Non-destructive classification of unlabeled cells: Combining an automated benchtop magnetic resonance scanner and artificial intelligence.

In order to treat degenerative diseases, the importance of advanced therapy medicinal products has increased in recent years. The newly developed treatment strategies require a rethinking of the appropriate analytical methods. Current standards are missing the complete and sterile analysis of the product of interest to make the drug manufacturing effort worthwhile. They only consider partial areas of the sample or product while also irreversibly damaging the investigated specimen. Two-dimensional T1 / T2 MR relaxometry meets these requirements and is therefore a promising in-process control during the manufacturing and classification process of cell-based treatments. In this study a tabletop MR scanner was used to perform two-dimensional MR relaxometry. Throughput was increased by developing an automation platform based on a low-cost robotic arm, resulting in the acquisition of a large dataset of cell-based measurements. Two-dimensional inverse Laplace transformation was used for post-processing, followed by data classification performed with support vector machines (SVM) as well as optimized artificial neural networks (ANN). The trained networks were able to distinguish non-differentiated from differentiated MSCs with a prediction accuracy of 85%. To increase versatility, an ANN was trained on 354 independent, biological replicates distributed across ten different cell lines, resulting in a prediction accuracy of up to 98% depending on data composition. The present study provides a proof of principle for the application of T1 / T2 relaxometry as a non-destructive cell classification method. It does not require labeling of cells and can perform whole mount analysis of each sample. Since all measurements can be performed under sterile conditions, it can be used as an in-process control for cellular differentiation. This distinguishes it from other characterization techniques, as most are destructive or require some type of cell labeling. These advantages highlight the technique's potential for preclinical screening of patient-specific cell-based transplants and drugs.

Safety and Effectiveness of Advanced Retrieval Techniques for Inferior Vena Cava Filters Compared with Standard Retrieval Techniques: A Systematic Review of the Literature and Meta-Analysis.

PURPOSE: To investigate the pooled safety and effectiveness of advanced retrieval techniques for inferior vena cava (IVC) filters compared with standard retrieval techniques through a systematic review of the literature and meta-analysis. MATERIALS AND METHODS: A systematic search of retrievable IVC filters between 1980 and 2020 was conducted. Studies were included if both standard and advanced retrieval techniques were utilized in the same cohort, retrieval success rates and adverse event rates were described for each technique, and advanced techniques were employed after the failure of standard techniques. Study heterogeneity was assessed by the I2 statistic. The outcomes included retrieval success rates and adverse event rates for standard and advanced retrieval techniques. RESULTS: Of 1,631 articles, 21 (1%) studies met inclusion criteria. The study heterogeneity was high with an I2 of 98%. The pooled random-effects outcomes included an overall standard retrieval success rate of 76% (95% confidence interval [CI], 65%-84%), with minor and major adverse event rates of 1% (95% CI, 0%-1%) and 1% (95% CI, 0%-1%), respectively. The overall pooled advanced retrieval success rates were 90% (95% CI, 82%-94%), with minor and major adverse event rates of 5% (95% CI, 2%-9%) and 4% (95% CI, 2%-6%), respectively. The standard retrievals were 16% less likely (risk ratio) to be successful (95% CI, 32% less likely to 4% more likely; P = .11). The major and minor adverse event rates were 88% and 84% less likely in standard retrievals compared with advanced retrievals, respectively (95% CI, 86%-94%; P < .0001; 95% CI, 70%-91%; P < .0001). CONCLUSIONS: Advanced retrieval techniques for IVC filters permit a higher retrieval success rate with low adverse event rates in cases of standard retrieval failure.

Simultaneous measurements of 3D wall shear stress and pulse wave velocity in the murine aortic arch.

