1. Graphene-coating of NiTi
<ol>
 <li>The graphene samples used in this study were grown on copper (Cu) substrates using the chemical vapor deposition technique, and subsequently transferred to 4.5 mm2NiTi substrates.</li>
 <li>Cu foils (1 cm x 1 cm) were placed in a 1 in. quartz tube furnace and heated to 1,000 &deg;C in the presence of 50 sccm of H2 and 450 sccm of Ar.</li>
 <li>Next, methane (1 and 4 sccm) was introduced into the furnace at different flow rates for 20-30 min. Th...

<ol>
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No conflicts of interest declared.

Biocompatibility and cytotoxicity: The chemical vapor deposition (CVD) method yielded polycrystalline graphene samples that mimicked Cu crystal grains, as shown in Figure 1a. We employed Raman spectroscopy to confirm the presence of monolayer (few layer) graphene on 1 sccm (4 sccm) samples (see Figure 1b). Clearly, 1 sccm (4 sccm) samples exhibit intense (relatively weaker) G' band indicative of monolayer (few layer) graphene. Figure 1c shows an atomic forc...

The past three decades have witnessed discovery of novel materials-based therapies and devices for disease treatments and diagnostics. Novel alloy materials such as nitinol (NiTi) and stainless steel are often used in biomedical implant manufacturing due to their superior mechanical properties.1-3 However, numerous challenges remain due to exogenous material cytotoxicity, bio- and hemo-compatibility. The metallic nature of these alloys results in poor bio- and hemocompatibility due to metal leaching, lack of cell adhesion, proliferation, and thrombosis when it comes in contact with flowing blood (such as catheters, blood vessel grafts, vascular stents, artificial heart valves etc.). 1, 4, 5 The interaction of proteins or living cells with the implant surface can lead to a strong immunological response and the ensuing cascade of biochemical reactions may adversely affect the device functionality. Therefore, it is pertinent to achieve control over the interactions between biomedical implants and its surrounding biological environment. Surface modification is often employed to reduce or prevent the adverse physiological response originating from the implant material. An ideal surface coating is expected to have high adhesion strength, chemical inertness, high smoothness, and good hemo- and biocompatibility. Previously, numerous materials including diamond-like carbon (DLC), SiC, TiN, TiO2 and many polymer materials have been tested as bio-compatible implant surface coatings. 1, 6-23 However, these materials are still unable to meet all of the functional criteria for a suitable implant surface coating.
The discovery of atom thick layer of sp2 carbon, known as graphene, has opened doors for the development of novel multifunctional materials. Graphene is expected to be an ideal candidate for implant surface coating since it is chemically inert, atomically smooth and highly durable. In this Letter, we investigate the viability of graphene as a surface coating for biomedical implants. Our studies show that the graphene coated nitinol (Gr-NiTi) meets all of the functional criteria, and additionally supports excellent smooth muscle and endothelial cell growth leading to better cell proliferation. We also find that the serum albumin adsorption on Gr-NiTi is higher than fibrinogen. Importantly, (i) our detailed spectroscopic measurements confirmed the lack of charge transfer between graphene and fibrinogen suggesting that graphene coating inhibits platelet activation by implants, (ii) graphene coatings do not exhibit any significant in vitro toxicity for endothelial and smooth muscle cell lines confirming their biocompatibility, and (iii) graphene coatings are chemically inert, durable and impermeable in flowing blood environment. These hemo-and biocompatible properties, along with high strength, chemical inertness and durability, render graphene coatings as an ideal surface coating.

