Bone Trabecular Separation Research Articles

RANKL Inhibition: A Novel Strategy to Decrease Femoral Head Deformity After Ischemic Osteonecrosis

A novel therapeutic strategy to decrease the development of femoral head deformity after ischemic osteonecrosis was studied in a large animal model of total head infarction. RANKL inhibition through exogenous osteoprotegerin administration significantly decreased pathologic bone resorption and deformity during repair of the infarcted head. Legg-Calvé-Perthes disease (LCPD) is a juvenile form of osteonecrosis of the femoral head that can produce permanent femoral head deformity (FHD) and premature osteoarthritis. The development of FHD in LCPD is closely associated with the repair process, characterized by a predominance of bone resorption in its early stage that produces a fragmented appearance and collapse of the femoral head. We present here a novel strategy to preserve the femoral head structure after ischemic osteonecrosis based on inhibition of interaction between RANK and RANKL using exogenous administration of osteoprotegerin (OPG-Fc) in a large animal model of ischemic osteonecrosis. Ischemic osteonecrosis was surgically induced in 18 male piglets by placing a ligature tightly around the right femoral neck to disrupt the blood flow to the right femoral head. Two weeks after the induction of total head infarction, OPG-Fc or saline was administered subcutaneously to nine animals per group for 6 weeks. The contralateral, normal (left) femoral heads from the animals treated with saline served as normal, nondisease controls. All animals were killed at 8 weeks when severe FHD has been previously shown to occur because of the repair process dominated by osteoclastic bone resorption. Radiographic, histomorphometric, and immunohistochemical assessments were performed. Radiographic assessment showed significantly better preservation of the femoral head structure in the OPG-Fc group compared with the saline group. Epiphyseal quotient (the ratio of epiphyseal height to diameter) was significantly higher in the OPG-Fc group (0.41 +/- 0.09) compared with the saline group (0.24 +/- 0.08, p < 0.001). Histomorphometric assessment revealed a significant reduction in the number of osteoclasts present in the OPG-Fc group (5.9 +/- 5.3mm(-2)) compared with the saline group (39.6 +/- 13.8 mm(-2), p < 0.001). Trabecular bone volume, number, and separation were significantly better preserved in the OPG-Fc group compared with the saline group (p < 0.001). No significant difference in femoral length was observed between the OPG-Fc and saline groups. Immunostaining revealed the presence of OPG-Fc only within the blood vessels, with no apparent staining of bone matrix or trabecular bone surfaces. To our knowledge, this is the first study to show that RANKL inhibition decreases bone resorption and FHD after ischemic osteonecrosis. Because RANKL inhibitors do not bind to bone, their effects on resorption are reversible as the drug is cleared from circulation. The reversible nature of RANKL inhibitors is very appealing for treating pediatric bone diseases such as LCPD, where the resorptive stage of the disease lasts for 1-2 years.

Trabecular structure of the condyle of the jaw joint in young and mature sheep: A comparative histomorphometric reference

Much of the literature regarding arthrotic changes in the temporomandibular joint (TMJ) is based on the assumption, rather than the demonstration, that joint degeneration is pathologically and biochemically similar to that which has been described for other arthrodial joints. Understanding such changes is axiomatic of an understanding of the specific histomorphometric structure of the normal TMJ, in particular the condyle. Unfortunately, very little has been established about the trabecular bone patterns in the mandibular condyle as it develops. As a consequence of the obvious practical difficulties in investigations of the human TMJ, the sheep has been variously used as an animal model. In order to augment a fuller characterisation of this animal model, this study focuses on the quantitative histomorphometries of the trabeculae in the mandibular condyles of young and mature sheep. Quantitative histomorphometric analyses of condylar trabeculae were performed on histological sections prepared from mature and young sheep condyles. Lateral, central and medial sagittal sections, and anterior and posterior coronal sections of the condyle were analysed using a Quantimet 500MC image analysis system that had been programmed to provide structural index values of trabecular bone volume, surface, thickness, separation and number. Analysis of histoquantitation data revealed a significant concordance in bone structural index values between lateral, central and medial regions in young and mature sheep as well as anterior and posterior regions in young sheep. Moreover, there was little significant variation between similar regions in the respective age groups. This study provides the first comparative histomorphometric quantitative analysis of the trabeculae in the mandibular condyle of both young and mature sheep. The findings of this study reinforce the appropriateness of the sheep TMJ as a model in previous experimental studies of the bony architecture of the condyle.

Optimization of MR-Relaxometry for BMD-measurements and its Correlation with DEXA.

