Masafumi Kashii

Long-term results of cervical myelopathy due to ossification of the posterior longitudinal ligament with an occupying ratio of 60% or more.

Study Design. Retrospective study. Objective. We sought to determine the long-term outcomes of laminoplasty versus anterior decompression and fusion in the treatment of cervical myelopathy caused by ossification of the posterior longitudinal ligament (OPLL) and to ascertain what factors should be considered in selecting appropriate surgical procedure. Summary of Background Data. There are little data about long-term results of cervical myelopathy due to OPLL with an occupying ratio 60% or more. Methods. We retrospectively studied 27 patients having OPLL with an occupying ratio 60% or more and a follow-up period of at least 2 years. Clinical outcome was evaluated using Japanese Orthopaedic Association scores and recovery rates (≥75%, excellent; 50%–74%, good; 25%–50%, fair; and <25%, poor). Results. The mean age and the mean duration of follow-up were 57 years and 10.2 years. The mean Japanese Orthopaedic Association score was 9.3 before surgery and 12.4 at the final follow-up examination. There were 15 patients in the laminoplasty group (LAM group) and 12 patients in the anterior decompression and fusion group (ADF group). The ADF group had a significantly better recovery rate at final evaluation (53% vs. 30%; P = 0.04), a longer duration of surgery (314 vs. 128 min; P < 0.01), and greater blood loss (600 vs. 240 mL; P < 0.01) than did the LAM group. In the LAM group, 4 patients with excellent or good results had a significantly larger degree of cervical lordosis (30°vs. 10°; P = 0.002) than others. Conclusion. The ADF group had a significantly better recovery rate than the LAM group, although the degree of surgical invasiveness was high. ADF is generally recommended for OPLL with an occupying ratio 60% or more. Level of Evidence: 3

A genome-wide association study identifies susceptibility loci for ossification of the posterior longitudinal ligament of the spine

Ossification of the posterior longitudinal ligament of the spine (OPLL) is a common spinal disorder among the elderly that causes myelopathy and radiculopathy. To identify genetic factors for OPLL, we performed a genome-wide association study (GWAS) in ∼8,000 individuals followed by a replication study using an additional ∼7,000 individuals. We identified six susceptibility loci for OPLL: 20p12.3 (rs2423294: P = 1.10 × 10−13), 8q23.1 (rs374810: P = 1.88 × 10−13), 12p11.22 (rs1979679: P = 4.34 × 10−12), 12p12.2 (rs11045000: P = 2.95 × 10−11), 8q23.3 (rs13279799: P = 1.28 × 10−10) and 6p21.1 (rs927485: P = 9.40 × 10−9). Analyses of gene expression in and around the loci suggested that several genes are involved in OPLL etiology through membranous and/or endochondral ossification processes. Our results bring new insight to the etiology of OPLL.

Alendronate treatment promotes bone formation with a less anisotropic microstructure during intramembranous ossification in rats

There are safety concerns regarding administration of bisphosphonates to children. Little is known about the effects of bisphosphonates on bone matrix organization during bone modeling. The present study examined the effects of alendronate (ALN) on bone matrices formed by intramembranous ossification in the appendicular growing skeleton. ALN was administered to 1-week-old Sprague–Dawley rats at a dose of 0, 35, or 350 µg/kg/week for 4 or 8 weeks. The position of femoral diaphysis formed exclusively by intramembranous ossification was identified, and cross sections of cortical bone at this position were analyzed. Bone mineral density (BMD) and geometric parameters were evaluated using peripheral quantitative computed tomography. The preferential orientation degree of biological apatite (BAp) crystals in the bone longitudinal direction, which shows the degree of bone matrix anisotropy, was evaluated using microbeam X-ray diffraction analysis. We analyzed bone histomorphometrical parameters and performed bone nanomechanical tests to examine the material properties of newly developing cortical bone. The preferential orientation degree of BAp crystals significantly decreased in 35 µg/kg/week ALN-treated groups compared with vehicle-treated groups, although there were no significant differences in BMD between the two groups. The periosteal mineral apposition rate significantly increased in the 35 µg/kg/week ALN-treated group. We found a high negative correlation between bone matrix anisotropy and the regional periosteal mineral apposition rate (r = −0.862, P < 0.001). Nanomechanical tests revealed that 35 µg/kg/week ALN administration caused deterioration of the material properties of the bone microstructure. These new findings suggest that alendronate affects bone matrix organization and promotes bone formation with a less anisotropic microstructure during intramembranous ossification.

