Investigating the importance of left atrial compliance on fluid dynamics in a novel mock circulatory loop.

The left atrium (LA) hemodynamic indices hold prognostic value in various cardiac diseases and disorders. To understand the mechanisms of these conditions and to assess the performance of cardiac devices and interventions, in vitro models can be used to replicate the complex physiological interplay between the pulmonary veins, LA, and left ventricle. In this study, a comprehensive and adaptable in vitro model was created. The model includes a flexible LA made from silicone and allows distinct control over the systolic and diastolic functions of both the LA and left ventricle. The LA was mechanically matched with porcine LAs through expansion tests. Fluid dynamic measures were validated against the literature and pulmonary venous flows recorded on five healthy individuals using magnetic resonance flow imaging. Furthermore, the fluid dynamic measures were also used to construct LA pressure-volume loops. The in vitro pressure and flow recordings expressed a high resemblance to physiological waveforms. By decreasing the compliance of the LA, the model behaved realistically, elevating the a- and v-wave peaks of the LA pressure from 12 to 19 mmHg and 22 to 26 mmHg, respectively, while reducing the S/D ratio of the pulmonary venous flowrate from 1.5 to 0.3. This model provides a realistic platform and framework for developing and evaluating left heart procedures and interventions.

A sustainable and regioselective synthesis of Hemi-bis(monoacylglycero)phosphates and bis(diacylglycero)phosphates.

A sustainable and green approach was developed for the scalable synthesis of uncommon naturally occurring phospholipid species, Hemi-bis(monoacylglycero)phosphates (Hemi-BMPs) and bis(diacylglycero)phosphates (BDPs) via the phospholipase D (PLD) mediated transphosphatidylation. PLD from Streptomyces sp. showed great substrate promiscuity for both phospholipids from different biological sources, and alcohol donors with diverse regiochemistry; monoacylglycerols with diverse fatty acyl structures (C12-C22), affording 74-92 wt% yields in 2 h. Experimental results demonstrated that the reaction rate is rather independent of phosphatidyls but to a large extent governed by the size, shape and regiolocation of fatty acyls incorporated on the glycerol backbone, particularly for the regio-isomers of bulky diacylglycerols (Sn-1,3 or Sn-1,2), which displays great diversity. In addition, a plausible mechanism is proposed based on molecular simulations for an elaborated explanation of the reaction thermodynamic and kinetic favorability toward the synthesis of Hemi-BMPs and BDPs.

Biomechanical comparison of porcine mitral leaflets with porcine small intestinal submucosa extracellular matrix.

Porcine small intestinal submucosa extracellular matrix (SIS-ECM) used for cardiac valve repair has shown conflicting clinical outcomes with respect to calcification and failure. This may be related to differences in biomechanical properties of the material compared with the host site. The aim of this study was to compare the biomechanical properties of porcine mitral valve leaflets with SIS-ECM. Fresh porcine anterior and posterior mitral leaflet samples were cut radially and circumferentially. Similarly, 2- and 4-layered SIS-ECM were cut in orthogonal directions: length and width. Samples were subjected to a uniaxial tensile test or a dynamic mechanical analysis. Results show that the load of the porcine anterior circumferential leaflet was 39.5 N (2.4-48.5 N), which was significantly higher compared with the 2-layered length SIS-ECM which was 7.5 N (7-7.9 N), and the 4- layered length SIS-ECM which was 7.5 N (7.1-8.1 N) (p < 0.001). The load of the posterior circumferential leaflet was 9.7 N (8.3-10.7 N), which is still significantly higher when compared with the two versions of SIS-ECM. The degree of anisotropy (i.e. the ratio between circumferential-radial and width-length properties) was higher for the anterior- (ratio: 19) and posterior leaflet (ratio: 6) than the 2-layered (ratio: 5.1) and 4-layered SIS-ECM (ratio: 1.9). Especially 2-layered SIS-ECM more closely resembles the posterior mitral leaflet than the anterior mitral leaflet tissue and would be more suitable as a repair material in this position. Additionally, the anisotropic properties of mitral leaflets and SIS-ECM underscore the importance of correct orientation of the implant to ensure optimal reconstruction.

Printed dry electrode for neuromuscular electrical stimulation (NMES) for e-textile.

