Electrochemical sensor for monitoring levels of oxygen and nitrogen reactive species to benefit ageing research

Lead Research Organisation: University of Brighton
Department Name: Sch of Pharmacy & Biomolecular Sciences

Abstract

This proposal is focused on the development of a novel microelectrode that can be tailored for any specific chemical post biological and chemical modification. These microelectrodes will be made using carbon nanotube composite material, which allows the means to have a varied electrode area. These sensors can be made using two different strategies, where they can be held in either glass capillaries or plastic pipettes, allowing for sensors to be rigid or flexible. This is a rapid improvement compared to conventionally made carbon based electrodes.
These sensors will be modified for the detection of various reactive oxygen and nitrogen species (ROS/RNS). These chemicals are of specific interest as they play key biological roles. They have been implicated in the ageing process and are the basis of many theories associated with the ageing process. At present, only total ROS/RNS have been detected with current technology, as the fluorescent dyes that are used offer poor specificity. This device will be made to benefit researchers working on ageing and is anticipated to increase the impact of these studies.
Measurements will be made using microelectrode arrays, which will be fabricated using boron-doped diamond. These microelectrode arrays will be modified for detection of ROS/RNS. Our microelectrodes are advantageous as they allow simultaneous measurements of ROS/RNS over various locations.
The ROS/RNS microelectrode and MEAs will be tested on two applications associated with ageing. The first will be to characterise the changes in ROS/RNS levels over various regions of old and young isolated neurones. The second application will be to look at ROS/RNS changes in cell lines that enter senescence. Measurements will be carried out in comparison with fluorescent dyes.
Finally to allow access to this new sensor technology for bioscience researchers, we will host a workshop that will allow scientist to learn about how the sensor functions and how it can be utilised for monitoring ROS/RNS levels from biological samples.
Overall this new sensing strategy will provide impact to studies associated with ageing, but with the main electrode having the ability to be modified for any specific analyte, the long-term benefits can be for various sectors and fields of importance. It is envisioned that this microelectrode can be tailored for detection of key analytes for major priority areas such as food security, understanding animal physiology and healthy ageing.

Technical Summary

This proposal is aimed at developing a radical new approach for the fabrication of carbon based microelectrodes. Such electrodes can be made, where the electrode size can be modified to any specific size, and also allows for the means to create rigid or flexible devices based on the application. This will be used alongside a novel microelectrode array (MEA) that will be fabricated using boron-doped diamond. Both the microelectrodes and MEA will be modified to allow for dynamic detection of various reactive oxygen and nitrogen species (ROS/RNS). ROS/RNS were chosen as the analyte to modify this sensor as they have been implicated in the ageing process. These sensors will be aimed initially towards bioscience researchers focusing on understanding the process of ageing. The MEA and microelectrode will be assessed on two specific applications. The ROS/RNS microelectrode will be from both division competent and senescent cells, whilst the MEA will be used to simultaneously record levels of various ROS/RNS from anatomical regions of an isolated neurone. Most important, the working of the sensor will be disseminated to researchers working on ageing through a hands-on workshop, which will provide scientists will information on how the microelectrode works and the level of information they can obtain from this.

Planned Impact

We anticipate that the results of this work will have important impact for several groups including the public health sector, commercial sector and the third sector. We will engage in a range of activities to maximise the impact of our research in these groups in the following ways:

The Public Health Sector: The developed device will have limited impact to the public health sector for the duration of the proposal. However the device will be aimed initially toward a major area of research, which is the ageing population. In the short to medium term, the device will provide impact on ageing research studies and thus will provide vital insight on the ageing process. It is anticipated in the long-term that these studies will yield new knowledge that can direct better pharmacological interventions. These developments will have a great impact on the public healthcare service, as the ageing population is anticipated to have a great economic impact on the healthcare service.

