Intercalated Degrees for Dental Students

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I have no idea what this isAn intercalated degree involves an extra year of study, which is inserted between the years of the undergraduate dental course, leading to the qualification of BMedSci.

It is an opportunity to obtain a deeper understanding of one of the subjects introduced in the dental course, and will allow students to undertake a substantial research project. Research forms an essential part of a student´s dental training, and this degree presents an early opportunity to develop research skills and potential.

An intercalated degree will also considerably enhance a student´s CV, and broaden employment prospects. For those interested in pursuing an academic career, it is even more worthwhile.

Although BMedSci students do very diverse projects it is expected that all students will have common aims and objectives (learning outcomes) and that these will be fostered in the mandatory short course and in the supervisory element of the programme as well as the students’ own self-directed study.

Meet our Ambassadors

These Ambassadors are former students who have successfully obtained their BMedSci degrees in the recent past. All of them had the opportunity to present their work at regional and National meetings with some students going on to win the ‘Junior Colgate Prize’ which is the most recognised and respected undergraduate research award in the country.

These Ambassadors have volunteered to help and guide future prospective BMedSci students or any students interested in research or academia. Feel free contact them via email for an informal chat from the student perspective.


Chris Plataischris

When did I do my BMedSci

I started my BMedSci in 2014, after my 2nd year of the BDS course.

My project

My project focused on a group of developmental genes known as the Hox genes and explored their role in head and neck squamous cell carcinoma. I specifically investigated the consequences of inhibiting the essential Hox cofactor, Pbx, and sought to determine whether this has potential as a targeted anti-cancer therapy. This project required a range of research techniques including cell culture, qPCR, Western blotting, cytotoxicity assays and flow cytometry.

Presentations

During the BMedSci year I presented my research in progress to the staff and students of the Dental School at the Friday lunchtime speaker slot. Since then, I have also presented my findings as a poster at the Guy’s Hospital INSPIRE event.

What I gained from my BMedSci

The BMedSci has been a great opportunity to get a taste for academia; it can be seen as a chance to independently explore a topic of interest in depth, and gives you the freedom to direct your own research project. In addition to learning the laboratory skills necessary for scientific research, I have also developed transferable skills such as critical appraisal, academic writing and time management. Furthermore, working in a laboratory is a sociable experience and gives you the opportunity to both learn from and teach a number of different people - an experience that has improved my ability to communicate relatively complicated information.


Paul Hankinsonstudent 2

When did I do my BMedSci

I completed my BMedSci project between my 2nd and 3rd BDS in the year 2013/2014

My project

I was investigating the role of a single gene in the development of oral cancer. Specifically looking at a potential role in the immortalisation process. It involved using a wide variety of research methods and skills such as cell culture, qPCR, western blotting, immunohistochemistry, transient transfections as well as flow cytometry.

Presentations

So far I have presented my work in Cardiff for the BSODR meeting and Guy’s hospital for the INSPIRE meeting. I also have the opportunity to present my work at the IADR meeting in Seoul in 2016.

Honours or prizes

Following my presentation at the BSODR meeting I won the junior Colgate prize. As part of this prize I have been given the opportunity to present my work in South Korea in 2016.

What I gained from my BMedSci

Completing a BMedSci has allowed me to develop many new skills as well as enhance others already gained. It has also provided a wealth of opportunities outside the normal BDS curriculum. For example as part of the writing process you will read many scientific papers vastly expanding you experience in this area. This is very useful latter in the BDS course and for postgraduate study and development. I have also gained knowledge and practise of several basic research methods which are transferable to other projects, such as during a PhD.

PHankinson1@sheffield.ac.uk


Sam McCarthystudent 1

When did I do my BMedSci?

2013-14, between 2nd and 3rd BDS

My project

My project attempted to investigate the dental pulp as a source of Schwann cells for future use in cell-conduit based therapies for nerve regeneration following peripheral nerve injuries. It involved a range of techniques, including primary tissue culture and immunocytochemistry, in order to see if dental pulp tissue could potentially offer a population of Schwann cells that could be directly cultured from the tissue itself or indirectly cultured through differentiation of dental pulp stem cells.

This qualitative study established primary culture techniques, from rat sciatic nerve and dental pulp, in a laboratory with little previous experience of this methodology. My results will facilitate future investigations into dental pulp derived Schwann cells, both directly and indirectly derived, and the incorporation of these Schwann cells into conduits to assess improved regeneration capacity in an animal model compared to current sub-optimal techniques.

Presentations

As well as presenting to staff and postgraduate students at the school of clinical dentistry, I also presented my work at the INSPIRE event at Guy's Hospital London, a meeting looking at how to encourage cross-over between clinical and academic dentistry; encouraging dental students to participate in research. I also presented a poster on my research at the 2016 ABAOMS conference in Sheffield.

