Gregory L. Burns

Magnetic field enhancement of antibiotic activity in biofilm forming Pseudomonas aeruginosa

Device related infection initiated by biofilm bacteria are often difficult to resolve with antimicrobial therapy. Study results indicate that application of static magnetic fields may enhance the activity of gentamicin against biofilm forming Pseudomonas aeruginosa adherent to a polymer substrate. Results indicate a maximal reduction of 86.5 +/- 7.2% (n = 6) in the number of adherent viable bacteria compared with a control for samples exposed to a 5 gauss (G) magnetic field and gentamicin. The effect appears to be limited to magnetic fields between 5 and 20 G. Experiments using glass, Chronoflex (Polymedica, Golden, CO), Biomer (Ethicon, Somerville, NJ), and polystyrene substrate showed that the effect was independent of substrate surface. Autoradiograms from In111 uptake experiments showed that bacteria colonizing the substrate surface were significantly reduced in samples subjected to a magnetic field and gentamicin.

Preclinical evaluation of the abdominal aortic counterpulsation device

A valveless, single-orifice abdominal aortic counterpulsation device (AACD) was implanted retroperitoneally on the abdominal aorta and was pumped continuously, providing aortic diastolic augmentation for 45 +/- 4.9 days in four calves (group A). The hematocrit, lactate dehydrogenase (LDH), bilirubin, blood urea nitrogen, and creatinine were obtained before implantation and weekly thereafter. Biocompatibility data were compared to those obtained from 11 calves (group B) that received a total artificial heart (TAH) and were electively terminated 50.0 +/- 19.5 days after implantation. The hematocrit values in the first week were 20.9 +/- 11.5% and 39.8 +/- 11.5% below control values for group A and group B, respectively (p less than 0.02); in the sixth week they were 5.1 +/- 14.6% above control values (group A) and 22.6 +/- 9.0% below control values (group B) (p less than 0.05). LDH did not change in group A, while in group B it was constantly about 100% above control values. Autopsy revealed one to three infarcts 1 to 3 mm in diameter in 63% of the kidneys in group A, while in all of group B kidneys there were multiple infarcts of more than 10 mm. After the induction of left ventricular (LV) failure, the AACD decreased the LV end-diastolic pressure (EDP) by 21.2% (p less than 0.005) and the aortic (AO) EDP by 18% (p less than 0.005). It increased the endocardial viability ratio by 300% (p less than 0.0005), and the cardiac index by 66.9% (p less than 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)

Markers of thromboembolization in a bovine ex vivo left ventricular assist device model.

The production of blood microemboli (BME) was studied using an ex vivo exteriorized left ventricular assist device (LVAD) model in calves. Each of eight calves received a series of three LVADs, each operating for 24 hr. Blood microemboli were measured directly by a laser (624 nm and 828 nm) light scattering microemboli detection (LSMD) system through the LVAD outflow cannula and by constant pressure filtration (CPF) of blood samples from the LVAD outflow cannula. Hematologic parameters were also measured. After LVAD removal, perivalvular thrombi were evaluated using polar coordinate mapping. The average LSMD and CPF results correlated. For example, in one series of three calves, one ventricle exhibited significantly greater thrombogenesis than did the other ventricles, as indicated by both the LSMD and CPF results. In a series of five calves, one calf developed an abnormally high activated thromboplastin time (APTT), even in the absence of heparin. For two of the three ventricles tested in that calf, microemboli concentration (CPF), Factor XII activity, level of fibrin degradation products (FDP), and accumulated thrombus were significantly lower than for the other calves. The whole blood viscosity (WBV, at 230 s-1) in this calf also decreased to lower values than were seen with the other calves.