The aim of this study was to optimize MRI conventional protocols for BMD measurements using MR-Relaxometry in systems not facilitated with special multi echo protocols. Since, cortical and trabecular bone separation can not be performed in DEXA, so the results might lead to erroneous interpretation of BMD values. One method for bone quality determination is MR relaxometry that derives R2(=1/T2), R2*(=1/T2*) and R2'(=R2*-R2). This study was performed by 1.5T MRI system(Picker Vista-Q800), an uniformity phantom(1.25gr/l CuSO4, with T2=200ms for calibration), a body RF-Coil, 7 normal, 7 osteopenia, 7 osteoporosis volunteers and Lunar DEXA system(DPX-MD). To determine R2*and R2, multi GE and SE protocols with different TE/TR were used. Then in phantom and in coronal section of femoral-neck, relaxation rates were compared with BMD. The slope of neperian-logarithm of signal vs. TE in GE as -R2*used for protocol optimization. Therefore, for phantom calibration, optimized GE parameters of TE=13.42/18/26.8 ms, TR=800ms and ST=8mm used for the measurement of R2*, while, the measurement of R2 required the optimized SE parameters of TE=30/60/90/120ms, TR=800ms and ST=8mm, with CV(R2*)=2.96%, CV(R2)=3%, respectively. In volunteers for SE, TE of 36/54/63/72ms and TR=800ms were used, while, for GE the TEs/TR were the same as those of phantom study. R2*and R2' showed a significant positive correlation with BMD, r=0.62(p<0.05) & r=0.62(p<0.05) respectively. Finally, in accordance with DEXA values, the results showed that MR-Relaxometry is a proper tool for BMD-measurements in femoral-neck. Also it may be used as a complement method for DEXA failure in BMD-assessments.

Effects of 17 ?-estradiol on the expression of interstitial collagenases-8 and ?13 (MMP-8 and MMP-13) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in ovariectomized rat osteoblastic cells

Estrogen plays an important role in maintaining normal bone metabolism via the direct or indirect regulation of bone cells. Osteoblastic cells, as the target cells of estrogen, can secrete multiple matrix metalloproteinases (MMPs) that participate in bone remodeling. It has been demonstrated that bone loss induced by estrogen deficiency is closely related to the abnormal expression of multiple MMPs in osteoblastic cells. However, the regulating action of estrogen on the expression of interstitial collagenases MMP-8 and MMP-13 in osteoblastic cells in vivo remains unclear. We used an ovariectomized osteoporotic rat model to analyze the changes in the histomorphometric parameters of bone after and without treatment with 17beta-estradiol (E(2)); We also used immunohistochemistry and in situ hybridization to observe changes in the expression of mRNA and the proteins MMP-8, MMP-13 and TIMP-1 in osteoblastic cells in rat proximal tibia. In this study, we found that in the ovariectomized rat the expression of MMP-13 mRNA and protein increased markedly, whereas the expression of MMP-8 and TIMP-1 mRNA and protein did not change significantly. Our analysis showed that the expression of MMP-13 protein was correlated positively to bone trabecular separation, osteoid surface area, and negatively to trabecular numbers and the percentage of trabecula bone volume/total tissue volume. Our results suggest that MMP-13 plays an important role in estrogen deficiency-induced bone loss, while estrogen can inhibit bone resorption and reduce bone turnover rate by down-regulating the expression of MMP-13 in osteoblastic cells.

Trabecular structure assessment in lumbar vertebrae specimens using quantitative magnetic resonance imaging and relationship with mechanical competence.

The purpose of this study was to use quantitative magnetic resonance imaging (MRI; high-resolution [HR] and relaxometry) to assess trabecular bone structure in lumbar vertebrae specimens and to compare these techniques with bone mineral density (BMD) in predicting stress values obtained from mechanical tests. Fourteen vertebral midsagittal sections from lumbar vertebrae L3 were obtained from cadavers (aged 22-76 years). HR images with a slice thickness of 300 microm and an in-plane spatial resolution of 117 microm2 x 117 microm2 were obtained. Transverse relaxation time T2' distribution was measured by using an asymmetric spin-echo (ASE) sequence. Traditional morphometric measures of bone structure such as apparent trabecular bone fraction (app. BV/TV), apparent trabecular bone number (app. Tb.N), apparent trabecular bone separation (app. Tb.Sp), and apparent trabecular bone thickness (app. Tb.Th) as well as the directional mean intercept length (MIL) were calculated. Additionally, BMD measurements of these sections were obtained by dual-energy X-ray absorptiometry (DXA) and biomechanical properties such as directional stress values (to fracture) were determined on adjacent specimens. With the exception of T2', all morphological parameters correlated very well with age, BMD, and stress values (R between 0.79 and 0.92). However, in the direction perpendicular to the magnetic field, T2' values enhanced the adjusted R2 correlation value with horizontal (M/L) stress values in addition to BMD from 0.70 to 0.91 (p < 0.05).

