Accuracy of Breast Magnetic Resonance Imaging Compared to Mammography in the Preoperative Detection and Measurement of Pure Ductal Carcinoma In Situ: A Retrospective Analysis.

RATIONALE AND OBJECTIVES: Ductal carcinoma in situ (DCIS) hinders imaging detection due to multifocal appearance and discontinuous growth. Preoperative determination of its extent is therefore challenging. Aim of this study was to investigate the additional benefit of breast magnetic resonance imaging (MRI) to mammography (MG) in the diagnosis of DCIS according to size and grading. MATERIALS AND METHODS: Retrospective analysis of 295 patients with biopsy-proven, pure DCIS. Mean patient age was 57.0 years (27-87 years). All patients obtained MG. Additional MRI was performed in 41.7% (123/295). Mammographic breast density, background parenchymal enhancement (BPE), tumor size and grading were analysed. Tumor size on MG and MRI were compared to histopathological size of the surgical specimen. RESULTS: Mean tumor size was 39.6 mm. DCIS was occult on MG in 24.4% (30/123) and on MRI in 1.6% (2/123). Size was underestimated by 4.6 mm (mean) mammographically. DCIS was high grade in 54.5% (67/123), intermediate grade in 40.7% (50/123) and low grade in 4.9% (6/123). MG was exact regarding tumor size in low grade DCIS, underestimated intermediate grade DCIS by 1 mm (median) and high grade DCIS by 10.5 mm. MRI overestimated low grade DCIS by 1 mm (median), was exact regarding intermediate grade DCIS and underestimated high grade DCIS by 1 mm. BPE did not influence tumor detection and measurement. CONCLUSION: MRI outperforms MG in the detection and size estimation of DCIS and can reduce positive margin rates.

Ultrasound-Assisted Tumor Surgery in Breast Cancer - A Prospective, Randomized, Single-Center Study (MAC 001).

PURPOSE: Breast-conserving therapy is associated with a risk of tumor-involved margins. For intraoperative orientation, non- palpable or indistinctly palpable lesions are wire-marked prior to surgery. Ultrasound-guided surgery has the potential to reduce the number of tumor-involved margins. In the MAC 001 trial we evaluated ultrasound-guided breast-conserving surgery compared to wire-guided surgery with regard to free tumor margins, duration of surgery and resection volume. MATERIALS AND METHODS: In this randomized, prospective, single-center controlled trial, patients with ductal invasive breast cancer were recruited for either ultrasound-guided or wire localization surgery. Primary outcomes were tumor-free resection margins, the reoperation rate and the resection volume in each group. The results were analyzed by intention to treat. The trial was registered under ClinicalTrials.gov NCT02222675. RESULTS: 56 patients were assessed, and 47 patients were evaluated in the trial. 93 % (25/27) of the patients in the ultrasound arm had an R0 reoperation compared to 65 % (13/20) in the wire localization control arm. This result was statistically significant (p = 0.026). No statistical difference was found for the resection volume or the duration of surgery between the two arms. No major complication was seen in either arm. CONCLUSION: Ultrasound-assisted breast surgery significantly increases the possibility of tumor-free margins and therefore reduces the risk of reoperations. Breast surgeons should be trained in ultrasound and ultrasound should be available in every breast surgery operating room.

Prediction of Non-sentinel Lymph Node Metastases After Positive Sentinel Lymph Nodes Using Nomograms.