Muscle atrophy is a well-known consequence of immobilization and critical illness, leading to prolonged rehabilitation and increased mortality. In this study, we develop a solution to preserve muscle mass using customized biocompatible neuromuscular electrical stimulation (NMES) device. Commercially available NMES solutions with gel-based electrodes often lead to skin irritation. We demonstrate the printing of conducting electrodes on a compressive stocking textile that can be used for more than seven days without observing any inflammation. This solution consists of a dry and biocompatible electrode directly integrated into the textile with good mechanical compatibility with skin (Young's modulus of 0.39 MPa). The surface roughness of the underlying substrate plays a significant role in obtaining good print quality. Electrochemical Impedance Spectroscopy (EIS) analysis showed that the printed electrode showed better performance than the commercial ones based on a matched interfacial performance and improved series resistance. Furthermore, we investigated our NMES solution in a hospital setting to evaluate its effectiveness on muscle atrophy, with promising results.

Electrospun nanofiber mesh with connective tissue growth factor and mesenchymal stem cells for pelvic floor repair: Long-term study.

Pelvic organ prolapse (POP) affects many women, with an estimated lifetime risk of surgical intervention of 18.7%. There is a need for alternative approaches as the use of synthetic nondegradable mesh was stopped due to severe adverse events, and as current methods for pelvic floor repair have high POP recurrence rates. Thus, we hypothesized that electrospun degradable meshes with stem cells and growth factor were safe and durable for the long term in elderly rats. In an abdominal repair model, electrospun polycaprolactone (PCL) meshes coated with connective tissue growth factor (CTGF)/PEG-fibrinogen (PF) and rat mesenchymal stem cells were implanted in elderly female rats and removed after in average 53 weeks (53-week group). Collagen amount and production were quantified by qPCR and Western blotting. Moreover, histological appearance and biomechanical properties were evaluated. Results were compared with previous results of young rats with identical mesh implanted for 24 weeks (24-week group). The 53-week group differed from the 24-week group in terms of (1) reduced collagen III, (2) strong reduction in foreign body response, and (3) altered histological appearance. We found comparable biomechanical properties, aside from higher, not significant, mean tissue stiffness in the 53-week group. Lastly, we identified mesh components 53 weeks after implantation. This study provides new insights into future POP repair in postmenopausal women by showing how CTGF/PF-coated electrospun PCL meshes with stem cells exhibit sufficient support, biocompatibility, and no mesh-related complications long term in an abdominal repair model in elderly rats.

Designer phospholipids - structural retrieval, chemo-/bio- synthesis and isotopic labeling.

Phospholipids are unique and versatile molecules, essential in a variety of biological systems. Moreover, their diverse structures and amphiphilic properties endorse their indispensable and unparalleled roles in research and industrial-related applications. However, in most cases of applications, naturally occurring phospholipids are either deficit in structural variety or insufficient in quantity; therefore, novel methods must be developed for the synthesis of new molecules or modification of natural structures. To identify sustainable and environmentally friendly approaches, this work reviews the latest progress in the acquisition of structurally defined phospholipids (designer phospholipids) from natural resources, including structural retrieval, redesign and synthesis of designer phospholipids via chemo-/enzymatic approaches. This review additionally highlights the opportunity to use biological systems to direct the production of specific phospholipid species through genetic engineering via defined metabolic pathways, and functionalization of natural phospholipids through synthetic modifications: substitutions, removals or additions of specific functional groups. A particular focus is given to the establishment of chemical and biological systems for the synthesis of isotopically labelled phospholipids for biomedical applications. The application of green chemistry principles in semi-synthesis of phospholipids including extended use of greener biocatalysts and diatomaceous earth and reduced use of hazardous and toxic solvents is also summarized.

Left atrial appendage occlusion with the Amulet device: to tug or not to tug?