PURPOSE: Wall shear stress (WSS) and pulse wave velocity (PWV) are important parameters to characterize blood flow in the vessel wall. Their quantification with flow-sensitive phase-contrast (PC) cardiovascular magnetic resonance (CMR), however, is time-consuming. Furthermore, the measurement of WSS requires high spatial resolution, whereas high temporal resolution is necessary for PWV measurements. For these reasons, PWV and WSS are challenging to measure in one CMR session, making it difficult to directly compare these parameters. By using a retrospective approach with a flexible reconstruction framework, we here aimed to simultaneously assess both PWV and WSS in the murine aortic arch from the same 4D flow measurement. METHODS: Flow was measured in the aortic arch of 18-week-old wildtype (n = 5) and ApoE-/- mice (n = 5) with a self-navigated radial 4D-PC-CMR sequence. Retrospective data analysis was used to reconstruct the same dataset either at low spatial and high temporal resolution (PWV analysis) or high spatial and low temporal resolution (WSS analysis). To assess WSS, the aortic lumen was labeled by semi-automatically segmenting the reconstruction with high spatial resolution. WSS was determined from the spatial velocity gradients at the lumen surface. For calculation of the PWV, segmentation data was interpolated along the temporal dimension. Subsequently, PWV was quantified from the through-plane flow data using the multiple-points transit-time method. Reconstructions with varying frame rates and spatial resolutions were performed to investigate the influence of spatiotemporal resolution on the PWV and WSS quantification. RESULTS: 4D flow measurements were conducted in an acquisition time of only 35 min. Increased peak flow and peak WSS values and lower errors in PWV estimation were observed in the reconstructions with high temporal resolution. Aortic PWV was significantly increased in ApoE-/- mice compared to the control group (1.7 ± 0.2 versus 2.6 ± 0.2 m/s, p < 0.001). Mean WSS magnitude values averaged over the aortic arch were (1.17 ± 0.07) N/m2 in wildtype mice and (1.27 ± 0.10) N/m2 in ApoE-/- mice. CONCLUSION: The post processing algorithm using the flexible reconstruction framework developed in this study permitted quantification of global PWV and 3D-WSS in a single acquisition. The possibility to assess both parameters in only 35 min will markedly improve the analyses and information content of in vivo measurements.

Biomimetic Mineralization Promotes Viability and Differentiation of Human Mesenchymal Stem Cells in a Perfusion Bioreactor.

In bone tissue engineering, the design of 3D systems capable of recreating composition, architecture and micromechanical environment of the native extracellular matrix (ECM) is still a challenge. While perfusion bioreactors have been proposed as potential tool to apply biomechanical stimuli, its use has been limited to a low number of biomaterials. In this work, we propose the culture of human mesenchymal stem cells (hMSC) in biomimetic mineralized recombinant collagen scaffolds with a perfusion bioreactor to simultaneously provide biochemical and biophysical cues guiding stem cell fate. The scaffolds were fabricated by mineralization of recombinant collagen in the presence of magnesium (RCP.MgAp). The organic matrix was homogeneously mineralized with apatite nanocrystals, similar in composition to those found in bone. X-Ray microtomography images revealed isotropic porous structure with optimum porosity for cell ingrowth. In fact, an optimal cell repopulation through the entire scaffolds was obtained after 1 day of dynamic seeding in the bioreactor. Remarkably, RCP.MgAp scaffolds exhibited higher cell viability and a clear trend of up-regulation of osteogenic genes than control (non-mineralized) scaffolds. Results demonstrate the potential of the combination of biomimetic mineralization of recombinant collagen in presence of magnesium and dynamic culture of hMSC as a promising strategy to closely mimic bone ECM.

Accuracy and Reproducibility of Linear and Angular Measurements in Virtual Reality: a Validation Study.

