In vitro association of type III collagen with elastin and with its solubilized peptides.

Elastin fibres or elastin solubilized peptides associated with type III collagen, under physiological conditions, to form a gel-like substance. The gel cohesion could be markedly improved either by desiccation process (gently air-dried rather than lyophilized) or by addition of adhesive connective proteins such as fibronectin, laminin and type IV collagen. They conferred to the material the composition and the structural appearance of typical basement membrane-like structure. Their properties enable us to investigate the biomaterial in several domains.

Microencapsulation of isolated pituitary cells by polyacrylamide microlatex coagulation on agarose beads.

Microlatex beads of homogenous size were made by polymerization of a mixture of acrylamide/bisacrylamide dispersed in a microemulsion. The microlatex was aggregated by dilution of the microemulsion in acrylamide solutions. The aggregates were then coagulated by polymerization at the interfaces of agarose beads circulating in a capillary tube containing paraffin oil. Biocompatibility was tested on isolated pituitary cells microencapsulated by this procedure.

Control and isotopic quantification of affinity of antithrombin III for heparin-like surfaces.

Heparin-like materials, characterized by a defined superficial density of functional groups which activate antithrombin III (AT III), when in contact with blood specifically inhibit thrombin as soon as it appears. This paper describes an isotopic method to estimate this density and to visualize the distribution of the affinity sites concerned, both directly with AT III labelled with 125Iodine and indirectly with an anti AT III monoclonal antibody labelled with 111Indium.

Experimental evaluation of a collagen-coated vicryl mesh as a dural substitute.

Dural substitutes must provide immediate restitution of a membranous covering for the brain without inducing any adverse reaction in the host or provoking adhesions to underlying nervous tissues. Ideally, the material should disappear completely and be replaced by tissues similar to the dura mater. In this study, parietal dural defects were created in 12 beagle dogs and closed with a vicryl mesh prosthesis made watertight by a film of bovine collagen. Clinical and biological tolerances were satisfactory. There was one case of early local infection. Gross and microscopic examinations performed between 7 days and 9 months after implantation showed degradation of the prosthetic mesh and connective tissue growth into the implant mimicking dura mater as early as 15 days after implantation. There was no attendant inflammatory reaction or cortical adhesions or other adverse reactions. The authors conclude that collagen-embedded vicryl mesh provides satisfactory biological function and compatibility when used as a substitute for dura mater in the dog.

ESCA studies on heparin-like materials used for extracorporeal shunts.

Amino-acids sulfamide substituted polystyrene surfaces exhibit an heparin-like activity. Similar treatments were achieved on small diameter tubings made of polystyrene grafted on polyethylene. Electron spectroscopy for Chemical Analysis (ESCA) appeared as the suitable method for a chemical characterization of the surface (approximately 50-100 A depth) to be in contact with circulating blood. These tubings were implanted as extracorporeal shunts on dogs. The variations of local concentrations of labelled platelets, red cells and fibrinogen were recorded in situ. Depending on the conditions of preparation and implantation, a slight adhesion of platelet was observed. By using such type of materials with an improved mechanical compliance, small diameter antithrombogenic tubings could be developed, provided the chlorosulfonation prior to amino-acid grafting is mild.

An ex vivo original test using radiotracers for evaluating haemocompatibility of tubular biomaterials.

The haemocompatibility of a vascular prosthesis can be estimated as the result of its interaction with blood components. The authors describe an ex vivo canine shunt for evaluating isotopic haemocompatibility in blood-wall interactions. Methods employing radioisotopic tracers can be used to dynamically monitor the adsorption of labelled blood cells and proteins on different biomaterial surfaces. This ex vivo test should enable materials to be assessed for quality according to two thrombogenic criteria: (i) number of adhered platelets mm-2 s-1; (ii) quantity of adsorbed fibrinogen expressed as microgram mm-2 s-1, which would provide the basis for a scale of haemocompatibility.

In situ polymerization of a microencapsulating medium round living cells.

A new process for microencapsulating living cells is described. Pancreatic islet cells in solution in agarose were extruded from a syringe into a paraffin-oil-containing medium. A rigid interface around each bead was produced when the agarose gelled. A polymerizable mixture of monomers containing 30% of acrylamide and 1.5% of bisacrylamide was then added to these beads. Polymerization was initiated by a photochemical process using riboflavin as a photosensitizer, which enabled use of oxygenated solutions. As other polymerizations initiated by an oxido-reduction reaction, this method did not require heating, and could thus be carried out at ambient temperature. The oil phase near the agarose beads effectively prevented any rise in temperature, and also led to formation of an acrylamide concentration gradient in the aqueous phase, which reduced the potential cytotoxicity of the reaction. Radioimmunoassay of insulin liberated from encapsulated islets of Langerhans demonstrated a good viability of the cells.