PURPOSE: Left atrial appendage occlusion (LAAO) involves a "tug test," in which implanters pull on the device delivery cable to ensure stable occluder placement. The aim of this study was to evaluate the recommendation to perform the tug test, by comparing forces exerted on the device during deployment and subsequent tug test. A secondary objective was to simulate forces experienced on left atrial appendage tissue by placement of a 20-mm device. METHODS: The AMPLATZER™ Amulet™ device was used for occlusion. A force transducer recorded forces in the delivery cable during deployment and tug test in 23 patients. Four patients were excluded due to improper transducer placement or technical errors in data collection. For a 20-mm device, the force imparted on the circumferential contact with left atrial appendage wall tissue was simulated in a computational model, using the measured externally applied forces as inputs. RESULTS: For devices < 25-mm in diameter, disc deployment force (mean ± standard deviation) was 1.72 ± 0.43 N, and tug force was 1.01 ± 0.59 N. For devices ≥ 25 mm in diameter, disc deployment force was 2.96 ± 0.57 N, and tug force was 1.04 ± 0.24 N. The increase in disc deployment force compared with tug test force was statistically significant for small devices (< 25 mm; p = 0.049) and large devices (≥ 25 mm; p < 0.001). CONCLUSIONS: Increased force applied on the AMPLATZER™ Amulet™ device during disc deployment compared with during tug test was statistically significant, suggesting that the tug test is redundant in most cases for checking device stability.

Effects of Dilution Systems in Olfactometry on the Recovery of Typical Livestock Odorants Determined by PTR-MS.

The present study provides an elaborate assessment of the performance of olfactometers in terms of odorant recovery for a selection of odorants emitted from livestock houses. The study includes three different olfactometer dilution systems, which have been in use at accredited odor laboratories. They consist of: (i) a custom-built olfactometer made of glass tubes, (ii) a TO8 olfactometer, and (iii) an Olfacton dilution system based on a mass flow controller. The odorants include hydrogen sulfide, methanethiol, dimethyl sulfide, acetic acid, butanoic acid, propanoic acid, 3-methylbutanoic acid, 4-methylphenol, and trimethylamine. Furthermore, n-butanol, as the reference gas in the European standard for olfactometry, EN13725, was included. All measurements were performed in real time with proton-transfer-reaction mass spectrometry (PTR-MS). The results show that only dimethyl sulfide was almost completely recovered in all cases, while for the remaining compounds, the performance was found to vary significantly (from 0 to 100%) depending on the chemical properties of the compounds, the concentration levels, the pulse duration, and the olfactometer material. To elucidate the latter, the recovery in different locations of the TO8 olfactometer and in tubes of different materials, that is, poly-tetrafluoroethylene (PTFE), perfluoroalkoxy (PFA), stainless steel and SilcoTek-coated steel, were tested. Significant saturation effects were observed when odorants were in contact with stainless steel.

Simulation of heterogeneous molecular delivery in tumours using µCT reconstructions and MRI validation.

PRIMARY OBJECTIVE: Utilizing the detailed vascular network obtained from micro-computed tomography (µCT) to establish a mathematical model of the temporal molecular distribution within a murine C3H mammary carcinoma. PROCEDURES: Female CDF1 mice with a C3H mammary carcinoma on the right rear foot were used in this study. Dynamic information for each tumour was achieved by Dynamic Contrast Enhanced-Magnetic Resonance Imaging (DCE-MRI) on a 16.4 T system. Detailed morphologic information on the tumour vasculature was obtained by ex vivo µCT and compared to CD34 immunohistochemical staining of tissue sections. The reconstructed vascular network served as origin for the diffusion (described by the apparent diffusion coefficient) within the tumour (the restricted volume described by the interstitial volume fraction derived from DCE-MRI). The resulting partial differential equation was solved using Finite-Element and a combined mathematical graph describing molecular distribution within the tumour was obtained. RESULTS: The established molecular distribution model predicted a heterogeneous distribution throughout the tumour related to the layout of the vascular network. Central tumour section concentration-time curves estimated from the established molecular distribution model were compared with physical measurements obtained by DCE-MRI of the same tumours and showed excellent correlation. CONCLUSIONS: A mathematical model describing temporal molecular distribution based on detailed vascular network structures was established and compared to DCE-MRI. The improved morphological insight will enhance future studies of heterogeneous tumours.

Three-Dimensional Polydopamine Functionalized Coiled Microfibrous Scaffolds Enhance Human Mesenchymal Stem Cells Colonization and Mild Myofibroblastic Differentiation.