The purpose of this experimental study is to validate linear and angular measurements acquired in a virtual reality (VR) environment via a comparison with the physical measurements. The hypotheses tested are as follows: VR linear and angular measurements (1) are equivalent to the corresponding physical measurements and (2) achieve a high degree of reproducibility. Both virtual and physical measurements were performed by two raters in four different sessions. A total of 40 linear and 15 angular measurements were acquired from three physical objects (an L-block, a hand model, and a dry skull) via the use of fiducial markers on selected locations. After both intra- and inter-rater reliability were evaluated using inter-class coefficient (ICC), equivalence between virtual and physical measurements was analyzed via paired t test and Bland-Altman plots. The accuracy of the virtual measurements was further estimated using two one-sided tests (TOST) procedure. The reproducibility of virtual measurements was evaluated via ICC as well as the repeatability coefficient. Virtual reality measurements were equivalent to physical measurements as evidenced by a paired t test with p values of 0.413 for linear and 0.533 for angular measurements and Bland-Altman plots in all three objects. The accuracy of virtual measurements was estimated to be 0.5 mm for linear and 0.7° for angular measurements, respectively. Reproducibility in VR measurements was high as evidenced by ICC of 1.00 for linear and 0.99 for angular measurements, respectively. Both linear and angular measurements in the VR environment are equivalent to the physical measurements with high accuracy and reproducibility.

Vascularised human skin equivalents as a novel in vitro model of skin fibrosis and platform for testing of antifibrotic drugs.

OBJECTIVES: Fibrosis is a complex pathophysiological process involving interplay between multiple cell types. Experimental modelling of fibrosis is essential for the understanding of its pathogenesis and for testing of putative antifibrotic drugs. However, most current models employ either phylogenetically distant species or rely on human cells cultured in an artificial environment. Here we evaluated the potential of vascularised in vitro human skin equivalents as a novel model of skin fibrosis and a platform for the evaluation of antifibrotic drugs. METHODS: Skin equivalents were assembled on a three-dimensional extracellular matrix by sequential seeding of endothelial cells, fibroblasts and keratinocytes. Fibrotic transformation on exposure to transforming growth factor-ß (TGFß) and response to treatment with nintedanib as an established antifibrotic agent were evaluated by quantitative polymerase chain reaction (qPCR), capillary Western immunoassay, immunostaining and histology. RESULTS: Skin equivalents perfused at a physiological pressure formed a mature, polarised epidermis, a stratified dermis and a functional vessel system. Exposure of these models to TGFß recapitulated key features of SSc skin with activation of TGFß pathways, fibroblast to myofibroblast transition, increased release of collagen and excessive deposition of extracellular matrix. Treatment with the antifibrotic agent nintedanib ameliorated this fibrotic transformation. CONCLUSION: Our data provide evidence that vascularised skin equivalents can replicate key features of fibrotic skin and may serve as a platform for evaluation of antifibrotic drugs in a pathophysiologically relevant human setting.

Albumin-Bilirubin and Platelet-Albumin-Bilirubin Grades Accurately Predict Overall Survival in High-Risk Patients Undergoing Conventional Transarterial Chemoembolization for Hepatocellular Carcinoma.

PURPOSE: To evaluate albumin-bilirubin (ALBI) and platelet-albumin-bilirubin (PALBI) grades in predicting overall survival in high-risk patients undergoing conventional transarterial chemoembolization for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: This single-center retrospective study included 180 high-risk patients (142 men, 59 y ± 9) between April 2007 and January 2015. Patients were considered high-risk based on laboratory abnormalities before the procedure (bilirubin > 2.0 mg/dL, albumin < 3.5 mg/dL, platelet count < 60,000/mL, creatinine > 1.2 mg/dL); presence of ascites, encephalopathy, portal vein thrombus, or transjugular intrahepatic portosystemic shunt; or Model for End-Stage Liver Disease score > 15. Serum albumin, bilirubin, and platelet values were used to determine ALBI and PALBI grades. Overall survival was stratified by ALBI and PALBI grades with substratification by Child-Pugh class (CPC) and Barcelona Liver Clinic Cancer (BCLC) stage using Kaplan-Meier analysis. C-index was used to determine discriminatory ability and survival prediction accuracy. RESULTS: Median survival for 79 ALBI grade 2 patients and 101 ALBI grade 3 patients was 20.3 and 10.7 months, respectively (P < .0001). Median survival for 30 PALBI grade 2 and 144 PALBI grade 3 patients was 20.3 and 12.9 months, respectively (P = .0667). Substratification yielded distinct ALBI grade survival curves for CPC B (P = .0022, C-index 0.892), BCLC A (P = .0308, C-index 0.887), and BCLC C (P = .0287, C-index 0.839). PALBI grade demonstrated distinct survival curves for BCLC A (P = 0.0229, C-index 0.869). CPC yielded distinct survival curves for the entire cohort (P = .0019) but not when substratified by BCLC stage (all P > .05). CONCLUSIONS: ALBI and PALBI grades are accurate survival metrics in high-risk patients undergoing conventional transarterial chemoembolization for HCC. Use of these scores allows for more refined survival stratification within CPC and BCLC stage.