The Commercial Private Sector: The sensor developed in the proposed study encompasses new strategies for the fabrication of microelectrodes. Thus the device itself and components of the device may need to be protected. Intellectual property (IP) arising from this project will be protected and exploited by the University of Brighton Business Development and Enterprise Department. The University is well networked with industry and has a history of cooperation with the Pharmaceutical and Biotech Industries. We will be working closely with partners from these sectors to maximise the economic and social impact of this project.
Initially in the short term we will provide a range of sensors for bioscience researchers who have already expressed an interest in our work. This device will be made available for scientists not only working in ageing research but for researchers with an interest in monitoring analytes in a wide range of disciplines. Other sensors have been developed within the group for a gastrointestinal researcher, so we have a trac record for providing electrochemical devices for bioscience researchers. To increase the outreach of our device, we will work alongside industrial links, which have a track record for the robust production of the sensor devices.

Third Sector: We will endeavour to disseminate our work within relevant third sector organisations (Age UK). RGF is on the Board of Governors for the local Age UK group, and thus will be able to disseminate our findings.

General Public.
The PI and CoIs have already been actively engaged in presenting research to the public and will continue to do so with regards to the findings from the proposed project.
MSY has previously given public lectures to Age (UK), and his work has been widely publicised via both the local (Argus) and national press (e.g. Times Higher Supplement). His recent work has also been published on both national (South Today) and international (Normandy) television. In addition MSY and BAP have compiled a short film on their research, which is available through YouTube and the University of Brighton's website. BAP is an active STEM Ambassador and would use this role to highlight the proposed research.
 
Description Within this project we have developed a sensor device and methodology that was capable for the detection of multiple reactive oxygen and nitrogen species in biological tissues. The sensor was evaluated to show the changes in these important molecules which have been implicated with ageing in the central nervous system of an invertebrate model system. The design of the sensor allows this approach to modified towards any given geometry and therefore provides high flexibility in applications for its use. We were fully able to meet the objectives of this proposal as we developed the device and showcased its potential at the British Society for Research into Ageing annual conference, where we presented data from the device. This was followed up by a one-day workshop event which allowed participants to gain an in depth understand on how the sensor worked and gain hands on experience of sensing.
Exploitation Route The findings of this work have provided the platform for a novel sensing platform that can be used by anyone keen to monitor these chemicals in a robust fashion. We have already worked with acedemic collaborators to use this device for the monitoring multiple reactive oxygen and nitrogen species from cancer tissue and evaluate the efficacy of chemotherapy, which was funded by CRUK. We are also working with other researchers who are keen to use this approach for monitoring changes in the cardiovascular system.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description Our finding have been used to support researcher who wish to monitor ROS/RNS in their investigations. We have made the technology available and promoted this through the leading conference for ageing researchers and anticipate supporting investigations and studies to raise impact of ageing research. We have already worked with acedemic collaborators to use this device for the monitoring multiple reactive oxygen and nitrogen species from cancer tissue and evaluate the efficacy of chemotherapy, which was funded by CRUK.
First Year Of Impact 2015
Sector Healthcare,Pharmaceuticals and Medical Biotechnology
Impact Types Economic

 
Title Development of ROS sensors 
Description Development of a reactive oxygen/nitrogen species sensor for monitoring from single cells and tissues. This was created to offer a new tool to benefit research for ageing science 
Type Of Material Physiological assessment or outcome measure 
Year Produced 2014 
Provided To Others? Yes  
Impact This tool has been used to study ROS/RNS signalling levels in breast cancer cells and invertebrates 
 
Description BigBang STEMfest event 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Attended an event to highlight the issues associated with food in Physiology in the local area.

We highlighted the functions and importance of the digestive tract and how this alters and our limited understanding of this process to school children.

Beneficiaries: School children and member of the public

Contribution Method: The research directed our finding on GI function and thus allowed us to show novel understanding about the relationship between signalling and motility.

The research directed our finding on GI function and thus allowed us to show novel understanding about the relationship between signalling and motility.
Year(s) Of Engagement Activity 2012,2013,2014,2015
 
Description Conference Presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Presentation of key science at an international conference.
Year(s) Of Engagement Activity 2012,2013,2014,2015
 
Description Conference Presentation - Pittcon 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Type Of Presentation paper presentation
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Presentation of an oral abstract and symposium presentation at the leading analytical chemistry conference.

None
Year(s) Of Engagement Activity 2006,2007,2008,2009,2010,2011,2012,2013,2014,2015