Electrospinning has been widely applied for tissue engineering due to its versatility of fabricating extracellular matrix (ECM) mimicking fibrillar scaffolds. Yet there are still challenges such as that these two-dimensional (2D) tightly packed, hydrophobic fibers often hinder cell infiltration and cell-scaffold integration. In this study, polycaprolactone (PCL) was electrospun into a grounded coagulation bath collector, resulting in 3D coiled microfibers with in situ surface functionalization with hydrophilic, catecholic polydopamine (pDA). The 3D scaffolds showed biocompatibility and were well-integrated with human bone marrow derived human mesenchymal stem cells (hMSCs), with significantly higher cell penetration depth compared to that of the 2D PCL microfibers from traditional electrospinning. Further differentiation of human mesenchymal stem cells (hMSCs) into fibroblast phenotype in vitro indicates that, compared to the stiff, tightly packed, 2D scaffolds which aggravated myofibroblasts related activities, such as upregulated gene and protein expression of α-smooth muscle actin (α-SMA), 3D scaffolds induced milder myofibroblastic differentiation. The flexible 3D fibers further allowed contraction with the well-integrated, mechanically active myofibroblasts, monitored under live-cell imaging, whereas the stiff 2D scaffolds restricted that.

Ultraporous nanofeatured PCL-PEO microfibrous scaffolds enhance cell infiltration, colonization and myofibroblastic differentiation.

In the field of tissue engineering, integration of micro-porosity, nano-topogaphical features and weattability into one three-dimensional (3D) scaffold remains a challenge. The extracellular matrix (ECM) mimicking feature of electrospun fibers endows them wide applications in tissue engineering. However, the tight-packing of electrospun submicron fibers hinder cell infiltration and further colonization. In this study, we fabricated hydrophilic, micro-porous scaffolds with nano-topographical cues by one-step electrospinning, and investigated NIH3T3 fibroblasts cell infiltration, colonization and myofibroblastic differentiation. The hierarchical porosity enhanced cell infiltration and proliferation significantly. Besides, the nano-topography influenced the cell actin distribution and cell morphology that stimulated myofibroblastic differentiation in a drastically different manner from that of traditional solid, smooth electrospun fibers, which may hold great potential in reconstructing tissues that require strong contractile forces.

Surface-modified functionalized polycaprolactone scaffolds for bone repair: in vitro and in vivo experiments.

A porcine calvaria defect study was carried out to investigate the bone repair potential of three-dimensional (3D)-printed poly-ε-caprolactone (PCL) scaffolds embedded with nanoporous PCL. A microscopic grid network was created by rapid prototyping making a 3D-fused deposition model (FDM-PCL). Afterward, the FDM-PCL scaffolds were infused with a mixture of PCL, water, and 1,4-dioxane and underwent a thermal-induced phase separation (TIPS) followed by lyophilization. The TIPS process lead to a nanoporous structure shielded by the printed microstructure (NSP-PCL). Sixteen Landrace pigs were divided into two groups with 8 and 12 weeks follow-up, respectively. A total of six nonpenetrating holes were drilled in the calvaria of each animal. The size of the cylindrical defects was h 10 mm and Ø 10 mm. The defects were distributed randomly using following groups: (a) NSP-PCL scaffold, (b) FDM-PCL scaffold, (c) autograft, (d) empty defect, (a1) NSP-PCL scaffold + autologous mononuclear cells, and (a2) NSP-PCL scaffold + bone morphogenetic protein 2. Bone volume to total volume was analyzed using microcomputed tomography (µCT) and histomorphometry. The µCT and histological data showed significantly less bone formation in the NSP-PCL scaffolds in all three variations after both 8 and 12 weeks compared to all other groups. The positive autograft control had significantly higher new bone formation compared to all other groups except the FDM-PCL when analyzed using histomorphometry. The NSP-PCL scaffolds were heavily infiltrated with foreign body giant cells suggesting an inflammatory response and perhaps active resorption of the scaffold material. The unmodified FDM-PCL scaffold showed good osteoconductivity and osseointegration after both 8 and 12 weeks.

A novel nano-structured porous polycaprolactone scaffold improves hyaline cartilage repair in a rabbit model compared to a collagen type I/III scaffold: in vitro and in vivo studies.

