The Influence of Nanofiller Shape and Nature on the Functional Properties of Waterborne Poly(urethane-urea) Nanocomposite Films.

A series of waterborne polycarbonate-based poly(urethane-urea) nanocomposite films were prepared and characterized. An isocyanate excess of 30 mol% with respect to the hydroxyl groups was used in the procedure, omitting the chain-extension step of the acetone process in the dispersion preparation. The individual steps of the synthesis of the poly(urethane-urea) matrix were followed by nuclear magnetic resonance (NMR) spectroscopy. The nanofillers (1 wt% in the final nanocomposite) differed in nature and shape. Starch, graphene oxide and nanocellulose were used as representatives of organic nanofillers, while halloysite, montmorillonite, nanosilica and hydroxyapatite were used as representatives of inorganic nanofillers. Moreover, the fillers differed in their shape and average particle size. The films were characterized by a set of methods to obtain the tensile, thermal and surface properties of the nanocomposites as well as the internal arrangement of the nanoparticles in the nanocomposite film. The degradation process was evaluated at 37 °C in a H2O2 + CoCl2 solution.

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.

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.

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.

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.

Adhesion and growth of vascular smooth muscle cells in cultures on bioactive RGD peptide-carrying polylactides.

The surface of poly(L-lactide) (PLLA) films deposited on glass coverslips was modified with poly(DL-lactide) (PDLLA), or 1:4 mixtures of PDLLA and PDLLA-b-PEO block copolymers, in which either none, 5% or 20% of the copolymer molecules carried a synthetic extracellular matrix-derived ligand for integrin adhesion receptors, the GRGDSG oligopeptide, attached to the end of the PEO chain. The materials, perspective for vascular tissue engineering, were seeded with rat aortic smooth muscle cells (11,000 cells/cm(2)) and the adhesion, spreading, DNA synthesis and proliferation of these cells was followed on inert and bioactive surfaces. In 24-h-old cultures in serum-supplemented media, the number of cells adhering to the PDLLA-b-PEO copolymer was almost eight times lower than that on the control PDLLA surface. On the surfaces containing 5% and 20% GRGDSG-PEO-b-PDLLA copolymer, the number of cells increased 6- and 3-fold respectively, compared to the PDLLA-b-PEO copolymer alone. On PDLLA-b-PEO copolymer alone, the cells were typically round and non-spread, whereas on GRGDSG-modified surfaces the cell spreading areas approached those found on PDLLA, reaching values of 991 microm(2) and 611 microm(2) for 5% and 20% GRGDSG respectively, compared to 958 microm(2) for PDLLA. The cells on GRGDSG-grafted copolymers were able to form vinculin-containing focal adhesion plaques, to synthesize DNA and even proliferate in a serum-free medium, which indicates specific binding to the GRGDSG sequences through their adhesion receptors.

Adsorption of poly(ethylene oxide)-block-polylactide copolymers on polylactide as studied by ATR-FTIR spectroscopy.

In this study, the adsorption of amphiphilic poly(ethylene oxide)-block-polylactide (mPEO-PLA) copolymers from a selective solvent onto a polylactide surface was studied as a method of polylactide surface modification and its effect on nonspecific protein adsorption was evaluated. A series of well defined mPEO-PLA copolymers was prepared to investigate the effect of copolymer composition on the resulting PEO chain density and on the surface resistance to protein adsorption. The copolymers contained PEO blocks with molecular weights ranging between 5600 and 23,800 and with 16-47 wt% of PLA. The adsorption of both the copolymers and bovine serum albumin was quantified by attenuated total reflection FTIR spectroscopy (ATR-FTIR). In addition to the adsorbed copolymer amount, its actual composition was determined. The PEO chain density on the surface was found to decrease with the molecular weight of the PEO block and to increase with the molecular weight of the PLA block. The adsorbed copolymers displayed the ability to reduce protein adsorption. The maximum reduction within the tested series (by 80%) was achieved with the copolymer containing PEO of MW 5600 and a PLA block of the same MW.

Biodegradable copolymers carrying cell-adhesion peptide sequences.

Amphiphilic block copolymers are used to create bioactive surfaces on biodegradable polymer scaffolds for tissue engineering. Cell-selective biomaterials can be prepared using copolymers containing peptide sequences derived from extracellular-matrix proteins (ECM). Here we discuss alternative ways for preparation of amphiphilic block copolymers composed of hydrophobic polylactide (PLA) and hydrophilic poly(ethylene oxide) (PEO) blocks with cell-adhesion peptide sequences. Copolymers PLA-b-PEO were prepared by a living polymerisation of lactide in dioxane with tin(II)2-ethylhexanoate as a catalyst. The following approaches for incorporation of peptides into copolymers were elaborated. (a) First, a side-chain protected Gly-Arg-Gly-Asp-Ser-Gly (GRGDSG) peptide was prepared by solid-phase peptide synthesis (SPPS) and then coupled with delta-hydroxy-Z-amino-PEO in solution. In the second step, the PLA block was grafted to it via a controlled polymerisation of lactide initiated by the hydroxy end-groups of PEO in the side-chain-protected GRGDSG-PEO. Deprotection of the peptide yielded a GRGDSG-b-PEO-b-PLA copolymer, with the peptide attached through its C-end. (b) A protected GRGDSG peptide was built up on a polymer resin and coupled with Z-carboxy-PEO using a solid-phase approach. After cleavage of the delta-hydroxy-PEO-GRGDSG copolymer from the resin, polymerisation of lactide followed by deprotection of the peptide yielded a PLA-b-PEO-b-GRGDSG block copolymer, in which the peptide is linked through its N-terminus.