Complications common to ventricular assist device support are rare with 90 days of DeBakey VAD® support in calves

The DeBakey VAD® is a miniaturized, electromagnetically driven axial flow pump intended for long-term ventricular assist. Safety and performance data from six calves implanted with the complete DeBakey VAD® system are reported elsewhere; here we describe complications and necropsy findings for these same six animals, all of which survived 90 days. The study was conducted according to a uniform protocol, which included anticoagulation and antibiotic prophylaxis. Clinical complications tracked included bleeding, cardiovascular abnormalities (e.g., arrhythmias, tachycardia unrelated to pain, bradycardia), hemolysis, hepatic dysfunction, renal dysfunction, thromboembolism (neurologic or peripheral), or infection. Each adverse event was retrospectively categorized with regard to severity (mild, moderate, severe) and relationship to device. Clinical findings were confirmed by necropsy. There was no evidence of systemic infection, thromboembolism, hemolysis, or renal or hepatic dysfunction in these six animals during the study period. A single adverse event was noted in each of two of the calves. Both events were considered mild according to the predefined criteria. Bleeding related to the surgical implantation procedure and requiring reoperation occurred in one animal. The other animal had evidence of a superficial infection at the exit site of the cables on the left lateral thoracic wall; the infection did not extend into the thoracic cavity. Chronic, healed small renal infarct scars were present in several animals. Mild valvular endocardiosis was observed in two calves and mild fibroelastosis was present in the endocardium at the site of the inflow cannula in three calves; however, these lesions were not considered clinically significant. No other gross or histologic abnormalities were noted at necropsy. In conclusion, calves implanted with the complete DeBakey VAD® for 90 days demonstrated few complications and had no significant necropsy findings. Complications common to ventricular assist device (VAD) support (i.e., hemolysis, infection, bleeding, thromboembolism) were rare during long-term support (90 days) with the DeBakey VAD.

In vivo long-term evaluation of the Utah electrohydraulic total artificial heart.

An electrohydraulic total artificial heart (EHTAH) has been developed and evaluated by long-term in vivo studies. The EHTAH is composed of blood pumps with an interatrial shunt (IAS), an energy converter, and electronics. The EHTAH with external electronics was implanted in four calves weighing from 81-90 kg. Two animals died on the 1st and 5th post operative days, the third animal survived for 32 days, and the fourth for 159 days. The IAS was free of thrombus at autopsy in all animals. The longest surviving animal increased in size from a pre operative weight of 81 kg to 134 kg on day 144. Cardiac output ranged from 9.3 to 10.5 L/min, whereas right and left atrial pressures increased with the calfs growth from 4-10 to 16-20 mmHg and from 8-14 to 18-22 mmHg, respectively. The animal favorably tolerated up to 3.4 km/hr of treadmill exercise, both hemodynamically and metabolically. The elevation of atrial pressures during treadmill exercise was significantly alleviated by employing an automatic control mode. It is concluded that the device has the potential to be a totally implantable system for permanent use.

Quantitative bacterial analysis of porous, fabric, and smooth non-blood contacting implant surfaces and their tissue interfaces in a 169 day pneumatic total artificial heart animal recipient.

All long-term total artificial heart (TAH) survivals are subject to sepsis. Survival can be prolonged, but the source of the infection cannot be eliminated with any known course of antibiotics or treatment regimen. Sambo, a U-100 pTAH calf, survived 169 days. At week 6, he became septic, growing a Pseudomonas species (Ps). Weekly blood cultures were intermittently positive until week 13 when they became continuously positive until his demise, from a ruptured left ventricular pumping diaphragm. Spatially specific porous silicone rubber (SSP) was used for surface modifications on the drive lines and as cuffs around the Dacron TAH graft to large vessel anastomoses. This gave an excellent opportunity to examine two types of porous implants surfaces (Dacron grafts and SSP) to the smooth Biomer ventricular surfaces with their respective adjoining tissue interfaces for bacterial colonization. Nine tissue samples and 13 implant surfaces were processed with Costertons quantitative bacterial techniques. The largest numbers of bacteria (> 10(6)/cm2 Ps.) were grown from the smooth ventricular surface and in the cul-de-sac where the SSP delaminated from the driveline (two smooth implant surfaces in contact but without tissue apposition). The Dacron grafts were intermediate in bacterial concentrations and SSP surface modified drivelines and tissues were sterile. In this model, the more intimate biointegration found in the porous implants showed improved bacterial resistance in a chronically infected pTAH. The more completely biointegrated and neo-vascularized porosity SSP was the only implant surface and opposing implant tissue interface sampled to remain sterile.