Nonenhanced ECG-gated quiescent-interval single shot MRA: image quality and stenosis assessment at 3 tesla compared with contrast-enhanced MRA and digital subtraction angiography.

PURPOSE: To evaluate the diagnostic accuracy of a nonenhanced electrocardiograph-gated quiescent-interval single shot MR-angiography (QISS-MRA) at 3 Tesla with contrast-enhanced MRA (CE-MRA) and digital subtraction angiography (DSA) serving as reference standard. MATERIALS AND METHODS: Following institutional review board approval, 16 consecutive patients with peripheral arterial disease underwent a combined peripheral MRA protocol consisting of a large field-of-view QISS-MRA, continuous table movement MRA, and an additional time-resolved MRA of the calves. DSA correlation was available in eight patients. Image quality and degree of stenosis was assessed. Sensitivity and specificity of QISS-MRA was evaluated with CE-MRA and DSA serving as the standards of reference and compared using the Fisher exact test. RESULTS: With the exception of the calf station, image quality with QISS-MRA was rated statistically significantly less than that of CE-MRA (P < 0.05, P = 0.17, and P = 0.6, respectively). A greater percentage of segments were not accessible with QISS-MRA (19.5-20.1%) in comparison to CE-MRA (10.9%). Relative to DSA, sensitivity for QISS-MRA was high (100% versus 91.2% for CE-MRA, P = 0.24) in the evaluated segments; however, specificity (76.5%) was substantially less than that of CE-MRA (94.6%, P = 0.003). CONCLUSION: Overall image quality and specificity of QISS-MRA at 3T are diminished relative to CE-MRA. However, when image quality is adequate, QISS-MRA has high sensitivity and, thus, has potential use in patients with contraindications to gadolinium.

Perfusable Tissue Bioprinted into a 3D-Printed Tailored Bioreactor System.

Bioprinting provides a powerful tool for regenerative medicine, as it allows tissue construction with a patient's specific geometry. However, tissue culture and maturation, commonly supported by dynamic bioreactors, are needed. We designed a workflow that creates an implant-specific bioreactor system, which is easily producible and customizable and supports cell cultivation and tissue maturation. First, a bioreactor was designed and different tissue geometries were simulated regarding shear stress and nutrient distribution to match cell culture requirements. These tissues were then directly bioprinted into the 3D-printed bioreactor. To prove the ability of cell maintenance, C2C12 cells in two bioinks were printed into the system and successfully cultured for two weeks. Next, human mesenchymal stem cells (hMSCs) were successfully differentiated toward an adipocyte lineage. As the last step of the presented strategy, we developed a prototype of an automated mobile docking station for the bioreactor. Overall, we present an open-source bioreactor system that is adaptable to a wound-specific geometry and allows cell culture and differentiation. This interdisciplinary roadmap is intended to close the gap between the lab and clinic and to integrate novel 3D-printing technologies for regenerative medicine.

A 3D bioreactor model to study osteocyte differentiation and mechanobiology under perfusion and compressive mechanical loading.

