Quantitative differentiation of minimal-fat angiomyolipomas from renal cell carcinomas using grating-based x-ray phase-contrast computed tomography: An ex vivo study.

BACKGROUND: The differentiation of minimal-fat-or low-fat-angiomyolipomas from other renal lesions is clinically challenging in conventional computed tomography. In this work, we have assessed the potential of grating-based x-ray phase-contrast computed tomography (GBPC-CT) for visualization and quantitative differentiation of minimal-fat angiomyolipomas (mfAMLs) and oncocytomas from renal cell carcinomas (RCCs) on ex vivo renal samples. MATERIALS AND METHODS: Laboratory GBPC-CT was performed at 40 kVp on 28 ex vivo kidney specimens including five angiomyolipomas with three minimal-fat (mfAMLs) and two high-fat (hfAMLs) subtypes as well as three oncocytomas and 20 RCCs with eight clear cell (ccRCCs), seven papillary (pRCCs) and five chromophobe RCC (chrRCC) subtypes. Quantitative values of conventional Hounsfield units (HU) and phase-contrast Hounsfield units (HUp) were determined and histogram analysis was performed on GBPC-CT and grating-based attenuation-contrast computed tomography (GBAC-CT) slices for each specimen. For comparison, the same specimens were imaged at a 3T magnetic resonance imaging (MRI) scanner. RESULTS: We have successfully matched GBPC-CT images with clinical MRI and histology, as GBPC-CT presented with increased soft tissue contrast compared to absorption-based images. GBPC-CT images revealed a qualitative and quantitative difference between mfAML samples (58±4 HUp) and oncocytomas (44±10 HUp, p = 0.057) and RCCs (ccRCCs: 40±12 HUp, p = 0.012; pRCCs: 43±9 HUp, p = 0.017; chrRCCs: 40±7 HUp, p = 0.057) in contrast to corresponding laboratory attenuation-contrast CT and clinical MRI, although not all differences were statistically significant. Due to the heterogeneity and lower signal of oncocytomas, quantitative differentiation of the samples based on HUp or in combination with HUs was not possible. CONCLUSIONS: GBPC-CT allows quantitative differentiation of minimal-fat angiomyolipomas from pRCCs and ccRCCs in contrast to absorption-based imaging and clinical MRI.

Dynamic CTA in Native Kidneys Using a Multiphase CT Protocol-Potential of Significant Reduction of Contrast Medium.

RATIONALE AND OBJECTIVE: The objective of this study was to assess an optimized renal multiphase computed tomography angiography (MP-CTA) protocol regarding reduction of contrast volume. MATERIALS AND METHODS: Thirty patients underwent MP-CTA (12 phases, every 3.5 seconds, 80 kV/120 mAs) using 30 mL of contrast medium. The quality of MP-CTA was assessed quantitatively measuring vessel attenuation, image noise, and contrast-to-noise ratio. MP-CTA was evaluated qualitatively regarding depiction of vessels, cortex differentiation, and motion artifacts (grades 1-4, 1 = best). Mean effective radiation dose was registered. Results were compared to standard renal computed tomography angiography (CTA) (80 mL). Student t test was applied, if variables followed normal distribution. For other variables, nonparametric Mann-Whitney U test was used. RESULTS: All acquisitions were successfully performed, and no patient had to be excluded from the study. MP-CTA enabled high attenuation (aorta: 503 ± 91 HU, renal arteries: 450 ± 73 HU/456 ± 72 HU) at adequate image noise (13.7 ± 1.5) and good contrast-to-noise ratio (34.2 ± 10.2). Good attenuation of renal veins was observed (286 ± 43 HU/282 ± 42 HU). Arterial enhancement was significantly higher compared to renal CTA (aorta: 396 ± 90 HU, renal arteries: 331 ± 74 HU/333 ± 80 HU; P < .001). MP-CTA protocol enabled good image quality of renal arteries (1.5 ± 0.6) and veins (1.7 ± 0.6). Cortex differentiation and motion artifacts were ranked 1.8 ± 0.8 and 1.6 ± 0.8. The mean effective radiation dose was 9 mSv (MP-CTA). CONCLUSIONS: Compared to standard renal CTA, the renal MP-CTA enabled the significant reduction of contrast volume and simultaneously provided a significantly higher arterial attenuation.