Trabecular bone microarchitecture, bone mineral density, and vertebral fractures in male osteoporosis.

Some studies have indicated that the risk of fragility fractures in men increases as bone mineral levels decrease, but there is an overlap in the bone mineral density (BMD) measurements between patients with or without fractures. Furthermore, it has been suggested that the biomechanical competence of trabecular bone is dependent not only on the absolute amount of bone present but also on the trabecular microarchitecture. In the present study, 108 men (mean age 52.1 years) with lumbar osteopenia (T score < -2.5) were recruited to examine the relationships between BMD, architectural changes in trabecular bone, and the presence of vertebral fractures. Lumbar BMD was assessed from L2 to L4 in the anteroposterior view with dual-energy X-ray absorptiometry. At the upper left femur, hip BMD was measured at the transcervical site. Spinal X-ray films were analyzed independently by two trained investigators, and vertebral fracture was defined as a reduction of at least 20% in the anterior, middle, or posterior vertebral height. Transiliac bone biopsy specimens were obtained for all patients. Histomorphometric studies were performed on an image analyzer, and the following parameters were determined: trabecular bone volume (BV/TV), trabecular thickness (Tb.Th), number (Tb.N), and separation (Tb.Sp), interconnectivity index (ICI), characterization of the trabecular network (node count and strut analysis), and star volume of the marrow spaces. Spinal radiographs evidenced at least one vertebral crush fracture in 62 patients (group II) and none in 46 patients (group I). After adjusting for age, body mass index, and BMD, there were no significant differences between the two groups in BV/TV, Tb.Th, or star volume. In contrast, the mean values of ICI, free end-to-free end struts (FF/TSL), and Tb.Sp were significantly higher, whereas Tb.N and node-to-node struts (NN/TSL) were lower in patients with at least one vertebral fracture. Logistic regression analysis showed that only ICI, FF/TSL, NN/TSL, and Tb.N were significant predictors of the presence of vertebral fracture: odds ratios for an alteration of 1 SD ranged from 1.7 (1.0-3.2) for NN/TSL to 3.2 (1.1-10.1) for ICI. Patients with at least three vertebral fractures (n = 23) were categorized as "multiple fractures." The results of logistic regression showed that spine BMD, BV/TV, and all architectural parameters were significant predictors of multiple vertebral fractures: odds ratios for an alteration of 1 SD ranged from 2.2 (1.1-4.6) for star volume to 3.7 (1.4-9.7) for ICI. These results strongly suggest that bone trabecular microarchitecture is a major and independent determinant of vertebral fractures in middle-aged men with osteopenia.

Minocycline prevents the decrease in bone mineral density and trabecular bone in ovariectomized aged rats.

In the current study, we examined the effects of minocycline, on the osteopenia of ovariectomized aged rats. Old female rats were randomly divided into five groups: sham, ovariectomized control and ovariectomized treated with minocycline, 17beta-estradiol, or both agents. Bone samples were collected 8 wk after the treatment. Ovariectomy reduced bone mineral density of the whole femur and at the condylar, distal metaphyseal and head-neck-trochanter regions 10%-19% and the loss of bone density was prevented by treatment with minocycline or 17beta-estradiol. Histomorphometric analysis of distal femur showed ovariectomy reduced the trabecular bone area, the trabecular bone number, trabecular bone thickness and increased the trabecular bone separation. The microanatomic structure of trabecular bone also showed that the number of nodes, node to node, cortical to node, node to free end was reduced by ovariectomy. Treatment with minocycline attenuated the effect of ovariectomy on trabecular bone in aged animals. In contrast, cortical bone was not affected by ovariectomy or minocycline treatment. The effect of minocycline on bone turnover was also examined. Minocycline increased osteoid surface, mineralizing surface, mineral apposition rate, bone formation rate and reduced eroded surface. We have therefore concluded that the modest increase in bone mineral density and the improvement in the trabecular bone status noted in minocycline treated ovariectomized aged rats is likely due to an increase in bone formation coupled with a decrease in bone resorption.