BACKGROUND/AIM: Only 30-50% of patients with sentinel lymph node (SLN) metastases present with further axillary lymph node metastases. Therefore, up to 70% of patients with positive SLN are overtreated by axillary dissection (AD) and may suffer from complications such as sensory disturbances or lymphedema. According to the current S3 guidelines, AD can be avoided in patients with a T1/T2 tumor if breast-conserving surgery with subsequent tangential irradiation is performed and no more than two SLNs are affected. Additionally, use of nomograms, that predict the probability of non-sentinel lymph node (NSLN) metastases, is recommended. Therefore, models for the prediction of NSLN metastases in our defined population were constructed and compared with the published nomograms. PATIENTS AND METHODS: In a retrospective study, 2,146 primary breast cancer patients, who underwent SLN biopsy at the University Women's Hospital in Tuebingen, were evaluated by dividing the patient group in a training and validation collective (TC or VC). Using the SLN-positive TC patients, three models for the prediction of the likelihood of NSLN metastases were adapted and were then validated using the SLN-positive VC patients. In addition, the predictive power of nomograms from Memorial Sloan Kettering Cancer Center (MSKCC), Stanford, and the Cambridge model were compared with regard to our patient collective. RESULTS: A total of 2,146 patients were included in the study. Of these, 470 patients had positive SLN, 295 consisted the training collective and 175 consisted the validation collective. In a regression model, three variants - with 11, 6 and 2 variables - were developed for the prediction of NSLN metastases in our defined population and compared to the most frequently used nomograms. Our variants with 11 and with 6 variables were proven to be a particularly suitable model and showed similarly good results as the published MSKCC nomogram. CONCLUSION: Our developed nomograms may be used as a prediction tool for NSLN metastases after positive SLN.

Contrast-enhanced spectral mammography in patients with MRI contraindications.

Background Contrast-enhanced spectral mammography (CESM) is a novel breast imaging technique providing comparable diagnostic accuracy to breast magnetic resonance imaging (MRI). Purpose To show that CESM in patients with MRI contraindications is feasible, accurate, and useful as a problem-solving tool, and to highlight its limitations. Material and Methods A total of 118 patients with MRI contraindications were examined by CESM. Histology was obtained in 94 lesions and used as gold standard for diagnostic accuracy calculations. Imaging data were reviewed retrospectively for feasibility, accuracy, and technical problems. The diagnostic yield of CESM as a problem-solving tool and for therapy response evaluation was reviewed separately. Results CESM was more accurate than mammography (MG) for lesion categorization (r = 0.731, P < 0.0001 vs. r = 0.279, P = 0.006) and for lesion size estimation (r = 0.738 vs. r = 0.689, P < 0.0001). Negative predictive value of CESM was significantly higher than of MG (85.71% vs. 30.77%, P < 0.0001). When used for problem-solving, CESM changed patient management in 2/8 (25%) cases. Superposition artifacts and timing problems affected diagnostic utility in 3/118 (2.5%) patients. Conclusion CESM is a feasible and accurate alternative for patients with MRI contraindications, but it is necessary to be aware of the method's technical limitations.

Longitudinal Computed Tomography Monitoring of Pelvic Bones in Patients With Breast Cancer Using Automated Bone Subtraction Software.

OBJECTIVE: The aim of this study was to optimize computed tomography (CT) surveillance of skeletal metastases in patients with breast cancer through the use of osseous subtraction maps between baseline and follow-up examinations created by a novel software algorithm. The new postprocessing algorithm segments the original bone followed by image intensity-based rigid alignment creating gray-shaded maps that highlight focal or diffuse loss or increase in bone attenuation. MATERIALS AND METHODS: Institutional review board was obtained for this retrospective data evaluation. A total of 33 consecutive patients (31 female; 2 male; mean age, 59.13 ± 12.68 years; range, 32-81 years) with breast cancer were included, who underwent 143 standardized baseline and follow-up CT examinations between February 2014 and June 2016. We classified bone metastases into lytic, sclerotic, and mixed osseous lesions. Any new osteolysis inside a known sclerotic lesion and enlargement of pre-existing sclerotic lesions were considered to represent progressive disease (PD), whereas no change was classified as stable disease (SD). Results were compared additionally with the course of the disease considering the entire skeleton and other involved organs. Software-created automated bone subtraction maps were compared with conventional CT interpretations of axial 5-mm and coronal 1-mm reformatted images. Region of interest measurements were used to quantify new lesions. Results were validated by clinical and CT follow-up. Reading time was evaluated. RESULTS: Skeletal metastases were present in 17/33 (51%) patients (9 sclerotic, 2 lytic, 6 mixed) at baseline. The use of bone subtraction maps resulted in an overall change of response classification into PD in 9/33 (8.1%) patients. Compared with conventional CT evaluation, the bone subtraction maps disclosed 123 new or enlarging sclerotic and 32 new lytic metastases in 23/33 (30.9%) examinations. Mean attenuation of new bone lesions (sclerotic or lytic) significantly increased or decreased (P < 0.01) in all patients. Bone attenuation in pelvic areas without evident metastatic disease significantly increased in patients with PD (P = 0.019), whereas there was no change in SD (P = 0.076). Lesion-based sensitivity, specificity, accuracy, positive predictive values, and negative predictive values were 98.7%, 79.5%, 94.5%, 95.1%, and 94.5%, respectively. Interobserver agreement was good (κ = 0.80; P = 0.077). Reading time was significantly faster for the bone subtraction maps versus 5-mm axial images (P < 0.001). CONCLUSIONS: Longitudinal bone subtraction maps increase the accuracy and efficiency of CT diagnosis of skeletal metastases in patients with breast cancer.

