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2.
Clin Hemorheol Microcirc ; 71(4): 451-461, 2019.
Article in English | MEDLINE | ID: mdl-30248048

ABSTRACT

Endovascular repair of aortic aneurysms (EVAR) has become an established treatment option currently applied in an increasing numbers of patients with aortic aneurysms. Advantages include reduced surgical trauma, procedural time, intensive care unit and hospital lengths of stay, blood loss as well as morbidity and mortality.The optimal imaging modalities in EVAR follow-up as well as the appropriate intervals between these follow-ups remain subject of controversial discussion. Objective of this study was the evaluation of the realistic radiation exposure and risk estimate postop EVAR treatment.Of the follow-ups required according to the surveillance schedule during the first year post-EVAR, only 68.3% were actually implemented. Of those required from the second year onwards, an average of 70% was actually performed. During the observation period, each patient underwent a mean of 4.3 CTAs. The median ED calculated from all CTAs was 24. 5 mSv. The minimum and maximum cumulative EDs for the entire observation period were 55 mSv and 310 mSv, respectively.


Subject(s)
Aortic Aneurysm, Abdominal/radiotherapy , Radiation Exposure/adverse effects , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Risk Factors , Time Factors , Treatment Outcome
3.
Int J Vasc Med ; 2017: 9763075, 2017.
Article in English | MEDLINE | ID: mdl-29250445

ABSTRACT

Patients who undergo endovascular repair of aortic aneurysms (EVAR) require life-long surveillance because complications including, in particular, endoleaks, aneurysm rupture, and graft dislocation are diagnosed in a certain share of the patient population and may occur at any time after the original procedure. Radiation exposure in patients undergoing EVAR and post-EVAR surveillance has been investigated by previous authors. Arriving at realistic exposure data is essential because radiation doses resulting from CT were shown to be not irrelevant. Efforts directed at identification of factors impacting the level of radiation exposure in both the course of the EVAR procedure and post-EVAR endovascular interventions and CTAs are warranted as potentially modifiable factors may offer opportunities to reduce the radiation. In the light of the risks found to be associated with radiation exposure and considering the findings above, those involved in EVAR and post-EVAR surveillance should aim at optimal dose management.

4.
J Cardiothorac Surg ; 11(1): 120, 2016 Aug 04.
Article in English | MEDLINE | ID: mdl-27488025

ABSTRACT

BACKGROUND: Infection is a main cause of morbidity and mortality after heart surgery, with multi-resistant pathogens increasingly representing a challenge. Daptomycin provides bactericidal activity against gram-positive organisms that are resistant to standard treatment including vancomycin. METHODS: A cohort of cardiac surgical patients, treated with daptomycin for major infection at two tertiary care centers, were retrospectively studied with a particular focus on the type of infection, causative pathogens and co-infections, daptomycin dosage, adverse events and outcome in order to provide evidence for the efficiency and safety of daptomycin in a distinct high-risk patient population. RESULTS: Sixty-five patients (87.7 % males, 60.4 ± 13.5 years) who had undergone aortic surgery (20.0 %), ventricular assist device (VAD) implantation (21.5 %), combined procedures (21.5 %), coronary artery bypass grafting (12.3 %), isolated valve surgery (15.4 %) and heart transplantation (7.7 %) were diagnosed with catheter-related infection (26.1 %), valve endocarditis (18.8 %), sternal wound (13.0 %), VAD-associated (11.6 %), cardiac implantable electrophysiological device (CIED)-associated (4.1 %), respiratory tract (4.3 %), bloodstream (4.3 %) and other infection (4.3 %). In 13.0 %, no focus of infection was identified though symptoms of severe infection were present. The most frequent pathogens were Staphylococcus epidermidis (30.4 %), Staphylococcus aureus (23.1 %) and Enterococcus species (10.1 %). Daptomycin doses ranging from 3 mg/kg every 48 h to 10 mg/kg every 24 h were administered for 15.4 ± 11.8 days. 87.0 % of the cases were classified as success, 7.2 % as treatment failure and 5.8 as non-evaluable. Adverse events were limited to one case of mild and one case of moderate neutropenia with recovery upon termination of treatment. CONCLUSION: Daptomycin proved safe and effective in major infection in high-risk cardiac surgical patients.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Cardiac Surgical Procedures/adverse effects , Daptomycin/therapeutic use , Gram-Positive Bacterial Infections/drug therapy , Gram-Positive Bacterial Infections/microbiology , Aged , Female , Gram-Positive Bacterial Infections/etiology , Humans , Male , Middle Aged , Retrospective Studies , Treatment Outcome
5.
Open Cardiovasc Med J ; 10: 148-57, 2016.
Article in English | MEDLINE | ID: mdl-27499818