PURPOSE: To develop a nano-structured porous polycaprolactone (NSP-PCL) scaffold and compare the articular cartilage repair potential with that of a commercially available collagen type I/III (Chondro-Gide) scaffold. METHODS: By combining rapid prototyping and thermally induced phase separation, the NSP-PCL scaffold was produced for matrix-assisted autologous chondrocyte implantation. Lyophilizing a water-dioxane-PCL solution created micro and nano-pores. In vitro: The scaffolds were seeded with rabbit chondrocytes and cultured in hypoxia for 6 days. qRT-PCR was performed using primers for sox9, aggrecan, collagen type 1 and 2. In vivo: 15 New Zealand White Rabbits received bilateral osteochondral defects in the femoral intercondylar grooves. Autologous chondrocytes were harvested 4 weeks prior to surgery. There were 3 treatment groups: (1) NSP-PCL scaffold without cells. (2) The Chondro-Gide scaffold with autologous chondrocytes and (3) NSP-PCL scaffold with autologous chondrocytes. Observation period was 13 weeks. Histological evaluation was made using the O'Driscoll score. RESULTS: In vitro: The expressions of sox9 and aggrecan were higher in the NSP-PCL scaffold, while expression of collagen 1 was lower compared to the Chondro-Gide scaffold. In vivo: Both NSP-PCL scaffolds with and without cells scored significantly higher than the Chondro-Gide scaffold when looking at the structural integrity and the surface regularity of the repair tissue. No differences were found between the NSP-PCL scaffold with and without cells. CONCLUSION: The NSP-PCL scaffold demonstrated higher in vitro expression of chondrogenic markers and had higher in vivo histological scores compared to the Chondro-Gide scaffold. The improved chondrocytic differentiation can potentially produce more hyaline cartilage during clinical cartilage repair. It appears to be a suitable cell-free implant for hyaline cartilage repair and could provide a less costly and more effective treatment option than the Chondro-Gide scaffold with cells.

Computational fluid dynamics simulation of a-v fistulas: from MRI and ultrasound scans to numeric evaluation of hemodynamics.

PURPOSE: A-v anastomosis entails dramatic changes in hemodynamic conditions, which may lead to major alterations to the vessels involved; primarily dilatations and devastating stenoses. Wall shear stress is thought to play a key role in the remodeling of the vessels exposed to abnormal levels and oscillating wall shear stress. In this study we sought to develop a framework suitable for thorough in vivo analyses of wall shear stress and vessel morphology of a-v fistulas in patients. METHODS: Using ultrasound and magnetic resonance imaging (MRI) transverse image stacks from six patient a-v fistulas were obtained. From the image stacks three-dimensional geometries of the patient fistulas were created using dedicated segmentation software. Geometries of three a-v fistulas were imported into finite element software in order to perform fluid flow simulations of blood flows and frictional forces on the vessel walls in the a-v fistulas. Boundary conditions for the simulations were obtained using both a MRI phase contrast and an ultrasound Doppler technique. RESULTS: The segmentation of the six fistulas of very different age and morphology (two end-to-side and four side-to-side) showed the ability of the approach to create geometries of various fistula morphologies. Simulations of the three fistulas showed an instant picture of the present status of the exposure to different levels of wall shear stress and the morphological status in the vessel remodeling process. CONCLUSION: The study demonstrated the capability of the CFD framework to analyze patient a-v fistulas on a regular basis using both MRI and ultrasound-based approaches.

Characterisation of internal morphologies in electrospun fibers by X-ray tomographic microscopy.

Electrospun fabrics for use in, for example, tissue engineering, wound dressings, textiles, filters and membranes have attracted a lot of attention due to their morphological nanoscale architectures which enhance their physical properties. A thorough detailed internal morphological study has been performed on electrospun polystyrene (PS) fibers produced from dimethylformamide (DMF) solutions. Investigations by transmission electron microscopy (TEM) and thorough studies for the first time by synchrotron based X-ray tomographic microscopy (XTM) revealed that the individual electrospun PS fibers and beads have a graded density and in some cases even an internal porous structure.

Longitudinal distribution of mechanical stresses in carotid plaques of symptomatic patients.