Implantability, Complications, and Follow-Up After Transjugular Intrahepatic Portosystemic Stent-Shunt Creation With the 6F Self-Expanding Sinus-SuperFlex-Visual Stent.

BACKGROUND: The transjugular intrahepatic portosystemic stent-shunt (TIPSS) builds a shortcut between the portal vein and a liver vein, and represents a sophisticated alternative to open surgery in the management of portal hypertension or its complications. OBJECTIVES: To describe clinical experiences with a low-profile nitinol stent system in TIPSS creation, and to assess primary and long-term success. PATIENTS AND METHODS: Twenty-six patients (5 females, 21 males; mean age 54.6 years) were treated using a low-profile 6F self-expanding sinus-SuperFlex-Visual stent system. The indication for TIPSS creation was refractory bleeding in 9 of the 26 patients, refractory ascites in 18 patients, and acute thrombosis of the portal vein confluence in one patient. Portosystemic pressure gradients before and after TIPSS, periprocedural and long-term complications, and the time to orthotopic liver transplantation (OLT) or death were recorded. RESULTS: The portosystemic pressure gradient was significantly reduced, from 20.9 ± 6.3 mmHg before to 8.2 ± 2.3 mmHg after TIPSS creation (P < 0.001). Procedure-related complications included acute tract occlusion (n = 2), liver hematoma (n = 1), hepatic encephalopathy (n = 1), and cardiac failure (n = 1). Three of the 26 patients had late-onset TIPSS occlusion (at 12, 12, and 39 months after TIPSS creation). Three patients died within one week after the procedure due to their poor general condition (multiorgan failure, acute respiratory distress syndrome, necrotizing pancreatitis, and aspiration pneumonia). Another four patients succumbed to their underlying advanced liver disease within one year after TIPSS insertion. Seven patients underwent OLT at a mean time of 9.4 months after TIPSS creation. CONCLUSION: The sinus-SuperFlex-Visual stent system can be safely deployed as a TIPSS device. The pressure gradient reduction was clinically sufficient to treat the patients' symptoms, and periprocedural complications were due to the TIPSS procedure per se rather than to the particular stent system employed in this study.

Labeling Human Melanoma Cells With SPIO: In Vitro Observations.

OBJECTIVES: To use the superparamagnetic iron oxide (SPIO) contrast agent Resovist (±transfection agent) to label human melanoma cells and determine its effects on cellular viability, microstructure, iron quantity, and magnetic resonance imaging (MRI) detectability. MATERIALS AND METHODS: Human SK-Mel28 melanoma cells were incubated with Resovist (±liposomal transfection agent DOSPER). The cellular iron content was measured, and labeled cells were examined at 1.5 T and 3.0 T. The intracellular and extracellular distributions of the contrast agent were assessed by light and electron microscopy. RESULTS: The incubation of melanoma cells with SPIO does not interfere with cell viability or proliferation. The iron is located both intracellularly and extracellularly as iron clusters associated with the exterior of the cell membrane. Despite thorough washing, the extracellular SPIO remained associated with the cell membrane. The liposomal transfection agent does not change the maximum achievable cellular iron content but promotes a faster iron uptake. The MRI detectability persists for at least 7 days. CONCLUSION: The transfection agent DOSPER facilitates the efficient labeling of human metastatic melanoma cells with Resovist. Our findings raise the possibility that other Resovist-labeled cells may collect associated extracellular nanoparticles. The SPIO may be available to other iron-handling cells and not completely compartmentalized during the labeling procedure.

Therapeutic angiographic procedures: differences in dose area product between analog image intensifier and digital flat panel detector.

