Cold bone defect on granulocytes labelled with technetium-99m-HMPAO scintigraphy: significance and usefulness for diagnosis and follow-up of osteoarticular infections.

We wished to determine the frequency and significance of cold bone defect on granulocytes labelled with technetium-99-m-hexamethylpropyleneamine oxime (99mTc-HMPAO-PMN) in non-spinal bone infection. Cold bone defect was investigated as part of a retrospective review during a 2-y period. Patients who had possible osteoarticular infection underwent bone scintigraphy combined with 99mTc-HMPAO-PMN for diagnosis and follow-up. Osteomyelitis was confirmed by isolation of the responsible pathogen. Among 210 patients who had possible infection, 17 (8%) demonstrated a cold bone defect. The site of cold bone defect was for all patients the hip. All 17 patients had proven bacterial orthopaedic hardware-related infection. The single causative micro-organism was staphylococcus. Whatever the outcome, cold bone defect was constant regardless of follow-up equal to or longer than 18 months. These data suggest that this uncommon scintigraphic pattern is an indication of an infectious process similar to increased uptake.

Can magnetic resonance imaging be the key technique to visualize and investigate endovascular biomaterials?

OBJECTIVE: Magnetic resonance imaging (MRI) is an established modality in clinical use but may be potentially underutilized to visualize and investigate biomaterials. As its use is totally contraindicated only for ferromagnetic devices, it was employed to visualize deployment, biofonctionality, healing, and biodurability of a commercially available endovascular device, namely the Medtronic-AVE AneuRx. The quality of the observations coupled with the absence of ionizing radiations are likely to make this technique an attractive imaging modality in the future. METHOD: The potential benefits of the MRI technique were investigated in a GE Vectra-MR 0.5T MRI for the Medtronic-AVE AneuRx endovascular prosthesis, under different conditions: undeployed i.e., inserted in the delivery cartridge as received from the manufacturer (step 1), deployed in a mock glass-aneurysm tube (step 2), and as a pathological explant harvested at the autopsy of a patient (step 3). The device was submitted to X-rays for examination in addition to MRI. At step 3, the device was further investigated with light microscopy and scanning electron microscopy (SEM) together with X-ray diffraction. RESULTS: The device which was inserted and pleated in the delivery cartridge did not demonstrate any significant observation either in MRI or in X-rays. When it was deployed in the mock aneurysmal glass tube, light artefacts were associated with the T2 weighed FSE images around the Nitinol whereas X-rays gave images of indisputable interest. Similar results were noted using the explanted device. Very high contrasts were obtained with T1 whereas T2 images were almost defect free. The X-rays allowed to accurate imaging of the Nitinol skeleton but were poor to discriminate between the different tissues. Pathology observations using light microscopy were not really challenged, as the magnetic resonance imaging was performed using a 0.5T machine. DISCUSSION: The benefits of magnetic resonance imaging as a quality control technique to examine an endovascular device within its cartridge remains ill defined. Similarly, the role of conventional X-rays is unknown. The observation of devices fully deployed in a mock aneurysmal glass-tube under MRI are potentially useful but X-rays images allowed better definition. The MRI examination of the explanted device does permit observations related to the healing of the device that might be obtained in vivo and, thus offers new avenues for the follow-up of implanted devices. The pathological investigations brought additional informations about the tissues and the corrosion of the Nitinol. However, it is unlikely that MRI will permit detailed analysis of the biomaterials and in particular the corrosion process of the stents. CONCLUSION: These early observations of the follow-up of devices using MRI warrant further investigation. The absence of ionizing radiation with MRI makes this technique particularly attractive. As there is no emission of ionizing radiation associated with magnetic resonance, it is recommended that further investigation using this environment friendly technique for the follow-up of devices made of biomaterials that are MRI compatible.

[Bone remodeling assessment after total hip replacement]. / Evaluation biologique et densitométrique du remodelage osseux induit par l'arthroplastie totale de hanche.

