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Diseases of small diameter blood vessels encompass the largest portion of cardiovascular diseases, with over 4.2 million people undergoing autologous vascular grafting every year. However, approximately one third of patients are ineligible for autologous vascular grafting due to lack of suitable donor vasculature. Acellular extracellular matrix (ECM) scaffolds derived from xenogeneic vascular tissue have potential to serve as ideal biomaterials for production of off-the-shelf vascular grafts capable of eliminating the need for autologous vessel harvest. A modified antigen removal (AR) tissue process, employing aminosulfabetaine-16 (ASB-16) was used to create off-the-shelf small diameter (< 3 mm) vascular graft from bovine saphenous vein ECM scaffolds with significantly reduced antigenic content, while retaining native vascular ECM protein structure and function. Elimination of native tissue antigen content conferred graft-specific adaptive immune avoidance, while retention of native ECM protein macromolecular structure resulted in pro-regenerative cellular infiltration, ECM turnover and innate immune self-recognition in a rabbit subpannicular model. Finally, retention of the delicate vascular basement membrane protein integrity conferred endothelial cell repopulation and 100% patency rate in a rabbit jugular interposition model, comparable only to Autograft implants. Alternatively, the lack of these important basement membrane proteins in otherwise identical scaffolds yielded a patency rate of only 20%. We conclude that acellular antigen removed bovine saphenous vein ECM scaffolds have potential to serve as ideal off-the-shelf small diameter vascular scaffolds with high in vivo patency rates due to their low antigen content, retained native tissue basement membrane integrity and preserved native ECM structure, composition and functional properties. The use of autologous vessels for the treatment of small diameter vascular diseases is common practice. However, the use of autologous tissue poses significant complications due to tissue harvest and limited availability. Developing an alternative vessel for use for the treatment of small diameter vessel diseases can potentially increase the success rate of autologous vascular grafting by eliminating complications related to the use of autologous vessel and increased availability. This manuscript demonstrates the potential of non-antigenic extracellular matrix (ECM) scaffolds derived from xenogeneic vascular tissue as off-the-shelf vascular grafts for the treatment of small diameter vascular diseases. [Display omitted] [ABSTRACT FROM AUTHOR]

Chronic venous disease (CVD) is the most common reported chronic condition in the United States, affecting more than 25 million Americans. Regardless of its high occurrence, current therapeutic options are far from ideal due to their palliative nature. For best treatment outcomes, challenging cases of chronic venous insufficiency (CVI) are treated by repair or replacement of venous valves. Regrettably, the success of venous valve transplant is dependent on the availability of autologous venous valves and hindered by the possibility of donor site complications and increased patient morbidity. Therefore, the use of alternative tissue sources to provide off-the-shelf venous valve replacements has potential to be extremely beneficial to the field of CVI. This manuscript demonstrates the capability of producing off-the-shelf fully functional venous valved extracellular matrix (ECM) scaffold conduits from bovine saphenous vein (SV), using an antigen removal (AR) method. AR ECM scaffolds maintained native SV structure-function relationships and associated venous valves function. Conversely, SDS decellularization caused significant changes to the collagen and elastin macromolecular structures, resulting in collagen fibril merging, elimination of fibril crimp, amalgaming collagen fibers and fragmentation of the inner elastic lamina. ECM changes induced by SDS decellularization resulted in significant venous valve dysfunction. Venous valved conduits generated using the AR approach have potential to serve as off-the-shelf venous valve replacements for CVI. Retention of the structure and composition of extracellular matrix (ECM) proteins within xenogeneic scaffolds for tissue engineering is of crucial importance, due to the undeniable effect ECM proteins can impose on repopulating cells and function of the resultant biomaterial. This manuscript demonstrates that alteration or elimination of ECM proteins via commonly utilized decellularization approach results in complete disruption of venous valve function. Conversely, retention of the delicate ECM structure and composition of native venous tissue, using an antigen removal tissue processing method, results in preservation of native venous valve function. Image, graphical abstract [ABSTRACT FROM AUTHOR]

