ABSTRACT
Catheter-induced thrombosis is a major contributor to infectious and mechanical complications of biomaterials that lead to device failure. Herein, a dualfunction submicron textured nitric oxide (NO)-releasing catheter was developed. The hemocompatibility and antithrombotic activity of vascular catheters were evaluated in both 20 h in vitro blood loop and 7 d in vivo rabbit model. Surface characterization assessments via atomic force microscopy show the durability of the submicron pattern after incorporation of NO donor S-nitroso-N-acetylpenicillamine (SNAP). The SNAP-doped catheters exhibited prolonged and controlled NO release mimicking the levels released by endothelium. Fabricated catheters showed cytocompatibility when evaluated against BJ human fibroblast cell lines. After 20h in vitro evaluation of catheters in a blood loop, textured-NO catheters exhibited a 13-times reduction in surface thrombus formation compared to the control catheters, which had 83% of the total area covered by clots. After the 7 d in vivo rabbit model, analysis on the catheter surface was examined via scanning electron microscopy, where significant reduction of platelet adhesion, fibrin mesh, and thrombi can be observed on the NO-releasing textured surfaces. Moreover, compared to relative controls, a 63% reduction in the degree of thrombus formation within the jugular vein was observed. Decreased levels of fibrotic tissue decomposition on the jugular vein and reduced platelet adhesion and thrombus formation on the texture of the NO-releasing catheter surface are indications of mitigated foreign body response. This study demonstrated a biocompatible and robust dual-functioning textured NO PU catheter in limiting fouling-induced complications for longer-term blood-contacting device applications. STATEMENT OF SIGNIFICANCE: Catheter-induced thrombosis is a major contributor to infectious and mechanical complications of biomaterials that lead to device failure. This study demonstrated a robust, biocompatible, dual-functioning textured nitric oxide (NO) polyurethane catheter in limiting fouling-induced complications for longer-term blood-contacting device applications. The fabricated catheters exhibited prolonged and controlled NO release that mimics endothelium levels. After the 7 d in vivo model, a significant reduction in platelet adhesion, fibrin mesh, and thrombi was observed on the NO-releasing textured catheters, along with decreased levels of fibrotic tissue decomposition on the jugular vein. Results illustrate that NO-textured catheter surface mitigates foreign body response.
Subject(s)
Catheters , Nitric Oxide , S-Nitroso-N-Acetylpenicillamine , Animals , Rabbits , Nitric Oxide/metabolism , Humans , S-Nitroso-N-Acetylpenicillamine/pharmacology , S-Nitroso-N-Acetylpenicillamine/chemistry , Thrombosis/pathology , Materials Testing , Cell Line , Platelet Adhesiveness/drug effects , Disease Models, AnimalABSTRACT
The Pennsylvania State University (PSU) Child Pump, a centrifugal continuous-flow ventricular assist device (cf-VAD), is being developed as a suitable long-term implantable device for pediatric heart failure patients between 10 and 35 kg, body surface area (BSA) of 0.5-1.2 m2, 1-11 years of age, and requiring a mean cardiac output of 1.0-3.5 L/min. In-vitro hydraulic and hemodynamic performances were evaluated on a custom mock circulatory loop with ovine blood. Normalized index of hemolysis (NIH) was evaluated under four conditions: 1) 8,300 rpm, 3.5 L/min, ΔP = 60 mm Hg, 2) 8,150 rpm, 5.1 L/min, ΔP = 20 mm Hg, 3) 8,400 rpm, 3.2 L/min, ΔP = 70 mm Hg, and 4) 9,850 rpm, 5.0 L/min, ΔP = 80 mm Hg, resulting in normalized index of hemolysis = 0.027 ± 0.013, 0.015 ± 0.006, 0.016 ± 0.008, and 0.026 ± 0.011 mg/dl, respectively. A mock fit study was conducted using a three-dimensional printed model of a 19 kg patient's thoracic cavity to compare the size of the PSU Child Pump to the HeartMate3 and the HVAD. Results indicate the PSU Child Pump will be a safer, appropriately sized device capable of providing the given patient cohort proper support while minimizing the risks of blood trauma as they wait for a transplant.