BACKGROUND: Radiation exposure remains an unceasing concern in angiographic procedures. Modern angiography machines such as analog image intensifiers (AII) or the new flat panel detectors (FPD) aim at a further dose reduction. PURPOSE: To present dose area products (DAP) in a broad spectrum of therapeutic angiographic procedures, comparing an AII to an FPD angiography system. MATERIAL AND METHODS: A total of 999 peripheral therapeutic angiography procedures performed with an FPD (n = 562) and an AII system (n = 437) were evaluated. DAP, fluoroscopy time, and patients' body mass index (BMI) were recorded. Interventions were classified into five main groups: percutaneous transluminal angioplasty (PTA); PTA and stent placement; intra-arterial thrombolysis; embolization procedures; and specialized interventions. RESULTS: DAP values in therapeutic angiographic procedures were significantly higher when performed with the FPD compared to the AII system. The increase of the FPD versus AII system was 100.1% for PTA, 39.9% for PTA and stent placement, 187% for intra-arterial thrombolysis, 31.3% for embolization procedures, and 361% for specialized interventions. These differences persisted after standardizing DAP values to the geometric mean fluoroscopy duration of each procedure. Fluoroscopy times were shorter in all interventions performed at the FPD as compared to the AII system. DAPs increased with higher BMI, but the DAP increase of both systems with elevated BMI was variable, depending on the individual intervention. CONCLUSION: In therapeutic angiographic procedures, the FPD system required higher DAPs despite shorter fluoroscopy times as compared to an AII system. Better ergonomics and speediness of the FPD system may be advantageous in the emergency setting.

Quantitative Assessment of Iron-Labeled Stem-Cell Adhesion at the Vessel Wall in a Vascular Flow Model: Correlation of T2*-Weighted Imaging at 3 T and Histology.

PURPOSE: To evaluate the distribution of superparamagnetic iron oxide (SPIO)-labeled cells in a perfused segment of a porcine artery and to estimate the number of adherent cells by means of magnetic resonance (MR) imaging. MATERIALS AND METHODS: Six vessel specimens (diameters between 0.8 and 1.2 cm) were placed in a bioreactor system, and 2 × 10(4) to 1 × 10(6) SPIO-labeled endothelial colony-forming cells were injected into the artery within the perfused reactor. The area of resulting signal extinctions at the inner wall of the vessels was quantified on MR images by using a high-resolution T2*-weighted sequence with a slice-by-slice approach. After imaging, the labeled cells were quantified histologically. RESULTS: The total iron load of each cell was 56.5 pg ± 14.4. In the applied range of 2 × 10(4) to 1 × 10(6) cells per vessel, the area of iron-induced signal extinction at the vessel wall on T2*-weighted imaging corresponded to the histologically detected cell number (r = 0.98, P < .001). CONCLUSIONS: A correlation between the area of signal extinction and the number of labeled cells at the vessel wall was found. This might help to evaluate dose rates in further clinical applications of intravascular cell-based therapies.

Cell Cycle Regulation of Smooth Muscle Cells--Searching for Inhibitors of Neointima Formation: Is Combretastatin A4 an Alternative to Sirolimus and Paclitaxel?

PURPOSE: To compare the effects of sirolimus, paclitaxel, and combretastatin A4 (CA4) on regulatory proteins of the cell cycle in proliferating smooth muscle cells (SMCs). MATERIALS AND METHODS: Human aortic SMCs were treated with sirolimus, paclitaxel, and CA4 at 5 × 10(-9) mol/L. After 1 day, half of the cells were harvested (DAY1 group). The treatment medium of the other half was replaced with culture medium on day 4, and those cells were harvested on day 5 (DAY5 group). Cyclins D1, D2, E, and A and cyclin-dependent kinase (CDK) inhibitors p16, p21, and p27 were detected by Western blot technique. Quantification was performed by scanning densitometry of the specific bands. RESULTS: In the DAY1 group, treatment with sirolimus resulted in decreased intracellular levels of cyclins D2 and A (P < .05). Increased D cyclins and reduced levels of cyclins E and A (P < .05) in the DAY5 group indicated a permanent G1/S block by sirolimus. Paclitaxel led to only slight alterations of cyclin and CDK inhibitor expression (P > .05). In the DAY1 group, CA4 decreased intracellular levels of cyclins D2, E, and A (P < .05). Despite recovery effects in the DAY5 group (increase of cyclins D1, D2, and A compared with DAY1 group; P < .05), the upregulation of the CDK inhibitor p21, increased D cyclins, and decreased cyclins E and A (P < .05) are compatible with a G1 arrest. CONCLUSIONS: CA4 is a stronger inhibitor of the SMC cycle than sirolimus or paclitaxel and may represent an alternative for drug-eluting stents in atherosclerotic luminal stenosis. The effect of CA4 on neointima formation should be evaluated further.

