Effect of electrode size and spacing on electrograms: Optimized electrode configuration for near-field electrogram characterization.

BACKGROUND: Detailed effects of electrode size on electrograms (EGMs) have not been systematically examined. OBJECTIVES: We aimed to elucidate the effect of electrode size on EGMs and investigate an optimal configuration of electrode size and interelectrode spacing for gap detection and far-field reduction. METHODS: This study included 8 sheep in which probes with different electrode size and interelectrode spacing were epicardially placed on healthy, fatty, and lesion tissues for measurements. Between 3 electrode sizes (0.1 mm/0.2 mm/0.5 mm) with 3 mm spacing. As indices of capability in gap detection and far-field reduction, in different electrode sizes (0.1 mm/0.2 mm/0.5 mm) and interelectrode spacing (0.1 mm/0.2 mm/0.3 mm/0.5 mm/3 mm) and the optimized electrode size and interelectrode spacing were determined. Compared between PentaRay and the optimal probe determined in study 2. RESULTS: Study 1 demonstrated that unipolar voltage and the duration of EGMs increased as the electrode size increased in any tissue (P < .001). Bipolar EGMs had the same tendency in healthy/fat tissues, but not in lesions. Study 2 showed that significantly higher gap to lesion volume ratio and healthy to fat tissue voltage ratio were provided by a smaller electrode (0.2 mm or 0.3 mm electrode) and smaller spacing (0.1 mm spacing), but 0.3 mm electrode/0.1 mm spacing provided a larger bipolar voltage (P < .05). Study 3 demonstrated that 0.3 mm electrode/0.1 mm spacing provided less deflection with more discrete EGMs (P < .0001) with longer and more reproducible AF cycle length (P < .0001) compared to PentaRay. CONCLUSION: Electrode size affects both unipolar and bipolar EGMs. Catheters with microelectrodes and very small interelectrode spacing may be superior in gap detection and far-field reduction. Importantly, this electrode configuration could dramatically reduce artifactual complex fractionated atrial electrograms and may open a new era for AF mapping.

Direction-aware mapping algorithms have minimal impact on bipolar voltage maps created using high-resolution multielectrode catheters.

INTRODUCTION: Direction-aware mapping algorithms improve the accuracy of voltage mapping by measuring the maximal voltage amplitude recorded in the direction of wavefront propagation. While beneficial for stationary catheters, its utility for roving catheters collecting electrograms (EGMs) at multiple angles is unknown. OBJECTIVE: To compare the directional dependence of bipolar voltage amplitude between stationary and roving catheters. METHODS: In 10 swine, a transcaval ablation line with a gap was created. The gap was mapped using an array catheter (Optrell™; Biosense Webster). In Step 1, the array was kept stationary over the gap, and four voltage maps were created during activation of the gap from superior, inferior, septal, and lateral directions. In Step 2, four additional maps were created; however, the catheter was allowed to move with points acquired at multiple angles. In Step 3, the gap was remapped; however, bipoles were computed using a direction-aware mapping algorithm. RESULTS: In a stationary catheter position, bipolar voltage distribution was influenced by the direction of activation with maximal differences obtained between orthogonal directions 32% (13%-53%). However, roving the catheter produced similar bipolar voltage maps irrespective of the direction of activation 11% (5%-18%). A direction-aware mapping algorithm was beneficial for reducing the directional dependence of voltage maps created by stationary catheters but not by roving catheters. CONCLUSION: The directional dependency of bipolar voltage amplitude is greatest when the catheter is stationary. However, when the catheter is allowed to rove and collect EGMs at multiple angles as occurs clinically, the directional dependence of bipolar voltage is minimal.

A novel octaray multielectrode catheter for high-resolution atrial mapping: Electrogram characterization and utility for mapping ablation gaps.