ABSTRACT

AIM: Intraoperative allogeneic blood product transfusion (ABPT) in cardiac surgery is associated with worse overall outcome, including mortality. The objective of this study was to evaluate the ABPTs in minimalized extracorporeal cardiopulmonary (MECC(TM)) compared with standard open system on-pump coronary revascularization. METHODS: Data of 156 patients undergoing myocardial revascularization between September 2008 and September 2010 were reviewed. 83 patients were operated by the MECC technique and 73 were treated by standard extracorporeal circulation (sECC). ABPT and overall early postoperative complications were analyzed. RESULTS: Operative mortality and morbidity were similar in both groups. ABPT in the MECC group was significantly lower than in the sECC group both intraoperatively (7.2 vs. 60.3% of patients p<0.001) and during the first five postoperative days (19.3 vs. 57.5%; p<0.001). "Skin to skin"- (214 ± 45 vs. 232 ± 45 min; p=0.012), cardiopulmonary bypass (CPB) - (82 ± 25 vs. 95 ± 26 min; p=0.014), and X-clamp- times (50 ± 16 vs. 56 ± 17 min; p=0.024) were significantly lower in the MECC group than in the sECC group. Length of ICU (intensive care unit) - and hospital stay were also significantly lower in the MECC group vs. the sECC group (26.7 ± 20.2 vs. 54.5 ± 68.9 h; p<0.001, and 12.0 ± 4.1 vs. 14.5 ± 4.6 days; p<0.001). CONCLUSION: Application of MECC as on-pump coronary artery bypass graft (CABG) results in significantly lower ABPT as well as shorter ICU and in-hospital stay. In order to achieve these benefits of MECC autologous retrograde priming, Bispectral index (BIS) monitoring, intraoperative cell salvage, meticulous hemostasis and strict peri- and postoperative volume management are crucial.

6.
J Cardiothorac Surg ; 11(1): 108, 2016 Jul 15.
Article in English | MEDLINE | ID: mdl-27422642

ABSTRACT

BACKGROUND: In patients with left ventricular non-compaction (LVNC), implantation of a left ventricular assist device (LVAD) may be performed as a bridge to transplantation. In this respect, the particular characteristics of the left ventricular myocardium may represent a challenge. CASE PRESENTATION: We report a patient with LVNC who required urgent heart transplantation for inflow cannula obstruction nine months after receiving a LVAD. LVAD parameters, echocardiography and examination of the explanted heart suggested changes of left ventricular configuration brought about by LVAD support as the most likely cause of inflow cannula obstruction. CONCLUSIONS: We conclude that changes experienced by non-compacted myocardium during LVAD support may give rise to inflow cannula obstruction and flow reduction. Presence of LVNC mandates tight surveillance for changes in LV configuration and LVAD flow characteristics and may justify urgent transplantation listing status.