What I gained from my BMedSci

Undertaking a BMedSci allowed me to gain a whole range of skills that will make me a better dentist as well as additional skill sets that open my options and may prove to be useful throughout my career. On top of the practical lab skills, I am now comfortable in tackling research papers and critiquing evidence which is a huge help in both clinical and academic dentistry. The experience of working on a project where nobody knows the answers and you have to find them out for yourself is challenging but hugely enjoyable, as well as making the larger projects and independence of later BDS years feel more natural. The extra clinical time during the BMedSci is also useful, adding to the confidence you gain…as well as being useful for taking the pressure off targets! I also really enjoyed gaining an insight ‘behind the scenes’ of a department I might be interested my career in; being in neuroscience you’re surrounded by clinical and semi-clinical oral surgeons and academics. It’s also been great to get to know a different year as I rejoined the BDS and really wasn’t a problem getting back into the swing of things.

Overall, it’s not an easy year and it’s quite a culture shift at first but I have no doubt it’s made me a better dentist and given me skills I’ll use in the future hopefully working at the interface between clinical and academic dentistry.

SJMccarthy1@sheffield.ac.uk


Mary Connollystudent 3

When did I do my BMedSci

I was the first for many years to undertake a BMedSci degree as a Sheffield BDS student between third and fourth BDS years (in July 2009 - July 2010).

My project

My project was an oral microbiology research project on the ‘role of outer membrane protein A in porphyromonas gingivalis biofilm formation and host cell interaction’ with supervisors Dr G.Stafford and Prof. I. Douglas. I dedicated my year to investigating the role of a major outer membrane protein (also found on the E.coli bacterium) on this key periodontal pathogen, and how it affects its ability to form biofilms and adhere to host cells in periodontal pathogenesis. My biggest achievement was the creation of an anaerobic P.gingivalis mutant, which took a great deal of time and hard work.

Presentations

I have presented at several national conferences as an oral presentation entrant, which include:
- British Society of Dental Research in Sheffield, Sept. 2011 (where I presented twice).
- BDA/Dentsply Student Clinician Awards in Enniskerry, Republic of Ireland, Feb. 2011.
- National Intercalator’s Conference in Bristol, Sept. 2010 (where I was the first dental student to present).
I also presented my findings locally at the microbiology journal club run by the department and at the Friday seminar session held in the School of Clinical Dentistry, during the year 2010.

Since completing my own project, my work has been continued and additionally presented as a poster by current University of Sheffield PhD student, Kate Naylor at:
- European Association of Microbiology 2014 workshop, Aarhus, June. 2014.
- British Society of Dental Research in Bath, Sept. 2013.
(Kate also presented recently using some of the ‘ground work’ from my original project, at the Oral Microbiology and Immunology group Amsterdam, Feb. 2015).

Honours or prizes

I achieved a 2:1 degree with honours and I was selected to represent the University of Sheffield at many prestigious conferences and the BDA/Dentsply Student Clinician Awards in 2011.

What I gained from my BMedSci

Taking a year out of my dental studies to undertake a year in research was a tough decision, but I would thoroughly recommend it to anyone looking at the prospect of an academic/ hospital career pathway. I gained an appreciation for the research that goes into developing a greater understanding of the main dental diseases for clinicians. From my own research I have a better understanding of oral microbiology, genetics and cell biology, as well as different types of research and techniques.

Undoubtedly I feel I gained a great deal from my BMedSci, which is regularly asked about at recent job interviews. Since graduating I have successfully secured hospital training placements including recently Paediatric DCT at the Birmingham Children’s Hospital, with a view to looking at specialist training in the future.

connollymhm@gmail.com


Hannah Cranehannah crane

When did I do my BMedSci: I studied for my BMedSci in 2010-2011.

My project: My project involved looking at the role of a specific signalling molecule in head and neck cancer. Working on this project enabled me to meet new people and develop my time management and writing skills. Initially I was apprehensive, but I really enjoyed the year and I am glad I made the decision to do a BMedSci.

Presentations and honours: My research gave me the chance to present my work at the PER/IADR conference in Finland. From this I won the Junior Colgate prize which has given me the fantastic opportunity to go to Seattle in February 2013.

What I gained from this year: Overall I found the year very beneficial and I have learnt many transferable skills which have come in useful during the BDS course. It is a really worthwhile year.

Email: mda08hc@sheffield.ac.uk


Chris ChathamChristopher Chatham

When did I do my BMedSci: I commenced with my BMedSci after finishing 2nd year of BDS, re-entering 3rd BDS a year later.

My project: The title of my project was the ‘The role of lymphotactin in oral mucosal wound healing’. Essentially, my year was spent conducting a follow on study from a previous PhD student, Dr. A Khurram, in the same department of the dental school (oral and maxillofacial pathology). He made a novel discovery that the receptor for the aforementioned lymphotactin, existed in the oral mucosa, and did copious amounts of lab work which suggested it may play a role in wound healing in the oral cavity. My study aimed to test this hypothesis by creating wound healing conditions inside a laboratory environment and trying to modify the rate at which it occurred with the addition of lymphotactin. I also undertook immunohistochemistry where I stained tissue sections for both lymphotactin and its receptor.

Presentations: I presented my work twice in poster format, at the Pathological Society of Great Britain and Northern Ireland conference in Sheffield and also the International Association of Dental Research in Helsinki.