Osteocytes perceive and process mechanical stimuli in the lacuno-canalicular network in bone. As a result, they secrete signaling molecules that mediate bone formation and resorption. To date, few three-dimensional (3D) models exist to study the response of mature osteocytes to biophysical stimuli that mimic fluid shear stress and substrate strain in a mineralized, biomimetic bone-like environment. Here we established a biomimetic 3D bone model by utilizing a state-of-art perfusion bioreactor platform where immortomouse/Dmp1-GFP-derived osteoblastic IDG-SW3 cells were differentiated into mature osteocytes. We evaluated proliferation and differentiation properties of the cells on 3D microporous scaffolds of decellularized bone (dBone), poly(L-lactide-co-trimethylene carbonate) lactide (LTMC), and beta-tricalcium phosphate (ß-TCP) under physiological fluid flow conditions over 21 days. Osteocyte viability and proliferation were similar on the scaffolds with equal distribution of IDG-SW3 cells on dBone and LTMC scaffolds. After seven days, the differentiation marker alkaline phosphatase (Alpl), dentin matrix acidic phosphoprotein 1 (Dmp1), and sclerostin (Sost) were significantly upregulated in IDG-SW3 cells (p = 0.05) on LTMC scaffolds under fluid flow conditions at 1.7 ml/min, indicating rapid and efficient maturation into osteocytes. Osteocytes responded by inducing the mechanoresponsive genes FBJ osteosarcoma oncogene (Fos) and prostaglandin-endoperoxide synthase 2 (Ptgs2) under perfusion and dynamic compressive loading at 1 Hz with 5 % strain. Together, we successfully created a 3D biomimetic platform as a robust tool to evaluate osteocyte differentiation and mechanobiology in vitro while recapitulating in vivo mechanical cues such as fluid flow within the lacuno-canalicular network. STATEMENT OF SIGNIFICANCE: This study highlights the importance of creating a three-dimensional (3D) in vitro model to study osteocyte differentiation and mechanobiology, as cellular functions are limited in two-dimensional (2D) models lacking in vivo tissue organization. By using a perfusion bioreactor platform, physiological conditions of fluid flow and compressive loading were mimicked to which osteocytes are exposed in vivo. Microporous poly(L-lactide-co-trimethylene carbonate) lactide (LTMC) scaffolds in 3D are identified as a valuable tool to create a favorable environment for osteocyte differentiation and to enable mechanical stimulation of osteocytes by perfusion and compressive loading. The LTMC platform imitates the mechanical bone environment of osteocytes, allowing the analysis of the interaction with other cell types in bone under in vivo biophysical stimuli.

Fat confounds the observed apparent diffusion coefficient in patients with hepatic steatosis.

PURPOSE: Triglyceride signal contained in peaks near the water peak remains unsuppressed by conventional fat suppression techniques used in diffusion-weighted imaging. In this work, we investigated the dependence of the apparent diffusion coefficient (ADC) on liver fat content and whether it is confounded by fat signal. METHODS: 43 patients underwent liver diffusion-weighted imaging (b = 0, 500 s/mm(2)) and single-voxel MR-spectroscopy. Proton density fat-fraction (PDFF; range 0.23-34.5%) was measured from MR-spectroscopy. A theoretical model was developed to account for the effects of fat on observed ADC, and used to correct the ADC. Linear correlation analysis was performed to assess the relationship between PDFF and ADC before and after correction. RESULTS: Linear correlation analysis showed an inverse dependence between observed ADC and PDFF before correction (r(2) = 0.132; P = 0.017), and no dependence after correction (r(2) = 0.033; P = 0.24). CONCLUSION: The observed decrease in ADC in patients with fatty liver is, at least in part, artifactual due to residual fat signal near the water peak.

Impact of time-resolved MRA on diagnostic accuracy in patients with symptomatic peripheral artery disease of the calf station.

