title: Application of Controlled Shear Stresses on the Erythrocyte Membrane as a New Approach to Promote Molecule Encapsulation.
abstract: Human red blood cells (RBCs) have a remarkable capacity to undergo reversible membrane swelling. Resealed erythrocytes have been proposed as carriers and bioreactors to be used in the treatment of various diseases. This work is aimed at developing a setup allowing the encapsulation of test molecules into erythrocytes by inducing reversible pore formation on the RBC membrane through the application of controlled mechanical shear stresses. The designed setup consists of two reservoirs connected by a glass capillary. Each reservoir is connected to a compressor; during the tests, the reservoirs were in turn pressurized to promote erythrocyte flow through the capillary. The setup was filled with a suspension of erythrocytes, phosphate buffer, and FITC-dextran. Dextran was chosen as the diffusive molecule to check membrane pore dimensions. Samples of the suspension were withdrawn at scheduled times while the setup was operating. Flow cytometry and stereo-optical microscopy analyses were used to evaluate the erythrocyte dextran uptake. The setup was shown to be safe, well controlled, and adjustable. The outcomes of the experimental tests showed significant dextran uptake by RBCs up to 8%. Microscopy observations highlighted the formation of echinocytes in the analyzed samples. Erythrocytes from different donors showed different reactions to mechanical stresses. The experimental outcomes proved the possibility to encapsulate test molecules into erythrocytes by applying controlled mechanical shear stresses on the RBC membrane, encouraging further studies.
authors.affiliation: Department of Chemistry, Materials and Chemical Engineering "Giulio Natta,", Politecnico di Milano, Milan, Italy. giustina.casagrande@polimi.it.
authors.forename: Giustina
authors.initials: G
authors.lastname: Casagrande
authors.affiliation: Service of Immunohematology & Transfusion Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy.
authors.forename: Flavio
authors.initials: F
authors.lastname: Arienti
authors.affiliation: Service of Immunohematology & Transfusion Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy.
authors.forename: Arabella
authors.initials: A
authors.lastname: Mazzocchi
authors.affiliation: Service of Immunohematology & Transfusion Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Taverna
authors.affiliation: Service of Immunohematology & Transfusion Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy.
authors.forename: Fernando
authors.initials: F
authors.lastname: Ravagnani
authors.affiliation: Department of Chemistry, Materials and Chemical Engineering "Giulio Natta,", Politecnico di Milano, Milan, Italy.
authors.forename: MariaLaura
authors.initials: M
authors.lastname: Costantino
mesh_terms: D000328:Adult; D003911:Dextrans / Q000008:administration & dosage / Q000737:chemistry; D004058:Diffusion; D004337:Drug Carriers / Q000737:chemistry*; D065546:Drug Liberation; D004910:Erythrocyte Membrane / Q000737:chemistry*; D004912:Erythrocytes / Q000166:cytology*; D016650:Fluorescein-5-isothiocyanate / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000737:chemistry; D006801:Humans; D013314:Stress, Mechanical
chemical_list: D003911:Dextrans; D004337:Drug Carriers; C015219:fluorescein isothiocyanate dextran; D016650:Fluorescein-5-isothiocyanate
keywords: Drug carriers; Erythrocytes; Mechanical stress; Molecule encapsulation; Red cell membrane; Shear stress