Subject(s)
Cardiomyopathies/physiopathology , Heart Failure/etiology , Heart Ventricles/physiopathology , Heart-Assist Devices , Adult , Cardiomyopathies/complications , Cardiomyopathies/surgery , Echocardiography , Heart Failure/physiopathology , Heart Failure/surgery , Heart Transplantation , Heart Ventricles/surgery , Humans , Male , Myocardium
7.
Article in English | MEDLINE | ID: mdl-26846598

ABSTRACT

Because aortic occlusion is arguably one of the most dangerous aortic manipulation maneuvers during cardiac surgery in terms of perioperative ischemic neurological injury, the purpose of this investigation is to assess the structural mechanical impact resulting from the use of existing and newly proposed occluders. Existing (clinically used) occluders considered include different cross-clamps (CCs) and endo-aortic balloon occlusion (EABO). A novel occluder is also introduced, namely, constrained EABO (CEABO), which consists of applying a constrainer externally around the aorta when performing EABO. Computational solid mechanics are employed to investigate each occluder according to a comprehensive list of functional requirements. The potential of a state of occlusion is also considered for the first time. Three different constrainer designs are evaluated for CEABO. Although the CCs were responsible for the highest strains, largest deformation, and most inefficient increase of the occlusion potential, it remains the most stable, simplest, and cheapest occluder. The different CC hinge geometries resulted in poorer performance of CC used for minimally invasive procedures than conventional ones. CEABO with a profiled constrainer successfully addresses the EABO shortcomings of safety, stability, and positioning accuracy, while maintaining its complexities of operation (disadvantage) and yielding additional functionalities (advantage). Moreover, CEABO is able to achieve the previously unattainable potential to provide a clinically determinable state of occlusion. CEABO offers an attractive alternative to the shortcomings of existing occluders, with its design rooted in achieving the highest patient safety. Copyright © 2016 John Wiley & Sons, Ltd.


Subject(s)
Aorta/physiology , Aorta/surgery , Balloon Occlusion/instrumentation , Balloon Occlusion/methods , Models, Cardiovascular , Adult , Animals , Balloon Occlusion/adverse effects , Computer Simulation , Humans , Middle Aged , Sheep
8.
J Cardiothorac Surg ; 10: 99, 2015 Jul 17.
Article in English | MEDLINE | ID: mdl-26183430

ABSTRACT

Device-related infections in recipients of left ventricular assist devices (LVAD) have been recognized as a major source of morbidity and mortality. They require a high level of diagnostic effort as part of the overall burden resulting from infectious complications in LVAD recipients. We present a multi-allergic patient who was treated for persistent sterile intrathoracic abscess formation and pericardial empyema following minimally invasive LVAD implantation including use of a sheet of e-polytetrafluoroethylene (ePTFE) membrane to restore pericardial integrity. Sterile abscess formation and pericardial empyema recurred after surgical removal until the ePTFE membrane was removed, suggesting that in disposed patients, ePTFE may be related to sterile abscess formation or sterile empyema.


Subject(s)
Abscess/etiology , Empyema/etiology , Heart-Assist Devices/adverse effects , Pericardium , Polytetrafluoroethylene/adverse effects , Thoracic Cavity , Abscess/diagnosis , Adult , Empyema/diagnosis , Humans , Male , Tomography, X-Ray Computed
9.
Transpl Infect Dis ; 17(5): 737-43, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26224318

ABSTRACT

Necrotizing pulmonary aspergillosis and Aspergillus device infection are rare and have potentially fatal complications after left ventricular assist device (LVAD) implantation. To date, few cases of patients surviving Aspergillus device infection have been published, with survival reported only after device removal. We present a patient implanted with an LVAD in whom necrotizing pulmonary aspergillosis with device involvement was successfully treated by segmentectomy and prolonged antifungal treatment without device exchange or removal. Similar cases in the literature were searched for and are discussed in view of the severity of this complication.