Graphical abstract Abstract Antigenicity remains the primary barrier towards expanding the use of unfixed xenogeneic biomaterials in clinical applications. An unfixed xenogeneic biomaterial devoid of antigenicity, with maintained structural and mechanical integrity, has potential to overcome the limitations of current clinically utilized glutaraldehyde-fixed xenogeneic biomaterials, such as heart valve bioprostheses. Unfortunately, the threshold level of residual antigenicity necessary to overcome graft-specific immune responses in unfixed xenogeneic tissue has yet to be determined. Furthermore, little information is known regarding the extent to which in vitro disruption of native ECM properties, resulting from decellularization or antigen removal procedures, are tolerated following in vivo implantation. This manuscript demonstrates that humoral adaptive immune responses are more sensitive to residual xenogeneic biomaterial antigen content than are cell-mediated adaptive responses. Critically, the threshold for tolerable residual antigenicity is identified, with removal of ≥92% of lipophilic antigens required to reduce adaptive immune responses to levels equivalent to glutaraldehyde fixed tissue. Finally, the results demonstrated that the innate immune system tolerates minor changes in protein organization provided that molecular structure is maintained. Antigen removed xenogeneic biomaterials achieving these in vitro success criteria induce in vivo adaptive and innate tolerance, while modulating pro-regenerative constructive remodeling. Statement of Significance Removal of antigenic components from candidate xenogeneic biomaterials is the primary success criteria for development of extracellular matrix (ECM) scaffolds in tissue engineering applications. Currently, the threshold level of residual biomaterial antigenicity required to overcome recipient graft-specific adaptive immune responses is unknown. Additionally, the extent to which the innate immune response tolerates changes to the native ECM, resulting from the ECM scaffold production process, has yet to be determined. This manuscript not only establishes the threshold for tolerable residual antigenicity, but also demonstrates that deviations in protein organization are tolerated by the innate immune system, provided macromolecular structure remains intact. In doing so, we provide the foundation of an immunologically-acceptable unfixed xenogeneic biomaterial for use in clinical applications. [ABSTRACT FROM AUTHOR]

Cardiac extracellular matrix (cECM) scaffolds are promising biomaterials for reconstructive surgery applications since they possess the structure/function properties of native tissue. Production of cECM scaffolds has been achieved using decellularization approaches, which commonly employ denaturing detergents, such as sodium dodecyl sulfate (SDS). Our antigen removal (AR) method has been shown to remove cellular and nonmyocyte components, while preserving cECM scaffold structure/function relationships. Here, we demonstrate that more human mesenchymal stem cells (MSCs) invaded AR scaffolds compared to SDS controls. Additionally, AR scaffolds stimulated a constructive remodeling response similar to allograft controls, and were transformed to adipose tissue in a xenogeneic rat to mouse subpannicular in vivo model. Conversely, SDS scaffolds showed a chronic inflammatory response that worsened throughout the 12-wk time course preventing constructive remodeling and mirroring the response seen towards xenogeneic tissue. AR scaffolds and xenogeneic controls recellularized with murine MSCs (mMSCs) were also implanted to assess whether mMSCs would offer any additive benefit in overcoming residual scaffold-specific immune responses. Paradoxically, recellularization resulted in chronic inflammatory response in AR-recellularized scaffolds. We conclude that AR cECM scaffolds represent a promising biomaterial, which is accepted by the recipient as self in origin and fosters implantation site appropriate regenerative responses. Statement of Significance We demonstrated that an antigen-removal (AR) approach utilizing principles of differential solubility for production of a xenogeneic rat cardiac extracellular matrix scaffold results in improved recellularization efficiency with human and mouse mesenchymal stem cells (MSCs) in vitro . Furthermore, we tested the immune response to AR scaffolds versus allograft and xenograft controls with or without MSC recellularization using a rat to mouse subcutaneous model. We showed that AR scaffolds and allograft controls resulted in significant adipose tissue transformation after 12 weeks. Paradoxically, MSCs had a positive impact in the immune response to xenografts, but had the opposite effect in AR scaffolds, resulting in chronic inflammatory response, which might be attributed to a change of their phenotype following recellularization into scaffolds. [ABSTRACT FROM AUTHOR]