What I gained from my BMedSci: This is best answered when I consider why I undertook an extra year out of my BDS degree – to gain an insight into research, and if it is a career path I would like to follow in the future. To this end, I definitely gained a lot from my BMedSci year. The year was varied and I was kept busy throughout the year either in the lab, writing up or attending the ethics or statistics courses (which the medical school runs). It is a must year for anyone who has a genuine interest in an academic career. I also gained skills in appraising evidence; useful in the rest of the BDS degree from assignments to clinical application of dental materials.

Email: mda08cc@sheffield.ac.uk


Dom SmithDom Smith

When did I do my BMedSci: I started my BMedSci in September 10 and completed it in July '11, having taken a year out between my second and third years.

My project: My project looked into the role of a gene called the RE-1 Silencing Transcription Factor (REST) in the progression of Head and Neck Squamous Cell Carcinoma (HNSCC). REST has been implicated as a tumour suppressor in an array of epithelial-derived tumours, such as Breast, Lung and Colon; however its role in HNSCC progression is unknown. We looked at RNA levels of REST in a number of different oral cell lines - from Normal Oral Keratinocytes, to Dysplastic cell lines, SCCs and also a metastatic cell line to see if there was any pattern in REST expression and cancer progression. However, no significant pattern was seen. We then artificially raised the expression levels of REST in one cell line and knocked it down in another cell line to study effect on cell behaviour, such as adhesion to fibronectin, migration and proliferation. Ultimately we showed that raising REST levels increased adhesion to fibronectin and reduced cell migration and knocking down REST levels had the opposite effect .
On the back of these results, we then began to look into possible mechanistic pathways for these changes, using a combination of bioinformatic databases and substantial literature searches. A few potential targets of REST were found and shown to be altered in the REST++ and REST-- samples; however more investigation needs to be done to see if these targets were responsible for the changes seen in cell behaviour.

Posters and presentations: I presented the findings of my research in the friday lunchtime speaker slot in May 2011, in a twenty minute talk to academic staff in the faculty. The poster ArRESTing Oral Cancer Development, which I presented at the events outlined in the next section.

Honours and prizes: I won the Junior Colgate Prize at the BSODR in Sheffield, September 2011, for which I won a trip to Brazil to represent the BSODR in the Hatton Prize and present my findings at the IADR in Iguazu Falls, June 2012.
I also represented Sheffield University in the BDA/Dentsply Student Clinician Awards at the Royal Horseguards Hotel in London, January 2012.

What I gained from my BMedSci: I gained a great idea of how academic life works at the University, along with a really good skills set for any future career in hospital or academic research.
As part of the course, you undertake lectures in Ethics and Statistics alongside the medical students who are also doing a BMedSci - through this I made some great friends, who I will definitely be keeping in touch with once I leave Sheffield.

Email: mda08djs@sheffield.ac.uk


thomas evansThomas Evans

When did I do my BMedSci: I carried out my BmedSci in 2010/11 between 2nd and 3rd years of the BDS degree.

My project: I worked in the oral medicine department on the oral fungus ‘Candida albicans’. I worked to help establish a model of Candida albicans infection in Zebrafish embryos, including real-time in vivo visualisation. In addition to this I looked at the adhesion of Candida
albicans to endothelial cells under conditions mimicking blood vessels.

Presentations: Evans, T. (2011, September 14). Using Zebrafish To Study Candida albicans-Mediated Systemic Candidiasis in vivo. Oral Session presented at the BSODR/IADR Congress, Sheffield. Evans, T. (2012, September 13). Investigating O-linked manan-mediated adhesion of Candida albicans to endothelium. Oral Session presented at the PER/IADR Congress, Helsinki.

Honours / prizes
• BMedSci degree with a first.
• Commended in the Junior Colgate Prize for presentation at British society of Oral and Dental research 2011 “Using Zebrafish To Study Candida albicans-Mediated Systemic Candidiasis in vivo”

What I gained from my BMedSci: I feel that my problem solving skills improved, and my confidence to tackle problems independently.Two skills that were improved upon were long-form writing as well as the ability to present work with clarity, and these have already proved useful both away from academia, as well as within the BDS degree. I also gained a deeper appreciation for
research methodologies and a propensity to ask deeper questions, not to mention the degree, prize and the travel!

Email: mda08te@sheffield.ac.uk


Joe WhiteJoe White

When did I do my BMedSci: 2011-2012 between my 2nd and 3rd BDS years

My project: I investigated the expression of a potential Tumour Supressor gene in Oral Cancer. Tumour Supressor genes are switched off in cancer and are therefore usually genes which control the cell cycle and tissue growth. This gene could therefore have had the potential to be a target for chemotherapy. My work was purely lab based, using cell cultures and whole tissue biopsy specimens. I performed analysis of protein expression, gene expression and attempted to induce expression of my gene in cancerous cells to see what would happen.

Presentations: I have presented at the all expenses paid BDA/Dentsply Undergraduate Research Conference in Marylebone, London

A paper is currently being written containing some of my work.

Honours or prizes: I have presented at the British Society of Oral and Dental Research Conference in Bath, UK where I won the Junior Colgate Award for best undergraduate research presentation.

