SCARAB

Sheffield Collaboratorium for Antimicrobial Resistance And Biofilms (SCARAB) brings doctors, scientists and engineers together to find new ways of fighting biofilm formation.

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Harmful bacteria form complex communities, called biofilms, as a normal part of the infection process. If these form in human tissues, it can lead to long-term (chronic) infection, with increasing levels of resistance to normal antibiotics. We have created a facility (SCARAB) that brings doctors, scientists and engineers together to find new ways of fighting biofilm formation. Pharmaceutical companies and NHS researchers are invited to contribute new ways of testing their therapies in infected tissues, leading to the faster development of the urgently needed therapies for chronic infections in the growing numbers of elderly and diabetic patients now living in the UK.

What hinders the UK’s ability to tackle antimicrobial resistance (AMR)?

Under-appreciation of the role biofilms contribute to AMR prevents stewardship of existing antimicrobials:

No existing facility for the study of biofilms in human tissues
Cost, technological and scientific barriers due to the lack of a platform for screening of novel AM.

SCARAB will help achieve the three aims of the UK’s five year AMR Strategy

Improve the knowledge and understanding of AMR
Conserve and steward the effectiveness of existing treatments
Stimulate the development of new antibiotics, diagnostics and novel therapies.

Our solution

An advanced platform for higher throughput screening of novel AM which simulates accurately biofilm infections in physiologically relevant conditions in 3D tissue models.

UK 5 year antimicrobial resistance strategy

Research

SCARAB brings together equipment and expertise to investigate two complex, interdependent systems: biofilms and human tissues. The new drugs and treatments for infections that are necessary because of the rise of Antimicrobial Resistance (AMR) must work in the face of this complexity and SCARAB is an essential infrastructure component for their development.
Biofilms are known to protect microorganisms from antibiotics, due to limited penetration, slowed growth, adaptive responses and 'persister' cells. Similarly, the location of an infection within a complex tissue can afford a degree of protection to the organisms concerned. Biofilm research currently involves a number of excellent research groups in Universities across the UK as well as industrial partners and charity organisations, as illustrated below. By providing experimentally-tractable systems of biofilm infection of skin, bladder, oral mucosa and the cornea, SCARAB will enable academia and industry to more rapidly progress through the phases of drug development to combat AMR. In particular, we expect that animal usage will be reduced if human tissue models are more easily available, with higher throughput and greater biological accuracy of the tissue models making subsequent animal studies better targeted and with improved knowledge of potential toxicity.

Tissue Engineering - a strength at Sheffield

Development of bacterially infected cornea organ cultures. Pinnock, A., Shivshetty, N., Roy, S. et al. Ex vivo rabbit and human corneas as models for bacterial and fungal keratitis. Graefes Arch Clin Exp Ophthalmol (2017) 255:333–342.
Initial projects will use Staphylococcus aureus, Streptococcus pneumoniae and Pseuodmonas aeurginosa, organisms that are already frequently used and well-understood in Sheffield. We expect to progress to the use of other organisms (and more complex mixtures) in collaboration with academic collaborators inside and outside of Sheffield, and in response to the demands of industrial partners. We also expect to diversify the range of tissue models available.

Existing projects (e.g. anti-microbial block polymers, anti-adhesive peptides, ultrasound disruption) which have so far been tested only on planktonic bacteria will be fast-tracked through SCARAB.

Development of bacterially infected cornea organ cultures. Pinnock, A., Shivshetty, N., Roy, S. et al. Ex vivo rabbit and human corneas as models for bacterial and fungal keratitis. Graefes Arch Clin Exp Ophthalmol (2017) 255:333–342.

People

Professor Peter Monk is chair of SCARAB. His research interests in inflammation as well as host-pathogen interactions, Professor Monk has successfully collaborated with tissue engineers to test a novel anti-bacterial adhesion strategy in a human skin model.

Dr Esther Karunakaran is the Stakeholder Liaison and day-to-day manager of SCARAB. She is a molecular microbiologist with experience of a systems biology approach to environmental biofilms.

Professor Sheila MacNeil is an internationally renowned tissue-engineer and she leads on the use of tissue culture infection models.

Dr Lynda Partridge leads on immunology and therapeutics development. She is an immunologist who has worked with Monk and MacNeil on infections in engineered skin.

Dr Annette Taylor has a background in complex systems modelling and leads the computational modelling arm of SCARAB.

Facilities

Our state-of-the-art equipment is housed in a dedicated SCARAB laboratory in Chemical and Biological Engineering. Additional specialized equipment is available in laboratories in Materials Science and Molecular Biology and Biotechnology. The tissue and bacterial growth facilities are in three separate rooms to allow for simultaneous investigations to be carried out, in aerobic and micro-aerobic/anaerobic conditions. We also host state-of-art supporting facilities dedicated for AMR research including a Multitron Pro incubation shaker, Heracell 160i incubators, Class II cabinets and -80°C freezers.

Bioflux 1000 with Zeiss Axio Observer Microscope

The Bioflux system allows live cell analysis under shear flow. The Bioflux 1000z incorporates microfluidic technology with a pneumatic controller for precisely controlled shear flow assays. A dedicated Zeiss inverted microscope is used for high resolution, time lapse imaging of samples. The BioFlux 1000z provides high-throughput screening of the interaction of cells in the 3D tissue culture models with the pathogenic bacterial biofilms under physiologically relevant conditions.

Leica TCS SP8 Confocal Microscope

The Leica TCS SP8 confocal microscope offers three-dimensional imaging of live cells with resolution down to 140 nm. The Leica TCS SP8 is a modular platform allowing for the visualisation of internal biofilm structures and the interactions with cells in engineered human tissues.

Apogee Micro-Plus Flow Cytometer

The Apogee Micro-Plus flow cytometer is suitable for the rapid detection and analysis of small particles (0.2 - 100 um) including extracellular vesicles, bacteria and nanoparticles. This device is used for the analysis of bacteria from disrupted biofilms and for the characterisation of extracellular vesicles released from infected tissues, a common feature of immune responses.

iXR Raman Spectrometer

The iXR Raman Spectrometer reveals not only chemical composition but also sample matrix morphology. The iXR was developed for integration with other analytical tools. Employing a fibre optic probe with moveable spectrophotometer (Raman in a box) allows the chemical and structural analysis of biofilm formations in-situ. In addition, a specialised Raman objective (60x) can be used to analyse single cells.

Thermo Scientific™ Varioskan LUX™ and Tecan Spark Microplate Reader

The Varioskan LUX™ and Tecan Spark® 20M Microplate Readers are versatile instruments for biological assays exploiting absorbance, fluorescence and luminescence detection methods. Both have environmental controls for real-time analyses.

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