Yusuke Kozai

The effect of simvastatin-loaded polymeric microspheres in a critical size bone defect in the rabbit calvaria

The present study describes the development of a microsphere capsule based on polylactide-co-glycolide (PLGA) loaded with simvastatin that was subsequently incorporated into synthetic bone cement. The osteogenic effect of simvastatin-loaded bone cement was in a critical sized defect in vivo to test the hypothesis the biologic response would be different depending on the dosage of simvastatin applied to bone cement. Our results showed that simvastatin loaded PLGA microspheres can be successfully obtained through O/W emulsion/solvent evaporation method with appropriate morphologic characteristics and high encapsulation efficiency for incorporation in bone cements. The biodegradable characteristic of the microspheres successfully presented a slow release and the duration of the release lasted for more than 1 month. The in vivo experiment revealed that the microspheres containing simvastatin significantly enhanced bone formation in the rabbit calvaria critical size defect.

The influence of nano hydroxyapatite coating on osseointegration after extended healing periods

OBJECTIVE Studies observing early wound healing periods around dental implants demonstrate an implants ability to enhance osseointegration, the bone-implant interactions for extended healing periods though have not been thoroughly studied. METHODS Twenty threaded titanium alloy (Ti6Al4V, Grade 5) implants were inserted bilaterally, half prepared to impart stable hydroxyapatite nanoparticles onto a sand blasted and acid etched surface (HA) and half with a non-coated control surface with only heat treatment (HT), into eighteen rabbit femurs. At 12 weeks, the bone-implant blocks were retrieved for micro computed tomography (μCT), histologic processing and histomorphometric evaluation. RESULTS The bone-to-implant contact for the entire threaded portion of the implant revealed 57.1% (21.0) for the HT group and 38.8% (17.7) for the HA group with a total bone area within the threads 72.5% (13.9) (HT) and 59.7% (12.5) (HA). The 3D reconstructed μCT image corresponded to the histomorphometric results. SIGNIFICANCE It is suggested that multiple factors such as the change in topography and chemistry may have influenced the outcomes.

PROLONGED ENDOCHONDRAL BONE HEALING IN SENESCENCE IS SHORTENED BY LOW-INTENSITY PULSED ULTRASOUND IN A MANNER DEPENDENT ON COX-2

To test whether mechanical loading produces faster healing in aged mice, fractured femurs of aged 1-year-old mice were subjected to low-intensity pulsed ultrasound (LIPUS), a treatment that is routinely used to help heal fractures in humans. Cyclooxygenase-2 knockout mice (COX-2(-/-)), which lack an immediate early mediator of mechanical stimulation, were also studied by histochemistry, microcomputed tomography and quantitative polymerase chain reaction to determine the role of COX-2. The healing in the aged COX-2(-/-) mice is slow during the endochondral bone remodeling (>30 d), a period generally prolonged in senescence. For aged wild-type mice, LIPUS halved the endochondral phase to about 10 d, whereas that was not the case for aged COX-2(-/-) mice, which showed no apparent shortening of the prolonged endochondral-phase healing time. Injecting prostaglandin E(2) receptor agonists, however, rescued the COX-2(-/-) callus from insensitivity to LIPUS. In conclusion, COX-2 is a limiting factor in the delayed endochondral bone healing and is induced by LIPUS, which normalizes healing rate to the wild-type level.

Osteoarthritic Changes of the Patellofemoral Joint in STR/OrtCrlj Mice Are the Earliest Detectable Changes and may be Caused by Internal Tibial Torsion