<table border="1">
 <tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Reagent</td>
 </tr>
 <tr>
 <td>Dulbecco's Modified Eagle Medium</td>
 <td>ATCC</td>
 <td>30-2002</td>
 <td></td>
 </tr>
 <tr>
 <td>Thiazolyl blue tetrazolium bromide</td>
 <td>Sigma-Aldrich</td>
 <td>M2128</td>
 <td></td>
 </tr>
 <tr>
 <td>CellTiter 96 Aqueous One solution cell proliferation assay (MTS)</td>
 <td>Promega</td>
 <td>G3582</td>
 <td></td>
 </tr>
 <tr>
 <td>Dimethyl sulfoxide</td>
 <td>Sigma-Aldrich</td>
 <td>D8418</td>
 <td></td>
 </tr>
 <tr>
 <td>36.5% formaldehyde</td>
 <td>Sigma-Aldrich</td>
 <td>F8775</td>
 <td></td>
 </tr>
 <tr>
 <td>Triton X-100</td>
 <td>Sigma-Aldrich</td>
 <td>T8787</td>
 <td></td>
 </tr>
 <tr>
 <td>Alexafluor 488 phalloidin</td>
 <td>Life Technologies</td>
 <td>A12379</td>
 <td></td>
 </tr>
 <tr>
 <td>VECTASHIELD mounting medium with DAPI</td>
 <td>Vector Laboratories</td>
 <td>H-1200</td>
 <td></td>
 </tr>
 <tr>
 <td>Human serum albumin</td>
 <td>Sigma-Aldrich</td>
 <td>A9511</td>
 <td></td>
 </tr>
 <tr>
 <td>Human fibrinogen</td>
 <td></td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Tris/Glycine/SDS</td>
 <td>Bio-Rad</td>
 <td>161-0732</td>
 <td></td>
 </tr>
 <tr>
 <td>Ready Gel Tris-HCl Gel</td>
 <td>Bio-Rad</td>
 <td>161-1158</td>
 <td></td>
 </tr>
 <tr>
 <td>Acetic acid</td>
 <td>Sigma-Aldrich</td>
 <td>45726</td>
 <td></td>
 </tr>
 <tr>
 <td>SYPRO Red</td>
 <td>Life Technologies</td>
 <td>S-6653</td>
 <td></td>
 </tr>
 <tr>
 <td>Protein low BCA assay</td>
 <td>Lamda Biotech</td>
 <td>G1003</td>
 <td></td>
 </tr>
 <tr>
 <td>Precision Plus Protein Kaleidoscope Standard</td>
 <td>Bio-Rad</td>
 <td>161-0375</td>
 <td></td>
 </tr>
 <tr>
 <td>Immun-Blot PVDF membrane</td>
 <td>Bio-Rad</td>
 <td>162-0177</td>
 <td></td>
 </tr>
 <tr>
 <td>Blotting grade blocker non-fat dry milk</td>
 <td>Bio-Rad</td>
 <td>170-6404XTU</td>
 <td></td>
 </tr>
 <tr>
 <td>Anti-actin antibody produced in rabbit</td>
 <td>Sigma-Aldrich</td>
 <td>A2066</td>
 <td></td>
 </tr>
 <tr>
 <td>BM Chemiluminescence Western Blotting kit (mouse/rabbit)</td>
 <td>Roche Applied Science</td>
 <td>11520709001</td>
 <td></td>
 </tr>
 <tr>
 <td>RIPA buffer</td>
 <td>Sigma-Aldrich</td>
 <td>R0278</td>
 <td></td>
 </tr>
 <tr>
 <td>NiTi (51% Ni, 49% Ti)</td>
 <td>Alfa-Aesar</td>
 <td>44953</td>
 <td></td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>Horiba JobinYvon</td>
 <td>Raman spectrometer</td>
 <td>Dilor XY 98</td>
 <td></td>
 </tr>
 <tr>
 <td>Nikon</td>
 <td>Confocal microscope</td>
 <td>Eclipse TI microscope</td>
 <td></td>
 </tr>
 <tr>
 <td>Thermoscientific</td>
 <td>Plate reader</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Bio-Rad</td>
 <td>Power supply</td>
 <td> 164-5050</td>
 <td>PowerPac basic power supply</td>
 </tr>
 <tr>
 <td>Bio-Rad</td>
 <td>Electrophoresis cell</td>
 <td>165-8004</td>
 <td>Mini-PROTEAN tetra cell </td>
 </tr>
 <tr>
 <td>Bio-Rad</td>
 <td>Gel holder cassette</td>
 <td> 170-3931</td>
 <td>Mini gel holder cassette</td>
 </tr>
 </tbody>
</table>

graphene coatings for biomedical implants

Atomically smooth graphene as a surface coating has potential to improve implant properties. This demonstrates a method for coating nitinol alloys with nanometer thick layers of graphene for applications as a stent material. Graphene was grown on copper substrates via chemical vapor deposition and then transferred onto nitinol substrates. In order to understand how the graphene coating could change biological response, cell viability of rat aortic endothelial cells and rat aortic smooth muscle cells was investigated. Moreover, the effect of graphene-coatings on cell adhesion and morphology was examined with fluorescent confocal microscopy. Cells were stained for actin and nuclei, and there were noticeable differences between pristine nitinol samples compared to graphene-coated samples. Total actin expression from rat aortic smooth muscle cells was found using western blot. Protein adsorption characteristics, an indicator for potential thrombogenicity, were determined for serum albumin and fibrinogen with gel electrophoresis. Moreover, the transfer of charge from fibrinogen to substrate was deduced using Raman spectroscopy. It was found that graphene coating on nitinol substrates met the functional requirements for a stent material and improved the biological response compared to uncoated nitinol. Thus, graphene-coated nitinol is a viable candidate for a stent material.

<img src="/files/ftp_upload/50276/50276fig1.jpg" alt="Figure 1"/> 
 Figure 1. a) CVD grown polycrystalline graphene on Cu foils mimics the metal crystal grains (scale bar: 10 &mu;m). b) Raman spectrum of 1 sccm (4 sccm) graphene shows intense (relatively weaker) G' band indicating monolayer (few layer) nature of as-prepared graphene. c) AFM image of graphene transferred on to NiTi shows a roughness of ~5 nm. Scale bar = 500 nm.
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Watch this Scientific Journal Video about Graphene Coatings for Biomedical Implants at JoVE.com

Graphene Coatings for Biomedical Implants

Graphene offers potential as a coating material for biomedical implants. In this study we demonstrate a method for coating nitinol alloys with nanometer thick layers of graphene and determine how graphene may influence implant response.

Atomically smooth graphene as a surface coating has potential to improve implant properties. This demonstrates a method for coating nitinol alloys with ...

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21.1K visualizações.  Clemson University. Graphene oferece um potencial como um material de revestimento para implantes biomédicos. Neste estudo, demonstrar um método para ligas de revestimento de nitinol com camadas de grafeno nanômetro de espessura e determinar como o grafeno pode influenciar a resposta do implante.