It is known that total hip arthroplasties (THA) lead to adaptative remodeling changes resulting in periprosthetic bone loss. DEXA is recognized as the most precise and accurate method for quantifying bone mineral density (BMD) in humans. The present study compares over two years after THA, DEXA data to those of urinary deoxypyridinoline (uDPYR), a pyridinium crosslink of bone collagen fibrils proven to be a reliable bone resorption marker. 41 patients (21 postmenopausal female, 20 male) underwent cemented THA. Urinary excretion of DPYR was determined using ACS : 180 SE (Bayer Diagnostics) and normalized for creatinine excretion while periprosthetic BMD (g/cm2) was measured (QDR 4500, Hologic), at post-operative day, 3 months, 1 year and 2 years after surgery. The 7 periprosthetic subregions (R1-R7) of the Gr en method are the regions of interest for evaluating bone remodeling process. uDPYR showed a significant decrease between postoperation and 1 year: 10.6 0.80 vs 4.8 0.6 nmol/mmol, p < 0.0001 (Wilcoxon Test for paired samples and statistical significance accepted at p < 0.05) and non significant variation between 1 and 2 years. Between post-operation and 3 months global and regional BMDs decrease significantly (p < 0.04) followed by an increase in distal BMD (R3, R4, R5). During the second year BMD increases also for other regions. At 2 years BMD in distal regions is recovered except in the proximal R7 when comparing post-operation and 2 years, pattern consistent with literature. Thus, a discrepancy is observed between uDPYR and DMO results that does not allow us to use a bone resorption urinary marker to monitore local bone periprosthetic remodeling.

A novel potential application for 99mTc-HMPAO: endothelial cell labeling for in vitro investigation of cell-biomaterial interactions.

UNLABELLED: Good adherence of endothelial cells (ECs) seeded on vascular prostheses and cell retention under flow conditions are important factors to consider in the use of functionalized prostheses in vascular surgery. Because 111In-oxine radiolabeling presents disadvantages, we wondered whether, because of its well-known physical properties, 99mTc-hexamethyl propyleneamine oxime (HMPAO or exametazime) could be used. METHODS: The cytotoxicity of unlabeled HMPAO and 99mTc-HMPAO at increasing concentrations and activities was tested on monolayers of the EC line EA-hy-926. The influence of temperature and time on tracer incorporation into cells was also tested. The optimal labeling conditions were applied to evaluate the retention of ECs seeded on polyester grafts under flow conditions by gamma camera detection. RESULTS: The activity of 10 MBq/10(6) cells corresponding to 4.5 microg/10(6) cells of unlabeled HMPAO, applied for 3 h at 37 degrees C (cellular uptake = 18%), was the best compromise between the maintenance of cell viability and metabolic activity and efficient detection by the gamma camera. Spontaneous leakage was observed and analyzed by high-performance liquid chromatography. A cell loss of 13% after 180-min exposure to shear stress was obtained. CONCLUSION: Our data thus indicate the feasibility of using such a radiolabeling technique to investigate EC-biomaterial interactions.

Optimization of a new scintillation gas detector used to localize electrons emitted by 99mTc.

UNLABELLED: We have developed a scintillation gas detector to localize electrons emitted by 99mTc. This type of detector allows direct quantification of images and so provides a clear advantage over autoradiographic film. We have optimized the device to give an image spatial resolution that closely approximates that of typical autoradiographic film. To improve this resolution, it was necessary to select only low-energy electrons (2 and 15 keV) and to devise novel detection and localization techniques for the ionizing particles. METHODS: A parallel-plate proportional avalanche chamber is subject to a uniform electrical field and amplifies the number of released electrons through collisions of ionizing particles in the gas mixture. Light emitted by the gas scintillator during the avalanche process is collected by a highly intensified charge coupled device camera. The centroid of each resulting light distribution is calculated, resulting in a quantitative mapping of the sample's activity. Insertion of the sample within the gas volume improves the efficiency and so provides a method that is both very sensitive and linear. RESULTS: We have shown that in a parallel-plate structure, the application of a high electrical field to the surface of the sample and the selection of appropriate light spots, according to their morphology, can overcome localization errors due to the particles' trajectories. We have obtained a resolution of the order of 30 microm, using electrons from 99mTc. CONCLUSION: This detection technique allows considerable improvement in image resolution. This "electron camera" is a serious rival to existing autoradiographic techniques, because it provides certain other advantages, including direct quantification, linearity, high dynamic range and low noise levels. Thus, new perspectives are made available in quantitative double tracer autoradiography, because electrons can be selected for imaging as a function of their energy.