INTRODUCTION: Multielectrode mapping catheters improve the ability to map within the heterogeneous scar. A novel Octaray catheter with eight spines and 48 electrodes may further improve the speed and resolution of atrial mapping. The aims of this study were to (1) establish the Octaray's baseline mapping performance and electrogram (EGM) characteristics in healthy atria and to (2) determine its utility for identifying gaps in a swine model of atrial ablation lines. METHODS AND RESULTS: The right atria of eight healthy swine were mapped with Octaray and Pentaray catheters (Biosense Webster, Irvine, CA) before and after the creation of ablation lines with intentional gaps. Baseline mapping characteristics including EGM amplitude, duration, number of EGMs, and mapping time were compared. Postablation maps were created and EGM characteristics of continuous lines and gaps were correlated with pathology. Compared with Pentaray, the Octaray collected more EGMs per map (2178 ± 637 vs 1046 ± 238; P < 0.001) at a shorter mapping duration (3.2 ± 0.79 vs 6.9 ± 2.67 minutes; P < 0.001). In healthy atria, the Octaray recorded lower bipolar voltage amplitude (1.96 ± 1.83 mV vs 2.41 ± 1.92 mV; P < 0.001) while ablation gaps were characterized by higher voltage amplitude (1.24 ± 1.12 mV vs 1.04 ± 1.27 mV; P < 0.001). Ablation gaps were similarly identified by both catheters (P = 1.0). The frequency of "false gaps," defined as intact ablation lines with increased voltage amplitude was more common with Pentaray (6 vs 2) and resulted from erroneous annotation of far-field EGMs. CONCLUSION: The Octaray increases the mapping speed and density compared with the Pentaray catheter. It is as sensitive for identifying ablation gaps and more specific for mapping intact ablation lines.

Catheter-based endomyocardial delivery of mesenchymal precursor cells using 3D echo guidance improves cardiac function in a chronic myocardial injury ovine model.

The administration of bone marrow-derived stem cells may provide a new treatment option for patients with heart failure. Transcatheter cell injection may require multi-imaging modalities to optimize delivery. This study sought to evaluate whether endomyocardial injection of mesenchymal precursor cells (MPCs) could be guided by real-time 3D echocardiography (RT3DE) in treating chronic, postinfarction (MI) left ventricular (LV) dysfunction in sheep. Four weeks after induction of an anterior wall myocardial infarction in 39 sheep, allogeneic MPCs in doses of either 25 × 10(6) (n = 10), 75 × 10(6) (n = 9), or 225 × 10(6) (n = 10) cells or nonconditioned control media (n = 10) were administered intramyocardially into infarct and border zone areas using a catheter designed for combined fluoroscopic and RT3DE-guided injections. LV function was assessed before and after injection. Infarct dimension and vascular density were evaluated histologically. RT3DE-guided injection procedures were safe. Compared to controls, the highest dose MPC treatment led to increments in ejection fraction (3 ventricula 3% in 225M MPCs vs. -5 ± 4% in the control group, p < 0.01) and wall thickening in both infarct (4 ± 4% in 225M MPCs vs. -3 ± 6% in the control group, p = 0.02) and border zones (4 ± 6% in 225M MPCs vs. -8 ± 9% in the control group, p = 0.01). Histology analysis demonstrated significantly higher arteriole density in the infarct and border zones in the highest dose MPC-treated animals compared to the lower dose or control groups. Endomyocardial implantation of MPCs under RT3DE guidance was safe and without observed logistical obstacles. Significant increases in LV performance (ejection fraction and wall thickening) and neovascularization resulted from this technique, and so this technique has important implications for treating patients with postischemic LV dysfunction.

Transcatheter injection-induced changes in human bone marrow-derived mesenchymal stem cells.

Human mesenchymal stem cells (hMSC) are being administered by direct intramyocardial (IM) injection into patients with myocardial dysfunction with an objective to improve clinical status. However, surprisingly little attention has been directed to qualifying hMSC functionality beyond simple viability. In particular, the transit of hMSCs through a small-caliber needle lumen, the final fluidic pathway for all IM injection devices, may be especially prone to inducing unwarranted effects on cell function. This study evaluated the changes in clonogenicity, gene expression, and cytokine secretion that may be induced in hMSC (20 million/ml) by injection through a 26-gauge Nitinol needle at two different flow rates compared to noninjected control samples. Results indicated that hMSC viability and colony forming unit (CFU) formation was not altered by changes in injection rate, although a trend toward lower titers was noted at the higher flow rate, for the specific batch of hMSCs studied. The gene expression and cytokine analysis data suggest that delivering a suspension of MSCs through narrow lumen needles may marginally alter certain gene expression programs, but that such in vitro effects are transient and not translated into measurable differences in protein production. Gene expression levels of four cytokines (bFGF, SDF-1, SCF, VEGF) were significantly different at 400 microl/min, and that of all cytokines were significantly different at 1600 microl/min when compared to controls (p < 0.05). These changes were less pronounced (statistically insignificant for most cases, p > 0.05) and, in certain instances directionally opposite, at 72 h. However, no differences in the amounts of secreted bFGF, VEGF, or TGF-beta were detectable at either of the two time points or flow rates. We infer that intramyocardial administration by transcatheter techniques is unlikely to interfere with the machinery required for cell replication or secretion of regulatory and other growth factors, which are the mainstays of MSC contribution to cardiac tissue repair and regeneration.

