Chantal Kang

Characterization of Dendritic Cells Subpopulations in Skin and Afferent Lymph in the Swine Model

Transcutaneous delivery of vaccines to specific skin dendritic cells (DC) subsets is foreseen as a promising strategy to induce strong and specific types of immune responses such as tolerance, cytotoxicity or humoral immunity. Because of striking histological similarities between human and pig skin, pig is recognized as the most suitable model to study the cutaneous delivery of medicine. Therefore improving the knowledge on swine skin DC subsets would be highly valuable to the skin vaccine field. In this study, we showed that pig skin DC comprise the classical epidermal langerhans cells (LC) and dermal DC (DDC) that could be divided in 3 subsets according to their phenotypes: (1) the CD163neg/CD172aneg, (2) the CD163highCD172apos and (3) the CD163lowCD172apos DDC. These subtypes have the capacity to migrate from skin to lymph node since we detected them in pseudo-afferent lymph. Extensive phenotyping with a set of markers suggested that the CD163high DDC resemble the antibody response-inducing human skin DC/macrophages whereas the CD163negCD172low DDC share properties with the CD8+ T cell response-inducing murine skin CD103pos DC. This work, by showing similarities between human, mouse and swine skin DC, establishes pig as a model of choice for the development of transcutaneous immunisation strategies targeting DC.

Steady state pig dendritic cells migrating in skin draining pseudo-afferent lymph are semi-mature

Dendritic cells (DC) in peripheral tissues are considered as immature cells that mature and migrate towards lymph nodes upon stimulation with pathogens. This commonly accepted paradigm is challenged by the fact that tolerance to peripheral self antigen is controlled by mature DC and that DC collected from afferent lymph draining different tissues from several species, in the absence of pathogen signaling, were inconsistently found to be either at a mature or semi-mature state. In order to better define the maturation state of DC that migrate in lymph in absence of pathogen stimulation, we compared skin lymph DC to resident and LPS (lipopolysaccharide)-activated skin DC thanks to the establishment of a mini-pig model of lymph duct cannulation. Based on their co-stimulatory molecules expression and endocytotic capacities, pig lymph skin DC were found at an intermediate state of maturation between resident and LPS-activated skin DC and were fully capable of allogeneic T cell stimulation. Furthermore, lymph skin DC could be further matured by LPS or influenza stimulation. Thus, using the pig skin model which is relevant to human, we show that skin-derived DC constantly migrate at an intermediate state of maturation that can be further enhanced upon appropriate stimulation.

Kinetics of endothelialization of the multilayer flow modulator and single-layer arterial stents.

The multilayer flow modulator (MFM; Cardiatis, Isnes, Belgium) is a self-expandable mesh of braided cobalt alloy wires, used for treatment of aortic and peripheral aneurysms. To further improve our understanding of this novel technology, the endothelialization kinetics of the MFM was investigated and compared with those of two marketed single-layer stents. Five porcine animal models were used in which a total of 19 stents were implanted in the iliac and carotid arteries between one and five weeks before sacrifice. All 19 stents were successfully delivered. For all devices, nonsignificant signs of inflammation or thrombosis were noted, and there was no evidence of local intolerance. The MFM developed a thin layer of endothelial cells earlier and was associated with less neointimal development than the two single-layer stents. A differing phenomenon of integration was also revealed and hypothesized as endothelialization from adhesion of circulating endothelial progenitor cells, as well as adhesion from the arterial wall, and also by the differences in trauma exposed to the arterial wall.

Assessment of Biocompatibility of the Multilayer Flow Modulator withDiffering Thread Designs

Background: The Multilayer Flow Modulator (MFM) (Cardiatis, Isnes, Belgium) is a self-expandable mesh of cobalt alloy wires used for the treatment of aortic aneurysms. The impact of design thread count and duration of implantation on the biocompatibility of the MFM in porcine animal models was assessed in this study. Methods: Eight mini-piglets received 26 MFM devices (12 with 56 threads, 14 with 80-96 threads) in the iliac, carotid, and renal arteries. Animals were sacrificed and specimens explanted at 1, 3, and 6 months, at which time histological and ultrastructural analyses were carried out. Results: The MFM was successfully deployed in 25 of the 26 implanted cases. The 56 thread devices were well tolerated locally and yielded fewer signs of inflammation and neo intimal hyperplasia. Percentage stenosis was 16.9% ± 5.1% for the 56 thread devices versus 33.4% ± 10.2% for the 80-96 thread devices (p=0.001) at 3 months, and 21.7% ± 9.9% for the 56 thread devices versus 33.6% ± 12.4% for the 80-96 thread devices (p=0.004) at 6 months. The 5 devices selected for SEM examination were well deployed, integrated into the vessel wall and endothelialized, and had patent side branches. Conclusions: No significant stenosis formation or inflammatory response was recorded in any of the implanted animals. The 80-96 thread devices elicited a greater intra-arterial response than the 56 thread devices, although the values for both groups remained within the normal range for stented carotid, renal, or iliac arteries. Further preclinical and clinical studies will extend assessment of the long-term safety and effectiveness of the MFM

RFID implantable pressure sensor for the follow-up of abdominal aortic aneurysm stented

An abdominal aortic aneurysm (AAA) is a dilatation of the aorta at the abdominal level, the rupture of which is a life threatening complication with an 80% mortality rate. Even though those devices keep improving, the failure rate of the endovascular treatment is due to persisting pressure into the excluded aneurysmal sac. Since 2005, several integrated sensors have been designed for the follow-up of the AAA treated by a stent. Solutions are based on the use of a single sensor. Thrombus in the excluded AAA can modify the field of pressure when leaks appeared and a network of sensors should be used. We present in this paper the ENDOCOM project that aims to design an implantable pressure sensor that can be used in a network configuration. To validate the new materials, we developed a framework composed of in vitro experiments and in vivo tests on large animal model. Numerical modeling has been investigated from the experimental data to determine the optimal position of sensor. Some results of those different parts are shown in this paper.