I will present my work at the International Association of Dental Research Conference in Cape Town, South Africa in Summer 2014 where I will be competing for the Junior Hatton Award

What I gained from my BMedSci: I gained a thorough appreciation for academic dentistry. It has improved by scientific understanding of oral pathology, oral medicine, oral surgery, cell biology, microbiology and biochemistry. I have learned a massive variety of crucial biological lab skills and not just pipetting! I feel confident in reading and critically analysing research papers. I have a good working understanding of statistics and statistical analysis. I have gained valuable work experience and an award that will hopefully make me stand out from the crowd when applying for jobs. I have gained greater clinical experience by having an extra year treating patients which has helped my clinical confidence and patient management. I have gained experience of what a non-clinical PhD would entail should I decide to do this at a later date.
I am getting my elective trip paid for as it coincides with a conference I am being sent to. I gained more friends by moving down a year and by working with PhD and Masters students in the department.

I got to have TWO GRADUATIONS!!!

Email: JBWhite1@sheffield.ac.uk.

FAQs

Programme aims:

  • To provide an enhanced knowledge and understanding of biomedical research and its methods

  • To develop skills in research evaluation, communication and ethics

  • To allow students to apply the above through an extended research project

Programme outcomes:

  • The place of research in dentistry

  • Current dental research and its methods

  • The conduct of research in accordance with correct research methodologies and procedures

  • The importance of conducting research in accordance with up-to-date ethical guidelines and policies

  • The fundamental principles of designing research projects and protocols

Academic and intellectual skills:

  • Design a research project in accordance with appropriate research methodologies and ethical principles

  • Exercise independent judgment and critical thinking

  • Apply basic statistical methods to data evaluation and interpretation

  • Present work orally and in writing to an academic audience

  • Where their project requires it, carry out practical experiments and tasks in a laboratory setting in accordance with health and safety guidelines

  • Produce a well-structured and substantial dissertation to present the results of their research project

  • Conduct an extensive literature review using relevant sources

Transferable skills:

  • Apply good time-management skills to structure their work and meet deadlines

  • Effectively use a wide range of IT packages for a variety of tasks

  • Work independently on a project

  • Display good written and oral communication skills

  • Understand and apply basic statistical methods

  • Self-direct their learning

Taught Element:

Students undertaking the BMedSci are required to attend a taught element to the programme. The taught element consists of:

  • Mandatory Short Course

  • Introduction to Statistics Course

The Introductory Short Course

The Introductory Short Course runs for 3 days at the start of the BMedSci Programme (21 August 2013) and provides an introduction to conducting a research project, including research ethics and governance and guidance on writing dissertations. Students are issued with the timetable for this course via email/ Minerva

Ethics Course

Allie Gartland and Mark Wilkinson run a half-day symposium on research ethics. This will run in September. After this course, students are required to complete an ethics assessment which constitutes 10% of the final degree mark. The Ethics Course Leads will provide separate information on the ethics teaching and assessment.

Statistics Course

The Introduction to Statistics Course, organised by Dr Steven Julious in ScHARR, will run over 8 weeks starting on 30th September 2013. It will consist of twice-weekly lectures, tutorials and practical sessions, including guidance on SPSS. A separate timetable for this course will be available on Minerva. During this course, students will be issued with their statistics assignment.

Summative Assessment

The three elements comprising the summative assessment of the BMedSci are:

  • Statistics assignment worth 20% of the final mark.

  • Ethics assignment worth 10% of the final mark.

  • Dissertation worth 70% of the final mark.

External employment:

In accordance with the guidelines set out in the University’s Students’ Charter, students are reminded that they are advised to undertake no more than 16 hours of external work per week in addition to that required by their programme of study.

Contact us

BMedSci Programme Lead

Professor Fiona Boissonade
f.boissonade@sheffield.ac.uk

0114 271 7964/7850

BMedSci Administrative Officer

Leyna Wright
leyna.wright@sheffield.ac.uk

0114 271 7801

BMedSci Student Ambassadors (Former BMedSci students)

Hannah Crane
mda08hc@sheffield.ac.uk

Chris Chatham
mda08cc@sheffield.ac.uk

Dom Smith
mda08djs@sheffield.ac.uk

Tom Evans
mda08te@sheffield.ac.uk

Joe White
JBWhite1@sheffield.ac.uk.

BMedSci Projects for 17/18

Click on the titles below for complete details of each project

Development of an innervated 3D tissue-engineered oral mucosa model

Supervisors: Professor FM Boissonade, Dr C Murdoch and Dr HE Colley

Background:

Over the last decade our understanding of epithelial and stromal cell interaction has been greatly enhanced by the development of a wide range of 3D tissue-engineered constructs including models of skin and oral mucosa. These 3D tissue-engineered models can be used for a wide variety of purposes such as studies of toxicity, wound healing, tumour biology, and other skin disease. In addition to their uses for in vitro investigations, they are also beginning to deliver benefit in the clinic (usually in small-scale reconstructive surgery procedures). A number of cell types have been incorporated into these systems, thus allowing a closer approximation to the conditions found in human skin and oral mucosa in vivo. These cell types include keratinocytes, dermal fibroblasts and a range of immune cells. However to date there are very few models of skin or oral mucosa that incorporate neuronal cells.

