapatite fabricated to match the defect and implanted with combined use of computer-aided design, computer-aided manufacturing, and computer-assisted surgery systems: a feasibility study in a canine model.

BACKGROUND: Currently, regenerative repair of large bone defects that result from bone tumor resection or severe trauma is a challenging issue because of the limited regenerative potential of bone and treatment modalities. The aim of this study was to achieve repair of large bone defects to the original three-dimensional (3D) anatomical state by combining computer-aided technologies and local delivery of bone morphogenetic protein (BMP) in a canine model. METHODS: Computed tomography (CT) images of the pelvic bone of each dog were obtained, and an imaginary spherical malignant bone tumor of 15-mm diameter was placed in the left ilium of a canine on the 3D CT image. Resection of the whole tumor with a 10-mm margin of healthy bone was planned preoperatively by using computer-aided design (CAD) software. In addition, an image of the implant to be used to fill the resulting bone defect was constructed on the computer image. A porous hydroxyapatite (HA) implant identical to the imaged bone defect was made by shaving a tetragonal porous apatite block (40 × 20 × 10 mm) with a computer-aided manufacturing system operated by using the CT-image data of the bone defect obtained from the CAD system. To resect the iliac bone as planned preoperatively on the 3D CT image, computer-aided surgery was performed using the CT data. The defect was filled with the HA implant fabricated as described and coated with a putty carrier either with BMP-2 (BMP group, n = 6) or without BMP-2 (control group, n = 6). RESULTS: In the BMP group, new bone formation was noted around each implant on CT images at 3 weeks after surgery and was remodeled to restore the original anatomy of the ilium on serial CT images. At 12 weeks, the implant was enclosed within new bone, and histological analysis revealed bone formation on and within the implant. Little bone formation was noted in the control group. CONCLUSIONS: This new method may enable efficacious and precise regenerative repair of large bone defects without bone grafting.

Conventional or navigated total knee arthroplasty affects sagittal component alignment.

PURPOSE: Navigation systems have recently been developed to achieve highly reliable prosthetic alignment in total knee arthroplasty (TKA). However, component alignment in the sagittal plane is less reliable than that in the coronal plane even when navigation systems are used. Previous reports examining men showed differences in targeted sagittal prosthetic alignments of TKA achieved using the conventional technique and various navigation systems. However, there have been few studies examining the use of this technique in women, who are the primary candidates for TKA. METHODS: Radiographs of the entire lower extremity were obtained from 20 elder women while standing and sagittal prosthetic alignments in TKA were planned using the conventional technique as well as three navigation approaches to establish reference frames, and the observations were compared. RESULTS: Sagittal alignments simulated using the radiographs for the conventional technique and navigation systems differed within a mean of 3.2° ± 1.7° (mean ± SD) to 6.3° ± 2.0°. The use of different reference points on the distal femoral condyles (insertion point of the intramedullary rod, center of the anteroposterior direction of the femoral condyles, and most distal point of the femoral condyles) for each navigation system resulted in differences in the sagittal plane up to 3.0° ± 1.5°. The use of navigation systems resulted in a more hyperextended position between the femoral and tibial components compared to that for the conventional technique. CONCLUSIONS: Targeted sagittal prosthetic alignments of TKA achieved using the conventional technique and various navigation systems differed in elderly women. The use of different reference points on the distal femoral condyle for navigation systems resulted in different alignment in the sagittal plane. This study showed that alignment tendencies are similar in men and women. LEVEL OF EVIDENCE: Prognostic studies, IV.

Repair of critical long bone defects using frozen bone allografts coated with an rhBMP-2-retaining paste.

BACKGROUND: Massive frozen stocked allogeneic bone grafts are often used to reconstruct large bone defects caused by trauma or tumor resections. However, the long-term failure rate of such massive allografts was reported to be 25% because of infection, fracture, and nonunion. In this study, we evaluated the ability of a recombinant human bone morphogenetic protein (rhBMP)-2-retaining paste to promote the osteogenic potential of frozen stocked allogeneic bone grafts to repair intercalated femoral shaft defects in a rat model. METHODS: After confirming the transplantation intolerance between two rat strains (Wistar and Lewis) by skin transplantation from Lewis rats to Wistar rats, an 8-mm-long bone segment was removed from the Wistar rats, and a frozen stocked allograft coated with the rhBMP-2-retaining paste from the Lewis rats was placed into the defect and subjected to intramedullary fixation with an 18-gauge injection needle pin. The allografted femurs were evaluated by radiographic, histologic, and biomechanical examinations at specified time points. RESULTS: The results revealed successful repair of critical-size cortical bone defects by implanting frozen stocked allografts coated with the rhBMP-2-retaining synthetic biodegradable carrier paste from an immunologically intolerant host. CONCLUSIONS: This experimental study suggest that allogeneic bone grafting in combination with rhBMP-2 and its local delivery system may represent an innovative approach to the reconstruction of bone defects.