Portosystemic pressure reduction achieved with TIPPS and impact of portosystemic collaterals for the prediction of the portosystemic-pressure gradient in cirrhotic patients.

PURPOSE: The portosystemic pressure gradient is an important factor defining prognosis in hepatic disease. However, noninvasive prediction of the gradient and the possible reduction by establishment of a TIPSS is challenging. A cohort of patients receiving TIPSS was evaluated with regard to imaging features of collaterals in cross-sectional imaging and the achievable reduction of the pressure gradient by establishment of a TIPSS. METHODS: In this study 70 consecutive patients with cirrhotic liver disease were retrospectively evaluated. Patients received either CT or MR imaging before invasive pressure measurement during TIPSS procedure. Images were evaluated with regard to esophageal and fundus varices, splenorenal collaterals, short gastric vein and paraumbilical vein. Results were correlated with Child stage, portosystemic pressure gradient and post-TIPSS reduction of the pressure gradient. RESULTS: In 55 of the 70 patients TIPSS reduced the pressure gradient to less than 12 mmHg. The pre-interventional pressure and the pressure reduction were not significantly different between Child stages. Imaging features of varices and portosystemic collaterals did not show significant differences. The only parameter with a significant predictive value for the reduction of the pressure gradient was the pre-TIPSS pressure gradient (r = 0.8, p<0.001). CONCLUSIONS: TIPSS allows a reliable reduction of the pressure gradient even at high pre-interventional pressure levels and a high collateral presence. In patients receiving TIPSS the presence and the characteristics of the collateral vessels seem to be too variable to draw reliable conclusions concerning the portosystemic pressure gradient.

Comparison of visibility for four self-expanding nitinol bare stents in vitro.

BACKGROUND: Sufficient radiopacity of stents is a prerequisite for safe interventions and minimization of the radiation dose for the patient and the interventionist. Modern nitinol stents are considered less radiopaque compared to formerly used stents. PURPOSE: To evaluate the objective detection rate (ODR) and the subjective radiopacity score (SRS) of four self-expanding nitinol stents with their markers on a phantom human pelvis. MATERIAL AND METHODS: We evaluated the ODR (as a percentage of correctly identified stents) and the SRS (on a scale from 0 = not visible to 4 = excellent visibility) for four self-expanding nitinol stents (SinusSuperflex, SMART, Luminexx, Zilver) with 8 mm diameter and 40 mm length. Stents were placed on a phantom human pelvis and images of the stents were taken in four different positions (right and left lumbosacral joint and near the right and left limbus acetabuli) using the following modes: spotfilm, pulsed fluoroscopy (4, 7.5, 15, and 30 pulses/min) and at three different digital magnification modes. Dose area products (DAPs) were assessed. RESULTS: ODR and SRS, respectively, were significantly increased for the SMART stent compared to all other tested stents (P < 0.05): SMART 93.53% and 2.43, SinusSuperflex 90.81% and 2.21, Luminexx 90.39% and 2.20, and Zilver 89.28% and 2.21. ODR was significantly reduced in position 3 where the bone overlap was more pronounced for all stents (detection rates 77.14-79.56%). An increase in magnification significantly improved the ODR and SRS for all stents (70.33-99.25% and 1.07-3.28, respectively, P < 0.05). Increased pulsing frequency did not improve the ODR of the various stents but did increase the DAP. CONCLUSION: The SMART stent had the best overall performance. In the presence of bone overlap, all self-expanding nitinol stents had poor results. Increased pulsing frequency did not improve ODR or SRS but did increase the DAP. Use of digital magnification modes had no effect on DAP increasing ODR and SRS.

Comparison of digital flat-panel detector and conventional angiography machines: evaluation of stent detection rates, visibility scores, and dose-area products.

