Takatoshi Okuda

The slow resorption with replacement by bone of a hydrothermally synthesized pure calcium-deficient hydroxyapatite

A newly developed calcium-deficient hydroxyapatite composed of rod-shaped particles synthesized by the hydrothermal method (HHA) and stoichiometric hydroxyapatite (SHA) synthesized by the sintering method was used for in vivo implantation and in vitro culture systems to compare these biological responses. In the rabbit femur, implanted HHA was slowly resorbed and about 80% of the implant remained 24 weeks after implantation; however, up to 72 weeks after implantation, most of the implanted HHA was resorbed. The implanted SHA was unresorbed throughout the experimental period, but degradation by the invasion of newly formed bone was seen at 72 weeks after implantation. Bone histomorphometry showed that the volume of newly formed bone and the number of osteoclasts in the implanted region were significantly higher in HHA than in SHA 24 weeks after implantation. In vitro culture of C2C12 cells with the induction of osteoblastic phenotypes using recombinant bone morphogenetic protein-2 showed similar cell density and the induction of alkaline phosphatase activity between the cells on HHA and SHA discs. In vitro osteoclastogenesis of HHA and SHA discs using bone marrow macrophages and recombinant receptor activator of nuclear factor-kappaB ligand showed higher TRAP activity of osteoclasts cultured on HHA discs. These results showed that slow biodegradability did not always correlate to final replaceability in bone tissue, and suggested that the activity of osteoclasts correlated to the bone-forming activity of osteoblasts.

Stimulatory effect of hydrothermally synthesized biodegradable hydroxyapatite granules on osteogenesis and direct association with osteoclasts

Calcium-deficient hydroxyapatite (HA) granules with a unique spherical shape were prepared using an applied hydrothermal method. Spherical stoichiometric HA granules were also prepared by normal sintering and both granules were used for implantation into rat tibiae to compare the biological responses to each implant. Twelve and 24 weeks after implantation, the volume of calcium-deficient HA granules was significantly less than that of stoichiometric HA granules, and the biodegradability of calcium-deficient HA granules was confirmed. The larger number of osteoclasts, larger osteoblast surface and larger bone volume in the implanted area of calcium-deficient HA than those of stoichiometric HA suggested that osteoclastic resorption of calcium-deficient HA affected osteogenesis in that area. To analyze the direct contribution of osteoclasts to osteogenesis, C2C12 multipotent myoblastic cells, which have the potential to differentiate into osteoblasts in the presence of bone morphogenetic protein 2, were cultured with supernatants of osteoclasts cultured on calcium-deficient HA, stoichiometric HA, beta-tricalcium phosphate disks or plastic dishes, or bone marrow macrophages cultured on plastic dishes. Supernatants of osteoclasts but not bone marrow macrophages stimulated the expression of Runx2 and osteocalcin in C2C12 cells in concert with bone morphogenetic protein 2. The expression of alkaline phosphatase was stimulated with supernatants of osteoclasts cultured on ceramic disks. These results suggested that osteoclasts produced certain soluble factors which stimulated osteoblastic differentiation and they were thought to be associated with the induction of a larger osteoblast surface and bone volume in the animals implanted with calcium-deficient HA granules.

Magnetic resonance imaging evaluation of patients with idiopathic scoliosis: a prospective study of four hundred seventy-two outpatients.

Study Design. A prospective study of magnetic resonance imaging findings in outpatients with idiopathic scoliosis. Objective. The purpose of this study was to determine the prevalence of neural axis abnormalities in outpatients with scoliosis and to analyze the characteristics of patients who had such abnormalities. Summary of Background Data. In previous studies, neural axis abnormalities were found in 2.9% to 37% of patients with idiopathic scoliosis. The current guidelines for MRI screening in scoliosis are valuable, and the proposed indications for performing MRI in the literature include early onset, atypical curvature, double thoracic curve (King type-5), rapid progression, male gender, and abnormal neurologic findings. Methods. A total of 472 outpatients with a primary diagnosis of idiopathic scoliosis were examined for neural axis abnormalities by magnetic resonance imaging. Logistic regression was used to determine significant predictors of neural axis abnormalities on MRI. Results. The incidence of neural axis abnormalities on MRI was 3.8% (18 of 472 patients). Among the 18 patients, 6 had a Chiari I malformation alone, 10 had a Chiari I malformation combined with syringomyelia, and 2 had a syringomyelia without Chiari I malformation. Male gender, patients younger than 11 years old, and abnormal superficial abdominal reflexes were significantly associated with the detection of neural axis abnormalities on MRI. Conclusion. We recommend routine use of MRI in male patients, younger than 11 years old, and abnormal superficial abdominal reflexes. Even if a patient has no specific indications for MRI, we recommend its routine use in preoperative planning.