X-Ray Dark-field Imaging to Depict Acute Lung Inflammation in Mice.

The aim of this study was to evaluate the feasibility of early stage imaging of acute lung inflammation in mice using grating-based X-ray dark-field imaging in vivo. Acute lung inflammation was induced in mice by orotracheal instillation of porcine pancreatic elastase. Control mice received orotracheal instillation of PBS. Mice were imaged immediately before and 1 day after the application of elastase or PBS to assess acute changes in pulmonary structure due to lung inflammation. Subsequently, 6 mice from each group were sacrificed and their lungs were lavaged and explanted for histological analysis. A further 7, 14 and 21 days later the remaining mice were imaged again. All images were acquired with a prototype grating-based small-animal scanner to generate dark-field and transmission radiographs. Lavage confirmed that mice in the experimental group had developed acute lung inflammation one day after administration of elastase. Acute lung inflammation was visible as a striking decrease in signal intensity of the pulmonary parenchyma on dark-field images at day 1. Quantitative analysis confirmed that dark-field signal intensity at day 1 was significantly lower than signal intensities measured at the remaining timepoints, confirming that acute lung inflammation can be depicted in vivo with dark-field radiography.

Qualitative and Quantitative Imaging Evaluation of Renal Cell Carcinoma Subtypes with Grating-based X-ray Phase-contrast CT.

Current clinical imaging methods face limitations in the detection and correct characterization of different subtypes of renal cell carcinoma (RCC), while these are important for therapy and prognosis. The present study evaluates the potential of grating-based X-ray phase-contrast computed tomography (gbPC-CT) for visualization and characterization of human RCC subtypes. The imaging results for 23 ex vivo formalin-fixed human kidney specimens obtained with phase-contrast CT were compared to the results of the absorption-based CT (gbCT), clinical CT and a 3T MRI and validated using histology. Regions of interest were placed on each specimen for quantitative evaluation. Qualitative and quantitative gbPC-CT imaging could significantly discriminate between normal kidney cortex (54 ± 4 HUp) and clear cell (42 ± 10), papillary (43 ± 6) and chromophobe RCCs (39 ± 7), p < 0.05 respectively. The sensitivity for detection of tumor areas was 100%, 50% and 40% for gbPC-CT, gbCT and clinical CT, respectively. RCC architecture like fibrous strands, pseudocapsules, necrosis or hyalinization was depicted clearly in gbPC-CT and was not equally well visualized in gbCT, clinical CT and MRI. The results show that gbPC-CT enables improved discrimination of normal kidney parenchyma and tumorous tissues as well as different soft-tissue components of RCCs without the use of contrast media.

X-ray Dark-field Radiography - In-Vivo Diagnosis of Lung Cancer in Mice.

Accounting for about 1.5 million deaths annually, lung cancer is the prevailing cause of cancer deaths worldwide, mostly associated with long-term smoking effects. Numerous small-animal studies are performed currently in order to better understand the pathogenesis of the disease and to develop treatment strategies. Within this letter, we propose to exploit X-ray dark-field imaging as a novel diagnostic tool for the detection of lung cancer on projection radiographs. Here, we demonstrate in living mice bearing lung tumors, that X-ray dark-field radiography provides significantly improved lung tumor detection rates without increasing the number of false-positives, especially in the case of small and superimposed nodules, when compared to conventional absorption-based imaging. While this method still needs to be adapted to larger mammals and finally humans, the technique presented here can already serve as a valuable tool in evaluating novel lung cancer therapies, tested in mice and other small animal models.