Comparison of enzymatic and non-enzymatic means of dissociating adherent monolayers of mesenchymal stem cells.

The dissociation of adherent mesenchymal stem cell (MSC) monolayers with trypsin and enzyme-free dissociation buffer was compared. A significantly lower proportion of viable cells were obtained with enzyme-free dissociation buffers compared to trypsin. Subsequently, the dissociated cells were re-seeded on new cell culture dishes and were subjected to the MTT assay 24 h later. The proportion of viable cells that reattached was significantly lower for cells obtained by dissociation with enzyme-free dissociation buffer compared to trypsin. Frozen-thawed MSC displayed a similar trend, yielding consistently higher cell viability and reattachment rates when dissociated with trypsin compared to enzyme-free dissociation buffer. It was also demonstrated that exposure of trypsin-dissociated MSC to enzyme-free dissociation buffer for 1 h had no significant detrimental effect on cell viability.

Electrostatic binding of nanoparticles to mesenchymal stem cells via high molecular weight polyelectrolyte chains.

Combining stem cell transplantation with nanoparticle-mediated delivery of drugs and pharmaceuticals is envisioned to be one of the next major developmental steps in regenerative medicine. However, a major challenge would be to keep nanoparticles co-localized with stem cells upon transplantation or transfusion in situ. Since nanoparticles are physically much smaller in size than cells and would not specifically bind to extracellular matrix, it is easier for them to disperse from the transplantation site via the blood circulation. Conjugating nanoparticles directly to the cell membrane can potentially interfere with cellular function by physically obstructing cell surface receptors from interacting with the extracellular matrix, various growth factors and cytokines and other cells. Moreover, drug-loaded nanoparticles may be internalized into the cytoplasm via endocytosis or phagocytosis, which may wreak damage on the cellular machinery, leading to impaired physiological function or cell death. A novel solution may be to utilize high molecular weight polyelectrolyte chains to electrostatically bind nanoparticles to cells. For this purpose, hyaluronan, poly-L-lysine and chitosan are of special interest, because these molecules are generally recognized to be biocompatible for application in various pharmaceutical and surgical products. This study investigated the use of these molecules to bind nanoparticles to mesenchymal stem cells (MSCs), and a novel technique of conjugating half the cell surface with nanoparticles through the use of polyelectrolyte chains was also developed. This would avoid blocking MSC interaction with cytokines, growth factors, extracellular matrix and other cells within the recipient tissue/organ upon delivery in situ.

Temperature and calcium ions affect aggregation of mesenchymal stem cells in phosphate buffered saline.

Bone marrow-derived mesenchymal stem cells (MSC) are being extensively studied as potential therapeutic agents for various diseases and have demonstrated tremendous promise to date. To reduce immunological and inflammatory reaction upon delivery of MSC in situ, the cells are often suspended in protein-free and nutrient-poor buffered saline solution at high titers and kept on ice (0 degrees C) until completion of the transplantation procedure. This study investigated the effects of suspending MSC (5 x 10(6) cells/mL) in phosphate buffered saline (PBS) with and without calcium, over a time course of 90 and 180 min, at temperatures of 0 and 37 degrees C. The results at 0 degrees C showed a small but significant decrease in cell viability within calcium-free PBS after 180 min, whereas no significant changes in cell viability were observed with PBS containing calcium. Additionally, it was observed that significant aggregation of MSC into cellular clumps occurred when incubated in PBS at 0 degrees C, with a higher degree of aggregation occurring under calcium-free conditions. By contrast at 37 degrees C, there was a more pronounced decrease in cell viability after 90 and 180 min, but lesser aggregation of MSC both in the presence and absence of calcium. The aggregation of MSC into cellular clumps could pose an embolic hazard if delivered into the arterial vasculature in cardiac applications, can clog-up injection or infusion catheters utilized for cell delivery during surgery, and can also possibly reduce the overall efficacy of transplantation therapy.

Targeted myocardial microinjections of a biocomposite material reduces infarct expansion in pigs.