Three-dimensional measurement of growth of ossification of the posterior longitudinal ligament

OBJECT Ossification of the posterior longitudinal ligament (OPLL) is a progressive disease that causes cervical myelopathy. Because 2D evaluation of ossification growth with plain lateral radiographs has limitations, the authors developed a unique technique to measure ossification progression and volume increase by using multidetector CT scanning. METHODS The authors used serial thin-slice volume data obtained by multidetector CT scanning in 5 patients. The mean patient age was 63 years, and the mean follow-up duration was 3.1 years. First, a 3D model of OPLL was semiautomatically segmented at a specific threshold. Then, a preoperative model of OPLL was superimposed on a postoperative model using voxel-based registration of the vertebral bodies. Progression and volume increase were measured using a digital viewer that was developed by the authors. Progression was visualized using a color-coded contour on the surface of the OPLL model. RESULTS All patients had progression of 0.5 mm or greater. The mean values concerning OPLL growth were as follows: maximum progression length, 4.7 mm; progression rate, 1.5 mm/year; volume increase, 1622 mm(3); volume expansion rate, 37%; and volume increase rate, 484 mm(3)/year. The accuracy of superimposition by voxel-based registration, defined as closeness to the true value, was less than 0.31 mm. For intraobserver reproducibility of the volume measurement, the mean intraclass correlation coefficient, root mean square error, and coefficient of variation were 0.987, 16.0 mm(3), and 1.7%, respectively. CONCLUSIONS Ossification of the posterior longitudinal ligament progresses even after surgery. Three-dimensional evaluation with the aid of CT scans is a useful and reliable method for assessing that growth.

Kinematics of the thoracic spine in trunk rotation: in vivo 3-dimensional analysis.

Study Design. In vivo 3-dimensional (3D) study of the thoracic spine. Objective. To demonstrate axial rotations (ARs) and coupled motions of the thoracic spine. Summary of Background Data. In vivo 3D kinematics of the thoracic spine in trunk rotation with intact thorax and soft tissues has not been well-known. There were no quantitative data of AR in the consecutive thoracic spinal segments. Patterns of coupled motion with AR have been controversial. Methods. Thirteen healthy volunteers underwent 3D computed tomography of the thoracic spine in 3 positions; neutral, right, and left maximum trunk rotation. Relative motions of vertebrae were calculated by automatically superimposing the vertebrae in a neutral position over images in rotational positions, using voxel-based registration. Motions were represented with 6 degrees of freedom by Euler angles and translations on the local coordinate system. Results. Mean (±SD) relative rotational angles of T1 with respect to L1 to 1 side were 24.9° ± 4.9° in maximum trunk rotation. AR of each thoracic segment with respect to the inferior adjacent vertebra to 1 side was 1.2° ± 0.8° at T1–T2, 1.6° ± 0.7° at T2–T3, 1.4° ± 0.9° at T3–T4, 1.6° ± 0.8° at T4–T5, 1.8° ± 0.7° at T5–T6, 1.9° ± 0.6° at T6–T7, 2.3° ± 0.7° at T7–T8, 2.5° ± 0.8° at T8–T9, 2.7° ± 0.6° at T9–T10, 2.6° ± 0.8° at T10–T11, 1.3° ± 0.7° at T11–T12, and 0.5° ± 0.4° at T12–L1. Significantly larger segmental AR was observed at the middle thoracic segments (T6–T11) than at the upper (T1–T6) and lower (T11–L1) segments. At the upper thoracic segments, coupled lateral bending with AR was observed in the same direction as AR. However, at the middle and lower thoracic segments, coupled lateral bending occurred both in the same and opposite directions. Conclusion. In vivo 3D ARs and coupled motions of the consecutive thoracic spinal segments in trunk rotation were investigated accurately for the first time.

Effect of Intermittent Administration of Teriparatide (Parathyroid Hormone 1-34) on Bone Morphogenetic Protein-Induced Bone Formation in a Rat Model of Spinal Fusion.