X-ray dark-field radiography facilitates the diagnosis of pulmonary fibrosis in a mouse model.

The aim of this study was to evaluate whether diagnosing pulmonary fibrosis with projection radiography can be improved by using X-ray dark-field radiograms. Pulmonary X-ray transmission and dark-field images of C57Bl/6N mice, either treated with bleomycin to induce pulmonary fibrosis or PBS to serve as controls, were acquired with a prototype grating-based small-animal scanner. Two blinded readers, both experienced radiologists and familiar with dark-field imaging, had to assess dark-field and transmission images for the absence or presence of fibrosis. Furthermore readers were asked to grade their stage of diagnostic confidence. Histological evaluation of the lungs served as the standard of reference in this study. Both readers showed a notably higher diagnostic confidence when analyzing the dark-field radiographs (p < 0.001). Diagnostic accuracy improved significantly when evaluating the lungs in dark-field images alone (p = 0.02) or in combination with transmission images (p = 0.01) compared to sole analysis of absorption images. Interreader agreement improved from good when assessing only transmission images to excellent when analyzing dark-field images alone or in combination with transmission images. Adding dark-field images to conventional transmission images in a murine model of pulmonary fibrosis leads to an improved diagnosis of this disease on chest radiographs.

The role of diffusion-weighted MRI in assessment of inflammatory bowel disease.

AIM: To evaluate the role of diffusion-weighted MRI (DW-MRI) in detecting and differentiating acute from chronic bowel inflammation in patients with Crohn's disease (CD). MATERIALS AND METHODS: MR-enteroclysis examinations with DW-MRI were reviewed from 24 patients with histologically proven CD. Segments of bowel were evaluated for acute and chronic inflammation in three different reviews of the MRI images: T2w alone, T2w + DWI, and T2w + CET1w. Mean ADC values of normal bowel segments, as well as bowel segments with acute and chronic inflammation were calculated and compared. Analyses of receiver-operating characteristic (ROC) curve were performed. RESULTS: Hundred and forty four bowel segments in total were reviewed. Inflammation was present in 45 segments. Acute inflammation was present in 31 segments, chronic inflammation in 14. 98 bowel segments showed no inflammatory activity. Sensitivity and specificity for differentiation between normal and inflamed bowel segments was 0.6, 0.67, and 0.80 on T2w, T2w + DWI, and T2w + CET1w datasets, respectively. Specificities for differentiation between normal and inflamed bowel segments were 0.96, 0.96, and 0.98. Sensitivities for differentiation between acute and chronically inflamed bowel segments were 0.85, 0.91, and 0.96, and specificities were 0.88, 0.89, and 1.0, respectively. The mean ADC value of normal bowel (2.18 ± 0.37 × 10(-3) mm(2)/s) was statistically significantly greater than the mean value of inflamed bowel segments (p < 0.001). The mean ADC value of acutely inflamed bowel segments was statistically significantly lower than that of chronically inflamed bowel segments (1.09 ± 0.18 × 10(-3) vs. 1.55 ± 0.21 × 10(-3) mm(2)/s) (p < 0.001). Estimated area under the ROC curve for the diagnosis of acute vs. chronic inflammation was 0.950. A threshold of ADC value of 1.41 × 10(-3) mm(2)/s was optimal for calculation of sensitivity and specificity. CONCLUSION: DW-MRI improves detection and differentiation of acute vs. chronic inflammatory changes of the bowel in patients with CD compared to T2w-images alone.

Dynamic Contrast-Enhanced Computed Tomography: A New Diagnostic Tool to Assess Renal Perfusion After Ischemia-Reperfusion Injury in Mice: Correlation of Perfusion Deficit to Histopathologic Damage.