Early biological fixation of porous implant coated with paste-retaining recombinant bone morphogenetic protein 2.

The aim of this study was to evaluate the period required for stable initial bone-implant fixation with recombinant human bone morphogenetic protein 2 (rhBMP-2) in the bone marrow of a rabbit model. The porous implants being coated with ß-tricalcium phosphate/polylactide-polyethylene glycol paste with 15, 30, or 60 µg of rhBMP-2 (n = 10) were implanted into animals in 3 experimental groups. In 2 control groups, the test implants were coated without rhBMP or no paste. In all groups, the implant was inserted for 3 and 6 weeks. At 3 weeks after implantation, the BMP-treated implants in the 2 lower dose groups had significantly more bone ingrowth to the implant surface than did the control groups, and the greatest effect occurred in the 30-µg rhBMP-2 group animals.

Three years of treatment with minodronate in patients with postmenopausal osteoporosis.

The objective of this study was to determine the safety and efficacy of long-term minodronate treatment in women with postmenopausal osteoporosis based on re-analysis of a phase III 2-year clinical trial with a 1-year extension. Women aged 55-80 years old with fragility fractures were enrolled and randomized to take 1 mg minodronate or placebo once a day in the original 2-year study. The subjects who completed the 2-year study were invited to participate in an additional 1-year extension in which all subjects were to receive minodronate. Finally, a total 380 subjects completed the extension study (186 from the placebo group and 194 from the minodronate group). Fracture results observed in the extension study were consistent with those observed in the first 2 years in minodronate group. In contrast, the placebo/minodronate group showed a decreased incidence of new vertebral fractures during year 3 compared to that in year 2. In the patients who received minodronate in the original 2-year study, lumbar bone mineral density (BMD) increased consistently during year 3 and bone turnover markers decreased within the first 6 months and remained constant thereafter over 3 years. Similar positive effects of minodronate on BMD and bone turnover markers occurred when therapy was initiated in the placebo/minodronate group. No new safety concerns observed during the extension period compared to the safety observations made during the 2-year study. It was concluded that daily administration of 1 mg oral minodronate is safe and well tolerated, and that the efficacy of this dose in reducing vertebral fracture risk in postmenopausal women over 2 years is sustained with continuing treatment.

Efficacy of interspinous process lumbar fusion with recombinant human bone morphogenetic protein-2 delivered with a synthetic polymer and ß-tricalcium phosphate in a rabbit model.

INTRODUCTION: As a powerful bone-inducing cytokine, rhBMP-2 has been used as a bone graft substitute in combination with animal-derived collagen to achieve interbody or posterolateral spinal fusion. Successful interspinous process fusion using rhBMP-2 in combination with synthetic carrier materials would offer a safe, minimally invasive spinal fusion option for the treatment of spinal disorders. The aims of the present study were to achieve interspinous process fusion by implanting rhBMP-2-retaining degradable material instead of bone grafting and to evaluate efficacy for vertebral stabilization. MATERIALS AND METHODS: A polymer gel (200 mg), ß-tricalcium phosphate powder (400 mg), and rhBMP-2 (0, 30, 60 or 120 µg) were mixed to generate a plastic implant, which was then placed during surgery to bridge the L5-6 interspinous processes of 58 rabbits. Control animals received implants either without rhBMP-2 or with autogenous bone chips from the iliac crest. L5-6 vertebrae were recovered 8 weeks postoperatively. Interspinous process fusion was evaluated by radiography, biomechanical bending test, intradiscal pressure (IDP) measurement, and histology. RESULTS: In bending tests, strength of fusion was significantly greater in BMP60 and BMP120 groups than in sham, BMP0, BMP30 or autogenous bone groups. IDP at L5-6 was significantly reduced in BMP60 and BMP120 groups compared to sham, BMP0, BMP30, and autograft groups. Histologically, coronal sections of the fusion mass showed a bone mass bridging both spinous processes. CONCLUSION: Solid interspinous process fusion was achieved in rabbit models by 8 weeks after implanting the biodegradable bone-inducing material. These results suggest a potential new less-invasive option without bone grafting for the treatment of lumbar disorders.