Subject(s)
Heart-Assist Devices/adverse effects , Prosthesis-Related Infections/diagnosis , Pulmonary Aspergillosis/diagnosis , Adult , Humans , Male
10.
Minerva Cardioangiol ; 63(2): 91-8, 2015 Apr.
Article in English | MEDLINE | ID: mdl-24941962

ABSTRACT

AIM: The aim of the study was to examine risks, implications and outcomes of coronary sinus (CS) lead extraction in patients with infections of cardiac resynchronization therapy (CRT) systems. METHODS: The study included 40 (65.5 ± 11.1 years; 80% male) transvenous CS lead extraction procedures performed between 2000-2011. Nine (22.5%) patients suffered from infection and included one sepsis (11.1%), two (22.2%) of lead and valve endocarditis, and four (44.4%) cases of pocket infection. CS lead extraction in the infection subgroup was performed between 14 days and more than five years after the last CIED-related surgical procedure. RESULTS: Totally 42 CS and 35 non-CS leads were extracted. Leads extracted in the infection subgroup were significantly longer in situ (49.7 ± 30.7 months) compared to the non-infection subgroup (19.2 ± 28.6 months). Extraction in infected patients required more aggressive methods and longer exposure to radiation than non-infected. Procedural success without major complications was achieved in all patients. Minor post-procedural complications occurred in four (44.4%) of the infected and one (3.2%) of the non-infected patients and were surgical-related in three cases. Overall hospitalization times were significantly longer for the infection than for the non-infection subgroup (21.4 ± 15 versus 9.6 ± 6.9 days). CONCLUSION: Our results support the concept of complete CIED-system removal in CIED-associated infection, regardless of whether or not infection appears to be limited to the generator pocket site, despite risk of heart failure, patient frailty and a high level of comorbidity. An interdisciplinary approach encompassing appropriate diagnostic, procedural and safety standards allows CS lead extraction in this high-risk subpopulation to be performed with excellent outcomes and low complication rates.


Subject(s)
Cardiac Resynchronization Therapy/adverse effects , Device Removal/methods , Electrodes, Implanted/adverse effects , Aged , Coronary Sinus , Device Removal/adverse effects , Endocarditis/epidemiology , Endocarditis/etiology , Female , Heart Valve Diseases/epidemiology , Heart Valve Diseases/etiology , Hospitalization/statistics & numerical data , Humans , Length of Stay , Male , Middle Aged , Retrospective Studies , Sepsis/epidemiology , Sepsis/etiology , Time Factors
11.
Transplant Proc ; 46(9): 2953-6, 2014 Nov.
Article in English | MEDLINE | ID: mdl-25420800

ABSTRACT

BACKGROUND: Polyclonal anti-thymocyte globulins (ATGs) and anti-CD25 antibodies are agents used for induction of immunosuppression in solid-organ transplantation. We aimed to investigate the effect of different regimens of these immunosuppressive induction agents on transendothelial migration of peripheral blood mononuclear cells (PBMC) and evaluated the endothelial apoptosis after treatment. METHODS: Human microvascular endothelial cells were either activated with tumor necrosis factor-α/interferon-γ or not and further treated with 25 or 125 µg/mL ATG (Thymoglobulin, Sanofi-Aventis, Germany) for 2 hours or 24 hours, or with 5 µg/mL Basiliximab (Simulect, Novartis, Germany) for 2 hours or 24 hours. PBMC were either activated with phytohaemagglutinin (PHA) or not and further treated with 25 or 125 µg/mL ATG or with 5 µg/mL Basiliximab for 2 h and then used for transendothelial migration assays. Apoptosis of endothelial cells was detected by means of Annexin-V staining after 2-hour incubation with either 25 or 125 µg/mL ATG or 5 µg/mL Basiliximab. RESULTS: Prophylactic 24-hour administration of ATG to naive endothelial cells without PBMC treatment reduced transendothelial migration. Prophylactic 24-hour administration of ATG and Basiliximab to naive endothelial cells after PBMC treatment with the same agents reduced the transendothelial migration after 24 hours. In both cases, no effect could be observed after 2-hour treatment. Basiliximab but not ATG showed a reduction of transmigration after 2-hour treatment of PBMCs without naive EC treatment. Apoptosis of endothelial cells after treatment increased in both cases, being in case of ATG dose-dependent, increasing from 1.2% after either 25 µg/mL ATG to 8.7% after 125 µg/mL ATG. CONCLUSIONS: Immunosuppressive induction agents modulate the endothelial activity in a dose- and time-dependent manner. Our results suggest that administration of induction agents over longer time periods could provide a potential benefit regarding endothelial immunomodulation. Increased doses may, however, show a deleterious effect on endothelial survival.


