ABSTRACT
Polyelectrolyte microcapsules (PEMCs) have been prepared by coating red blood cells with the polyelectrolytes poly(styrenesulfonate), poly(allylamine hydrochloride), and dextran sulfate applying the layer-by-layer technique with subsequent dissolution of the core. The capsule permeability for human serum albumin (HSA) was studied as a function of the ionic strength and pH by means of confocal microscopy. PEMCs produced with dextran sulfate and poly(allylamine hydrochloride) show a significant increase in permeability for HSA at salt concentrations over 1 mM. For PEMCs prepared with poly(styrenesulfonate) and poly(allylamine hydrochloride) the limiting salt concentration is 5 mM. No pH dependence for permeation was observed. A correlation between the permeation and adsorption of HSA on the PEMC walls was investigated. Finally, a mechanism for the permeability, combining electrostatic interactions, and the presence of pores in the polymer layers is presented confirmed by the considerable increase of permeation of charged molecules in the presence of salt and the permeation of neutral molecules regardless of the ionic strength.
Subject(s)
Biopolymers/pharmacokinetics , Capsules/chemistry , Polymers/chemistry , Dextran Sulfate/chemistry , Erythrocytes , Humans , Hydrogen-Ion Concentration , Microscopy, Atomic Force , Osmolar Concentration , Permeability , Polyamines/chemistry , Polyelectrolytes , Polystyrenes/chemistry , Serum Albumin/pharmacokineticsABSTRACT
The electrophoretic mobility of native and glutaraldehyde-fixed bovine, human, and horse red blood cells (RBC) was investigated as a function of ionic strength (5-150 mM) and concentration of 464 kDa dextran (2 and 3 g/dl); RBC aggregation in autologous plasma and in dextran solutions was also measured. In agreement with previous observations, human and horse RBC form stable rouleaux whereas bovine RBC do not aggregate in either plasma or in dextran 464 kDa solutions. Electrophoretic measurements showed a species-dependent adsorption and depletion of dextran that can be theoretically evaluated. Adsorption of polymer is not a prerequisite for RBC aggregation (bovine RBC show the highest amount of adsorbed dextran yet do not aggregate). Aggregate formation thus occurs as long as the Gibbs free energy difference, given by the osmotic pressure difference between the bulk phase and the polymer-depleted region between two RBC, is larger than the steric and electrostatic repulsive energy contributed by the macromolecules present on the RBC surface. With increasing bulk-phase polymer concentration the depletion layer thickness decreases and the amount of adsorbed macromolecules increases, thereby resulting in an increase of the repulsive component of the interaction energy and decreased aggregation. We thus view electrophoretic measurements of RBC in various media as an important tool for understanding polymer behavior near the red cell surface and hence the mechanisms involved in RBC aggregation.
Subject(s)
Cattle/blood , Erythrocyte Aggregation , Horses/blood , Polymers/pharmacology , Adult , Animals , Blood Viscosity , Cell Culture Techniques , Dextrans/pharmacology , Electrophoresis , Erythrocyte Aggregation/drug effects , Erythrocytes/chemistry , Female , Humans , Male , N-Acetylneuraminic Acid/blood , Plasma , Species SpecificityABSTRACT
Microcapsules in the micrometer size range with walls of nanometer thickness are of both scientific and technological interest, since they can be employed as micro- and nano-containers. Liposomes represent one example, yet their general use is hampered due to limited stability and a low permeability for polar molecules. Microcapsules formed from polyelectrolytes offer some improvement, since they are permeable to small polar molecules and resistant to chemical and physical influences. Both types of closed films are, however, limited by their spherical shape which precludes producing capsules with anisotropic properties. Biological cells possess a wide variety of shapes and sizes, and, thus, using them as templates would allow the production of capsules with a wide range of morphologies. In the present study, human red blood cells (RBC) as well as Escherichia coli bacteria were used; these cells were fixed by glutardialdehyde prior to layer-by-layer (LbL) adsorption of polyelectrolytes. The growth of the layers was verified by electrophoresis and flow cytometry, with morphology investigated by atomic force and electron microscopy; the dissolution process of the biological template was followed by confocal laser scanning microscopy. The resulting microcapsules are exact copies of the biological template, exhibit elastic properties, and have permeabilities which can be controlled by experimental parameters; this method for microcapsule fabrication, thus, offers an important new approach for this area of biotechnology.
Subject(s)
Capsules , Drug Compounding/methods , Erythrocytes/ultrastructure , Escherichia coli/ultrastructure , Biotechnology , Electrolytes , Electrophoresis , Fluorescence , Humans , In Vitro Techniques , Microscopy, Atomic Force , Microscopy, Confocal , Microscopy, Electron, ScanningABSTRACT
The deformability and osmotic properties of hollow microcapsules were studied by means of the micropipette video microscopic technique. The microcapsules were prepared by consecutive multiple adsorption of the polyanion, poly(styrene sulphonate), and the polycation, poly(allylamine hydrochloride), onto melamine formaldehyde resin latex of 5 microm diameter, which was decomposed after completing the coating by transferring to hydrochloric acid of pH 1.1. The polyelectrolyte microcapsules reacted to micropipette suction with plastic deformation. If lipids are added to the polyelectrolyte layers, the capsules cannot be visibly deformed by micropipette suction up to 10(4) N/m2. However, plastic shrinking was observed if the stress was generated by the osmotic pressure of a sucrose solution of 10(6) N/m2.