OBJECTIVE: The purpose of this article is to evaluate the added diagnostic accuracy of time-resolved MR angiography (MRA) of the calves compared with continuous-table-movement MRA in patients with symptomatic lower extremity peripheral artery disease (PAD) using digital subtraction angiography (DSA) correlation. MATERIALS AND METHODS: Eighty-four consecutive patients with symptomatic PAD underwent a low-dose 3-T MRA protocol, consisting of continuous-table-movement MRA, acquired from the diaphragm to the calves, and an additional time-resolved MRA of the calves; 0.1 mmol/kg body weight (bw) of contrast material was used (0.07 mmol/kg bw for continuous-table-movement MRA and 0.03 mmol/kg bw for time-resolved MRA). Two radiologists rated image quality on a 4-point scale and stenosis degree on a 3-point scale. An additional assessment determined the degree of venous contamination and whether time-resolved MRA improved diagnostic confidence. The accuracy of stenosis gradation with continuous-table-movement and time-resolved MRA was compared with that of DSA as a correlation. Overall diagnostic accuracy was calculated for continuous-table-movement and time-resolved MRA. RESULTS: Median image quality was rated as good for 578 vessel segments with continuous-table-movement MRA and as excellent for 565 vessel segments with time-resolved MRA. Interreader agreement was excellent (κ = 0.80-0.84). Venous contamination interfered with diagnosis in more than 60% of continuous-table-movement MRA examinations. The degree of stenosis was assessed for 340 vessel segments. The diagnostic accuracies (continuous-table-movement MRA/time-resolved MRA) combined for the readers were obtained for the tibioperoneal trunk (84%/93%), anterior tibial (69%/87%), posterior tibial (85%/91%), and peroneal (67%/81%) arteries. The addition of time-resolved MRA improved diagnostic confidence in 69% of examinations. CONCLUSION: The addition of time-resolved MRA at the calf station improves diagnostic accuracy over continuous-table-movement MRA alone in symptomatic patients with PAD.

Translation of biophysical environment in bone into dynamic cell culture under flow for bone tissue engineering.

Bone is a dynamic environment where osteocytes, osteoblasts, and mesenchymal stem/progenitor cells perceive mechanical cues and regulate bone metabolism accordingly. In particular, interstitial fluid flow in bone and bone marrow serves as a primary biophysical stimulus, which regulates the growth and fate of the cellular components of bone. The processes of mechano-sensory and -transduction towards bone formation have been well studied mainly in vivo as well as in two-dimensional (2D) dynamic cell culture platforms, which elucidated mechanically induced osteogenesis starting with anabolic responses, such as production of nitrogen oxide and prostaglandins followed by the activation of canonical Wnt signaling, upon mechanosensation. The knowledge has been now translated into regenerative medicine, particularly into the field of bone tissue engineering, where multipotent stem cells are combined with three-dimensional (3D) scaffolding biomaterials to produce transplantable constructs for bone regeneration. In the presence of 3D scaffolds, the importance of suitable dynamic cell culture platforms increases further not only to improve mass transfer inside the scaffolds but to provide appropriate biophysical cues to guide cell fate. In principle, the concept of dynamic cell culture platforms is rooted to bone mechanobiology. Therefore, this review primarily focuses on biophysical environment in bone and its translation into dynamic cell culture platforms commonly used for 2D and 3D cell expansion, including their advancement, challenges, and future perspectives. Additionally, it provides the literature review of recent empirical studies using 2D and 3D flow-based dynamic cell culture systems for bone tissue engineering.

Establishing and testing a robot-based platform to enable the automated production of nanoparticles in a flexible and modular way.

