Difference between revisions of "Team:Pasteur Paris/Membrane"

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                 <h1 id="Introduction">INTRODUCTION</h1>
 
                 <h1 id="Introduction">INTRODUCTION</h1>
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                 <p><i>When manipulating genetically engineered organisms, it is crucial to guarantee the confinement of these organisms. In our case, we want the genetically modified bacteria to stay at the interface between the prosthesis and the external organic medium. At the same time, one of the main issues our project wants to tackle is the conduction of the neuron influx to the prosthesis. The answer to these questions came as a double solution: confinement of the bacteria by conductive nanoporous membranes. The membrane’s nanoporosity allows substances produced by our modified biofilm to pass through the membrane, but the bacteria remain confined. We tested the conductivity and biocompatibility of two types of membranes.</i></p>
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                 <p><i>When manipulating genetically engineered organisms, it is crucial to guarantee the confinement of these organisms. In our case, we want the genetically modified bacteria to stay at the interface between the prosthesis and the external organic medium. At the same time, one of the main issues our project wants to tackle is the conduction of the neuron influx to the prosthesis. The answer to these questions came as a double solution: confinement of the bacteria by conductive nanoporous membranes. The membrane’s nanoporosity allows substances produced by our modified biofilm to pass through the membrane, but the bacteria remain confined. We tested the conductivity and biocompatibility of two types of membranes.</i></p></div>
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                 <img src="https://static.igem.org/mediawiki/2018/d/df/T--Pasteur_Paris--Membrane-intro.png">
 
                 <img src="https://static.igem.org/mediawiki/2018/d/df/T--Pasteur_Paris--Membrane-intro.png">
 
                 <div class="legend"><b>Figure 1: </b>Bacteria + Conductive Nanoporous Membrane = Confined Bacteria</div>
 
                 <div class="legend"><b>Figure 1: </b>Bacteria + Conductive Nanoporous Membrane = Confined Bacteria</div>

Revision as of 15:56, 14 October 2018

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INTRODUCTION

When manipulating genetically engineered organisms, it is crucial to guarantee the confinement of these organisms. In our case, we want the genetically modified bacteria to stay at the interface between the prosthesis and the external organic medium. At the same time, one of the main issues our project wants to tackle is the conduction of the neuron influx to the prosthesis. The answer to these questions came as a double solution: confinement of the bacteria by conductive nanoporous membranes. The membrane’s nanoporosity allows substances produced by our modified biofilm to pass through the membrane, but the bacteria remain confined. We tested the conductivity and biocompatibility of two types of membranes.

Figure 1: Bacteria + Conductive Nanoporous Membrane = Confined Bacteria

Gold-coated membranes

Sterlitech Polycarbonate Gold-Coated Membrane Filters were the first membranes we tested. The pores have a diameter of 0.4 micrometer, which is small enough to confine E. coli bacteria, which diameter and size are respectively about 1 micrometer and 2 micrometers. These membranes were relatively easy to manipulate with a forceps because of their high flexibility.

Figure 2: Gold-Coated Membrane
Figure 3: Gold-Coated Membrane (Electron Microscope)

Polymer-coated membranes

The other membranes were Sterlitech Alumina Oxide Membrane Filters with 0.2 micrometer pores. Their higher rigidity compared to the gold-coated membranes led to several membranes being broken while manipulating them with a forceps. We used these membranes as a support for different conductive and biocompatible polymers: PEDOT:PSS (poly(3,4-ethylenedioxythiophene) polystyrene sulfonate), PEDOT:Cl and PEDOT:Ts.

Figure 4: Alumnia Oxyde Membrane in grey
Figure 5: Alumnia Oxyde Membrane in grey (electron microscope)

For PEDOT:PSS, an aqueous solution of PEDOT:PSS was prepared (Jikui Wang, Guofeng Cai, Xudong Zhu, Xiaping Zhou, Oxidative Chemical Polymerization of 3,4-Ethylenedioxythiophene and its Applications in Antistatic coatings, Journal of Applied Polymer Science, 2012, Vol. 124, 109-115 .) and alumina oxide membranes were dipped for 24 hours in this solution. Electron microscopy of the membranes before and after the experiment showed the deposit of a substance on their surface; however its nature hasn’t been tested.

Figure 6: PEDOT:PSS-coated membrane
Figure 7: PEDOT:PSS-coated membrane (electron microscope)

Vapor-phase polymerization of PEDOT:Cl and PEDOT:Ts (Alexis E. Abelow, Kristin M. Persson, Edwin W.H. Jager, Magnus Berggren, Ilya Zharov, Electroresponsive Nanoporous Membranes by Coating Anodized Alumina with Poly(3,4ethylenedioxythiophene) and Polypyrrole. 2014, 299, 190-197.) also induced a change in the surface of the membranes (its exact nature also hasn’t been verified).

Figure 8: PEDOT:TS (left) / PEDOT:CL (right) - coated membranes
Figure 9: PEDOT:CL - coated membrane (electron microscope)

Biocompatibility

The first issue to tackle for such an interface is its biocompatibility, so its ability to coexist with a living organism. Experiments in self-made PDMS culture wells with E. coli showed a low biocompatibility for the gold coated membrane, but an enhanced biocompatibility for the polymer-coated membranes.

Conductivity

The second criterion for a fully functional interface is its ability to conduct a neuron’s influx. Thus, conductivity measurements were made for signals of different frequencies on the membranes. Results showed excellent conductive properties for the gold-coated membranes and very good conductive properties for the polymer-coated membranes.

CONCLUSION

Biocompatible polymers like PEDOT:PSS represent ideal materials for engineering biocompatible and conductive interfaces, that are also relatively easy to produce, thus making them our preferred choice in our project. However, it is worth mentioning that we are totally aware of the fact that we can’t just expect neuron axons to bind to our interface and produce an electric signal. The electric signal transmitted by a nerve is heavily limited to the interior of the nerve by myelin covering the axon, and the signal transmitted by the axon is purely chemical. So it requires special electrodes, like Fine or Cuff electrodes, to detect an electric signal. We might explore these solutions in the continuation of our project to enhance our interface’s ability to transmit neuron signals.

Gold-coated membranes