OBJECTIVE: The objective of this study was to compare the performance and radiation doses of a flat-panel detector (FPD) angiography machine with an image intensifier (II) angiography machine. MATERIALS AND METHODS: Images of four nitinol stents (Sinus-SuperFlex, SMART, Luminexx, and Zilver stents) in a phantom of a human pelvis were acquired on an FPD system (Axiom Artis) and an II system (Fluorospot TOP) using the following modes: spot-film, continuous fluoroscopy (4, 7.5, 15, and 30 pulses/s), and three amplification modes. Objective stent detection rates and subjective radiopacity scores (scale: 0 [not visible] to 4 [excellent visibility]) were calculated. The radiation doses evaluated by the respective machines were compared. RESULTS: Over all modes and stents, the mean objective correct stent detection rates and mean subjective radiopacity scores were 89.49% and 1.81, respectively, for the Axiom Artis and 91.00% and 2.26 for the Fluorospot TOP. The stent detection rates over all modes for the SMART and Luminexx stents were better using the Axiom Artis machine (97.61% vs 93.55% and 98.28% vs 90.41%, respectively) and those for the Sinus-SuperFlex and Zilver stents were better using the Fluorospot TOP machine (90.83% vs 83.56% and 89.29% vs 80.50%). The subjective radiopacity scores of stent visibility were worse for the Axiom Artis than the Fluorospot TOP for all stents except the Luminexx stent (mean score, 2.34 vs 2.21, respectively). The objective stent detection rates and subjective radiopacity scores improved using the spot-film mode and with raising amplification, whereas increases in the fluoroscopy pulsing frequency did not improve stent detection rates or radiopacity scores for either machine. The radiation doses at continuous fluoroscopy were approximately 90% higher for the Axiom Artis than for the Fluorospot TOP (2.60 vs 1.41 µGy/m(2) at 30 pulses/s, respectively). CONCLUSION: The objective correct stent detection rates were similar for both machines with differences in detection for the respective stents. The subjective radiopacity scores were almost always better for the Fluorospot TOP machine. Also, the Axiom Artis machine generated approximately 90% higher radiation doses in fluoroscopy. For both machines, using a higher fluoroscopy pulsing frequency had no positive effect on objective correct stent detection rates or subjective radiopacity scores.

Decreased neointimal extracellular matrix formation in RAGE-knockout mice after microvascular denudation.

PURPOSE: To evaluate in vivo the role of RAGE (receptor for advanced glycated end products) in the development of restenosis and neointimal proliferation in RAGE-deficient knockout (KO) mice compared with wild-type (WT) mice in an animal model. MATERIALS AND METHODS: Sixteen WT and 15 RAGE-deficient mice underwent microvascular denudation of the common femoral artery under general anaesthesia. Contralateral arteries underwent a sham operation and served as controls. Four weeks after the intervention, all animals were killed, and paraformaldehyde-fixed specimens of the femoral artery were analysed with different stains (hematoxylin and eosin and Elastica van Gieson) and several different types of immunostaining (proliferating cell nuclear antigen, α-actin, collagen, von Willebrand factor, RAGE). Luminal area, area of the neointima, and area of the media were measured in all specimens. In addition, colony-formation assays were performed, and collagen production by WT smooth muscle cells (SMCs) and RAGE-KO SMCs was determined. For statistical analysis, P < 0.05 was considered statistically significant. RESULTS: Four weeks after denudation, WT mice showed a 49.6% loss of luminal area compared with 14.9% loss of luminal area in RAGE-deficient mice (sham = 0% loss) (P < 0.001). The neointima was 18.2 (*1000 µm(2) [n = 15) in the WT group compared with only 8.4 (*1000 µm(2) [n = 16]) in the RAGE-KO group. RAGE-KO SMCs showed significantly decreased proliferation activity and production of extracellular matrix protein. CONCLUSION: RAGE may be shown to play a considerable role in the formation of neointima leading to restenosis after vascular injury.

A dual-inhibition study on vascular smooth muscle cells with meclofenamic acid and ß-irradiation for the prevention of restenosis.