Prognostic factors for reduction of activities of daily living following osteoporotic vertebral fractures.

STUDY DESIGN: Prospective cohort study. OBJECTIVE: To elucidate the prognostic factors indicating reduced activities of daily living (ADL) at the time of the 6-month follow-up after osteoporotic vertebral fracture (OVF). SUMMARY OF BACKGROUND DATA: OVF has severe effects on ADL and quality of life (QOL) in elderly patients and leads to long-term deteriorations in physical condition. Many patients recover ADL with acceleration of bony union and spinal stability, but some experience impaired ADL even months after fracture. Identifying factors predicting reduced ADL after OVF may prove valuable. METHODS: Subjects in this prospective study comprised 310 OVF patients from 25 institutes. All patients were treated conservatively without surgery. Pain, ADL, QOL, and other factors were evaluated on enrollment and at 6 months. ADL were evaluated using the criteria of the Japanese long-term care insurance system to evaluate the degree of independence. We defined reduced ADL as a reduction of at least single grade at 6 months after fracture and investigated factors predicting reduced ADL after OVF, using uni- and multivariate regression analysis. RESULTS: ADL were reduced at 6 months after OVF in 66 of 310 patients (21.3%). In univariate analysis, age more than 75 years (P = 0.044), female sex (P = 0.041), 2 or more previous spine fractures (P = 0.009), presence of middle column injury (P = 0.021), and lack of regular exercise before fracture (P = 0.001) were significantly associated with reduced ADL. In multivariate analysis, presence of middle column injury (odds ratio [OR], 2.26; P = 0.022) and lack of regular exercise before fracture (OR, 2.49; P = 0.030) were significantly associated with reduced ADL. CONCLUSION: These results identified presence of middle column injury of the vertebral body and lack of regular exercise before fracture as prognostic factors for reduced ADL. With clarification and validation, these risk factors may provide crucial tools for determining subsequent OVF treatments. Patients showing these prognostic factors should be observed carefully and treated with more intensive treatment options.

Local delivery of siRNA using a biodegradable polymer application to enhance BMP-induced bone formation.

Small interfering RNA (siRNA) is useful tool for specific and efficient knockdown of disease-related genes. However, in vivo applications of siRNA are limited due to difficulty in its efficient delivery to target cells. In this study, we investigated the efficacy of a biodegradable hydrogel, poly-d,l-lactic acid-p-dioxanone-polyethylene glycol block co-polymer (PLA-DX-PEG), as a siRNA carrier. PLA-DX-PEG pellets with or without fluorescein-labeled dsRNA were implanted into mouse dosal muscle pouches. The cellular uptake of dsRNA surround the polymer was confirmed by fluorescent microscopy. The fluorescence intensity was dose-dependent of the dsRNA, and exhibited a time-dependent decrease. To investigate its biological efficiency, noggin (antagonoist to BMPs) gene-silencing with siRNA (siRNA/Noggin) was examined by the amount of suppression of BMP-2-induced noggin expression and the level of performance of BMP, indicated by ectopic bone formation. Noggin gene expression induced by BMP-2 was suppressed by addition of siRNA/Noggin to the implant, and the ectopic bone formation induced by implants with both BMP-2 and siRNA/Noggin was significantly greater than those induced by implants with BMP-2 alone. These results indicate the efficacy of local delivery of siRNAs by PLA-DX-PEG polymer, which intensified bone-inducing effects of BMP and promoted new bone formation by suppressing gene expression of Noggin.

Early onset of Runx2 expression caused craniosynostosis, ectopic bone formation, and limb defects.

RUNX2 is an essential transcription factor for osteoblast differentiation, because osteoblast differentiation is completely blocked in Runx2-deficient mice. However, it remains to be clarified whether RUNX2 is sufficient for osteoblast differentiation during embryogenesis. To address this issue, Runx2 transgenic mice were generated under the control of the Prrx1 promoter, which directs the transgene expression to mesenchymal cells before the onset of bone development. The transgene expression was detected in the cranium, limb buds, and the region from the mandible to anterior chest wall. The skull became small and the limbs were shortened depending on the levels of the transgene expression. Early onset of Runx2 expression in the cranial mesenchyme induced mineralization on E13.0, when no mineralization was observed in wild-type mice, and resulted in craniosynostosis as shown by the closure of sutures and fontanelles on E18.5. Col1a1 and Spp1 expressions were detected in the mineralized regions on E12.5-13.5. The limb bones were hypoplastic and fused, and ectopic bones were formed in the hands and feet. Col2a1 expression was inhibited but Col1a1 expression was induced in the limb buds on E12.5. In the anterior chest wall, ectopic bones were formed through the process of intramembranous ossification, interrupting the formation of cartilaginous anlagen of sternal manubrium. These findings indicate that RUNX2 is sufficient to direct mesenchymal cells to osteoblasts and lead to intramembranous bone formation during embryogenesis; Runx2 inhibits chondrocyte differentiation at an early stage; and that Runx2 expression at appropriate level, times and spaces during embryogenesis is essential for skeletal development.