Robotic systems facilitate relatively simple human-robot interaction for non-robot experts, providing the flexibility to implement different processes. In this context, shorter process times, as well as an increased product and process quality could be achieved. Robots short time-consuming processes, take over ergonomically unfavorable tasks and work efficiently all the time. In addition, flexible production is possible while maintaining or even increasing safety. This study describes the successful development of a dual-arm robot-based modular infrastructure and the establishment of an automated process for the reproducible production of nanoparticles. As proof of concept, a manual synthesis protocol for silica nanoparticle preparation with a diameter of about 200 nm as building blocks for photonic crystals was translated into a fully automated process. All devices and components of the automated system were optimized and adapted according to the synthesis requirements. To demonstrate the benefit of the automated nanoparticle production, manual (synthesis done by lab technicians) and automated syntheses were benchmarked. To this end, different processing parameters (time of synthesis procedure, accuracy of dosage etc.) and the properties of the produced nanoparticles were compared. We demonstrate that the use of the robot not only increased the synthesis accuracy and reproducibility but reduced the personnel time and costs up to 75%.

Unique osteogenic profile of bone marrow stem cells stimulated in perfusion bioreactor is Rho-ROCK-mediated contractility dependent.

The fate determination of bone marrow mesenchymal stem/stromal cells (BMSC) is tightly regulated by mechanical cues, including fluid shear stress. Knowledge of mechanobiology in 2D culture has allowed researchers in bone tissue engineering to develop 3D dynamic culture systems with the potential for clinical translation in which the fate and growth of BMSC are mechanically controlled. However, due to the complexity of 3D dynamic cell culture compared to the 2D counterpart, the mechanisms of cell regulation in the dynamic environment remain relatively undescribed. In the present study, we analyzed the cytoskeletal modulation and osteogenic profiles of BMSC under fluid stimuli in a 3D culture condition using a perfusion bioreactor. BMSC subjected to fluid shear stress (mean 1.56 mPa) showed increased actomyosin contractility, accompanied by the upregulation of mechanoreceptors, focal adhesions, and Rho GTPase-mediated signaling molecules. Osteogenic gene expression profiling revealed that fluid shear stress promoted the expression of osteogenic markers differently from chemically induced osteogenesis. Osteogenic marker mRNA expression, type 1 collagen formation, ALP activity, and mineralization were promoted in the dynamic condition, even in the absence of chemical supplementation. The inhibition of cell contractility under flow by Rhosin chloride, Y27632, MLCK inhibitor peptide-18, or Blebbistatin revealed that actomyosin contractility was required for maintaining the proliferative status and mechanically induced osteogenic differentiation in the dynamic culture. The study highlights the cytoskeletal response and unique osteogenic profile of BMSC in this type of dynamic cell culture, stepping toward the clinical translation of mechanically stimulated BMCS for bone regeneration.

Biomimetic Connection of Transcutaneous Implants with Skin.

Bacterial infection is a crucial complication in implant restoration, in particular in permanent skin-penetrating implants. Therein, the resulting gap between transcutaneous implant and skin represents a permanent infection risk, limiting the field of application and the duration of application. To overcome this limitation, a tight physiological connection is required to achieve a biological and mechanical welding for a long-term stable closure including self-healing probabilities. This study describes a new approach, wherein the implant is connected covalently to a highly porous electrospun fleece featuring physiological dermal integration potential. The integrative potential of the scaffold is shown in vitro and confirmed in vivo, further demonstrating tissue integration by neovascularization, extracellular matrix formation, and prevention of encapsulation. To achieve a covalent connection between fleece and implant surface, self-initiated photografting and photopolymerization of hydroxyethylmethacrylate is combined with a new crosslinker (methacrylic acid coordinated titanium-oxo clusters) on proton-abstractable implant surfaces. For implant modification, the attached fleece is directed perpendicular from the implant surface into the surrounding dermal tissue. First in vitro skin implantations demonstrate the implants' dermal integration capability as well as wound closure potential on top of the fleece by epithelialization, establishing a bacteria-proof and self-healing connection of skin and transcutaneous implant.

Renal BOLD-MRI does not reflect renal function in chronic kidney disease.