Subject(s)
Antibodies, Monoclonal/pharmacology , Antilymphocyte Serum/pharmacology , Apoptosis/drug effects , Endothelial Cells/drug effects , Immunosuppressive Agents/pharmacology , Leukocytes, Mononuclear/drug effects , Recombinant Fusion Proteins/pharmacology , Transendothelial and Transepithelial Migration/drug effects , Antibodies, Monoclonal/therapeutic use , Antilymphocyte Serum/therapeutic use , Basiliximab , Dose-Response Relationship, Drug , Graft Rejection/immunology , Graft Rejection/prevention & control , Humans , Immunosuppressive Agents/therapeutic use , Recombinant Fusion Proteins/therapeutic use , Time Factors
12.
Diabetologia ; 55(5): 1514-25, 2012 May.
Article in English | MEDLINE | ID: mdl-22327346

ABSTRACT

AIMS/HYPOTHESIS: We have previously found that the mass of perivascular adipose tissue (PVAT) correlates negatively with insulin sensitivity and post-ischaemic increase in blood flow. To understand how PVAT communicates with vascular vessels, interactions between perivascular, subcutaneous and visceral fat cells with endothelial cells (ECs) were examined with regard to inflammatory, metabolic and angiogenic proteins. To test for possible in vivo relevance of these findings, circulating levels of the predominant secretion product, hepatocyte growth factor (HGF), was measured in individuals carefully phenotyped for fat distribution patterns. METHODS: Mono- and co-cultures of human primary fat cells with ECs were performed. mRNA expression and protein production were studied using Luminex, cytokine array, RealTime Ready and ELISA systems. Effects of HGF on vascular cells were determined by WST assays. In patients, HGF levels were measured by ELISA, and the mass of different fat compartments was determined by whole-body MRI. RESULTS: In contrast with other fat cell types, PVAT cells released higher amounts of angiogenic factors, e.g. HGF, acidic fibroblast growth factor, thrombospondin-1, serpin-E1, monocyte chemotactic protein-1 and insulin-like growth factor-binding protein -3. Cocultures showed different expression profiles from monocultures, and mature adipocytes differed from pre-adipocytes. HGF was preferentially released by PVAT cells and stimulated EC growth and smooth muscle cell cytokine release. Finally, in 95 patients, only PVAT, not visceral or subcutaneous mass, correlated independently with serum HGF levels (p = 0.03; r = 0.225). CONCLUSIONS: Perivascular (pre-)adipocytes differ substantially from other fat cells with regard to mRNA expression and protein production of angiogenic factors. This may contribute to fat tissue growth and atherosclerotic plaque complications. Higher levels of angiogenic factors, such as HGF, in patients with increased perivascular fat mass may have pathological relevance.


Subject(s)
Adipocytes/metabolism , Hepatocyte Growth Factor/metabolism , Abdominal Fat/metabolism , Adult , Aged , Angiogenesis Inducing Agents/metabolism , Angiogenic Proteins/metabolism , Cells, Cultured , Coculture Techniques , Endothelial Cells/metabolism , Female , Gene Expression Profiling , Hepatocyte Growth Factor/blood , Humans , Male , Middle Aged , Subcutaneous Fat/metabolism , Young Adult
14.
J Thorac Cardiovasc Surg ; 131(6): 1323-30, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16733165