BACKGROUND: Left ventricular (LV) remodeling after myocardial infarction (MI) commonly causes infarct expansion (IE). This study sought to interrupt IE through microinjections of a biocompatible composite material into the post-MI myocardium. METHODS: MI was created in 21 pigs (coronary ligation). Radiopaque markers (2-mm diameter) were placed for IE (fluoroscopy). Pigs were randomized for microinjections (25 injections; 2- x 2-cm array; 200 microL/injection) at 7 days post-MI of a fibrin-alginate composite (Fib-Alg; fibrinogen, fibronectin, factor XIII, gelatin-grafted alginate, thrombin; n = 11) or saline (n = 10). RESULTS: At 7 days after injection (14 days post-MI), LV posterior wall thickness was higher in the Fib-Alg group than in the saline group (1.07 +/- 0.11 vs 0.69 +/- 0.07 cm, respectively, p = 0.002). At 28 days post-MI, the area within the markers (IE) increased from baseline (1 cm2) in the saline (1.71 +/- 0.13 cm2, p = 0.010) and Fib-Alg groups (1.44 +/- 0.23 cm2, p < 0.001). However, the change in IE at 21 and 28 days post-MI was reduced in the Fib-Alg group (p=0.043 and p=0.019). Total collagen content within the MI region was similar in the saline and Fib-Alg groups (12.8 +/- 1.7 and 11.6 +/- 1.5 microg/mg, respectively, p = NS). However, extractable collagen, indicative of solubility, was lower in the Fib-Alg group than the saline group (59.1 +/- 3.5 vs 71.0 +/- 6.1 microg/mL, p = 0.020). CONCLUSIONS: Targeted myocardial microinjection of the biocomposite attenuated the post-MI decrease in LV wall thickness and infarct expansion. Thus, intraoperative microinjections of biocompatible material may provide a novel approach for interrupting post-MI LV remodeling.

Astrocyte growth and glial cell line-derived neurotrophic factor secretion in three-dimensional polyethylene terephthalate fibrous matrices.

The ability of human astrocytes grown in nonwoven fibrous matrices to produce glial cell line-derived neurotrophic factor (GDNF) was studied. GDNF has the ability to selectively nourish and regenerate dopaminergic neurons and thus can provide a new treatment of Parkinson's disease. Compressed polyethylene terephthalate (PET) fabrics (porosity, 88.8%; mean pore diameter, 64 microm), treated with boiling NaOH, was effective in supporting high-density growth of astrocytes with stable GDNF production over the entire period of 18 days studied. Treatment of PET with NaOH renders the fiber surface more hydrophilic, thereby facilitating attachment and spreading of cells, whereas matrix compression allows cells to grow along and also between the fibers of these matrices to a higher density. The average production of GDNF by cells grown in these matrices (approximately 2 cm in diameter) was 21.7 pg/mL x day, with an average high concentration of 64.6 pg/mL, which is well above the effective concentration of 40 pg/mL. This work shows promise in culturing astrocytes in PET matrices as the first step in developing a potential implantable tissue-engineering device for treating patients with Parkinson's disease.

Fabrication of well-defined PLGA scaffolds using novel microembossing and carbon dioxide bonding.

A novel biologically benign technique was developed to produce three-dimensional tissue engineering scaffolds with well-defined structure. Photolithography was used to design and pattern a planar scaffold skeletal structure on a photoresist (SU-8), and a variety of microembossing processes including sacrificial layer embossing and bilayer embossing were developed to transfer the skeletal pattern to the poly(DL-lactide-co-glycolide) substrate as scaffold skeletons. Subcritical carbon dioxide was then introduced to assemble these skeletons to a three-dimensional scaffold at a low temperature. Compared with conventional scaffolds, which have a broad pore size distribution and varying pore geometry, these microfabricated scaffolds have a uniform and well-defined geometry and structure. This uniformity of structural parameters allows for the studies of cell attachment, spreading, and proliferation in scaffolds in a controlled and logical manner. The cytocompatibility of these microfabricated scaffolds was tested by seeding three different cell lines with different morphologies and growth patterns into these scaffolds. All three cell lines attached well to the scaffolds and grew to high densities as observed with scanning electron microscopy. This study demonstrates a controllable method to fabricate tissue scaffolds with a well-defined 3D architecture that can be used to better elucidate the effect of structure parameters such as pore geometry and pore size on tissue growth in 3D scaffolds.

Effect of seed culture on solid-state bioconversion of wheat straw by Phanerochaete chrysosporium for animal feed production.

The solid-state bioconversion of wheat straw by Phanerochaete chrysosporium for the production of animal feed was studied. This study was performed based on a central composite experimental design. The conditions of the seed culture most suitable for rapid induction of the ligninolytic activity of the fungus, when the seed culture is subsequently used for solid-state bioconversion of wheat straw, were determined. When the seed culture with an initial pH of 5.8 was grown under agitated conditions at 130 rpm in baffled flasks at 38 degrees C, it was predicted to give lignin degradation of 19.5% and cellulose degradation of 17.8%. A time profile study of the solid-state bioconversion of wheat straw indicated that the highest lignin and lowest cellulose degradation levels occurred on the sixth day of cultivation. The desirability coefficient for this process also passed through a maximum of 0.705 on the sixth day.