OBJECTIVE: The aim of this study was to investigate the value of dynamic contrast-enhanced computed tomography (CT) in the assessment of renal perfusion parameters after ischemia-reperfusion (I/R) injury in an experimental murine model. MATERIALS AND METHODS: Balb/cJ wildtype mice were subjected to 45 minutes (AKI45) or 60 minutes (AKI60) of unilateral warm I/R injury by clamping the pedicle of the right kidney. Two, 7, and 18 days after right I/R injury, renal blood flow (RBF), renal volume of distribution (RVD), and mean transit time were quantitatively assessed in the cortex of both kidneys by dynamic contrast-enhanced CT. Acute tubular injury (ATI) was assessed by histologic analysis using a semiquantitative sum score (score, 0-18) and correlated with RBF, RVD, and mean transit time. RESULTS: Histologic signs of ATI could be detected in the clamped kidneys in both groups already at day 2. Pathologic features of ATI worsened in AKI60 until day 18 (score, 7 ± 0), whereas mice in AKI45 group showed amelioration over time (score, 4 ± 2). Renal blood flow was significantly reduced in ischemic kidneys in AKI45 (287 ± 32 mL/100 mL per minute; P < 0.01) and AKI60 group (249 ± 73 mL/100 mL per minute; P < 0.01) as compared with that in healthy kidneys (402 ± 49 mL/100 mL per minute) on day 2. It decreased further at day 7 in both groups (AKI45: 165 ± 44 mL/100 mL per minute, P < 0.01; AKI60: 151 ± 72 mL/100 mL per minute, P < 0.05) and improved at day 18 in AKI45 (261 ± 11 mL/100 mL per minute, P < 0.05) and to a lesser degree in AKI60 (197 ± 52 mL/100 mL per minute, P > 0.05). Values of RVD paralleled RBF at all time points. Renal blood flow (r = -0.79; P < 0.01) and RVD (r = -0.8; P < 0.01) significantly correlated with the histological damage score (Spearman rank correlation). CONCLUSIONS: Dynamic contrast-enhanced CT is a noninvasive method to determine renal perfusion changes in acute kidney injury. It might be a valuable diagnostic tool to predict outcome or monitor treatment effects of renal I/R injury.

Dynamic contrast-enhanced magnetic resonance imaging measurements in renal cell carcinoma: effect of region of interest size and positioning on interobserver and intraobserver variability.

PURPOSE: The purpose of this study was to assess the influence of region of interest (ROI) size and positioning on perfusion and permeability parameters as well as on interobserver and intraobserver variability of dynamic contrast-enhanced (DCE-MRI) of primary renal cell carcinoma (RCC) and metastases. MATERIALS AND METHODS: Thirty-nine DCE-MRI examinations of 34 patients with primary RCC and 20 examinations of 9 patients with RCC metastases obtained at 1.5 T were evaluated. Pretreatment and posttreatment analysis with antiangiogenic therapy was performed in 4 patients with primary RCCs and 5 patients with metastases. The ROIs of the whole tumor (wROI), the circular edge (cROI), a user-defined arbitrary small region (sROI), and a semiautomated segmented ROI were independently defined by 2 readers on 1 slice on arterial phase DCE-MRI images or on parametric plasma-flow maps. Analysis with a 2-compartment exchange model provided 4 parameters: plasma flow (FP), plasma volume (vp), permeability-surface product (PS), and extravascular-extracellular volume (ve). Interobserver and intraobserver parameter correlations were calculated using the intraclass correlation coefficient, and within-subject variability were considered on the basis of the coefficient of variation. Differences in measurement values of variable ROI size were assessed with paired t test. RESULTS: Mean values of FP and vp with sROIs were significantly higher than those with wROI, cROI, and semiautomated segmented ROI placement in tumor or metastases. Values of ve showed no significant difference between ROI sizes. The highest interobserver and intraobserver correlation with 0.99/0.98 for metastases and 0.97/0.98 for primary RCCs, respectively, was observed for all parameters when defining wROIs on dynamic images. Perfusion parameters of wROI measurements for FP (dynamic, 0.97; parametric maps, 0.96) and vp (0.95/0.89) showed higher interobserver correlation than did permeability parameters ve (0.64/0.6) and PS (0.79/0.5) in primary RCCs. The wROIs showed also the lowest within-subject coefficients of variation for perfusion parameters FP and vp compared with cROI and sROIs in primary RCCs and metastases. CONCLUSIONS: The ROI size and positioning do substantially influence quantitative perfusion and permeability parameters in DCE-MRI. The best interobserver and intraobserver correlation can be obtained when defining a whole-tumor ROI. The perfusion parameters are the most reliable, whereas the permeability parameters are more susceptible to interobserver variability. No significant differences between placing ROIs on morphological or parametric images were observed.