ABSTRACT

BACKGROUND: Endovascular application of pulmonary heart valves has been recently introduced clinically. A tissue-engineering approach was pursued to overcome the current limitations of bovine jugular vein valves (degeneration and limited longevity). However, deployment of the delicate tissue-engineered valves resulted in severe tissue damage. Therefore the objective of this study was to prevent tissue damage during the folding and deployment maneuver. MATERIAL AND METHODS: Porcine pulmonary heart valves, small intestinal submucosa, and ovine carotid arteries were obtained from a slaughterhouse. After dissection and antimicrobial incubation, the valves were trimmed (removal of sinus and most of the muscular ring) to fit into the deployment catheter. The inside (in-stent group, n = 6) or outside (out-stent group, n = 6) of a nitinol stent was covered by an acellular small intestinal submucosa, and the valves were sutured into the stent. The valves were folded, tested for placement in the deployment catheter, and decellularized enzymatically. Myofibroblasts were obtained from carotid artery segments and seeded onto the scaffolds. The seeded constructs were placed in a dynamic bioreactor system and cultured for 16 consecutive days. After endothelial cell seeding, the constructs were folded, deployed, and processed for histology and surface electron microscopy. RESULTS: The valves opened and closed competently throughout the entire dynamic culture. Surface electron microscopy revealed an almost completely preserved tissue in the in-stent group. Stents covered with small intestinal submucosa on the outside, however, showed severe damage. CONCLUSION: This study demonstrates that small intestinal submucosa covering of the inside of a pulmonary valved stent can prevent stent strut-related tissue damage.


Subject(s)
Bioprosthesis/adverse effects , Heart Valve Prosthesis Implantation/methods , Heart Valve Prosthesis/adverse effects , Pulmonary Valve/surgery , Tissue Engineering , Animals , Cells, Cultured , Humans , Postoperative Complications/etiology , Postoperative Complications/prevention & control , Prosthesis Design , Stents
16.
Thorac Cardiovasc Surg ; 54(1): 39-41, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16485187

ABSTRACT

OBJECTIVE: Standard extracorporeal circulation (ECC) remains the staple procedure for cardiac surgeons. Despite modern membrane oxygenators and arterial filters micro bubbles are regularly detected in the arterial line. We investigated whether initial deairing of the venous drainage during connection can reduce the quantity and size of micro bubbles on the arterial side. METHODS: 12 patients underwent isolated coronary artery revascularization with conventional ECC using a two-stage venous catheter and an open, passive venous return into a reservoir. In 6 patients (Control) the venous catheter was routinely connected to the venous line, thereby accepting moderate incorporation of air. In another 6 patients (deaired) the catheter was connected avoiding any visible air entrapment. A bubble counter was used to detect the number and size of any micro bubbles in the arterial line of the ECC. The total number of bubbles as well as the number of bubbles of different sizes was assessed directly after initiation of ECC and during the first 60 sec. RESULTS: All patients had an uneventful surgery with a normal postoperative course. In the Control group a considerable number of bubbles of all sizes occurred initially and after 60 sec. In contrast, very few bubbles were detected in the deaired group. CONCLUSION: Incorporated venous air inevitably reaches the arterial side of the ECC. As deairing of the venous line is a simple and effective manoeuvre to significantly reduce the amount of micro bubbles on the arterial side, we recommend and perform routine deairing in all our patients.


Subject(s)
Coronary Artery Disease/surgery , Drainage , Embolism, Air/prevention & control , Extracorporeal Circulation/instrumentation , Microbubbles/adverse effects , Cardiac Surgical Procedures/instrumentation , Cardiopulmonary Bypass/instrumentation , Case-Control Studies , Catheters, Indwelling , Cerebrovascular Circulation , Coronary Artery Disease/physiopathology , Embolism, Air/etiology , Embolism, Air/physiopathology , Equipment Design , Equipment Safety , Extracorporeal Circulation/adverse effects , Filtration/instrumentation , Humans , Oxygenators, Membrane , Particle Size , Pulmonary Circulation , Pulmonary Veins/surgery , Treatment Outcome
17.
Biomaterials ; 26(5): 495-500, 2005 Feb.
Article in English | MEDLINE | ID: mdl-15276357