Value of captopril renal scintigraphy in hypertensive patients with renal failure.

UNLABELLED: The aims of this study were to show the value of captopril renal scintigraphy for detecting a renovascular cause in hypertensive patients with renal failure and to assess the ability to predict the beneficial effect of revascularization on renal function. METHODS: Thirty-eight patients with renal failure (mean glomerular filtration rate = 35 mL/min) underwent renal scintigraphy after injection of 99mTc-mercaptoacetyltriglycine. Baseline scintigraphy was performed, and the test was repeated 24 h later after oral administration of 50 mg captopril given 60 min before the test. RESULTS: In 5 of 6 patients with a renovascular cause for renal failure, and 2 of 3 patients with a probable arterial pathology, scintigraphy had a high probability. The result was indeterminate in the other 2 patients. In 5 of 11 patients with negative arteriography and 14 of 18 patients with probable absence of renovascular pathology, we found a low probability of functional renal artery stenosis. Six revascularization procedures were performed and were predictive of a beneficial effect in 5 patients. Time of peak activity was an effective predictor in each case. CONCLUSION: In hypertensive patients with renal failure, captopril renal scintigraphy can detect hemodynamic dysfunction downstream from a renal artery stenosis and can predict the beneficial effect of revascularization in some cases.

Influence of strontium on bone mineral density and bone mineral content measurements by dual X-ray absorptiometry.

The presence of Sr in bone influences bone mineral density (BMD) and content (BMC) measurements by dual-energy X-ray absorptiometry (DXA). This interaction is of interest, since strontium ranelate (S12911) demonstrated positive effects on bone metabolism in various animal models of osteoporosis, and is currently being evaluated for treatment of postmenopausal osteoporosis. The present in vitro study aimed to determine adjustment factors for DXA measurements of BMC and BMD at different Sr concentrations in order to estimate the corresponding values that would have been measured without Sr. A series of mixtures of Ca and Sr hydroxyapatites were prepared, with biologically relevant Sr/Ca ratios ranging from 0 to 3.5 mol/mol%, and a constant total concentration of divalent cations (145 mmol). The mixtures were conditioned in plastic dishes 4.5 cm in diameter, to obtain an areal density close to the human vertebral mineral density of 0.7-1.1 g/cm(2). DXA measurements of the mixtures were made with a wide range of different instruments and various acquisition modes. A direct linear relationship (r(2) > 0.99) was found between strontium content and overestimation of BMD and BMC. There were no significant differences in adjustment factors for BMC or BMD between the different machines or acquisition modes, and the presence of Sr in the water bath used to mimic soft tissues did not affect the accuracy and precision of the method. This demonstrates that reliable DXA determinations of BMD may be carried out in the presence of Sr, and may be interpreted in terms of calcium hydroxyapatite equivalent if the bone Sr content of the measured bone is known. The same adjustment factor (10% overestimation for 1 mol/mol% Sr) can be used for all presently available types of instrument and acquisition modes.

Evolution of the local calcium content around irradiated beta-tricalcium phosphate ceramic implants: in vivo study in the rabbit.

To evaluate whether dissolved calcium from tricalcium phosphate implants contributes to osseous wound healing in bone defects, the authors used nuclear radioactivated materials. Six months after irradiation, the calcium was still radioactive. Samples of the material were prepared and placed in rabbit condyles for 1, 3 and 9 months. Over time the condyles were retrieved and treated for histology or radiocounting. Measurements of the radioactivity of the slices and histomorphometry of the implants and surrounding tissues were performed. The authors observed that the radioactivity decreased regularly. Connective tissue had penetrated the pores and totally invaded the implants, first at the periphery of the implants, then inside the pores. Comparison of the results of radioactivity and histomorphometry suggest that part of the calcium from the implants was re-used specifically in the new osseous tissue.