BACKGROUND AND PURPOSE: Mechanical stress may contribute to plaque rupture in patients with carotid atherosclerosis. We determined longitudinal mechanical stresses in carotid atherosclerotic plaques and compared them with known markers of plaque vulnerability. METHODS: Nineteen symptomatic patients scheduled for carotid endarterectomy underwent carotid MRI with a multicontrast protocol to characterize plaque morphology and geometry. Longitudinal 2-dimensional computational models were generated from the MRI data, and the mechanical stresses were calculated. RESULTS: Peak longitudinal mechanical stresses occurred predominantly in the shoulder regions of the carotid plaque and correlated inversely with fibrous cap thickness (r(s)=-0.61; P=0.01), and increasing degrees of stenosis (r(s)=0.71; P=0.003). Peak stress levels were asymmetrically distributed longitudinally, with 50% occurring proximal to the maximal stenosis, 25% at the point of maximal stenosis, and 25% distal to the maximal stenosis. CONCLUSIONS: The peak longitudinal mechanical stresses in the fibrous caps of symptomatic patients with carotid atherosclerotic stenosis were located at known predilection sites for plaque rupture, suggesting that mechanical stresses may play a role in plaque destabilization.

Curvature of synthetic and natural surfaces is an important target feature in classical pathway complement activation.

The binding of Abs to microbial surfaces followed by complement activation constitutes an important line of defense against infections. In this study, we have investigated the relationship between complement activation and the binding of human IgM Abs to surfaces with different curvatures. IgM Abs to dextran were shown to activate complement potently on dextran-coated particles having a diameter around 250 nm, whereas larger (600 nm) particles were less potent activators. This selectivity regarding particle dimension was also found for complement activation by colloidal substances of microbial origin. Peptidoglycan (PGN) is the major chemical component in the cell wall of Gram-positive bacteria. Fragments of purified PGN with sizes of approximately 100 nm promoted complement activation effectively through the classical pathway. By contrast, larger or smaller fragments of PGN did not activate complement strongly. A careful analysis of PGN fragments released during planctonic growth of Staphylococcus aureus showed that these include curvatures that would permit strong IgM-mediated complement activation, whereas the curvature of intact cells would be less effective for such activation. Consistently, we found that the suspended PGN fragments were strong activators of complement through the classical pathway. We suggest that these fragments act as decoy targets for complement activation, providing protection for S. aureus against the host immune response to infection.

Differences in early osteogenesis and bone micro-architecture in anterior lumbar interbody fusion with rhBMP-2, equine bone protein extract, and autograft.

PURPOSE: To investigate the microstructural differences and responsible mechanisms in early bone formation in anterior lumbar interbody fusion (ALIF) in the spine using rhBMP-2 (INFUSE), equine bone protein extract (COLLOSS E) or autograft. METHODS: Twelve Danish female landrace pigs underwent a 3-level ALIF procedure at L3-6. PEEK interbody cages packed with rhBMP-2, COLLOSS E, or autograft were inserted. The animals were divided into two groups of six, and observed for four and eight weeks postoperatively. MicroCT was performed for evaluation of microstructure of the bone within the cage. A mathematical finite element model was developed to investigate the aqueous behavior within the cages when exposed to external compressive forces. RESULTS: At 4 weeks postoperative bone surface volume fraction (BS/TV) using rhBMP-2 was higher than with use of COLLOSS E and autograft, while trabecular thickness (Tb.Th.) was lower using rhBMP-2 at this time-point. At eight weeks BS/TV and trabecular number (Tb.N.) were still higher using rhBMP-2 than autograft and COLLOSS E. Connectivity density was significantly higher using rhBMP-2 than using autograft or COLLOSS E at both time-points. Between four- and eight-week time-points BV/TV and Tb.Th. rose while Tb.N. declined using rhBMP-2. The degree of anisotropy and the calculated amount of trabeculae with main direction along the spinal axis, were higher at four weeks using COLLOSS E. rhBMP-2 had the highest amount of trabeculae directed along the spinal axis at eight weeks. A change in main direction between four and eight weeks was observed for both autograft and rhBMP-2. The numerical results from the finite element model verify that significantly different flow pattern emerges as the boundary conditions are altered. At four weeks there was an evident correlation between trabecular orientation and flow pattern using rhBMP-2. CONCLUSION: This study reveals large differences in microstructure in the early osteogenesis and explains important mechanisms of early bone formation using rhBMP-2, COLLOSS E or autograft treatment. These differences might explain some of the unfortunate events reported such as edema, swelling, and excessive bone formation using different bone graft substitutes in spinal fusion procedures.