X-ray phase-contrast tomography of renal ischemia-reperfusion damage.

PURPOSE: The aim of the study was to investigate microstructural changes occurring in unilateral renal ischemia-reperfusion injury in a murine animal model using synchrotron radiation. MATERIAL AND METHODS: The effects of renal ischemia-reperfusion were investigated in a murine animal model of unilateral ischemia. Kidney samples were harvested on day 18. Grating-Based Phase-Contrast Imaging (GB-PCI) of the paraffin-embedded kidney samples was performed at a Synchrotron Radiation Facility (beam energy of 19 keV). To obtain phase information, a two-grating Talbot interferometer was used applying the phase stepping technique. The imaging system provided an effective pixel size of 7.5 µm. The resulting attenuation and differential phase projections were tomographically reconstructed using filtered back-projection. Semi-automated segmentation and volumetry and correlation to histopathology were performed. RESULTS: GB-PCI provided good discrimination of the cortex, outer and inner medulla in non-ischemic control kidneys. Post-ischemic kidneys showed a reduced compartmental differentiation, particularly of the outer stripe of the outer medulla, which could not be differentiated from the inner stripe. Compared to the contralateral kidney, after ischemia a volume loss was detected, while the inner medulla mainly retained its volume (ratio 0.94). Post-ischemic kidneys exhibited severe tissue damage as evidenced by tubular atrophy and dilatation, moderate inflammatory infiltration, loss of brush borders and tubular protein cylinders. CONCLUSION: In conclusion GB-PCI with synchrotron radiation allows for non-destructive microstructural assessment of parenchymal kidney disease and vessel architecture. If translation to lab-based approaches generates sufficient density resolution, and with a time-optimized image analysis protocol, GB-PCI may ultimately serve as a non-invasive, non-enhanced alternative for imaging of pathological changes of the kidney.

Dynamic contrast-enhanced magnetic resonance imaging assessment of kidney function and renal masses: single slice versus whole organ/tumor.

OBJECTIVES: The aim of this study was to compare single-slice and 3-dimensional (3D) analysis for magnetic resonance renography (plasma flow [FP], plasma volume [VP], and glomerular filtration rate [GFR]) and for dynamic contrast-enhanced magnetic resonance imaging (MRI) of renal tumors (FP, VP, permeability-surface area product), respectively. MATERIAL AND METHODS: We prospectively included 22 patients (43 kidneys with 22 suspicious renal lesions) and performed preoperative and postoperative imaging before and after partial nephrectomy, respectively. Of the 22 renal lesions, 15 turned out to be renal cell carcinoma and were included in the tumor analysis, altogether leading to 86 renal and 15 tumor MRI scans, respectively. Dynamic contrast-enhanced MRI was performed with a time-resolved angiography with stochastic trajectories sequence (spatial resolution, 2.6 × 2.6 × 2.6 mm3; temporal resolution, 2.5 seconds) at 3 T (Magnetom Verio; Siemens Healthcare Sector) after injection of 0.05 mmol/kg body weight Gadobutrol (Bayer Healthcare Pharmaceuticals). Analysis was performed using regions of interest encompassing a single central slice and the whole kidney/tumor, respectively. A 2-compartment model yielding FP, VP, GFR, or tumor permeability-surface area product was used for kinetic modelling. Modelling was performed based on relative contrast enhancement to account for coil-related inhomogeneity. Significance in difference, agreement, and goodness of fit of the data to the curve was assessed with paired t tests, Bland-Altman plots, and χ2 test, respectively. RESULTS: Bland-Altman analysis revealed a good agreement between both types of measurement for kidneys and tumors, respectively. Results between single-slice and whole-kidney regions of interest showed significant differences for Fp (single slice, 256.1 ± 104.1 mL/100 mL/min; whole kidney, 217.2 ± 92.5 mL/100 mL/min; P < 0.01). Regarding VP and GFR, no significant differences were observed. The χ2 test showed a significantly better goodness of fit of the data to the curve for whole kidneys (0.30% ± 0.18%) than for single slices (0.43% ± 0.26%) (P < 0.01). In contrast to renal assessment, tumor analysis showed no significant differences regarding functional parameters and χ test, respectively. CONCLUSION: In dynamic contrast-enhanced MRI of the kidney, both 3D whole-organ/tumor and single-slice analyses provide roughly comparable values in functional analysis. However, 3D assessment is considerably more precise and should be preferred if available.