There is now growing evidence that interaction between keratinocytes, fibroblasts and sensory neurones plays a significant role in both skin homeostasis and disease (eg wound healing, eczema and psoriasis). The neuropeptides calcitonin gene-related peptide (CGRP) and substance P have an established role in neurogenic inflammation and are thought to influence proliferation and morphogenesis of both keratinocytes and fibroblasts. Is it also well known that skin and oral mucosal cells produce a wide range of neurotrophic factors that have significant influence on neuronal development.

Aims:

To develop an innervated 3D tissue- engineered oral mucosa model, and study the effects of innervation on skin and mucosal morphology and response to injury.

Techniques:

A variety of cell culture techniques will be used. 3D oral mucosa and trigeminal ganglion neurones will be cultured using techniques developed in our laboratories. Successful co-culture will be determined by immunohistochemical identification of different cell types using methods that are well established in our laboratories.

References:

  • MacNeil S. Progress and opportunities for tissue-engineered skin. Nature 2007;445:874-880

  • Roggenkamp D, Kopnick S, Stab F et al. Epidermal nerve fibres modulate keratinocyte growth via neuropeptide signaling in an innervated skin model. J Investigative Dermatology 2013;133:1620-1628

  • Truzzi F, Marconi A, Pincelli C. Neurotrophins in healthy and diseased skin. Dermato-Endocrinology 2011;3:32-36

  • Yu XJ, Li CY, Xu YH, et al. Calcitonin gene-related peptide increases proliferation of human HaCaT keratinocytes by activation of MAP kinases. Cell Biol Int. 2009;33:1144-8

  • Colley HE, Eves PC, Pinnock A, Thornhill MH, Murdoch C. Tissue-engineered oral mucosa to study radiotherapy-induced oral mucositis. Int J Radiat Biol. 2013 Nov;89(11):907-14.

For further information please contact Professor F Boissonade on f.boissonade@sheffield.ac.uk

Targeting HOX/PBX interactions in head and neck cancer – potential for therapy?

Supervisors: Dr KD Hunter and Dr DW Lambert

Background:

Hox genes are a family of transcription factors which are involved in a wide range of developmental processes. Expression of many HOX genes are also de-regulated in cancer development and thus represent possible therapeutic targets1. In Dr Hunter’s laboratory, the expression of all 39 Hox genes has been investigated in a number of HNSCC cell lines2. The highest expression at each of the Hox loci was of HOXA4, HOXB9, HOXC9 and HOXD10, all of which are well recognised in the literature in other cancers. However these was marked variability between cell lines, particularly between primary tumour cells and metastases.

Novel small molecule inhibitors, developed in the lab of our collaborator Dr Richard Morgan, target the interaction between Hox proteins and PBX, a second transcription factor that serves as a co-factor for many Hox proteins3. As inhibiting Hox genes may be a possible therapy, we wish to assess the effect of these small molecules on the behaviour of a panel of HNSCC cells with differing Hox gene expression, using a variety of functional assays. This will determine how they influence the phenotype of cancer cells which express HOX genes at differing levels, reflecting the variable expression observed in head and neck tumours, and thereby providing information about the likely effectiveness of this drug in different patients.

Hypothesis: Novel small molecules will inhibit growth of HNSCC cells in a HOX gene-dependent manner

Experimental plan:

1. Assess the effect of a panel of candidate small molecule inhibitors of HOX-PBX interactions in a dose dependent manner, by:
a. Cytotoxicity assays (LDH/MTS) assay
b. Proliferation (EDU incorporation)
c. Apoptosis (Annexin V-PE detection)
2. Relate the effects seen to the gene expression data, selecting any Hox genes which correlate with sensitivity or resistance to HXR9
3. Over expression of a Hox gene from 2. to assess similar features as in 1., with and without the inhibitor.

References:

  • Shah and Sukumar, The Hox genes and their roles in oncogenesis. Nat Rev Cancer. 2010 May;10(5):361-71. Shah N1, Sukumar S.

  • Darda et al, The role of HOXB9 and miR-196a in head and neck squamous cell carcinoma, under review with PLOSOne

  • Morgan et al, Targeting HOX and PBX transcription factors in ovarian cancer. BMC Cancer. 2010; 10:89.

For further information please contact Dr K Hunter on k.hunter@sheffield.ac.uk or Dr Lambert on d.w.lambert@sheffield.ac.uk

Oral Histories of Oral Health and Healthcare

Supervisor: Professor B Gibson

Background:

Traditionally medicine and dentistry have been dominated by the biomedical perspectives were the voice of the patient has been secondary to clinical diagnosis and treatment planning. In this world view it has proven difficult to find ways to enable the patients’ perspective to be heard. Over the last thirty years there has thankfully been an increasing appreciation of the voices of patients in dentistry. This movement has involved the use of oral health related quality of life instruments in measuring the effectiveness of clinical interventions. It is now generally accepted that no clinical trial should take place without some form of assessment of oral health related quality of life. Not only this but the University of Sheffield has been at the forefront of giving patients a voice, we have contributed internationally to the development of new condition specific impact measures and our research on children has given them an increasing voice and role in oral health care.