ACL reconstruction using bone-tendon-bone graft engineered from the semitendinosus tendon by injection of recombinant BMP-2 in a rabbit model.

We attempted to generate a bone-tendon-bone structure by injecting human-type recombinant human bone morphogenetic protein-2 (rhBMP-2) into the semitendinosus tendon, and an anterior cruciate ligament (ACL) defect was reconstructed by grafting the engineered bone-tendon-bone graft. Two ossicles with a separation distance of 1 cm were generated within the left semitendinosus tendon of a rabbit 6 weeks after the injection of rhBMP-2 (15 µg at each site). The engineered bone-tendon-bone graft was transplanted in order to reconstruct the ACL by passing the graft through the bone tunnels. In the control group, the ACL was reconstructed with the semitendinosus tendon without BMP-2 using the same methods as those used in the experimental group. The animals were harvested at 4 or 8 weeks after surgery and examined by radiographic, histological, and biomechanical methods. In the experimental group, ossicles in the bone-tendon-bone graft were successfully integrated into the host bone of the femur and tibia. Histological analysis revealed that characteristic features identical to the normal direct insertion morphology had been restored. Biomechanical pull-out testing showed that the ultimate failure load and stiffness of the reconstructed ACL in the experimental group were significantly higher than those in the control group at both 4 and 8 weeks (p < 0.05). These results indicate the potential of regenerative reconstruction of the ACL, and the reconstruction resulted in the restoration of morphology and function equivalent to those of the normal ACL.

Characteristic radiographic or magnetic resonance images of fresh osteoporotic vertebral fractures predicting potential risk for nonunion: a prospective multicenter study.

STUDY DESIGN: Prospective multicenter study. OBJECTIVE: To identify radiographic or magnetic resonance (MR) images of fresh vertebral fractures that can predict a high risk for delayed union or nonunion of osteoporotic vertebral fractures (OVFs). SUMMARY OF BACKGROUND DATA: Vertebral body fractures are the most common fractures in osteoporosis patients. Conservative treatments are typically chosen for OVFs, and associated back pain generally subsides within several weeks with residual persistent deformity of the vertebral body. In some patients, OVF healing is impaired and correlated with prolonged back pain. However, assessments such as plain radiograph or MR images taken during the early phase to predict high risks for nonunions of OVFs and/or poor prognoses have not been identified. METHODS: A total of 350 OVF patients from 25 institutes were enrolled in this clinical study. Plain radiograph and MR images of the OVFs were routinely taken at enrollment at the respective institutes. The findings on the plain radiograph and MR images were classified after enrollment in the study. All the patients were treated conservatively without any surgical intervention. After a 6-month follow-up, the patients were classified into two groups, a union group and a nonunion group, depending on the presence of an intravertebral cleft on plain radiograph or MR images. The associations of the images from the first visit with those of the corresponding nonunions at the 6-month follow-up were analyzed by multivariate logistic regression to elucidate specific image characteristics that may predict a high risk for nonunion of OVFs. RESULTS: Forty-eight patients (49 vertebrae) among the 350 patients (363 vertebrae) were classified as nonunions, indicating a nonunion incidence of 13.5% for conventional conservative treatments for OVFs. The statistical analyses revealed that a vertebral fracture in the thoracolumbar spine, presence of a middle-column injury, and a confined high intensity or a diffuse low intensity area in the fractured vertebrae on T2-weighted MR images were significant risk factors for nonunion of OVFs. CONCLUSION: The results of this study revealed significant relationships between plain radiograph and MR images of acute phase OVFs and the incidence of nonunion. As these risk factors are defined more clearly and further validated, they may become essential assessment tools for determining subsequent OVF treatments. Patients with one or more of the earlier-described risk factors for nonunion should be observed carefully and provided with more intensive treatments.