Objective To determine if pimobendan, a phosphodiesterase III inhibitor and calcium sensitizer with positive survival benefits, has an effect on incidence of arrhythmias compared to placebo in small breed dogs with congestive heart failure (CHF) due to myxomatous mitral valve degeneration (MMVD). Animals Eight client-owned small breed dogs (<15 kg) with CHF due to MMVD. Methods A prospective double-blind randomized placebo-controlled crossover study design was used. Data were recorded at baseline and 2 weeks post-administration of placebo or pimobendan. Average heart rate and incidence of arrhythmia were determined from 24 h Holter analysis. Owners completed a quality of life (QOL) questionnaire at each time point and recorded sleeping respiratory rates (SRR). Mixed effects analysis of variance, with dog as the random variable was used to compare values obtained between baseline, placebo, and pimobendan. Results Compared to baseline, QOL scores were significantly improved following administration of either placebo or pimobendan ( p = 0.021 and p < 0.001, respectively). No significant differences in type or incidence of supraventricular or ventricular arrhythmia were identified. Average heart rate with pimobendan was significantly lower than baseline ( p < 0.001). Compared to baseline, SRR was significantly lower with pimobendan ( p = 0.004), and significantly different from placebo ( p = 0.045). Conclusions No significant difference between pimobendan and placebo was found on incidence of supraventricular or ventricular arrhythmia. The decrease in average heart rate and SRR may be reflective of superior heart failure control achieved with pimobendan therapy. [ABSTRACT FROM AUTHOR]

Abstract: Decades of research have been undertaken towards the goal of tissue engineering using xenogeneic scaffolds. The primary advantages associated with use of xenogeneic tissue-derived scaffolds for in vitro development of replacement tissues and organs stem from the inherent extracellular matrix (ECM) composition and architecture. Native ECM possesses appropriate mechanical properties for physiological function of the biomaterial and signals for cell binding, growth and differentiation. Additionally, xenogeneic tissue is readily available. However, translation of xenogeneic scaffold-derived engineered tissues or organs into clinical therapies requires xenoantigenicity of the material to be adequately addressed prior to implantation. Failure to achieve this goal will result in a graft-specific host immune rejection response, jeopardizing in vivo survival of the resultant scaffold, tissue or organ. This review explores (i) the appropriateness of scaffold acellularity as an outcome measure for assessing reduction of the immunological barriers to the use of xenogeneic scaffolds for tissue engineering applications and (ii) the need for tissue engineers to strive for antigen removal during xenogeneic scaffold generation. [Copyright &y& Elsevier]

Abstract: The ability of residual antigens on decellularized tissue to elicit the immune response upon implantation motivates development of a more rigorous antigen removal (AR) process for xenogeneic scaffold generation. Antigen removal strategies promoting solubilization of hydrophilic proteins (predominantly cytoplasmic) enhance the reduction of hydrophilic antigenicity in bovine pericardium (BP); however, the diversity of protein antigens within a tissue necessitates development of AR strategies capable of addressing a spectrum of protein antigen solubilities. In the present study, methods for promoting solubilization of lipophilic proteins (predominantly membrane) were investigated for their ability to reduce lipophilic antigenicity of BP when applied as a second AR step following our previously described hydrophilic AR method. Bovine pericardium following AR (BP-AR) was assessed for residual hydrophilic and lipophilic antigenicity, removal of known lipophilic xenoantigens, tensile properties, and extracellular matrix structure and composition. Facilitating hydrophile solubilization (using dithiothreitol and potassium chloride) followed by lipophile solubilization (using amidosulfobetaine-14 (ASB-14)), in a two-step sequential, differential AR strategy, significantly reduces residual hydrophilic and lipophilic antigenicity of BP-AR beyond that achieved with either one-step hydrophilic AR or decellularization using 1% (w/v) sodium dodecyl sulfate. Moreover, use of 1% (w/v) ASB-14 for lipophilic AR eliminates the two most critical known barriers to xenotransplantation (galactose-α(1,3)-galactose and major histocompatibility complex I)) from BP-AR without compromising the structure–function properties of the biomaterial. This study demonstrates the importance of a sequential, differential protein solubilization approach to reduce biomaterial antigenicity in the production of a xenogeneic scaffold for heart valve tissue engineering. [Copyright &y& Elsevier]