ABSTRACT

Multiphoton induced blue/green autofluorescence by near infrared femtosecond laser pulses has been used to selectively image intratissue elastic fibers in native and tissue engineered (TE) viable heart valves without any invasive tissue removal, embedding, fixation, and staining. Elastic fibers could be clearly distinguished from collagenous structures which emit ultraviolet/violet radiation when excited with intense ultrashort pulses due to second harmonic generation. Deep-tissue three-dimensional imaging of elastic fibers with submicron spatial resolution was performed by optical sectioning of heart valves using a multiphoton laser scanning microscope in connection with a tunable 80 MHz femtosecond laser source. The technology was used to diagnose extracellular matrix structures and cell resettlement of TE heart valves prior implantation. This novel non-invasive method opens the general possibility of high-resolution in situ imaging of elastic fibers, collagen structures and intracellular organelles in living intact tissues without staining.


Subject(s)
Elastic Tissue/ultrastructure , Imaging, Three-Dimensional/methods , Microscopy, Confocal/methods , Pulmonary Valve/ultrastructure , Tissue Engineering , Animals , Bioreactors , Carotid Arteries/cytology , Cells, Cultured/cytology , Collagen/ultrastructure , Cryoultramicrotomy , Elastic Tissue/chemistry , Endothelial Cells/ultrastructure , Extracellular Matrix/ultrastructure , Fibroblasts/ultrastructure , Fluorescence , Microscopy, Electron , Microscopy, Electron, Scanning , Sheep , Sus scrofa
18.
Matrix Biol ; 23(2): 113-25, 2004 May.
Article in English | MEDLINE | ID: mdl-15246110

ABSTRACT

Tissue engineering of heart valves utilizes biodegradable or metabolizable scaffolds for remodeling by seeded autologous cells. The aim of this study was to determine and compare extracellular matrix (ECM) formations, cellular phenotypes and cell location of native and tissue engineered (TE) valve leaflets. Ovine carotid arteries, ovine and porcine hearts were obtained from slaughterhouses. Cells were isolated from carotid arteries and dissected ovine, porcine and TE leaflets. TE constructs were fabricated from decellularized porcine pulmonary valves, seeded ovine arterial cells and subsequent 16 days dynamic in vitro culture using a pulsatile bioreactor. Native and TE valves were studied by histology (hematoxylin-eosin, resorcin-fuchsin, Movat pentachrome), NIR femtosecond multiphoton laser scanning microscopy and scanning electron microscopy (SEM). Cells of native and TE tissues were identified and localized by immunohistochemistry. Arterial, valvular and re-isolated TE-construct cells were processed for immunocytochemistry and Western blotting. ECM analysis and SEM revealed characteristical and comparable structures in native and TE leaflets. Most cells in native leaflets stained strongly positive for vimentin. Cells positive to alpha-smooth muscle actin (alpha-SMA), myosin and calponin were only found at the ventricular (inflow) side of ovine aortic and porcine pulmonary valve leaflets. Cells from TE constructs had a strong expression of vimentin, alpha-SMA, myosin, calponin and h-caldesmon throughout the entire leaflet. Comparable ECM formation and endothelial cell lining of native and TE leaflets could be demonstrated. However, immunostaining revealed significant differences between valvular cell phenotypes of native and TE leaflets. These results may be essential for further cardiovascular tissue engineering efforts.