The role of functional imaging in the era of targeted therapy of renal cell carcinoma.

Antiangiogenic therapies interacting with tumor-specific pathways have been established for targeted therapy of renal cell carcinoma (RCC). However, evaluation of tumor response based on morphologic tumor diameter measurements has limitations, as tumor shrinkage may lag behind pathophysiological response. Functional imaging techniques such as dynamic contrast-enhanced (DCE) ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI), unenhanced diffusion-weighted MRI (DW-MRI), and also metabolic imaging with positron emission tomography (PET) have the ability to assess physiological parameters and to predict and monitor therapy response. Assessment of changes in vascularity, cellularity, oxygenation, and glucose uptake with functional imaging during targeted therapy may correlate with progression-free survival and can predict tumor response or progression. In this review, we explore the potential of functional imaging techniques for assessing the effects of targeted therapy of RCC and as well review the reproducibility and limitations.

Optimization of contrast material delivery for dual-energy computed tomography pulmonary angiography in patients with suspected pulmonary embolism.

OBJECTIVES: To prospectively compare subjective and objective measures of image quality using 4 different contrast material injection protocols in dual-energy computed tomography pulmonary angiography (CTPA) studies of patients with suspected pulmonary embolism. MATERIALS AND METHODS: A total of 100 consecutive patients referred for CTPA for the exclusion of pulmonary embolism were randomized into 1 of 4 contrast material injection protocols manipulating iodine concentration and iodine delivery rate (IDR, expressed as grams of iodine per second): Iomeprol 400 at 3 mL/s (IDR = 1.2 gI/s), iomeprol 400 at 4 mL/s (IDR = 1.6 gI/s), iomeprol 300 at 5.4 mL/s (IDR = 1.6 gI/s), or iomeprol 300 at 4 mL/s (IDR = 1.2 gI/s). Total iodine delivery was held constant. Dual-energy CTPA of the lungs were acquired and used to calculate virtual 120 kV CTPA images as well as iodine perfusion maps. Attenuation values in the thoracic vasculature and image quality of virtual 120 kV CTPAs were compared between groups. Iodine perfusion maps were also compared by identifying differences in the extent of beam-hardening artifacts and subjective image quality. RESULTS: Protocols with an IDR of 1.6 gI/s provided the best attenuation profiles. CTPA image quality was greatest in the high concentration, high IDR (1.6 gI/s) protocol (P < 0.05 for all group comparisons) with no differences between the other groups (all P ≥ 0.05). Extent of beam-hardening artifacts and perfusion map image quality was significantly better using the high concentration, high IDR protocol as compared with all groups (P < 0.05 for all comparisons) and significantly worse using the low concentration, low IDR protocol as compared with all groups (all P ≥ 0.05); no difference was found between the high concentration, low IDR protocol and the low concentration, high IDR protocol (P = 0.73 for comparison of beam-hardening artifacts; P = 0.50 for comparison of perfusion map image quality). CONCLUSION: High iodine concentration and high IDR contrast material delivery protocols provide the best image quality of both CTPA and perfusion map images of the lung through high attenuation in the pulmonary arteries and minimization of beam-hardening artifacts.