Retrodental mass in rheumatoid arthritis.

Study Design: A retrospective investigation of the retrodental mass secondary to rheumatoid arthritis (RA). Objective: To propose a new classification of the retrodental mass in RA, and to evaluate their resorption processes and surgical procedures. Summary of Background Data: The retrodental mass secondary to RA has long been recognized as pannus formation. It is also known that pannus will disappear or radically reduce after stabilization of the atlantoaxial segment. The past reports, however, leave unanswered the following question; are there other types of mass with significantly different degeneration processes from the pannus? The need for anterior transoral decompression is still controversial. Methods: Eleven patients with retrodental masses in RA were retrospectively analyzed. They underwent posterior fusion without decompression for atlantoaxial subluxation and occipitocervical fusion with decompressive laminectomy of the atlas for vertical subluxation. All patients had neurological, radiologic, and magnetic resonance imaging (MRI) evaluations both before and after surgery. MRI study was performed preoperatively and at 1-month interval after surgery until the mass had disappeared or stopped further reduction. Results: This study identified 3 distinctive types in the rheumatoid retrodental mass on MRI. Type 1 displayed high intensity on T2 and low intensity on T1-weighted MRI or the pattern specific to pannus. Type 2 was identified with low intensity on T2-weighted MRI or pattern specific to pseudotumor. Type 3 displayed a combination of high and low intensity on T2-weighted images or indication that the mass was the mixture of pannus and pseudotumor. All the masses of types 1 and 3 disappeared within 1 month of surgery. The process in type 2 was found a few months slower. In all 11 cases, myelopathy improved postoperatively to the status before the emergence of the symptom. Conclusions: This article recognized 3 distinctive types of the retrodental mass in RA; type 1 (pannus), type 2 (pseudotumor), type 3 (mixed). It further concludes in all types of the retrodental mass associated with atlantoaxial subluxation secondary to RA, posterior fusion without decompression can achieve improvement of the myelopathy. In type 2 associated with vertical subluxation, on balance between calculable benefits and high risks of anterior transoral decompression, the authors prefer to opt for occipitocervical fusion with decompressive laminectomy of the atlas.

Porous Granules of β-Tricalcium Phosphate Composed of Rod-Shaped Particles

Porous granules of β-tricalcium phosphate (β-Ca3(PO4)2; β-TCP) were prepared from porous hydroxyapatite granules with calcium deficient composition synthesized by hydrothermal method. The β-TCP granules were composed of rod-shaped particles of about 10-20 µm in length. Rod-shaped particles were locked together to make micro-pores, and the size of micro-pores formed by tangling of rod-shaped particles was about 0.1-0.5 µm. The granule size, particle size, and micro-pore size could be controlled by our unique method. The porous granules of β-TCP must be suitable for the bone graft material and for scaffold of cultured bone.

Application of Newly Developed Globular-Shaped Granules of β-Tricalcium Phosphate for Bone Substitute

Globular-shaped beta-tricalcium phosphate (β-TCP) granules were synthesized using a unique dropping slurry method and the biological response was analyzed by implantation into the rat femur. Two kinds of globular-shaped β-TCP granules, composed of either rod-shaped particles or conventional non-rod-shaped particles were synthesized, implanted into the left femurs of 8-week-old female Wistar rats, and analyzed histologically. Commercially available β-TCP granules composed of conventional non-rod-shaped particles were also implanted as a control experiment. Four weeks after the operation, part of each implant was already resorbed and the resorbed area was replaced by newly formed bone tissue and bone marrow. Eight weeks after the operation, the resorption and replacement were advanced in each implant. At 12 weeks after the operation, residual globular-shaped β-TCP granules composed of rod-shaped particles were resorbed into compact petrotic bone tissue almost completely. The formation of less compact petrotic bone tissue was observed in specimens implanted with globular-shaped β-TCP granules composed of conventional non-rod-shaped particles. Commercially available β-TCP granules composed of conventional non-rod-shaped particles were mostly resorbed into trabecular bone and the formation of compact petrotic bone tissue was rarely observed. These data suggested that both the unique spherical shape and microstructure of β-TCP particles affected bone-forming activity after the operation.

Stimulation of Osteogenesis in Bone Defects Implanted with Biodegradable Hydroxyapatite Composed of Rod-Shaped Particles under Mechanical Unloading.