Surface analysis of an encapsulation membrane after its implantation in mini-pigs.

The biocompatibility of membranes aiming at being a part of a bioartificial pancreas has been tested. For that purpose, we have studied a polycarbonate membrane surface after its implantation in mini-pigs. The membranes were made hydrophilic by an argon plasma surface treatment followed by a dipping in a hydrophilic polymer solution. Two polymers were tested: polyvinylpyrrolidone (PVP) and hydroxypropylmethylcellulose (HPMC). To test their biocompatibility, an encapsulation device for pig Langerhans islets, with external membranes treated as described above, was implanted in different mini-pigs. The pigs received no further treatment. The devices were explanted after in vivo exposure and the membranes were analysed by XPS (x-ray photoelectron spectroscopy) and ToF-SIMS (time-of-flight secondary ion mass spectrometry). After this time, the substrate with the PVP or HPMC treatment was still detected on the different samples. The surface treatment signal, however, was attenuated. This is explained by the detection of other components partly covering the surface. XPS and ToF-SIMS analyses revealed the presence of biological molecules on the two faces of the membrane: the outside face in contact with the biological environment and the inside face in contact with the device. ToF-SIMS images show the inhomogeneity of the biological molecules on the membrane surface. In conclusion, biological molecules adhered to the encapsulation membrane surface after implantation but the surface treatments remained unaltered.

Synchronized Pulsatile Flow With Low Systolic Output From Veno-Arterial Extracorporeal Membrane Oxygenation Improves Myocardial Recovery After Experimental Cardiac Arrest in Pigs: SYNCHRONIZED PULSATILE FLOW

Circulatory failure following cardiac arrest (CA) requires catecholamine support and occasionally veno-arterial extracorporeal membrane oxygenation (vaECMO). VaECMO-generated blood flow is continuous and retrograde, increasing ventricular stroke work. Our aim was to assess the benefit of a device generating a pulsatile vaECMO flow synchronized with the heart rhythm lowering systolic vaECMO output on the left ventricular ejection fraction (LVEF) and pulmonary capillary pressure (Pcap) after CA. This experimental randomized study in pigs compared standard nonpulsatile vaECMO (control) with pulsatile synchronized vaECMO (study) group using a pulsatility-generating device. After sedation and intubation, ventricular fibrillation was induced by pacing. After 10-min ventricular fibrillation, cardiopulmonary resuscitation was performed for 20 min then vaECMO, defibrillation and 0.15 µg/kg/min intravenous epinephrine infusion were initiated. Hemodynamics, Pcap, LVEF by echocardiography and angiography were measured at baseline and every 30 min after the vaECMO start until vaECMO and epinephrine were stopped (at 120 min), and 30 min later. Baseline hemodynamics did not differ between groups; 120 min after vaECMO initiation, LVEF by echocardiography and angiography was significantly higher in the study than control group 55 ± 19% versus 34 ± 13% (P = 0.042), 50 ± 16% versus 33 ± 12% (P = 0.043), respectively. Pcap decreased from baseline by 4.2 ± 8.6 mm Hg in the study group but increased by 5.6 ± 5.9 mm Hg in the control group (P = 0.043). Thirty minutes later, LVEF remained higher in the study group 44 ± 7% versus 26 ± 11% (P = 0.008) while Pcap did not differ. A synchronized pulsatile device decreasing systolic output from vaECMO improved LVEF and Pcap in a pig model of CA and resuscitation.

Intrapulmonary artery balloon pulsation improves circulatory function after acute myocardial infarction in pigs

ABSTRACT Aim: To examine whether pulmonary artery balloon pulsation (PABP) could improve circulatory function in acute myocardial infarction (AMI) in pigs. Methods/results: Ten downsize pigs were sedated and ventilated. AMI was induced by inserting a plug into the left anterior descending artery. A pulsation balloon was placed in the pulmonary artery in all animals. In the treatment group (TG), pulsations began when life-threatening arrhythmia or > 30% drop in mean blood pressure (MBP) or > 40% decrease in cardiac output compared to baseline occurred. Pulsation rate was 120/min, independent of the heartbeat, maintained for 10 min. The control group (CG) received no pulsation. In the TG (n = 5), mean BP after the AMI improved by 7 ± 12 mmHg after 150 min while in the CG, MBP decreased by 17 ± 25 mmHg, P < 0.05; coronary perfusion pressure improved by 8 ± 7 mmHg in the TG but decreased by 15 ± 12 in the CG (P < 0.05). In the CG, cardiac output did not change but in the TG it improved from 3.5 ± 0.9 after the AMI to 4.2 ± 1.1 l/min 150 min after AMI (P < 0.05). The TG required 1.8 ± 0.4 electric shocks for ventricular fibrillation versus 0.8 ± 0.4 in the pulsation group (P < 0.05). Conclusion: PABP could be useful in the management of AMI due to improved mean arterial BP, coronary perfusion pressure, cardiac output and electrical stability. The mechanism of this effect remains to be determined.