"In vitro" dissolution of coral in peritoneal or fibroblast cell cultures.

Previous studies have shown that in vivo coral resorption involves a biphasic process: First, the edges of the coral block become powdery, then extracellular fluid and phagocytosis contribute to the dissolution of the crystals. The authors examined some types of cells that could be involved in phagocytosis, particularly the ability of both dermal fibroblasts and mouse-resident peritoneal cells to phagocytose and dissolve coral powder "in vitro". Radioactive coral was incubated for 24, 48, or 72 hrs with cells in the presence or absence of cytochalasin B (a phagocytic inhibitor) or chloroquine (a lysosomotropic agent). Furthermore, to specify the role of crystal cell contacts in the solubilization process, they incubated radioactive coral in conditioned media (obtained from two-day human fibroblastic or macrophagic cell culture in the presence or absence of non-radioactive coral) or at a distance from the cells using culture inserts. Measurements of the radioactivity in the different supernatants were performed. Transmission electron microscopy was carried out on the cells cultivated in the presence or absence of radioactive coral. The data suggest that both fibroblasts and macrophages dissolve the coral, and that the intracellular degradation in phagolysosomes is one of the mechanisms explaining coral powder dissolution.

Measurement of bone mineral density in mother-daughter pairs for evaluating the family influence on bone mass acquisition: a GRIO survey.

The relative influence of genetic and environmental determinants on bone mass is still unclear. Using an original multicentric mode of recruitment, based on absorptiometry current practice, the hypothesis of a familial predisposition to low bone mineral content was assessed. The study was based on dual-energy X-ray absorptiometry (DXA) measurements of lumbar and femoral neck bone mineral density (BMD), using daughters of women with a low BMD (case mothers). These BMD values were compared with those of control daughters of women with a normal BMD. Case mothers (n = 72) aged 54.3 +/- 4.8 years were recruited on the basis of a questionnaire and a vertebral Z-score < -2 SD. Their healthy daughters of more than 20 years (n = 77) aged 28.2 +/- 4.9 years had their vertebral and femoral BMD Z-score determined. The control groups were composed of mothers aged 54.1 +/- 4.7 years, paired by age +/- 2 years to the case mothers, and of their daughters of more than 20 years old, aged 27.7 +/- 5.8 years. For daughters, a significant difference was found between the mean vertebral Z-scores (-0.82 +/- 1.08 for cases and 0.01 +/- 1.14 for controls, p < 0.0001). The difference was in the same direction but was not statistically significant for mean femoral Z-scores (-0.58 +/- 1.15 for cases and -0.22 +/- 1.33 for controls, p < 0.073). These findings confirm the hypothesis of a familial predisposition to low BMD.

Comparison of two Hologic DXA systems (QDR 1000 and QDR 4500/A).

Bone mineral content is reliably measured by dual energy X-ray absorptiometry (DXA), if manufacturers' recommendations and quality control (QC) procedures are followed. Several phantoms (Hologic anthropomorphic spine phantom, the Groupe de Recherche et d'Informations sur les Osteoporoses (GRIO) test objects and the European semi-anthropomorphic phantoms) were used to evaluate reproducibility, linearity, accuracy and spatial resolution of two DXA devices in vitro. These parameters were also evaluated in vivo from measurements performed on 120 volunteer patients. It was found that when one device (a single beam monodetector QDR 1000) is replaced by another (a fan beam multidetector QDR 4500/A), the novel combination of procedures described here, ensures that the accuracy of DXA study results is maintained when both devices are used in succession for the same patient. To study the possible responses in clinical situations, the influence of bone environment (soft and adipose tissues) was also evaluated. In both systems, similar performances (in vitro coefficients of variation of 0.5%) were established. At extreme bone density values, slight differences in linearity were found, as well as differences in accuracy and spatial resolution. Lumbar spine and femoral neck measurements were performed with both systems in 120 volunteers, both measurements being made on the same day. The corresponding bone mineral density (BMD) values were highly correlated (r2 = 0.985 for lumbar spine and 0.948 for the femoral neck), and the mean BMD differences were 0.68% and 0.37% for each anatomical site, respectively. Although small, these differences add to the precision error of the method, which is near 1%. A calibration curve has to be obtained in order that both devices can be equally used in regular clinical study. We concluded that when a DXA system is replaced by a new one, appropriate QC procedures must be strictly observed.