[Preservation of the functional integrity of cellular material after cell microencapsulation by polymerization]. / Préservation de l'intégrité fonctionnelle d'un matériel cellulaire après enrobage par polymérisation.

A new process for microencapsulating living cells is described. A cross linked network was polymerized using acrylamide and bisacrylamide monomers round agarose-embedded cells in an hydrophobic medium. The reaction was initiated by excitation of Vit. B2 under visible irradiation. The absence of toxic effect was evaluated from the insulin secretion of encapsulated islets of Langerhans.

Crosslinking with diacrylylpiperazine (PIP) reduces activation of complement by polyacrylamide microcapsules.

Activation of the complement system in vitro by a series of microcapsules based on polylysine, alginate or polyacrylamide was determined. Least activation was observed with microcapsules whose outer layer was composed of polyacrylamide. Activation was further reduced using diacrylylpiperazine instead of bisacrylamide as crosslinker.

Topographic and dynamic radioisotopic investigations of interactions between biomaterials and blood components.

Functional investigations using radionuclides to study protein adsorption and platelet adhesion onto biomaterials are described. The authors' novel methods use radiotracers with a dynamic technique. This allows direct observation of the interaction between blood, or simpler biological substances, and artificial materials. Several radiotracers were used in this study, including 111In-platelets, 123I-fibrinogen, 123I-antithrombin III, and 99mTc-tagged red blood cells. The detectors employed were a semiconductor diode or gamma-camera equipped with special collimators. The acquisition and treatment of data were performed with an original device. These methods allowed precise comparisons, especially between platelet adhesion upon different materials in the form of tubes and also protein adsorption and desorption. The results are discussed in terms of materials to be used for work in vivo.

Agarose encapsulation of islets of Langerhans: reduced toxicity in vitro.

The physical and chemical properties of agarose have been exploited for encapsulation of islets of Langerhans. Formation of microcapsules by extrusion of a hydrophilic polymer (agarose) into a hydrophobic solution created an interface to which an immunoprotective membrane could be polymerized. The substances and procedure used were devoid of cellular toxicity. Islet function in vitro was found to be normal.

In vitro diffusion in polyacrylamide embedded agarose microbeads.

125I Sodium iodide, 125I insulin, 125I albumin, and 111indium IGG were employed to investigate release from, and penetration of different sized molecules into agarose/polyacrylamide microcapsules. The microcapsules were formed by photopolymerization of an acrylamide solution round agarose beads. The indium-chelated antibody gave a particular low background count. The different release times were explained in terms of differences in diffusion coefficient. By retarding in vitro penetration of antibodies, these microcapsules could be of value for the encapsulation of living cells in bioartificial organs.

In vitro evaluation of an epoxy resin's cytocompatibility using cell lines and human differentiated cells.

The cytocompatibility of a polyepoxy resin (Elf Aquitaine) has been studied using both cell lines and human differentiated cell cultures. The human models were gingival fibroblasts and bone osteoblasts, while the cell lines were Hela cells and 3T3 Balb/c cells. Basal cytocompatibility was assessed by estimation of the cell proliferation, total cell protein content, cell membrane sub-lysis, and cell attachment and spreading. Specific cytocompatibility concerning human osteoblasts, from both alveolar and trabecular bone, was determined by measuring the intracellular alkaline phosphatase activity. Resin colonization by the cells was studied by both TEM and SEM. The behaviour of the two cell lines reveals a significant level of discrepancy, whereas the behaviour of human cells, whatever the model, is comparable; however, osteoblasts look more sensitive. Moreover, the results show that this epoxy resin exhibits a moderate cytocompatibility which could be the result of the cytotoxicity of early released products, associated with the considerable surface roughness.

Biocompatibility of polyacrylamide microcapsules implanted in peritoneal cavity or spleen of the rat. Effect on various inflammatory reactions in vitro.

The biocompatibility of polyacrylamide microcapsules was investigated by implanting microcapsules in the peritoneal cavity or the spleen of rats. The capsules were retrieved every four weeks for twenty weeks. They remained isolated and free in the peritoneal cavity, but led to a slight inflammatory reaction in the spleen. These results were confirmed, in vitro, by determinations of chemiluminescence and interleukin I levels, and by evaluation of attachment and growth of human fibroblasts. The microcapsules did not enhance these inflammatory reactions, and they were a poor support for cellular proliferation.