BACKGROUND Although clinical bone morphogenetic protein (BMP) therapy is effective at enhancing bone formation in patients managed with spinal arthrodesis, the required doses are very high. Teriparatide (parathyroid hormone 1-34) is approved by the U.S. Food and Drug Administration to treat osteoporosis and is a potent anabolic agent. In this study, intermittent administration of parathyroid hormone 1-34 combined with transplantation of BMP was performed to elucidate the effect of parathyroid hormone 1-34 on the fusion rate and quality of newly formed bone in a rat model. METHODS A total of forty-eight male Sprague-Dawley rats underwent posterolateral lumbar spinal arthrodesis with one of three different treatments with recombinant human (rh) BMP-2: (1) 0 μg (control), (2) 2 μg (low dose), or (3) 50 μg (high dose). Each of the rhBMP-2 treatments was studied in combination with intermittent injections of either parathyroid hormone 1-34 (180 μg/kg/wk) or saline solution starting two weeks before the operation and continuing until six weeks after the operation. Osseous fusion was assessed with use of radiographs and a manual palpation test. Microstructural indices of the newly formed bone were evaluated with use of micro-computed tomography. The serum markers of bone metabolism were also quantified. RESULTS The fusion rate in the group treated with 2 μg of rhBMP-2 significantly increased (from 57% to 100%) with the administration of parathyroid hormone 1-34 (p < 0.05). The fusion rates in the other groups did not change significantly with the administration of parathyroid hormone 1-34. The bone volume density of the newly formed bone significantly increased in both the 2-μg and 50-μg rhBMP-2 treatment groups with the administration of parathyroid hormone 1-34 (p < 0.01). Micro-computed tomography scans of the newly formed bone clearly demonstrated an abundance of trabecular bone formation in the group treated with parathyroid hormone 1-34. In addition, serum levels of osteocalcin were significantly increased in the parathyroid hormone 1-34 treatment group. CONCLUSIONS Intermittent administration of parathyroid hormone 1-34 significantly increased fusion rates in the group treated with low-dose rhBMP-2, and it improved the quality of the newly formed bone in both the high and low-dose groups in a rat model of rhBMP-2-induced spinal fusion. CLINICAL RELEVANCE Our results suggest that the combined administration of rhBMP-2 and parathyroid hormone 1-34 may lead to efficient bone regeneration.

In vivo three-dimensional kinematics of the cervical spine during head rotation in patients with cervical spondylosis.

Study Design. Kinematics of the cervical spine during head rotation was investigated using 3-dimensional (3D) magnetic resonance imaging (MRI) in patients with cervical spondylosis (CS). Objective. To demonstrate in vivo 3D kinematics of the spondylotic cervical spine during head rotation. Summary of Background Data. Several in vivo studies have identified kinematic differences between normal and spondylotic subjects, but only two-dimensional flexion/extension motion has been investigated. Differences of in vivo 3D cervical motion during head rotation between normal and spondylotic subjects have yet to be clarified. Methods. Ten healthy volunteers (control group) and 15 patients with CS (CS group) underwent 3D MRI of the cervical spine with the head rotated to 5 positions (neutral, ±45° and ±maximal head rotation). Relative motions of the cervical spine were calculated by automatically superimposing a segmented 3D MRI of the vertebra in the neutral position over images for each position using volume registration. The 3D motions of adjacent vertebra were represented with 6 degrees of freedom by Euler angles and translations on the coordinate system. Results. Compared with the control group, the CS group showed significantly decreased mean axial rotation and mean coupled lateral bending at C5–C6 and C6–C7 and significantly increased mean coupled lateral bending at C2–C3 and C3–C4, although both the groups showed the same pattern of coupled motions. Conclusion. The in vivo 3D kinematics of the spondylotic cervical spine during head rotation was accurately depicted and compared with those of healthy cervical spines for the first time.

The bone morphogenetic protein-2/7 heterodimer is a stronger inducer of bone regeneration than the individual homodimers in a rat spinal fusion model

BACKGROUND CONTEXT Bone morphogenetic proteins (BMPs) are a group of dimeric growth factors that belong to the transforming growth factor super family and are capable of eliciting new bone formation. Previous studies have suggested that the coexpression of two different BMP genes in a cell can result in the production of BMP heterodimers that are more potent than homodimers. However, because of the difficulty in optimizing the level of BMP gene expression, the coexpression of two different BMP genes also produces BMP homodimers as a by-product. These homodimers could, in theory, interact with the heterodimers. PURPOSE To elucidate the effects of a BMP-2/7 heterodimer, which were investigated in depth using purified BMP-2/7 heterodimers, BMP-2 homodimers, and BMP-7 homodimers in a rat spinal fusion model. METHODS Bilateral posterolateral fusion at L4-L5 was performed in four different groups: control group animals were implanted with collagen carriers alone; BMP-7 group animals with collagen carriers+1 μg of BMP-7 homodimer; BMP-2 group animals with collagen carriers+1 μg of BMP-2 homodimer; and BMP-2/7 group animals with collagen carriers+1 μg of the BMP-2/7 heterodimer. The following assessments were performed: bone microstructural analysis of the fusion mass and tissue volume (TV) with microcomputed tomography (micro-CT); fusion assessment with manual palpation testing and three-dimensional CT images; and bone histomorphometrical analysis of the fusion mass. RESULTS The fusion scores, as determined by radiography, and the TV of the newly formed bone, as determined by micro-CT, were significantly higher in the BMP-2/7 heterodimer group than the other groups (p<.0001). The microstructural indices of the newly formed bone did not differ between the groups. Moreover, histologic analysis of the fused spines revealed that the formation of the trabecular bone bridging the transverse process was the highest in this group. CONCLUSIONS This study demonstrated that BMP-2/7 heterodimer is a stronger inducer of bone regeneration than BMP-2 or -7 homodimers. The use of a purified BMP-2/7 heterodimer may represent an efficient alternative to the current clinical use of BMP-2 or -7 homodimers. Further studies as to the side effects of BMP-2/7 heterodimer are required.