ABSTRACT
Segmented polyurethane (PU) block copolymers are widely used in implantable cardiovascular medical devices due to their good biocompatibility and excellent mechanical properties. More specifically, PU Biospan MS/0.4 was used in ventricular assist devices over the past decades. However, this product is being discontinued and it has become necessary to find an alternative PU biomaterial for application in cardiovascular devices. One important criterion for assessing cardiac biomaterials is blood compatibility. In this study, we characterized the surface properties of four medical-grade PU biomaterials: Biospan MS/0.4, BioSpan S, BioSpan 2F, and CarboSil 20 80A, including surface chemistry, topography, microphase separation structure and wettability, and then measured the blood plasma coagulation responses using bovine and human blood plasma. Results showed that BioSpan 2F contains high amounts of fluorine and has the lowest surface free energy while the other materials have surfaces with silicone present. An in vitro coagulation assay shows that these materials demonstrated improved blood coagulation responses compared to the polystyrene control and there were no significant differences in coagulation time among all PU biomaterials. The chromogenic assay showed all PU materials led to low FXII contact activation, and there were no significant differences in FXII contact activation, consistent with plasma coagulation responses.
Subject(s)
Polymers , Polyurethanes , Animals , Cattle , Humans , Polymers/chemistry , Polyurethanes/chemistry , Blood Coagulation , Biocompatible Materials/chemistry , Plasma/chemistry , Surface PropertiesABSTRACT
The loss of high molecular weight multimers (HMWM) of von Willebrand factor (vWF) in aortic stenosis (AS) and continuous-flow left ventricular assist devices (cf-LVADs) is believed to be associated with high turbulent blood shear. The objective of this study is to understand the degradation mechanism of HMWM in terms of exposure time (kinetic) and flow regime (dynamics) within clinically relevant pathophysiologic conditions. A custom high-shear rotary device capable of creating fully controlled exposure times and flows was used. The system was set so that human platelet-poor plasma flowed through at 1.75 ml/sec, 0.76 ml/sec, or 0.38 ml/sec resulting in the exposure time ( texp ) of 22, 50, or 100 ms, respectively. The flow was characterized by the Reynolds number (Re). The device was run under laminar (Re = 1,500), transitional (Re = 3,000; Re = 3,500), and turbulent (Re = 4,500) conditions at a given texp followed by multimer analysis. No degradation was observed at laminar flow at all given texp . Degradation of HMWM at a given texp increases with the Re. Re ( p < 0.0001) and texp ( p = 0.0034) are significant factors in the degradation of HMWM. Interaction between Re and texp , however, is not always significant ( p = 0.73).
Subject(s)
Heart-Assist Devices , von Willebrand Diseases , Humans , von Willebrand Factor/metabolism , Kinetics , Molecular WeightABSTRACT
BACKGROUND AND AIMS: Peripheral nerve injury is common with poor functional recovery and consequent high personal and societal costs. Sciatic nerve transection and assessment of recovery using sciatic functional index (SFI) are widely used. SFI is biologically limited as axonal misdirection of axons supplying flexors and extensors in the hindlimb, after nerve injury can lead to synkinetic innervation and function which does not correspond to the degree of axonal regeneration. METHODS: We reevaluated the use of traditional metrics such as print length (PL), toe spread (TS), and intermediate toe spread (ITS) as well as hock angle at mid-swing as approaches for determining recovery. We used two alternative approaches in discrete cohorts of rats following common peroneal crush injury, transection with repair and critical gap, using transection with ligation as a negative control. We compared walking track analysis (print) with digital capture and kinematics. RESULTS: PL, TS, and ITS varied as expected after injury. The traditional functional index for common peroneal injury using inked prints failed to describe recovery and we derived new indices to describe recovery (all R2 > 0.88, p < .0001) although pre-injury PFI was never attained by any of the models. Kinematic analysis identified hock angle at mid-swing as a useful predictor of recovery (p < .0001). INTERPRETATION: Using complementary approaches.