Summary: Background: The DIEAP flap has gained popularity for breast reconstruction worldwide. Despite DIEAP flap arterial anatomy being well known, venous congestion is still an important complication and the literature on DIEAP flap venous anatomy is lacking. Venous drainage is less predictable and research on venous perforators is of increasing interest. The aim of this study was to investigate the anatomical distribution, diameters and relationships between arterial and venous perforators and their source vessels using Duplex ultrasonography, in order to evaluate the potential benefits of pre-operative evaluation of DIEAP flap venous perforators. Methods: Prospectively, 140 patients undergoing DIEAP flap breast reconstruction, were pre-operatively assessed using Duplex ultrasonography for location and diameter of arterial and venous perforators, DIEA, DIEV, and SIEV. Perforators were plotted laterally and inferiorly from the umbilicus. Means were compared using t-tests. Pearson''s correlation coefficients were calculated. Results: We identified 702 arterial and 355 venous perforators in 280 lower hemi-abdomens. No venous perforators were identified in 9 (6.5%) patients; none on the right in 25 (17.9%) and none on the left in 36 (25.7%). Venous perforators were larger on the right (p = 0.031) but DIEV and SIEV diameters were not statistically different on either sides. The diameters of DIEA, DIEV, perforating arteries and veins were correlated, but unrelated to the diameter of the SIEV. When a medium/large perforator vein is identified first, there is a 93.5% chance of finding an associated medium/large perforator artery; this reduces to 69.8% when a medium/large perforator artery is first identified. Conclusions: There is no correlation between the sizes of perforator veins and DIEV, and the size of the SIEV. Our data suggests that first identifying a medium/large venous perforator increases the chances of finding a better suitable perforator complex. Pre-operative evaluation of venous perforators may be of great interest for its potential clinical benefits. [Copyright &y& Elsevier]

Abstract: Aortopulmonary window is a rare congenital anomaly, resulting from incomplete septation of the truncus arteriosus into the aorta and the pulmonary artery during embryogenesis. The utility of three-dimensional (3D) echocardiogram has not been previously reported in dogs with aortopulmonary windows. The authors demonstrate the diagnostic potentials of 3D echocardiogram and its ability to more completely define the anatomical features of aortopulmonary window in a dog when compared to two-dimensional imaging. Ultimately 3D echocardiogram was considered to provide information critical to determining the feasibility of beating heart surgical correction in this case. [Copyright &y& Elsevier]

Abstract: Objective: To describe the surgical technique and report outcome of dogs undergoing bioprosthesis valve replacement for severe tricuspid regurgitation (TR) secondary to congenital tricuspid valve dysplasia (TVD). Animals, materials and methods: Twelve client-owned dogs (19–43 kg) with TVD underwent tricuspid valve replacement with a bovine pericardial or porcine aortic bioprosthesis with the aid of cardiopulmonary bypass. Anticoagulation with warfarin was maintained for 3 months after surgery and then discontinued. Results: Ten of 12 (83.3%) dogs survived surgery and were discharged from the hospital. Seven dogs were alive with complete resolution of TR for a median period of 48 months (range 1–66 months) after surgery. Two dogs underwent euthanasia because of bioprosthesis failure due to inflammatory pannus at 10 and 13 months after surgery. Two dogs experienced valve thrombosis that was resolved by tissue plasminogen activator. One dog developed suspected endocarditis after surgery that was resolved with antibiotics. Serious cardiac complications included atrial fibrillation and flutter, right-to-left shunt through an uncorrected patent foramen ovale, complete atrioventricular block, and sudden cardiac arrest. Postoperative atrial fibrillation or flutter did not occur in 7 dogs treated prophylactically with oral amiodarone before surgery. Conclusions: Curative intermediate-term outcomes are possible in dogs undergoing open tricuspid valve replacement with a bioprosthesis. Prosthesis-related complications include inflammatory pannus, thrombosis, and endocarditis. Postoperative atrial fibrillation or flutter can be reduced or prevented by prophylactic preoperative treatment with amiodarone. Several identified complications are avoidable or can be reduced with increased awareness and experience with these techniques. [Copyright &y& Elsevier]

