Increasing Pulmonary Artery Pulsatile Flow Improves Hypoxic Pulmonary Hypertension in Piglets

Pulmonary arterial hypertension (PAH) is a disease affecting distal pulmonary arteries (PA). These arteries are deformed, leading to right ventricular ...

Institut national de la recherche agronomique

Small-sized apolipoprotein(a) [apo(a)] isoforms with high antifibrinolytic activity are frequently found in cardiovascular diseases, suggesting a role for apo(a) size in atherothrombosis. To test this hypothesis, we sought to characterize the lysine (fibrin)-binding function of isolated apo(a) of variable sizes.

Lp(a) particles mold fibrin-binding properties of apo(a) in size-dependent manner: a study with different-length recombinant apo(a), native Lp(a), and monoclonal antibody.

Most animal models of venous thrombosis involve acute thrombosis with hypercoagulability in small rodents. To better replicate human disease, we developed two models in the pig, a species similar to humans in size and in vascular and coagulation reactivity. One model involves de-endothelialisation with 50% or 80% stenosis and the other replacement of a venous segment by a Gore-Tex vascular prosthesis. Both models were tested with and without acute induced hypercoagulability (thromboplastin infusion). Thrombi obtained without thromboplastin infusion were composed of a multilayered platelet and a fibrin meshwork structure similar to that usually found in humans. With thromboplastin infusion, the thrombi were homogeneous fibrin structures imprisoning red blood cells. The high incidence of thrombosis obtained with the 80% stenosis model would be useful for studying anticoagulant treatments, whereas the low incidence with 50% stenosis would be useful for evaluating procoagulant effects of conditions or treatments. These new models shed further light on the development of venous thrombi under conditions similar to those seen in humans and may prove useful for investigating anticoagulant and procoagulant effects.

In vivo pig models of venous thrombosis mimicking human disease.

To describe a new type of stent consisting of a 3-dimensional (3D) braided tube made of 2 interconnected layers without any covering to treat a renal artery aneurysm.

Treatment of renal artery aneurysm with the multilayer stent.

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.

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

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 CD163(neg)/CD172a(neg), (2) the CD163(high)CD172a(pos) and (3) the CD163(low)CD172a(pos) 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 CD163(high) DDC resemble the antibody response-inducing human skin DC/macrophages whereas the CD163(neg)CD172(low) DDC share properties with the CD8(+) T cell response-inducing murine skin CD103(pos) 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.

Characterization of dendritic cells subpopulations in skin and afferent lymph in the swine model.

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.

Chantal Kang

Institut national de la recherche agronomique

Increasing Pulmonary Artery Pulsatile Flow Improves Hypoxic Pulmonary Hypertension in Piglets