Subject(s)
Bioartificial Organs , Extracellular Matrix/chemistry , Extracellular Matrix/metabolism , Heart Valves/cytology , Heart Valves/metabolism , Heart, Artificial , Tissue Engineering , Animals , Arteries/cytology , Arteries/metabolism , Cells, Cultured , Endothelial Cells/cytology , Endothelial Cells/metabolism , Fibroblasts/cytology , Fibroblasts/metabolism , Heart Valves/chemistry , Heart Valves/ultrastructure , Immunohistochemistry , Lasers , Microscopy, Fluorescence , Sheep, Domestic , Swine
19.
Tissue Eng ; 10(3-4): 611-20, 2004.
Article in English | MEDLINE | ID: mdl-15165477

ABSTRACT

At the time of implantation, tissue-engineered constructs should resemble native tissues as closely as possible. At present, histology and biochemical methods are commonly used to compare tissue-engineered constructs with native tissue. A ProteinChip system based on surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI) has been developed that allows visualization of complex protein profiles from biological samples. The aim of this study was to determine whether the ProteinChip system is a suitable tool with which to compare the protein expression profiles of tissue-engineered aortic blood vessels with native tissues. Tissue-engineered blood vessel substitutes were fabricated with poly-4-hydroxybutyrate scaffolds, ovine vascular cell seeding, and dynamic tissue culture conditions. Engineered, ovine aortic, and carotid tissues were homogenized and total protein was extracted. Samples were analyzed on ProteinChip arrays. Analysis yielded reproducible protein profiles from all samples. About 150 distinct protein peaks were detected. Comparative analysis with ProteinChip software revealed that the protein profiles from native aorta and native carotid arteries were similar whereas early tissue-engineered samples displayed more distinct deviations. In conclusion, ProteinChip system technology is rapid, reproducible, and highly sensitive in highlighting differentially expressed proteins in tissue-engineered blood vessel substitutes.


Subject(s)
Blood Vessels/metabolism , Gene Expression/physiology , Protein Array Analysis , Tissue Engineering , Animals , Aorta/metabolism , Carotid Arteries/metabolism , Gene Expression Profiling , Mass Spectrometry , Sheep
20.
Ann Biomed Eng ; 32(2): 212-22, 2004 Feb.
Article in English | MEDLINE | ID: mdl-15008369

ABSTRACT

Compared to native blood vessels, all clinically available blood vessel substitutes perform suboptimally. Numerous approaches to tissue engineer (TE) blood vessels have been pursued using different scaffold materials, cell types, and culture conditions. Several limitations however remain to be overcome prior to the potential application in the arterial system. This study aimed at tissue engineering viable ovine blood vessels suitable for implantation into the systemic circulation of sheep. In recent studies vascular smooth muscle cells (vSMC) were derived by an explant technique. However, in this study we show that homogenous populations of differentiated vSMC were only obtained by enzymatic dispersion as characterized by immunostaining for specific vSMC marker proteins. In contrast the explant method yielded predominantly less differentiated myofibroblast-like cells. Enzymatically derived vSMC were seeded onto P-4-HB scaffolds and incubated either in a pulsatile flow bioreactor or under static conditions. Dynamically cultured TE blood vessel substitutes showed confluent layered tissue formation and were completely water resistant. They displayed significantly increased ECM synthesis, DNA, and protein content as well as vSMC marker expression. Mechanical properties of bioreactor cultured TE blood vessels approached those of native aorta. In conclusion ovine, aortic blood vessel substitutes were successfully created using enzymatically derived vSMC, bioabsorbable scaffolds, and applied shear stress.


Subject(s)
Blood Vessels/growth & development , Cell Culture Techniques/methods , Endopeptidases/metabolism , Endothelial Cells/physiology , Mechanotransduction, Cellular/physiology , Muscle, Smooth, Vascular/growth & development , Tissue Engineering/methods , Animals , Bioreactors , Blood Vessels/cytology , Cell Differentiation/physiology , Cells, Cultured , Collagenases/metabolism , Elasticity , Endothelial Cells/cytology , Muscle Contraction/physiology , Muscle, Smooth, Vascular/cytology , Pancreatic Elastase , Sheep , Stress, Mechanical , Tensile Strength , Transplants
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