Influence of the structure of three corals on their resorption kinetics.

The aim of this study was to investigate the influence of the structure of corals on their resorption kinetics after implantation in subcutaneous areas. Three types of coral (Porites astreoides, Montastrea annularis and Dichocoenia stokesi) identical in composition but different in structure were implanted for periods of 1 and 2 months in subcutaneous sites in OF1 mice. The resorption of the implants was studied by means of qualitative (histology, scanning electron microscopy, fluorochrome labelling method) and quantitative approaches (gravimetric method). The results of the qualitative study revealed a process of irregular deterioration of the coral, linked to the detachment of crystals at the surface of the implant. The results of the quantitative study showed that the speed of resorption increases with the implantation time and the open porosity of the coral. These reactions are explained by the increase of the surface exchange area in contact with factors responsible for resorption: biological medium and cells. When considering the choice of coral as a bone substitute, these factors must be taken into account to allow the in situ maintenance of the implant over a sufficiently long period of time according to the clinical situation.

[Development of bone mineral density after cure of Cushing's syndrome]. / Evolution du contenu minéral osseux après guérison du syndrome de Cushing.

Both endogenous and exogenous glucocorticoid excess are well establish as causes of osteoporosis, however the reversibility of bone mass following the restauration of normal steroid levels is not well documented. In this longitudinal study, we mesured bone mineral density (expressed as Z-score) by dual-photon and X-ray absorptiometry of the lumbar spine (L2-L4) in 9 patients cured of Cushing's syndrome who were followed for the next 48 months (16-76). Initial Z-scores ranged from -2 to -1 standard-deviation (SD) in 6 patients consistent with osteopenia, and were below -3 SD in 2 patients consistent with osteoporosis. One patient developed lumbar spine fractures. There is no relationship between the severity of the Cushing's syndrome (assessed by the urinary free cortisol) and initial bone reduction (inital Z-score), nor between length of Cushing's symptoms and initial bone reduction. Our data show a marked variation (+74 +/- 9%) in bone mass in patients successfully treated for Cushing's syndrome. Seven patients completely recovered from steroid-induced osteoporosis, one patient partially recovered but remained osteopenic. One post-menopausal women presented several lumbar spine fractures despite successfull treatment of Cushing's syndrome. This longitudinal study confirms that if steroid-induced bone loss may improve substantially by cure of steroid excess even without other treatment, osteoporosis may worsen particularly in post-menopausal women. These results are important to take into account to properly manage patients with steroid-induced osteoporosis.

In corticosteroid-treated respiratory diseases, monofluorophosphate increases lumbar bone density: a double-masked randomized study.