Carnosine prevents apoptosis of glomerular cells and podocyte loss in STZ diabetic rats.

BACKGROUND/AIMS: We identified carnosinase-1 (CN-1) as risk-factor for diabetic nephropathy (DN). Carnosine, the substrate for CN-1, supposedly is a protective factor regarding diabetic complications. In this study, we hypothesized that carnosine administration to diabetic rats might protect the kidneys from glomerular apoptosis and podocyte loss. METHODS: We examined the effect of oral L-carnosine administration (1g/kg BW per day) on apoptosis, podocyte loss, oxidative stress, AGEs and hexosamine pathway in kidneys of streptozotocin-induced diabetic Wistar rats after 3 months of diabetes and treatment. RESULTS: Hyperglycemia significantly reduced endogenous kidney carnosine levels. In parallel, podocyte numbers significantly decreased (-21% compared to non-diabetics, p<0.05), apoptotic glomerular cells numbers increased (32%, compared to non-diabetic, p<0.05) and protein levels of bax and cytochrome c increased (175% and 117%). Carnosine treatment restored carnosine kidney levels, prevented podocytes loss (+23% compared to diabetic, p<0.05), restrained glomerular apoptosis (-34% compared to diabetic; p<0.05) and reduced expression of bax and cytochrome c (-63% and -54% compared to diabetics, both p<0.05). In kidneys of all diabetic animals, levels of ROS, AGEs and GlcNAc-modified proteins were increased. CONCLUSION: By inhibition of pro-apoptotic signaling and independent of biochemical abnormalities, carnosine protects diabetic rat kidneys from apoptosis and podocyte loss.

L-carnosine inhibits high-glucose-mediated matrix accumulation in human mesangial cells by interfering with TGF-ß production and signalling.

BACKGROUND: Transforming growth factor beta is recognized as a major cytokine in extracellular matrix (ECM) pathobiology as occurs in diabetic nephropathy. While experimental studies have advanced a protective role of carnosine for diabetic complications, a link between carnosine, TGF-ß and matrix accumulation remains to be elucidated. In the present study, we tested the hypothesis that L-carnosine inhibits TGF-ß production and signalling, thereby reducing hyperglycaemia-associated ECM accumulation. METHODS: Human mesangial cells (MC) were cultured in high-glucose (HG, 25 mM D-glucose) medium alone or in HG medium to which 20 mM L-carnosine was added. Collagen VI (Col6) and fibronectin (FN) deposition and messenger RNA expression were studied. In addition, TGF-ß production and activation of Smad1/5/8 (ALK1) and Smad2/3 (ALK5) pathways were assessed. RESULTS: Under HG conditions, deposition of Col6 and FN were increased 1.4- and 1.6-fold. This was significantly inhibited on the protein and messenger RNA level by L-carnosine. TGF-ß production increased under HG conditions but was completely normalized by addition of L-carnosine. Addition of exogenous TGF-ß could not overcome the effect of L-carnosine on Col6 and FN expression, indicating additionally interference with TGF-ß downstream signalling. Along the same line, L-carnosine reduced TGF-ß-mediated Smad2 phosphorylation, suggesting an inhibitory effect on ALK5 signalling. ALK1 signalling remained unchanged. Under HG conditions, pharmacologic inhibition of ALK5 prevented Col6 accumulation but did not change FN deposition. CONCLUSIONS: L-carnosine can modulate matrix accumulation in two ways. Firstly, inhibition of TGF-ß production might result in an overall inhibition of matrix accumulation and secondly, L-carnosine inhibits TGF-ß-induced matrix accumulation, most likely via inhibition of the ALK5 pathway.