PURPOSE: Restenosis is still one of the major limitations after angioplasty. A therapeutic treatment combining ß-irradiation and pharmacologic cyclooxygenase-2 inhibition was employed to study the impact on vascular smooth muscle cells (SMCs). MATERIALS AND METHODS: The effects of meclofenamic acid in combination with yttrium-90 ((90)Y) on cell growth, clonogenic activity, cell migration, and cell cycle distribution of human aortic SMCs were investigated. Treatment was sustained over a period of 4 days and recovery of cells was determined until day 20 after initiation. The hypothesis was that there is no difference between control and treated groups. RESULTS: A dose-dependent growth inhibition was observed in single and combined treatment groups for meclofenamic acid and ß-irradiation. Cumulative radiation dosage of 8 Gy completely inhibited colony formation. This was also observed for 200 µM meclofenamic acid alone or in combination with minor ß-irradiation dosages. Results of the migration tests showed also a dose dependency with additive effects of combined therapy. Meclofenamic acid 200 µM alone and with cumulative ß-irradiation dosages resulted in an increased G2/M-phase share. CONCLUSIONS: Incubating human SMCs with meclofenamic acid and (90)Y for a period of 4 d (ie, 1.5 half-life times) resulted in an effective inhibition of smooth muscle cell proliferation, colony formation, and migration.

Effects of magnetic resonance imaging contrast agents on human umbilical vein endothelial cells and evaluation of magnetic resonance imaging contrast media-triggered transforming growth factor-beta induction in dermal fibroblasts (HSF) as a model for nephrogenic systemic fibrosis.

RATIONALE AND OBJECTIVES: The objective of this study was to evaluate effects of 6 commercially available magnetic resonance contrast media (CM) on human umbilical vein endothelial cells (HUVEC) and the induction of transforming growth factor-beta (TGF-ß) in dermal fibroblasts (HSF) as a possible model for the pathogenesis of nephrogenic systemic fibrosis. METHODS: HUVECs were incubated with 10× and 20× of the molar standard blood concentration achieved with CM applications for magnetic resonance imaging examinations (10× and 20× concentration) for 24 hours using gadolinium-based CM Gadovist, Magnevist, Multihance, and Omniscan, as well as Teslascan (Manganese-based), and Resovist (Iron-based). Proliferation kinetics (PK), colony formation, and viability assays were performed. Additionally, human dermal fibroblasts (HSF) were incubated for 24 hours with 1× and 20× concentration in all 6 CM, and TGF-ß levels were assessed directly after the incubation period as well as on days 3 and 8 postincubation. RESULTS: HUVEC PK data show similar gains in cell numbers for all 6 CM in both concentration groups over the 17-day assessment period. Only cells incubated with Omniscan and Teslascan differed from the other groups on days 3 and 7 postincubation (P < 0.05). After day 7, a cell regain occurred in the Omniscan and Teslascan groups reaching the numbers of the other groups in sequel. Differences in colony formation were consistent with PK results with a statistically significant reduction in clonogenic activity for Teslascan and Omniscan in HUVEC cells, P < 0.05. No reduction in viability was seen for all groups and conditions. TGF-ß expression of HSF cells incubated with 1× concentration and all CM did not differ significantly from control cells for any point in time investigated. At 20× concentration directly after incubation, TGF-ß was significantly reduced for the Teslascan and Resovist group as 3 compared with control and all other CM groups, P < 0.05. On day 3 postincubation, only Resovist-incubated HSF cells showed a significant reduction of TGF-ß (1.614, standard deviations: 89) as compared with the control group (2.883, standard deviations: 30) and the other CM. TGF-ß was slightly reduced for all CM groups 8 days after incubation (not statistically significant, P > 0.05). CONCLUSIONS: After 24 hours of incubation with Omniscan and Teslascan (10× and 20× concentration), considerable short-term antiproliferative effects in HUVECs were observed. HSF cells (20× concentration) showed a reduction of TGF-ß for Resovist and Teslascan directly after incubation, whereas TGF-ß levels in HSF cells were slightly reduced for all CM 8 days after incubation. Therefore, TGF-ß-mediated proliferative effects on fibroblasts or on collagen synthesis potentially leading to nephrogenic systemic fibrosis may mainly be triggered by tissue monocytes and macrophages in the peripheral blood instead of dermal fibroblasts.

In vitro comparison of the antiproliferative effects of rhenium-186 and rhenium-188 on human aortic endothelial cells.