Subject(s)
Peripheral Nerve Injuries , Sciatic Nerve , Animals , Axons , Nerve Crush , Nerve Regeneration , Peroneal Nerve , Rats , Recovery of FunctionABSTRACT
Mechanical circulatory support for children under 6 years of age remains a challenge. This article describes the preclinical status and the results of recent animal testing with the Penn State Infant Left Ventricular Assist Device (VAD). The objectives have been to 1) demonstrate acceptably low thromboembolic risk to support Food and Drug Administration approval, 2) challenge the device by using minimal to no anticoagulation in order to identify any design or manufacturing weaknesses, and 3) improve our understanding of device thrombogenicity in the ovine animal model, using multicomponent measurements of the coagulation system and renal ischemia quantification, in order to better correlate animal results with human results.The Infant VAD was implanted as a left VAD (LVAD) in 18-29 kg lambs. Twelve LVAD and five surgical sham animals were electively terminated after approximately 30 or 60 days. Anticoagulation was by unfractionated heparin targeting thromboelastography R times of 2x normal (n = 6) or 1x normal (n = 6) resulting in negligible heparin activity as measured by anti-Xa assay (<0.1 IU/ml). Platelet inhibitors were not used.There were no clinically evident strokes or evidence of end organ dysfunction in any of the 12 electively terminated LVAD studies. The degree of renal ischemic lesions in device animals was not significantly different than that found in five surgical sham studies, demonstrating minimal device thromboembolism.In summary, these results in a challenging animal test protocol support the conclusion that the Penn State Infant VAD has a low thromboembolic risk and may allow lower levels of anticoagulation.
Subject(s)
Heart Failure/surgery , Heart-Assist Devices/adverse effects , Thromboembolism/prevention & control , Animals , Anticoagulants/therapeutic use , Blood Coagulation , Equipment Design , Female , Heart Failure/complications , Heparin , Humans , Infant , Male , Models, Animal , Sheep , Sheep, DomesticABSTRACT
Despite the prevailing use of the continuous flow left ventricular assist devices (cf-LVAD), acquired von Willebrand syndrome (AvWS) associated with cf-LVAD still remains a major complication. As AvWS is known to be dependent on shear stress (τ) and exposure time (texp ), this study examined the degradation of high molecular weight multimers (HMWM) of von Willebrand factor (vWF) in terms of τ and texp . Two custom apparatus, i.e., capillary-tubing-type degrader (CTD) and Taylor-Couette-type degrader (TCD) were developed for short-term (0.033 sec ≤ texp ≤ 1.05 s) and long-term (10 s ≤ texp ≤ 10 min) shear exposures of vWF, respectively. Flow conditions indexed by Reynolds number (Re) for CTD were 14 ≤ Re ≤ 288 with corresponding laminar stress level of 52 ≤ τ CTD ≤ 1042 dyne/cm2 . Flow conditions for TCD were 100 ≤ Re ≤ 2500 with corresponding rotor speed of 180 ≤ o ≤ 4000 RPM and laminar stress level of 50 ≤ τ TCD ≤ 1114 dyne/cm2 . Due to transitional and turbulent flows in TCD at Re > 1117, total stress (i.e., τ total = laminar + turbulent) was also calculated using a computational fluid dynamics (CFD) solver, Converge CFD (Converge Science Inc., Madison, WI, USA). Inhibition of ADAMTS13 with different concentration of EDTA (5 mM and 10 mM) was also performed to investigate the mechanism of cleavage in terms of mechanical and enzymatic aspects. Degradation of HMWM with CTD was negligible at all given testing conditions. Although no degradation of HMWM was observed with TCD at Re < 1117 ( τ total = 1012 dyne/cm2 ), increase in degradation of HMWM was observed beyond Re of 1117 for all given exposure times. At Re ~ 2500 ( τ total = 3070 dyne/cm2 ) with texp = 60 s, a severe degradation of HMWM (90.7 ± 3.8%, abnormal) was observed, and almost complete degradation of HMWM (96.1 ± 1.9%, abnormal) was observed with texp = 600 s. The inhibition studies with 5 mM EDTA at Re ~ 2500 showed that loss of HMWM was negligible (<10%, normal) for all given exposure times except for texp = 10 min (39.5 ± 22.3%, borderline-abnormal). With 10 mM EDTA, no degradation of HMWM was observed (11.1 ± 4.4%, normal) even for texp = 10 min. This study investigated the effect of shear stress and exposure time on the HMWM of vWF in laminar and turbulent flows. The inhibition study by EDTA confirms that degradation of HMWM is initiated by shear-induced unfolding followed by enzymatic cleavage at given conditions. Determination of magnitude of each mechanism needs further investigation. It is also important to note that the degradation of vWF is highly dependent on turbulence regardless of the time exposed within our testing conditions.