STR/ort mice develop a naturally occurring osteoarthritis (OA) of the knee joints. However, the evaluation of early OA changes has been difficult due to variability caused by gender, individual differences, and differences between the right and left lower limbs. The objective of this study was to analyze the variability of the early OA changes with age in STR/ort mice and to identify the cause of onset. A total of 115 STR/OrtCrlj mice aged 10–45 weeks were examined. In addition to conventional radiological and histological evaluation of the knee joints, histological sections were used to examine the patellofemoral, femorotibial, and growth plate cartilage under similar conditions. A morphological evaluation of tibiae, including micro-3-dimensional computed tomography, was performed. Radiological evaluation showed OA changes in the joints of mice over 35 weeks old and histological evaluation showed early OA changes in the femorotibial joints of mice over 26 weeks old. However, these changes were not common in all individuals. In contrast, most common and reproducible OA changes were observed in the bilateral patellofemoral joints of all individuals, and even in subjects ranging from 10 to 20 weeks of age. Morphological evaluations also demonstrated an abnormal tibial internal torsion that increased with age and was associated with medial patellar dislocation. In conclusion, the earliest histological OA change was observed in the patellofemoral joint prior to similar observations in the femorotibial joint. Internal tibial torsion may be a cause of OA in the patellofemoral joints, which leads to the development of medial femorotibial OA.

Nanosized Hydroxyapatite Coating on PEEK Implants Enhances Early Bone Formation : A Histological and Three-Dimensional Investigation in Rabbit Bone

Polyether ether ketone (PEEK) has been frequently used in spinal surgery with good clinical results. The material has a low elastic modulus and is radiolucent. However, in oral implantology PEEK has displayed inferior ability to osseointegrate compared to titanium materials. One idea to reinforce PEEK would be to coat it with hydroxyapatite (HA), a ceramic material of good biocompatibility. In the present study we analyzed HA-coated PEEK tibial implants via histology and radiography when following up at 3 and 12 weeks. Of the 48 implants, 24 were HA-coated PEEK screws (test) and another 24 implants served as uncoated PEEK controls. HA-coated PEEK implants were always osseointegrated. The total bone area (BA) was higher for test compared to control implants at 3 (p < 0.05) and 12 weeks (p < 0.05). Mean bone implant contact (BIC) percentage was significantly higher (p = 0.024) for the test compared to control implants at 3 weeks and higher without statistical significance at 12 weeks. The effect of HA-coating was concluded to be significant with respect to early bone formation, and HA-coated PEEK implants may represent a good material to serve as bone anchored clinical devices.

Bone apposition to laminin-1 coated implants: histologic and 3D evaluation

Laminin-1 has been reported as one of the factors responsible for the nucleation of calcium phosphates and, in vitro, has been reported to selectively recruit osteoprogenitors. This article focused on its in vivo effects, and evaluated the effect of laminin-1 local application on osseointegration. Polished cylindrical hydroxyapatite implants were coated with laminin-1 (test) and the bone responses in the rabbit tibiae after 2 and 4 weeks were evaluated and compared to the non-coated implants (control). Before the samples were processed for histological sectioning, they were three-dimensionally analysed with micro computed tomography (μCT). Both evaluation methods were analysed with regards to bone area around the implant and bone to implant contact. From the histologic observation, new bone formation around the laminin-1 coated implant at 2 weeks seemed to have increased the amount of supporting bone around the implant, however, at 4 weeks, the two groups presented no notable differences. The two-dimensional and three-dimensional morphometric evaluation revealed that both histologic and three-dimensional analysis showed some tendency in favour of the test group implants, however there was no statistical significance between the test and control group results.

Effects of citric acid administration on femoral trabecular structures in ovariectomized mice