What we wish to do is to take our work on giving patients a voice further by developing a new initiative to build an archive of oral history interviews. Our reasons for doing this are that patients’ voices can tell us a lot about the history and context of dentistry as it has changed through time. This voice can provide us with important sensitivity to the fact that no matter how much dentistry is innovating and changing over time that the experiences patients have are shaped by social and historical forces. Evidence demonstrates that the cohort experiences different groups of people have about oral health and health care has changed dramatically over time (Sussex et al., 2010). Someone who experienced dentistry in the 1930’s will have had a very different experience and set of expectations to someone who has experienced dentistry in the 1950’s. The tools of dentistry were radically different in just twenty years (Ettinger, 1993). What this means is that patients who come to the dentist have radically different memories and experiences of oral health and health care and to date there are very few studies systematically looking at these different experiences.

Aims:

The aim of this study is to explore oral histories of oral health and healthcare.

Methods:

Conducting and analysing oral history interviews with patients over the age of 70 years with a focus on social inequalities. We have all of the equipment and the student(s) who decide to do this will contribute to the development of a new archive of oral history interviews that will be held at the University of Sheffield. These interviews will subsequently be used in Undergraduate and Postgraduate teaching.

References:

  • Ettinger, R. (1993). Cohort differences among aging populations: A challenge for the dental professions. Special Care in Dentistry, 13, 19-26.

  • Sussex, P., Thomson, W., & Fitzgerald, R. (2010). Understanding the 'epidemic' of complete tooth loss among older New Zealanders. Gerodontology, 27, 85-95.

For further information please contact Dr B Gibson on b.j.gibson@sheffield.ac.uk

Elucidating the mechanism of extracellular vesicle release in oral cancer

Supervisor: Dr S Hunt

Background

Oral cancer mortality rates have increased by ~10% in the UK in the last decade. According to Cancer Research UK, there were 7316 new cases of oral cancer diagnosed and 2119 oral cancer related deaths in 2012. Sixty percent of patients present with late stage oral cancer, after the tumour has spread to produce regional metastases, as many early cancers are asymptomatic1. Extracellular vesicles (EVs) are increasingly recognised as a source of biomarkers for disease diagnosis and also as novel therapeutic targets. They are nanoscale sized, membrane enclosed vesicles that contain molecular cargo including nucleic acids and proteins. They are produced by all cells of the body and are present in biological fluids including saliva2. Indeed, the saliva of oral cancer patients contains up to 4-fold more EVs than that of healthy volunteers3. Cancer cells produce increased numbers of EVs with altered molecular cargo. These EVs can be taken up by cells that surround the tumour, which converts them to a pro-tumourigenic phenotype. A number of key proteins that are involved in the biogenesis of EVs have already been identified, including the components of the endosomal sorting complex required for transport (ESCRT)4. Certain members of the ESCRT (Hrs and TSG101) have also been shown to be overexpressed in cancer5. However, nothing is known about the mechanism of EV biogenesis in oral cancer. A better understanding of this process may lead to the identification of novel therapeutic targets for oral cancer treatment.

Hypothesis: ESCRT proteins are aberrantly expressed in oral cancer cells, leading to increased production and release of extracellular vesicles.

Research questions:

  • What is the expression of ESCRT members in normal oral keratinocytes compared to oral cancer cells? The expression of ESCRT members will be determined by QRT-PCR and western blot.

  • What is the cellular localisation of ESCRT proteins in normal keratinocytes compared to oral cancer cells? Immunofluoresence microscopy will be used to image the cellular location of ESCRT proteins to determine if their distribution is altered in oral cancer.

  • Does silencing of ESCRT gene expression reduce EV production by oral cancer cells in vitro? Transient siRNA knockdown of ESCRT gene expression will be confirmed by QRT-PCR and western blot. EV release will be assessed by tunable resistive pulse sensing (TRPS).

References:

  • Guneri, P., and Epstein, J.B. (2014). Late stage diagnosis of oral cancer: Components and possible solutions. Oral Oncology, 50: 1131-1136.

  • D’Souza-Schorey, C., and Clancy, J.W. (2012). Tumour-derived microvesicles: shedding light on novel microenvironment modulators and prospective cancer biomarkers. Genes & Development, 26: 1287-1299.

  • Sharma, S., et al. (2011). Quantitative nano-structural and single molecule force spectroscopy bio-molecular analysis of human saliva derived exosomes. Langmuir, 27(23): 14394-14400.

  • Colombo, M., et al. (2013). Analysis of ESCRT functions in exosome biogenesis, composition and secretion highlights the heterogeneity of extracellular vesicles. Journal of Cell Science, 126(24): 5553-5565.

  • Tu, C., et al. (2011). ESCRT proteins: Double-edged regulators of cellular signaling. Bioarchitecture, 1(1), 45–48.

For further information please contact Dr S Hunt on s.hunt@sheffield.ac.uk

Why do patients on Nicorandil therapy get non-healing ulcers?