Subject(s)
Heart-Assist Devices/adverse effects , von Willebrand Diseases/etiology , von Willebrand Factor/metabolism , Hemodynamics/physiology , Humans , Materials Testing , von Willebrand Diseases/bloodABSTRACT
Peripheral nerve possesses the inherent ability to regrow and recover following injury. However, nerve regeneration is often slow and incomplete due to limitations associated with the local microenvironment during the repair process. Manipulation of the local microenvironment at the site of nerve repair, therefore, represents a significant opportunity for improvement in downstream outcomes. Macrophages and Schwann cells play a key role in the orchestration of early events after peripheral nerve injury. We describe the production, characterization, and use of an injectable, peripheral nerve-specific extracellular matrix-based hydrogel (PNSECM) for promoting modulation of the local macrophage and Schwann cell responses at the site of nerve repair in a rodent model of sciatic nerve injury. We show that PNSECM hydrogels largely maintain the matrix structure associated with normal native peripheral nerve tissue. PNSECM hydrogels were also found to promote increased macrophage invasion, higher percentages of M2 macrophages and enhanced Schwann cell migration when used as a lumen filler in a rodent model of nerve gap repair using an inert nerve guidance conduit. These results suggest that an injectable PNSECM hydrogel can provide a supportive, bioactive scaffold which promotes repair of peripheral nerve in vivo. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 450-459, 2018.
Subject(s)
Extracellular Matrix/metabolism , Hydrogels/pharmacology , Nerve Regeneration/drug effects , Peripheral Nerve Injuries/physiopathology , Recovery of Function , Sciatic Nerve/metabolism , Animals , Cell Movement/drug effects , Dogs , Extracellular Matrix/ultrastructure , Female , Hindlimb/physiopathology , Macrophages/drug effects , Macrophages/metabolism , Organ Specificity , Peripheral Nerve Injuries/pathology , Rats, Sprague-Dawley , Recovery of Function/drug effects , Schwann Cells/drug effects , Schwann Cells/pathology , Sciatic Nerve/drug effectsABSTRACT
Reynolds shear stress (RSS) has served as a metric for the effect of turbulence on hemolysis. Forstrom (1969) and Sallam and Hwang (1984) determined the RSS threshold for hemolysis to be 50,000 and 4,000 dyne/cm, respectively, using a turbulent jet. Despite the order of magnitude discrepancy, the threshold by Sallam and Hwang has been frequently cited for hemolytic potential in blood pumps. We recreated a Sallam apparatus (SA) to resolve this discrepancy and provide additional data to be used in developing a more accurate hemolysis model. Hemolysis was measured over a large range of Reynolds numbers (Re) (Re = 1,000-80,000). Washed bovine red blood cells (RBCs) were injected into the free jet of phosphate buffered saline, and hemolysis was quantified using a percent hemolysis, Hp = h (100 - hematocrit [HCT])/Hb, where h (mg/dl) is free hemoglobin and Hb (mg/dl) is total hemoglobin. Reynolds shear stress was calculated using two-dimensional laser Doppler velocimetry. Reynolds shear stress of ≥30,000 dyne/cm corresponding to Re of ≥60,000 appeared to cause hemolysis (p < 0.05). This RSS is an order of magnitude greater than the RSS threshold that Sallam and Hwang suggested, and it is similar to Forstrom's RSS threshold. This study resolved a long-standing uncertainty regarding the critical values of RSS for hemolysis and may provide a foundation for a more accurate hemolysis model.
Subject(s)
Erythrocytes/cytology , Hematologic Tests/methods , Hemolysis/physiology , Stress, Mechanical , Animals , Blood Flow Velocity , Cattle , Hematocrit , Hemoglobins , Humans , Laser-Doppler Flowmetry/methodsABSTRACT
The Penn State Infant Ventricular Assist Device (VAD) is a 12-14 ml stroke volume pneumatically actuated pump, with custom Björk-Shiley monostrut valves, developed under the National Heart, Lung, and Blood Institute Pediatric Circulatory Support program. In this report, we describe the seven most recent chronic animal studies of the Infant VAD in the juvenile ovine model, with a mean body weight of 23.5 ± 4.1 kg. The goal of 4-6 weeks survival was achieved in five of seven studies, with support duration ranging from 5 to 41 days; mean 26.1 days. Anticoagulation was accomplished using unfractionated heparin, and study animals were divided into two protocol groups: the first based on a target activated partial thromboplastin time of 1.5-2 times normal, and a second group using a target thromboelastography R-time of two times normal. The second group required significantly less heparin, which was verified by barely detectable heparin activity (anti-Xa). In both groups, there was no evidence of thromboembolism except in one animal with a chronic infection and fever. Device thrombi were minimal and were further reduced by introduction of the custom valve. These results are consistent with results of adult VAD testing in animals and are encouraging given the extremely low levels of anticoagulation in the second group.