ObjectivesTo examine the effects of citric acid administration on trabecular structures.MethodsOvariectomized female ICR mice (n = 39), 8 weeks of age, were divided into four groups: ovariectomized control (OVX-control, n = 9, standard diet); citric acid (CIT, n = 10, citric acid 5 g/100 g diet); vitamin K2 (menaquinone 7; VK2, n = 10, vitamin K2 50 µg/100 g diet); and citric acid + vitamin K2 (CIT + VK2, n = 10, citric acid 5 g/100 g diet and vitamin K2 50 µg/100 g diet). After 12 weeks, the bone mass of the right femur was measured using peripheral quantitative computed tomography and the three-dimensional (3-D) trabecular structure of the right femur was assessed using microfocus computed tomography.ResultsThe bone mineral density was significantly increased in the VK2 group versus the OVX-control group (P < 0.05). In the 3-D trabecular structure analysis, skeletal perimeter and number were significantly greater in the CIT, VK2, and CIT + VK2 groups than in the OVX-control group. Skeletal separation, spacing, and trabecular bone pattern factor were significantly lower in the CIT, VK2, and CIT + VK2 groups than in the OVX-control group. Almost all parameters in node–strut analysis were significantly greater in the CIT, VK2, and CIT + VK2 groups than in the OVX-control group.ConclusionsThese results suggest that citric acid improved trabecular structures. However, simultaneous administration of citric acid and vitamin K2 did not have any additional effect.

Radiological Morphometric Analysis of the Mandibular Bone Structure after Ovariectomy in Mature Cynomolgus Monkeys

Abstract [Purpose] The effects of experimental osteoporosis on the trabecular bone structure of the mandible in cynomolgus monkeys were examined by radiological bone morphometric analysis. [Methods] Ovariectomy (OVX) was performed on twelve 10-year-old female cynomolgus monkeys, which were fed a controlled diet for 2 years. Twelve monkeys in a sham control group were fed under the same conditions. Using a microfocus tube and computed radiography, the removed mandibular bone samples were imaged by standardized magnification radiography, and two-dimensional digital imaging data were obtained. The structural parameters, such as skeletal area, perimeter, number, complexity, continuity and anisotropy, were measured using radiological bone morphometric analysis. The bone mineral density (BMD) was measured by dual energy X-ray absorptiometry. The width of the cortical bone was measured using magnified radiographic images. [Results] There were no significant differences between the OVX and the sham control groups in the skeletal structure indicated by the skeletal volume, number, width, perimeter, complexity, continuity, separation and spacing. However, there were significant differences between the two groups in the BMD of the mandibular body, cortical bone width, anisotropy and some parameters of the skeletal continuity. Among these parameters, the difference in the thinning of the cortical bone was most significant. [Conclusions] Using two-dimensional digital radiographic image data, this study suggests that the cortical bone width is more useful than the trabecular bone structure as the morphologic parameter for diagnosis of osteoporosis in the mandibular body.

Guided bone augmentation using a ceramic space-maintaining device

OBJECTIVE The purpose of this study was to evaluate 3-dimensionally whether vertical bone augmentation can be achieved using a hollow hydroxyapatite space-maintaining device in a rabbit calvarial model. Furthermore, different inner surface topographies, different permeabilities, and different porosities of the ceramic were tested to determine the optimal conditions for bone regeneration. STUDY DESIGN A total of 48 hollow domes made of hydroxyapatite in 4 different designs were placed subperiosteally on rabbit skull bone. The rabbits were humanely killed after 12 weeks, and the results were analyzed 3-dimensionally using micro-computed tomography. RESULTS The results suggest a larger production of bone volume when using an occlusive, dense hydroxyapatite space-maintaining device with a rough inner surface. CONCLUSIONS Hydroxyapatite space-maintaining devices permit new bone formation and osteoconduction within the dome.

Studies on the Relationship between Bone Strength and Bone Quality in Rats Fed with a Low-magnesium Diet

Abstract The relationship between bone strength and bone quality in rats fed with a low-magnesium (low-Mg) diet was examined. Twenty four-week-old male Wistar rats were divided into a control group (n = 10) and a low-Mg group (n = 10). Each group was fed with a conventional diet or a low-Mg diet (Mg, 6 mg/100 g diet) for 8 weeks, respectively. After the rats were sacrificed, bone strength, bone mineral content (BMC) and three-dimensional (3D) trabecular structure of the lumbar vertebra were measured, respectively. The results showed that the values of the BMC were almost the same between the control and the low-Mg diet groups. On the other hand, the bone strength of the low-Mg diet group was significantly lower than that of the control group (p