The efficacy of a monofluorophosphate-calcium combination (MFP-Ca) in increasing lumbar bone mineral density (BMD) was assessed in a prospective double-masked study. Patients (n = 35), who had been treated for 1 year or more with prednisone-equivalent doses > or = 7 mg/day for asthma or other respiratory diseases, were randomly assigned to receive twice a day, for 2 years, either one MFP-Ca tablet [100 mg sodium monofluorophosphate (13.2 mg F-) + 500.5 mg Ca2+] or one Ca tablet (500.5 mg Ca2+). BMD was measured from L2 to L4 using a dual photon absorptiometer. The eligible patients (7 premenopausal women, 21 men), who had no previous vertebral fractures and were aged 46.5 (21-65) years, had received 18 (7.5-60) mg prednisone-equivalent/day and had a mean lumbar BMD of 0.917 +/- 0.141 g/cm2 at baseline (MO); in these 28 patients, the mean increase in lumbar BMD at final assessment was significantly greater in the MFP-Ca group (p = 0.05; Mann-Whitney). There was also a significant difference after 2 years between the two groups (p = 0.05, ANOVA) in favour of MFP-Ca, with an increase in lumbar BMD of 11% (MFP-Ca) compared with 1% (Ca); thus, with MFP-Ca, lumbar BMD increased by an average of approximately 5.5%/year. There was no statistically significant difference between the two groups in doses of corticosteroids used during the 2 study years, rate of vertebral fractures, or frequency of side-effects (which were all minor). No bone fissure was observed. Thus, the daily dose of 200 mg monofluorophosphate (26.4 mg F-) combined with 1 g Ca2+ in patients with long-term corticosteroid-treated respiratory diseases appears to be a safe and efficient way of increasing lumbar BMD, suggesting that its use should be further studied in corticosteroid-induced osteoporosis.

Magnetization transfer on in vitro circulating blood: implications for time-of-flight MR angiography.

Effects of magnetization transfer were evaluated in vitro on circulating blood. Various velocities were tested from 0 to 72 cm/second. Decrease signal intensity caused by magnetization transfer effects was inversely proportional to inflow velocity. It reached 10% at very low velocities and disappeared at velocities higher than 30 cm/second.

A canine ex vivo shunt for isotopic hemocompatibility evaluation of a NHLBI DTB primary reference material and of a IUPAC reference material.

Factors determining the thrombogenic response to particular artificial surfaces were investigated ex vivo in a canine shunt model. Methods using radioisotopic tracers made it possible to dynamically monitor the deposition of labelled blood cells and proteins on a NHLBI.DTB primary reference material polydimethylsiloxane (PRM.PDMS) and on a IUPAC reference material polyvinyl chloride (IUPAC.PVC). On the one hand, leukocyte affinity tau s(leu) (number of deposited leukocytes mm-2s-1) was not significantly different between IUPAC.PVC (tau s(leu) = 1.2-2.5) and PRM.PDMS (tau s(leu) = 1.5-3.4) and the fibrinogen adsorption rate varied from 33 to 48.10(-5) micrograms mm-2s-1 for both these materials. On the other hand, platelet affinity tau s(plat) (number of deposited platelets mm-2s-1) was significantly different (p < 0.05) for IUPAC.PVC and PRM.PDMS (tau s(plat)PVC = 683 +/- 200 > tau s(plat)PDMS = 327 +/- 80). Scanning electron micrographs of adherent platelets, red cells and leukocytes after blood contact ex vivo were performed after each experiment. This preliminary work contributes not only to quantify the adsorption of different radiotracers, but also to evaluate the superficial distribution of the labelled biological species on the inner surface of the tested biomaterials.

Radioimmunodetection of rat and rabbit cartilage using a monoclonal antibody specific to link proteins.

Biodistribution analysis using [125I]Fab-6F3 specific to link proteins from human articular cartilage performed in rats by autoradiography showed a high concentration of radioactivity in all cartilaginous tissues. Preliminary immunoscintigraphic assays were performed in rabbits. Front and side view images of whole animals exhibited high uptake in cartilage tissue of the knee articulation, in the invertebral disk and the humeral head. This fixation was still detected 24 h post-injection, although high washout of radioactivity was observed.

Hemocompatibility of diamond-like carbon coating.

The new prosthetic heart valve that has been designed by FII Company and Pr. Baudet involves a new "composite" material: titanium alloy T16A14V coated with Diamond-like Carbon. The purpose of this study was to evaluate the in vitro hemocompatibility of this new material in terms of protein adsorption and platelet retention. The static protein adsorption test gave interesting results, particularly for the albumin assay (237%) compared to the results obtained with a silicone elastomer chosen as a reference; the fibrinogen quantity, adsorbed on the surface of the material was slightly higher than that adsorbed on the silicone surface. Platelets adhere quite twice as much as they do on the reference surface. Such investigations showed good hemocompatibility results and should initiate further studies.