Ludka Machova

Perivascular sirolimus-delivery system.

Autologous vein grafts are often used for treating damaged vessels, e.g. arteriovenous fistulas or arterial bypass conduits. Veins have a different histological structure from arteries, which often leads to intimal hyperplasia and graft restenosis. The aim of this study was to develop a perivascular sirolimus-delivery system that would release the antiproliferative drug sirolimus in a controlled manner. Polyester Mesh I was coated with purasorb, i.e. a copolymer of L-lactide and ɛ-caprolactone, with dissolved sirolimus; Mesh II was coated with two copolymer layers; the layer with dissolved sirolimus was overlaid with pure purasorb. This arrangement allowed sirolimus to be released for 6 and 4 weeks, for Mesh I and Mesh II, respectively. Mesh II released sirolimus more homogeneously, without the initial burst effect during the first week. However, the cumulative release curve was steeper at later time points than the curve for Mesh I. Both meshes inhibited proliferation of rat vascular smooth muscle cells during 14-day culture in vitro and preserved excellent cell viability. Newly developed sirolimus-releasing perivascular meshes are promising devices for preventing autologous graft restenosis.

Capillary zone electrophoresis with external radial electric field control of electroosmotic flow and its application to the separation of synthetic oligopeptides

Abstract A new way of applying external radial electric field for the control of electroosmotic flow in capillary zone electrophoresis (CZE) has been developed. It is based on a new type of low-conductivity coating of the outer capillary surface by a polyaniline dispersion in hydroxypropylcellulose and on the application of a set of three high-voltage power supplies to form a constant radial electric field across the capillary wall as far as possible along the capillary length. Two power supplies are connected to the ends of the outer low-conductivity coating and the third one is applied to the ends of the inner capillary compartment filled with background electrolyte. The difference of electric potentials at the inner and outer capillary surface determines the voltage of radial electric field across the capillary wall and affects the electrokinetic potential at the solid–liquid interface inside the capillary. The effect of magnitude and polarity of external radial electric field on the flow-rate of electroosmotic flow, on the migration times of charged analytes (speed of analysis) and on the separation efficiency and resolution of CZE separations of synthetic oligopeptides, diglycine, triglycine, dalargin and dalargin–ethylamide has been evaluated. Application of external radial electric field has proved to be an efficient tool for regulation of electroosmotic flow in CZE and for optimization of migration times and resolution of oligopeptides in their CZE separations and analyses.

Effective long-term immunosuppression in rats by subcutaneously implanted sustained-release tacrolimus pellet: Effect on spinally grafted human neural precursor survival

Achievement of effective, safe and long-term immunosuppression represents one of the challenges in experimental allogeneic and xenogeneic cell and organ transplantation. The goal of the present study was to develop a reliable, long-term immunosuppression protocol in Sprague-Dawley (SD) rats by: 1) comparing the pharmacokinetics of four different subcutaneously delivered/implanted tacrolimus (TAC) formulations, including: i) caster oil/saline solution, ii) unilamellar or multilamellar liposomes, iii) biodegradable microspheres, and iv) biodegradable 3-month lasting pellets; and 2) defining the survival and immune response in animals receiving spinal injections of human neural precursors at 6 weeks to 3 months after cell grafting. In animals implanted with TAC pellets (3.4 mg/kg/day), a stable 3-month lasting plasma concentration of TAC averaging 19.1 ± 4.9 ng/ml was measured. Analysis of grafted cell survival in SOD+ or spinal trauma-injured SD rats immunosuppressed with 3-month lasting TAC pellets (3.4-5.1 mg/kg/day) showed the consistent presence of implanted human neurons with minimal or no local T-cell infiltration. These data demonstrate that the use of TAC pellets can represent an effective, long-lasting immunosuppressive drug delivery system that is safe, simple to implement and is associated with a long-term human neural precursor survival after grafting into the spinal cord of SOD+ or spinal trauma-injured SD rats.

Modification of Polylactide Surfaces with Lactide-Ethylene Oxide Functional Block Copolymers: Accessibility of Functional Groups

Feasibility of using amphiphilic block copolymers composed of polylactide (PLA) and poly(ethylene oxide) (PEO) blocks for biomimetic surface modification of polylactide-based biomaterials for tissue engineering was investigated. PEO-b-PLA copolymers were deposited on the PLA surface from a solution in PEO-selective solvent. Copolymers with a neutral omega-methoxy end group of the PEO block (mPEO-b-PLA) were used to provide hydrophilic surface of PLLA, which exhibited suppressed nonspecific protein adsorption. Their analogues, containing biotin group at the end of PEO block (bPEO-b-PLA), were used as a model of functional copolymers, carrying a biomimetic group, for example, a cell-adhesion fibronectine-derived peptide sequence. The surface topography of functional groups on the modified surface and their accessibility for interaction with a protein receptor was investigated, taking advantage of specific biotin-avidin interaction, on surfaces modified with a combination of mPEO-b-PLA and bPEO-b-PLA copolymers. The accessibility of model biotin groups for interaction with their protein counterpart was proven through visualization of avidin or avidin-labeled nanospheres with atomic force microscopy.

Dip TIPS as a facile and versatile method for fabrication of polymer foams with controlled shape, size and pore architecture for bioengineering applications.

The porous polymer foams act as a template for neotissuegenesis in tissue engineering, and, as a reservoir for cell transplants such as pancreatic islets while simultaneously providing a functional interface with the host body. The fabrication of foams with the controlled shape, size and pore structure is of prime importance in various bioengineering applications. To this end, here we demonstrate a thermally induced phase separation (TIPS) based facile process for the fabrication of polymer foams with a controlled architecture. The setup comprises of a metallic template bar (T), a metallic conducting block (C) and a non-metallic reservoir tube (R), connected in sequence T-C-R. The process hereinafter termed as Dip TIPS, involves the dipping of the T-bar into a polymer solution, followed by filling of the R-tube with a freezing mixture to induce the phase separation of a polymer solution in the immediate vicinity of T-bar; Subsequent free-drying or freeze-extraction steps produced the polymer foams. An easy exchange of the T-bar of a spherical or rectangular shape allowed the fabrication of tubular, open- capsular and flat-sheet shaped foams. A mere change in the quenching time produced the foams with a thickness ranging from hundreds of microns to several millimeters. And, the pore size was conveniently controlled by varying either the polymer concentration or the quenching temperature. Subsequent in vivo studies in brown Norway rats for 4-weeks demonstrated the guided cell infiltration and homogenous cell distribution through the polymer matrix, without any fibrous capsule and necrotic core. In conclusion, the results show the “Dip TIPS” as a facile and adaptable process for the fabrication of anisotropic channeled porous polymer foams of various shapes and sizes for potential applications in tissue engineering, cell transplantation and other related fields.