PURPOSE: Rhenium-186 ((186)Re) and rhenium-188 ((188)Re) are promising radionuclides for the inhibition of restenosis after percutaneous transluminal angioplasty or other vascular interventions. Until now the maximal dose tolerance of endothelial cells has not been clearly known. MATERIALS AND METHODS: To characterize the effects of local irradiation treatment, human aortic endothelial cells (ECs) were incubated with different doses of (186)Re and (188)Re. Two days after plating, ECs received treatment for a period of 5 days. The total radiation doses applied were 1, 4, 8, 16, and 32 Gy. On days 1, 3, 5, 7, and 12 after initial rhenium incubation, cell growth, clonogenic activity, cell-cycle distribution, and cytoskeletal architecture were evaluated. RESULTS: From the first day on, a dose-dependent growth inhibition was observed. Cumulative doses of ≥32 Gy caused a weak colony formation and significant alterations in the cytoskeletal architecture. An increased fraction of cells in G2/M phase was seen for cumulative radiation doses of ≥16 Gy. Interestingly, there were no significant differences between (186)Re and (188)Re. CONCLUSION: Even for low dose rates of ß particles a dose-dependent proliferation inhibition of ECs is seen. Doses beyond 32 Gy alter the cytoskeletal architecture with possibly endothelial dysfunction and late thrombosis.

Effects of MRI contrast agents on human embryonic lung fibroblasts.

RATIONALE AND OBJECTIVES: The objective of this investigation was to evaluate 6 magnetic resonance contrast media (CM) with regard to their different effects on human embryonic lung fibroblasts (HEL-299). METHODS: Human embryonic fibroblasts (HEL-299) were incubated with 1x, 5x, 10x, and 20x of the normal molar blood concentration (1x, 5x, 10x, 20x conc.) reached through routine contrast media applications for MRI examinations. Four gadolinium-based CM, ie, Gadovist, Magnevist, Multihance, Omniscan, Teslascan (Manganese-based), and Resovist (Iron-based), with incubation periods over 4 hours and 24 hours were investigated. Proliferation kinetics, colony formation, and viability assays were performed after 4 and 24 hours of treatment. Apoptotic cells were quantified after tetramethylrhodamine ethyl ester staining following 24 hours of CM media incubation (20x conc.) by fluorescence activated cell sorting cytometry. Furthermore, immunofluorescence images with vimentin staining were obtained (20x conc., 24 hours treatment). Cell cycle analysis was performed after 24 hours of incubation and 20x conc. directly after incubation and 24 hours later (fluorescence activated cell sorting cytometry). RESULTS: The proliferation kinetics performed with 20x conc. revealed a persistent increase in cell numbers until day 11 for all CM without significant differences after 4 hours of incubation. A significant reduction in initial cell numbers was recorded in the 24-hours-group after 4 days of CM incubation with Magnevist, Multihance, Omniscan, and Teslascan. Solely cells incubated with Resovist and Gadovist failed to show decreased cell numbers when compared with the control group. However, a considerable cell regain occurred afterward reaching control-group levels on day 21. Colony numbers were significantly reduced (about 20%, respectively) with Magnevist at 10x and 20x conc., as well as Omniscan and Multihance at 20x conc. when compared with all other groups, P < 0.05. Cell-cycle distribution showed a reduction of S-phase cells for Magnevist, Omniscan, and Multihance (2.9%-10.5%) when compared with Gadovist, Resovist and Teslascan (16.7%-21.0%). Twenty-four hours after incubation, the percentiles of cells in S-phase were significantly increased for Magnevist, Omniscan, and Multihance (31.4%-38.5%) when compared with Gadovist, Resovist, and Teslascan (18.6%-26.8%), P < 0.05. Viability was not impaired by administration of any CM and no apoptosis was seen after tetramethylrhodamine ethyl ester staining at 24 hours of incubation. Cell morphology remained unchanged in vimentin-staining for all CM and conditioning regimens. CONCLUSIONS: No toxic effects on embryonic fetal lung fibroblasts were detectable after 4 and 24 hours of incubation in 6 MRI CM and 10x to 20x conc. in our setting. Antiproliferative effects, initially detected with Magnevist, Omniscan and Multihance, were rapidly compensated for.