Kinematics of the thoracic spine in trunk lateral bending: in vivo three-dimensional analysis

BACKGROUND CONTEXT In vivo three-dimensional kinematics of the thoracic spine in trunk lateral bending with an intact rib cage and soft tissues has not been well documented. There is no quantitative data in the literature for lateral bending in consecutive thoracic spinal segments, and there has not been consensus on the patterns of coupled motion with lateral bending. PURPOSE To demonstrate segmental ranges of motion (ROMs) in lateral bending and coupled motions of the thoracic spine. STUDY DESIGN In vivo three-dimensional biomechanics study of the thoracic spine. PATIENT SAMPLE Fifteen healthy male volunteers. OUTCOME MEASURES Computed analysis by using voxel-based registration. METHODS Participants underwent computed tomography of the thoracic spine in three supine positions: neutral, right maximum lateral bending, and left maximum lateral bending. The relative motions of vertebrae were calculated by automatically superimposing an image of vertebrae in a neutral position over images in bending positions, using voxel-based registration. Mean values of lateral bending were compared among the upper (T1-T2 to T3-T4), the middle-upper (T4-T5 to T6-T7), the middle-lower (T7-T8 to T9-T10), and the lower (T10-T11 to T12-L1) parts of the spine. RESULTS At lateral bending, the mean ROM (±standard deviation) of T1 with respect to L1 was 15.6°±6.3° for lateral bending and 6.2°±4.8° for coupled axial rotation in the same direction as lateral bending. The mean lateral bending of each spinal segment with respect to the inferior adjacent vertebra was 1.4°±1.3° at T1-T2, 1.3°±1.2° at T2-T3, 1.4°±1.3° at T3-T4, 0.9°±0.9° at T4-T5, 0.8°±1.0° at T5-T6, 1.1°±1.1° at T6-T7, 1.7°±1.2° at T7-T8, 1.3°±1.2° at T8-T9, 1.6°±0.7° at T9-T10, 1.8°±0.8° at T10-T11, 2.3°±1.0° at T11-T12, and 2.2°±0.8° at T12-L1. The smallest and the largest amounts of lateral bending were observed in the middle-upper and the lower parts, respectively. There was no significant difference in lateral bending between the upper and the middle-lower parts. Coupled axial rotation of each segment was generally observed in the same direction as lateral bending. However, high variability was found at the T2-T3 to T5-T6 segments. Coupled flexion was observed at the upper and middle parts, and coupled extension was observed at the lower part. CONCLUSIONS This study revealed in vivo three-dimensional motions of consecutive thoracic spinal segments in trunk lateral bending. The thoracolumbar segments significantly contributed to lateral bending. Coupled axial rotation generally occurred in the same direction with lateral bending. However, more variability was observed in the direction of coupled axial rotation at T2-T3 to T5-T6 segments in the supine position. These results are useful for understanding normal kinematics of the thoracic spine.

Femtosecond Laser Processing of Protein Crystals in Crystallization Drop

We have developed a novel processing technique for protein crystals using femtosecond laser irradiation. We call this technique the femtosecond-laser-induced cut and cleave operation (fs-CACO). By precisely controlling the laser fluence and the position of the laser focal point, we are able to perform accurate and reproducible processing of protein crystals with little damage without troublesome treatment such as the unsealing of vessels and removal of solutions surrounding the crystals. To our knowledge, this is the first report on processing protein crystals using ultra-short-pulse laser ablation. Fs-CACO will be a powerful tool for making problematic protein crystals suitable for X-ray diffraction measurements.