Renal blood oxygen level-dependent magnetic resonance imaging (BOLD-MRI) is a noninvasive fast technique to characterize renal function. Here we evaluated the impact of renal function on the relaxation rate (R2(*)) in the cortex and medulla to provide baseline data for further use of renal BOLD-MRI. This parameter was evaluated in 400 patients scheduled for abdominal imaging who underwent transversal blood oxygen level-dependent measurements with a multi-echo gradient-echo sequence with 12 echo times. The loss of phase coherence (T2(*)) maps were generated in which kidney regions of interest were selected to differentiate the medulla and cortex, and R2(*) was equated to 1/T2(*). Individual R2(*) values were, in turn, correlated to the eGFR (MDRD formula of 280 patients with available serum creatinine measurements), age, and gender each for 1.5 and 3.0 T field-strength scans of 342 patients. At both the field strengths, no significant differences in R2(*) of the cortex and medulla were found between patient gender, age, eGFR, or between different stages of chronic kidney disease determined using the KDOQI system. Thus, BOLD-MRI of a non-specific patient population failed to discriminate between the patients with various stages of chronic kidney disease.

Healthberry 865® and a Subset of Its Single Anthocyanins Attenuate Oxidative Stress in Human Endothelial In Vitro Models.

Oxidative stress and inflammation play a pivotal role in the development of cardiovascular diseases, an ever-growing worldwide problem. As a non-pharmacological approach, diet, especially a flavonoid-rich diet, showed promising results in the reduction of cardiovascular diseases and alleviation of their symptoms. In this study, in vitro systems based on human microvascular endothelial cells (hmvEC) and human umbilical cord endothelial cells (HUVEC) were established to determine the effect of Healthberry 865® (HB) and ten of its relating single anthocyanins on oxidative stress. Furthermore, five metabolites were used in order to examine the effect of anthocyanin's most common breakdown molecules. The results showed an effect of HB in both models after 24 h, as well as most of its single anthocyanins. Cyanidin-rutinoside, peonidin-galactoside, and petunidin-glucoside had a model-specific effect. For the metabolites, phloroglucinaldeyhde (PGA) showed an effect in both models, while vanillic acid (VA) only had an effect in HUVEC. When combined, a combination of several anthocyanins did not have a cumulative effect, except for combining glucosides in hmvEC. The combination of PGA and VA even revealed an inhibitive behavior. Overall, the study demonstrates the antioxidative effect of HB and several of its single anthocyanins and metabolites, which are partially model specific, and coincides with animal studies.

Optimization and Validation of a Custom-Designed Perfusion Bioreactor for Bone Tissue Engineering: Flow Assessment and Optimal Culture Environmental Conditions.

Various perfusion bioreactor systems have been designed to improve cell culture with three-dimensional porous scaffolds, and there is some evidence that fluid force improves the osteogenic commitment of the progenitors. However, because of the unique design concept and operational configuration of each study, the experimental setups of perfusion bioreactor systems are not always compatible with other systems. To reconcile results from different systems, the thorough optimization and validation of experimental configuration are required in each system. In this study, optimal experimental conditions for a perfusion bioreactor were explored in three steps. First, an in silico modeling was performed using a scaffold geometry obtained by microCT and an expedient geometry parameterized with porosity and permeability to assess the accuracy of calculated fluid shear stress and computational time. Then, environmental factors for cell culture were optimized, including the volume of the medium, bubble suppression, and medium evaporation. Further, by combining the findings, it was possible to determine the optimal flow rate at which cell growth was supported while osteogenic differentiation was triggered. Here, we demonstrated that fluid shear stress up to 15 mPa was sufficient to induce osteogenesis, but cell growth was severely impacted by the volume of perfused medium, the presence of air bubbles, and medium evaporation, all of which are common concerns in perfusion bioreactor systems. This study emphasizes the necessity of optimization of experimental variables, which may often be underreported or overlooked, and indicates steps which can be taken to address issues common to perfusion bioreactors for bone tissue engineering.