Supervisors: Dr K Hunter and Dr D Lambert

Background

The treatment of angina pectoris has been revolutionised by the development of a class of potassium channel opening drugs, one of which is Nicorandil1. This drug, loved by cardiologists and their patients, has markedly improved the quality of life of many patients with ischaemic heart disease. However, soon after the widespread prescribing of Nicorandil started, cases of mucosal ulceration started to surface. These ulcers, which can affect any part of the gastrointestinal tract, are painful and very slow to heal2. A significant number of these patients have oral ulcers.

The reasons for this are not clear. However, we have some preliminary evidence that the formation of granulation tissue is inhibited and that the proliferation of cells in the stroma of these ulcers is reduced. This would suggest some abnormality of fibroblasts or endothelial cells which compose this tissue which is so important in healing. Others have shown that Nicorandil affects the activation of fibroblasts by mediators which may be present in these wounds.

Hypothesis: Nicorandil reduces the activation of fibroblasts (and/or endothelial cells) which will delay the healing process.

Experimental plan:

Assess the effect of treating oral fibroblasts with nicorandil by activating the cells (using TGF-β1) and then, in the presence and absence of nicorandil measure:

  • Proliferation

  • Migration

  • Changes in β-actin expression

  • Contractility

Assess the effect of treating endothelial cells with nicorandil by, in the presence and absence of nicorandil, measuring:

  • Cell proliferation

  • Migration

  • Tube formation (how well the cells form blood vessels)

References:

  • Simpson D, Wellington K. Nicorandil: a review of its use in the management of stable angina pectoris, including high-risk patients. Drugs. 2004;64(17):1941-55.

  • Pisano U, Deosaran J, Leslie SJ, Rushworth GF, Stewart D, Ford I, Watson AJ. Nicorandil, Gastrointestinal Adverse Drug Reactions and Ulcerations: A Systematic Review. Adv Ther. 2016 Mar;33(3):320-44. doi: 10.1007/s12325-016-0294-9. Epub 2016 Feb 9.

  • Liou JY, Hong HJ, Sung LC, Chao HH, Chen PY, Cheng TH, Chan P, Liu JC. Nicorandil inhibits angiotensin-II-induced proliferation of cultured rat cardiac fibroblasts. Pharmacology. 2011;87(3-4):144-51. doi: 10.1159/000323555. Epub 2011 Feb 22.

For further information please contact Dr K Hunter on k.hunter@sheffield.ac.uk or Dr Lambert on d.w.lambert@sheffield.ac.uk

Biting the hand that feeds you: how periodontal pathogenic bacteria feed on your oral tissues

Supervisors: Dr G Stafford, Dr J Rafferty

Background:

Periodontitis is an inflammatory disease leading to tooth loss that affects over 700 million people worldwide, i.e. approximately 11% of the global population. It is a significant healthcare burden, particularly in the elderly where a link between quality of life in old-age and periodontal disease is well established. One of the bacteria causing periodontal disease- Tannerella forsythia is able to harvest a sugar present on human cell-surfaces and use it as a food source. This sugar is known as sialic acid and is the most common and important glycan component in humans, acting as a label of humanity but also defines the responsiveness of many immune pathways.

The mechanism by which Tannerella accesses this sugar is via action of enzymes that remove (‘bite’) it off of human cell surface proteins before the bacteria uptake the sugar via specific transport mechanisms. In Sheffield we have identified a range of these ‘feeding’ enzymes and in fact one of these, called NanS, is now sold by a biopharmaceutical analysis company due to its unique properties. However, the role of this unique enzyme in bacterial growth, cellular infection or its biochemistry and enzyme structure are not currently known.

Aims:

  • Investigate the importance of the NanS enzyme in infection in experiments using a strain of Tannerella which we have constructed that lacks a functional NanS enzyme

  • Investigate the biochemistry of NanS via in vitro enzyme assays of its constituent domains

Techniques:

  • Bacterial plaque assays- Bacterial are grown in artificial plaque systems using artificial saliva substrates

  • Host-pathogen interaction studies- in vitro infection assay to test role of NanS in infections

  • Immunofluoresence microscopy of sialic acid on human oral cells to test enzyme activity

  • In vitro enzyme assays to test activity of recombinant enzyme

  • X-ray crystallographic characterization of NanS- in collaboration with dept of Mol biology to establish atomic structure of NanS.

References:

  • Roy, S. et al. Beta-hexosaminidase activity of the oral pathogen Tannerella forsythia influences biofilm formation on glycoprotein substrates. FEMS Immunol. Med. Microbiol. 65, 116–20 (2012).

  • Douglas, C. W. I. et al. Physiological adaptations of key oral bacteria. Adv. Microb. Physiol. 65, 257–335 (2014).

  • Phansopa, C. et al. Characterization of a sialate-O-acetylesterase (NanS) from the oral pathogen Tannerella forsythia that enhances sialic acid release by NanH, its cognate sialidase. Biochem. J. 472, 157–67 (2015).

  • Phansopa, C. et al. Structural and functional characterization of NanU, a novel high-affinity sialic acid-inducible binding protein of oral and gut-dwelling Bacteroidetes species. Biochem. J. 458, 499–511 (2014).