Abstract: Objectives: The aims of this prospective study are to (1) generate normal ranges for interventricular and intraventricular mechanical synchrony in dogs, and (2) generate normal ranges for tissue Doppler imaging (TDI) velocity imaging and speckle tracking strain imaging assessment of segmental intraventricular mechanical synchrony in dogs. Animals: 10 prospectively recruited healthy dogs. Methods: Dogs were excluded if they had abnormal historical, physical examination, echocardiographic, ECG or systolic blood pressure findings. Interventricular mechanical synchrony was assessed using time difference between left and right ventricular pre-ejection periods. Intraventricular mechanical synchrony was assessed using both M-mode and color M-mode septal to posterior wall mechanical delay (SPWMD). Intraventricular segmental mechanical synchrony was assessed using both color TDI and speckle tracking strain analysis of segmental myocardial motion during systole and diastole. Results: All synchrony measures were found to be independent of age or body weight. Normal range for mechanical interventricular synchrony was found to be −10.2 to 12.6 ms. Assessment of mechanical intraventricular synchrony using either M-mode or color M-mode SPWMD was found to be associated with an extremely wide normal range, limiting clinical applicability. Normal ranges for segmental intraventricular mechanical synchrony assessed using either color TDI or speckle tracking were found to be comparable to those published for human subjects. Conclusions: Interventricular and intraventricular mechanical synchrony in dogs is independent of age and body weight. The normal ranges identified in this study form a basis for assessment of normal versus abnormal mechanical synchrony in canine cardiovascular disease patients. [Copyright &y& Elsevier]

Abstract: Objective: Document safety and efficacy of transvenous catheter occlusion of patent ductus arteriosus (PDA) over a wide range of ductal and patient sizes. Animals, materials and methods: Retrospective study of fifty-six consecutive dogs referred to Colorado State University with diagnosis of PDA. All cases utilized the transvenous approach, via the femoral vein. Occlusion was achieved using a coil (Flipper™) in dogs with PDA minimal dimension of ≤4 mm. Amplatzer® duct occluder or Amplatzer® vascular plugs were used in dogs with PDA minimal dimension >4 mm, or if a coil was unstable following deployment. Results: Ductal occlusion was achieved using the transvenous route alone in 54/56 dogs (96.4%). Occlusion was achieved using detachable Flipper™ coils in 39/42 dogs (92.9%) in which coil occlusion was attempted, with 38/39 dogs (97.4%) requiring only a single coil. In 16 dogs, occlusion was achieved using an Amplatzer® duct occluder or vascular plug. Post-procedure residual ductal flow was absent or only mild in 36/39 (92.3%) dogs in which it was assessed. Procedural mortality rate was 1.7%, and major complication rate 7.0%. Conclusion: Transcatheter occlusion via a transvenous approach is a safe and effective method for treating PDA in dogs and is useful for small dogs (<2.5 kg). [ABSTRACT FROM AUTHOR]

Abstract: Strong westerly winds that emanate from San Gorgonio Pass, the lowest point between Palm Springs and Los Angeles, California, dominate aeolian transport in the Coachella Valley of the western Sonoran Desert. These winds deposit sand in coppice dunes that are critical habitat for several species, including the state and federally listed threatened species Uma inornata, a lizard. Although wind directions are generally defined in this valley, the wind field has complex interactions with local topography and becomes more variable with distance from the pass. Local, dominant wind directions are preserved by growth patterns of Larrea tridentata (creosote bush), a shrub characteristic of the hot North American deserts, and ventifacts. Exceptionally long-lived, Larrea has the potential to preserve wind direction over centuries to millennia, shaped by the abrasive pruning of windward branches and the persistent training of leeward branches. Wind direction preserved in Larrea individuals and clones was mapped at 192 locations. Compared with wind data from three weather stations, Larrea vectors effectively reflect annual prevailing winds. Ventifacts measured at 24 locations record winds 10° more westerly than Larrea and appear to reflect the direction of the most erosive winds. Based on detailed mapping of local wind directions as preserved in Larrea, only the northern half of the Mission-Morongo Creek floodplain is likely to supply sand to protected U. inornata habitat in the Willow Hole ecological reserve. [Copyright &y& Elsevier]

Abstract: Valvular pulmonic stenosis has been successfully treated in dogs via minimally invasive transcatheter balloon dilation procedures for almost 20years. Supravalvular pulmonic stenosis however is not amenable to balloon dilation procedures. Although several surgical procedures are described for treatment of pulmonic stenosis, the morbidity and mortality of these procedures are significantly higher than for transcatheter approaches. Two cases of supravalvular pulmonic stenosis are presented, one acquired and one congenital, which were successfully treated with transcatheter intravascular stent placement. The procedure alleviated the stenotic lesion and resolved clinical signs. [Copyright &y& Elsevier]