Characterization of blood microemboli associated with ex vivo left ventricular assist devices in a bovine model.

An ex vivo left ventricular assist device (LVAD) model was used in calves to study the production of blood microemboli (BME) and to evaluate possible correlations between constant-pressure filtration (CPF) measurements of flow-resistant BME, light-scattering microemboli detection (LSMD), observable thrombus accumulation, and hematologic markers. Aortic LVAD cannulae were implanted through the chest wall in two calves, and each calf received a series of LVADs, each operating 1-4 days. Blood samples from the LVADs underwent CPF through 20 mu pore filters at 20 mm Hg to produce estimates of occlusive BME concentration. Laser (He-Ne) light was directed through the outflow cannula, and the differentially-scattered light was detected for computer estimation of microemboli size, volume, and frequency. Blood chemistry and coagulation parameters were also analyzed. Removed LVADs were examined with polar coordinate mapping of accumulated perivalvular thrombi. One ventricle produced significantly greater CPF results, LSMD results, and Factor XII levels. During the use of that ventricle, the CPF results increased slowly with time after LVAD connection, while the LSMD results and Factor XII levels increased immediately after connection, followed by a later decrease. This contrast is explainable in terms of the dynamic development of BME strength. The calf model appears useful for the study of thrombogenesis, and the CPF and LSMD methods of BME analysis complement each other, and other measurements.

A ventricular assist device powered by conditioned skeletal muscle

BACKGROUND We are developing and testing a new ventricular assist device (VAD) to be powered by conditioned skeletal muscle. METHODS To evaluate the VAD hardware and to develop a muscle training regimen, 8 calves have been used in studies in which the right latissimus dorsi muscle was employed. The experiments were carried out to an approximately 4-month duration. RESULTS There was significant conversion of type II (fast twitch) to type I (slow twitch) muscle fibers. This did not correlate well, however, with device performance. The device stroke volumes ranged from approximately 17 to 90 cc. This variability of outcome occurred despite the fact that identical hardware, surgical procedures, and training regimens were employed. CONCLUSIONS The results from the first eight studies lead us to speculate that perfusion may be important even when the muscle is working at pressures much lower than systemic blood pressure levels. In an attempt to augment tissue perfusion, we plan to investigate thermally induced angiogenesis as a possible mechanism for increasing blood flow to the tissue.

Apo-transferrin as a potent inhibitor of bacterial adhesion to biomaterials

Methods of inhibiting bacterial adhesion to medical implants and reducing device-associated infection are effectuated by administering an effective amount of apo-transferrin to an individual with such an implant. Preferably the apo-transferrin is administered by controlled release at or near the implant.

Immune response changes with blood pump use in calves.

Device-related bacterial infections are a significant deterrent to the long-term implantation of the total artificial heart (TAH). Device-related infections in the calf are caused by the same spectrum of organisms, over approximately the same time course, as those reported in man. We studied immunologic function in two groups of 3-4 month old, Holstein steer calves; four animals underwent cardiopulmonary bypass (CPB) only and seven calves underwent CPB and TAH implantation. There was a transient rise in total white blood cell counts and a transient fall in the hematocrit following the procedures in each group. Absolute numbers of neutrophils in the TAH group declined to near or below preoperative levels by the end of the 1 month study period. There was a transient increase in chemotactic response of isolated neutrophils to bovine C5a immediately following CPB and increased hydrogen peroxide production on postoperative day 3. Hydrogen peroxide levels were elevated in the TAH animals from postoperative day 14 to the end of the study period. Serum immunoglobulin levels of IgG, IgM, and IgA did not change significantly during the study period in the TAH group. IgG levels were elevated after week 3 in the CPB group.