The aim of this study was to evaluate the influence of mechanical unloading on the repair of bone defects with implantation of biodegradable bone substitutes. Spherical granules of biodegradable hydroxyapatite composed of rod-shaped particles (RHA) or beta-tricalcium phosphate composed of rod-shaped particles (RTCP) were implanted into a bone defect created in the distal end of the right femur of 8-week-old Wistar rats. Two, 6, 10, and 22 weeks after implantation, part of the sciatic nerve in the thigh was resected and exposed to mechanical unloading for 2 weeks. Then, 4, 8, 12 and 24 weeks after implantation, repair of the bone defect was analyzed. As a control, the bone defect without implantation of ceramic granules was also analyzed. Both RHA and RTCP tended to be reduced, but the reduction was not obvious during the experimental period. At 12 and 24 weeks after implantation, the amount of newly formed bone in the animal implanted with RHA was significantly greater than that at 4 weeks after implantation, but that in the animal implanted with RTCP or without implantation was not significantly different. The number of osteoclasts in the region implanted with RHA was significantly larger than that of the region implanted with RTCP or without implantation at 12 and 24 weeks. The activities of alkaline phosphatase in osteoblasts and tartrate-resistant acid phosphatase in osteoclasts were remarkably increased in the bone defects with implantation compared with those in the bone defects without implantation. These results suggested that RHA stimulated osteogenesis and osteoclastogenesis even after 2 weeks of mechanical unloading, and that RHA could be expected to improve the repair of bone defects in patients under the condition of skeletal unloading.

Osteoconductivity of hydrothermally synthesized beta-tricalcium phosphate composed of rod-shaped particles under mechanical unloading

Spherical beta-tricalcium phosphate (b-TCP) granules synthesized using a unique dropping slurry method expressed good osteoconductivity with prominent bone apposition and bioresorbability when implanted into the rat femur (Gonda et al., Key Eng. Mater. 361-363:1013-1016, 2008). The spherical b-TCP granules were implanted into the bone defect created in the distal end of the right femur of each 8-week-old female Wistar rat. To analyze performance of the spherical b-TCP granules as bone substitute in the bone with reduction in osteogenic potential, the right sciatic neurectomy was performed after implantation and the right hind limb was kept unloaded for 2 weeks before euthanization. Four weeks after implantation, some spherical b-TCP granules with resorption in part were surrounded by newly formed bone. Eight and 12 weeks after implantation, most of the residual b-TCP granules were embedded in newly formed bone, and total volume of the implant and newly formed bone was more than the other portions of the bone or the bone of control animals. Osteoclast activity in the implanted area was also higher than the other portions of the bone or the bone of control animals. Replacement of the intraosseous residual b-TCP granules for bone progressed at 12 weeks after implantation compared to those at 8 weeks after implantation. These data suggested that the spherical b-TCP granules stimulated osteogenesis and osteoclast activity of the unloaded bone.

Microstructure Designing of Porous β-Tricalcium Phosphate for Control of Reactions in the Bone

Porous materials of β-tricalcium phosphate (β-Ca3(PO4)2; β-TCP) were prepared from porous hydroxyapatite (Ca10(PO4)6(OH)2; HA) with calcium deficient composition of Ca/P molar ratio of 1.50 synthesized by hydrothermal method. The porous β-TCP was composed of rod-shaped particles of about 10-20 μm in length. Rod-shaped particles were locked together to make micro-pores, and the size of micro-pores formed by tangling of rod-shaped particles was about 0.1-0.5 μm. The particle size and micro-pore size could be controlled by our unique method. It must be suitable for the bone graft material and as the scaffold of cultured bone.

Porous Ceramics of β-Tricalcium Phosphate Composed of Rod-Shaped Particles is Highly Biodegradable and Expresses Prominent Osteoconductivity

Newly developed porous β-tricalcium phosphate composed of rod-shaped particles was grafted into distal end of the left femur of female Japan White rabbit. As a control, hydrixyapatite generated in the same size was grafted. In both implants, 350µm in diameter-sized holes were created vertically and horizontally. Three weeks after grafting, margin of the β-tricalcium phosphate was absorbed by osteoclasts and bone formation was seen near the absorbed region. Twenty-four weeks after grafting, more than 80% of the β-tricalcium phosphate was absorbed and newly formed bone was prominently observed. The hydroxyapatite was not remarkably absorbed even 24 weeks after grafting, and newly formed bone was observed only in the holes created in the implants. These results suggest that porous β-tricalcium phosphate composed of rod-shaped particles has highly biodegradable and osteoconductive nature, and appropriate to bone graft substitute.