  • Roy, S., Douglas, C. W. I. & Stafford, G. P. A novel sialic acid utilization and uptake system in the periodontal pathogen Tannerella forsythia. J. Bacteriol. 192, 2285–93 (2010).

For further information please contact Dr G Stafford on g.stafford@sheffield.ac.uk

Identification of human cellular targets of a bacterial pathogen Surface protein key to infection: from Omps to antimicrobials

Supervisors: Dr G Stafford, Dr C Murdoch

Background:

Recent work by the Stafford group has revealed the role of a key outer membrane surface proteins in infections by the keystone periodontal pathogen P. gingivalis. An orgamism that causes periondontal disease- a condition affecting 700 million pepols globally. This OmpA2 protein is part of a large family of virulence proteins in human pathogenic bacteria, and a Porphyromonas mutant lacking the OmpA2 gene is unable to form biofilms or infect human cells and furthermore one of its surface exposed protein loops is directly involved in interactions with oral epithelial cells in in vitro infection models. The aim of this project is to identify the OmpA2 human receptor, begin characterisation of its innate immune stimulatory capability and thus begin work aimed at developing these peptides as potential antimicrobials. The project will give broad training in eukaryotic and microbial culture, Immunoprecipitation, chromatography, basic bioinformatics, western blotting, ELISA, bacterial infection assay and immunofluorescence.

Aims:

1 - Identification of cell surface receptor for OmpA2 surface binding loops

  • Work in the lab has established that OmpA2 surface loop 4 is able to direct binding of inert beads to human oral epithelial cells (ref 4 above). However nothing is known of cellular receptor identity.

  • Using a combination of cross-linking and Streptavidin-biotin based chromatography/ Immunoprecipitation the student will attempt to identify human cellular proteins that interact with OmpA2-4 (using a scrambled version of the same peptide as negative control). Interacting partners will be identified by Mass Spectrometry (identified by collaborators in Chem-Biol Engineering or commercially).

  • If successfully identified these will be verified by Immunoblotting of eluted fractions and by assessing the effect of blocking antibodies on bacterial-human cell interactions in antibiotic protection assays.

2 - Investigation of innate immune responses to OmpA2

  • Despite having evidence of direction interaction of P. gingivalis with oral cell being OmpA2 dependent we have no information on host-cell responses. Using both our OmpA2-4 peptide and its scrambled control we will stimulate human oral epithelial cells with peptides (50ug/ml) and assess responses of innate immune response markers by Western blot (e.g. IL-8, IL-1-beta) and ELISA of supernatants or using Cytokine Bead Arrays (FACS based). In addition OmpA is known to act via TLRs in some spp, and we will use a TLR reporter (THP-1) monocyte cell line to assess its ability to stimulate this pathway via its Secreted Alkaline Phosphate activity.

3-Pathogenic cross-talk of OmpA2 binding peptides-

  • OmpA is a widespread bacterial surface protein of Gram-negative pathogens with homologues in many oral bacteria.

  • Using bioinformatics (structural prediction software- Phyre2), the student will identify analogous proteins to OmpA in other oral red-complex pathogens (T. forsythia, T.denticola) and model the structure to identify homologous surface loops to OmpA2-4 that we will have synthesized commercially before testing in fluorosphere binding assays and in inhibition of bacterial invasion assays.

Techniques to be employed:

  • ELISA, Cell culture, microbial culture, immunofluorescence, Immunoprecipitation, crosslinking, protein chromatography, basic bioinformatics

For further information please contact Dr G Stafford on g.stafford@sheffield.ac.uk or Dr C Murdoch on c.murdoch@sheffield.ac.uk

An in vitro models of human salivary gland tumours

Supervisors: Dr L Bingle and Dr A Khurram

Background:

Salivary gland tumours are uncommon but histologically heterogeneous and so are often very difficult to diagnose, even for experienced pathologists. They most commonly occur in the major glands but are also relatively frequently found in the minor glands associated with the palate. Very little is known about the pathogenesis of tumour growth and development but associations with the presence of novel fusion proteins and viral infections has been established.

Aim/Hypothesis:

The aim of this project is to establish in vitro models of human salivary gland tumours, as an in vitro model, replicating the complex glandular structure, would provide an extremely useful tool to investigate tumourigenesis and could also have potential as a first screen for the novel biomarkers desperately needed as prognostic/diagnostic tools.

Method/Experimental plan/Techniques:

We have already developed 3D models of normal, human salivary glands from excess surgical tissue. We will use information gathered through developing models of normal glands and apply to the growth of tumour models. Tissue from human salivary gland tumours will be used as a starting material to replicate, in vitro, the 3D in vivo structure of a tumour. An aim would be to develop a number of models with different cell phenotypes to reflect the heterogeneity of salivary gland tumours. These will allow us to determine the processes involved in the development of a tumour, particularly the role of fusion proteins and viruses. Functional assays will allow characterisation of the aggressiveness of tumours in relation to cell phenotype, heterogeneity and involvement of fusion proteins.

For further information please contact Dr L Bingle on l.bingle@sheffield.ac.uk