COMPOSITIONS AND METHODS FOR 3D PRINTED FIBROUS SCAFFOLDS WITH ANTIMICROBIAL PROPERTIES INCORPORATING GRAPHENE OXIDE AND POLY(E-CAPROLACTONE)

Abstract

A composition of Poly(e-caprolactone)—PLC—and Graphene Oxide (GO) for use in killing bacteria that cause infections in patients implanted with medical devices, for example Staphylococcus epidermidis and Escherichia coli. Also disclosed is a method for constructing PLC/GO fibers and fibrous scaffolds by additive manufacturing and wet spinning, employing the composition and for example 3D printing. The method and compositions can be developed to produce a fibrous scaffold in which fiber diameter and PLC/GO concentrations are such that GO sheets are incorporated but at the same time exposed at the polymer surface, coffering bactericidal properties to the material, while keeping biocompatibility. Also disclosed is a fibrous PLC/GO bactericidal scaffold and the implanted medical devices having such scaffold. The composition, method, scaffold and medical devices may be used to achieve PLC/GO scaffolds and medical devices with bactericidal properties that have reduced risk of implant-associated infections.

Claims

1. A composition of Poly(e-caprolactone)—PCL—and Graphene Oxide for use in killing bacteria that cause infections in patients implanted with medical devices comprising: a) A solvent consisting of Tetrahydrofuran (THF); b) Graphene Oxide (GO) at final concentration between 5% and 7.5% (w/w), most preferably 5% (w/w); c) Poly(e-caprolactone) at final concentration between 7.5% and 5% (w/v), most preferably 7.5% (w/v); d) A coagulation non-solvent consisting of ethanol.

2. The composition according to claim 1 wherein the said bacteria that causes infections in patients implanted with medical devices are selected from a list comprising Staphylococcus epidermidis and Escherichia coli.

3. The composition according to claim 1 wherein the said Poly(e-caprolactone) and Graphene Oxide polymer is constructed into a scaffold.

4. The composition according to claim 1 wherein the said Poly(e-caprolactone) and Graphene Oxide polymer is constructed into a fibrous scaffold.

5. A fibrous scaffold of Poly(e-caprolactone)—PCL—and Graphene Oxide for killing bacteria that cause infections in patients implanted with medical devices comprising: a) A solvent consisting of Tetrahydrofuran (THF); b) Graphene Oxide (GO) at final concentration between 5% and 7.5% (w/w), most preferably 5% (w/w); c) Poly(e-caprolactone) at final concentration between 5% and 7.5% (w/v), most preferably 7.5%; d) A coagulation non-solvent consisting of ethanol.

6. A-The fibrous scaffold according to claim 5 wherein the fibers have a diameter from 50 to 100 μm, most preferably of 100 μm.

7. The fibrous scaffold according to claim 4 wherein the said bacteria that causes infections in patients implanted with medical devices are selected from a list comprising Staphylococcus epidermidis and Escherichia coli.

8. The composition or the scaffold for use according to claim 1, wherein the said composition or scaffold is comprised in a medical device implanted in an animal or the human body.

9. A medical device comprising the composition or scaffold for use according to claim 1 wherein the said medical device is selected from the list consisting of surgical sutures, 3D scaffolds for tissue engineered implants as well as other non-limiting examples of implantable medical devices such as a stent, a vascular implant, a dental implant, and a bone implant.

10. A method for constructing a fibrous scaffold of Poly(e-caprolactone) and Graphene Oxide by additive manufacturing and wet-spinning of comprising the steps of: a) Dispersing graphene oxide (GO) in Tetrahydrofuran (THF), solvent at a final concentration of 5% w/w to 7.5% w/w, most preferably 5% w/w; b) Dissolving Poly(e-caprolactone)—PLC—in the previous mixture at a concentration of 7.5% to 15%, most preferably 7.5% c) Loading the previous mixture into a syringe or a 3D printer head d) Extruding the THF/PCL/GO mixture into a coagulation bath consisting of ethanol e) Rinsing PCL/GO scaffolds with ethanol and drying the scaffolds.

11. The method according to claim 10 wherein the said extrusion of THF/PCL/GO mixture is performed layer-by-layer in a 3D plotting machine or a 3D printer.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0064] FIG. 1: 3D model of the produced scaffolds.

[0065] Top view (where dx and dy are displayed) and cross-section view (where dz is shown and the staggering between layers is visible)

[0066] FIG. 2: Stereomicroscope images of obtained PCL scaffolds with 0%, 5% and 7.5% GO.

[0067] The lower lane represents zoomed regions of the upper lane images. Scale bar: top—5 mm; bottom (zoom)—500 μm.

[0068] PCL and composite PCL/GO fibrous scaffolds with approximately 1.2 cm×1.2 cm were printed, according to the 3D model previously disclosed in FIG. 1. The obtained 0% GO scaffolds (pristine PCL) are white, and the incorporation of GO within the polymeric matrix can be verified by the change in color as GO-containing scaffolds present a light brown (for 5% GO) and dark brown (for 7.5% GO) color. As can be observed, fibers are well defined and precisely plotted following the virtual design

[0069] FIG. 3: SEM analysis of the top views and cross-sections of PCL scaffolds with 0%, 5% and 7.5% GO.

[0070] Scale bar (from the top to the bottom row): top view—400 μm, 50 μm, 2 μm; bottom—200 μm, 50 μm.

[0071] FIG. 4: Antibacterial properties of PCL/GO fibrous scaffolds

[0072] S. epidermidis adhesion to PCL scaffolds with 0%, 5% and 7.5% GO, after 2 h and 24 h incubation in 10% v/v plasma supplemented TSB. Bacteria were stained with the LIVE/DEAD Backlight kit (ThermoFisher) and counted by confocal microscopy. Stacked bars graph with live and dead bacteria counting per 10.sup.4 μm.sup.2 of fiber is displayed. Statistically significant differences on the number of live and dead (*) bacteria compared to 0% GO scaffolds are indicated on top of the stacked bars (p=0.05; non-parametric Kruskal-Wallis test).

[0073] FIG. 5: In vitro biocompatibility assessment

[0074] Representative confocal microscopy images of HFF-1 human fibroblasts interaction with PCL scaffolds with 0%, 5% and 7.5% GO, after 1 day and 7 days of culture. Cells are identified by staining the nuclei with DAPI (DNA) and cytoskeleton (F-actin staining). The scaffold fibres can be identified by the faint-clear dashed lines. Images represent single plane projections of a 150 μm height z-stack. Scale bar: 100 μm.

[0075] The following references should be considered herewith incorporated in their entirety.

[0076] The following claims further set out embodiments of the invention.

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