Micro-patterned polymer substrates as novel models of epidermal wound healing

Lead Research Organisation: Queen Mary, University of London
Department Name: Blizard Institute of Cell and Molecular

Abstract

The proposed research will develop a novel in vitro model of wound healing within the skin. We will employ state-of-the-art micro-fabrication techniques to create engineered substrates for culturing human keratinocytes. This technology will allow us to precisely control multiple parameters of the wound environment. Specifically, we will determine how the composition, geometry, and stiffness of the wound influence cell behaviour. In addition, we will examine the molecular signalling pathways that regulate the cellular responses to these different environments. This project has the potential to provide significant insights into how human skin cells sense and respond to extrinsic signals during the wound healing process. Moreover, this model system may be a powerful tool for future cell biology studies, drug screening, and other types of translational research.

Technical Summary

The overall objective of the proposed research is to determine how human keratinocytes sense and respond to changes in their environment during wound healing. Micro-contact printing techniques will be used to create a novel in vitro model of epidermal wound healing. Micro-patterned substrates will consist of islands of protein resistant polymer brushes, which will serve as model 'wound beds', and will be surrounded by confluent human keratinocytes. Migration into the wound will be activated by coupling specific extracellular matrix proteins to the polymer brushes. This system will allow us to precisely control the size, shape, and composition of the wound bed. In addition, this assay will be used to investigate the role of serum response factor (SRF) signalling in keratinocyte mechano-sensing during the wound healing process. This work will be carried out in three specific aims and test two major hypotheses: (1) The composition and structure of the wound bed regulates re-epithelialization via altered migration and proliferation of the keratinocytes. (2) SRF mediates cellular responses to physical cues during epidermal wound healing.

Planned Impact

The main objective of the project is to improve our fundamental understanding of the normal wound healing process through the use of a novel in vitro system. While the immediate benefits of this knowledge will be to related academic communities, the model system may have long-term benefits to the UK economy and to public health and well-being. Our model of wound healing may be a useful assay for screening new compounds or bio-active molecules for treating chronic ulcers or skin diseases. As a tool for pharmaceutical companies, this model could benefit the UK and world economy. More importantly, the assay could aid the drug development process and lead to improved treatments for patients and overall public health. The knowledge generated by the proposed studies may also provide important information for wound management and tissue repair. Our studies will establish the basic role of physical cues in the normal wound healing process. This information may be useful to those in the tissue engineering field and provide design criteria for biomaterials and tissue scaffolds. Understanding the physical regulation of wound healing may also be important for healthcare professionals when applying wound dressings and sutures. Therefore, this project could benefit the economy by guiding the development of new tissue engineering and regenerative medicine products. Patients would also benefit from more effective products and treatments. Beyond the pharmaceutical and healthcare industries, this project may stimulate the materials and chemical industries. By providing a new application for micro-contact printing and polymer brushes, this project may lead to the development of new materials, chemicals, or processes that are more ideally suited for these types of experimental systems.

Publications

10 25 50
 
Description We have successfully developed a novel cell migration assay using micro-fabrication technologies. This platform has allowed us to control and study how interactions between cells and their environment regulate collective migration, a fundamental processes involved in wound repair. We are currently investigating how these extracellular cues impact cell polarity (i.e. directionality) and fate. In addition, further research to scale up this platform for high-content screening has uncovered novel epigenetic regulatory mechanisms controlling keratinocyte migration. These findings have important implications for wound healing and may represent new therapeutic targets for improving impaired healing.
Exploitation Route The technology developed during this project could be used in several different ways. First, it will likely lead to new knowledge and understanding of basic biological processes. Second, as a biotechnology tool it could be a valuable resource for drug testing and development. Finally, the specific findings of our studies could be used to guide the development of scaffolds for tissue engineering and wound repair.
Sectors Education,Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description Through this research grant we have developed a novel, micro-patterned assay for studying cell migration. This work has led to several key outputs, including knowledge dissemination through primary research articles and conference presentations, and securing further grant funding to develop this technology into a high-throughout format. In addition, this project has fostered public engagement activities through the development of an on-line learning resources about wound healing and tissue engineering.
First Year Of Impact 2012
Sector Digital/Communication/Information Technologies (including Software),Education
Impact Types Societal,Economic

 
Description Honor Fell Travel Award
Amount £1,000 (GBP)
Organisation Company of Biologists 
Sector Charity/Non Profit
Country United Kingdom
Start 06/2012 
End 07/2012
 
Description Queen Mary Innovation Award
Amount £10,000 (GBP)
Organisation Queen Mary University of London 
Sector Academic/University
Country United Kingdom
Start 01/2014 
End 09/2014
 
Description Research Project Grant
Amount £117,823 (GBP)
Organisation Dr Hadwen Trust (DHT) 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2015 
End 12/2016
 
Title Development of micro-patterned migration assay 
Description We have successfully developed a novel cell migration assay that allows us to precisely control migration over space and time. This technique involves micro-contact printing and a light activated coupling reaction to create dynamically adhesive surfaces. 
Type Of Material Technology assay or reagent 
Year Produced 2013 
Provided To Others? Yes  
Impact We are in the process of developing a new research collaboration with Dr. Simona Parinello at Imperial College, and this assay will be used in a recently awarded grant from the Dr. Hadwen Trust. 
 
Description Development of virtual experiment educational tool 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Schools
Results and Impact Through the Centre of the Cell and the X-plore Health initiative, we developed an on-line virtual experiment called 'Engineer Skin'. The aim was to teach secondary school children about the concepts of tissue engineering and how it could specifically be applied to skin repair.

None aware of yet.
Year(s) Of Engagement Activity 2012
URL http://www.xplorehealth.eu/en/media/engineer-skin
 
Description Invited seminar at Imperial College London 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Dr. Connelly gave an invited seminar to the Bioengineering Department at Imperial College London. Approximately 60 academics and students attended the talk, and it resulted in discussion and questions afterwards.
Year(s) Of Engagement Activity 2015
 
Description Invited seminar at King's College London 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact The talk led to valuable discussion and plans for future studies and potential collaborations.

None known yet
Year(s) Of Engagement Activity 2014
 
Description Invited seminar at Trinity College Dublin 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact I presented our previous and on-going research on micro-patterned models of wound healing to the Bioengineering Department at Trinity College Dublin. The seminar resulted in questions and discussion about the research and the development of new professional contacts in Ireland.
Year(s) Of Engagement Activity 2016
 
Description Invited seminar at University of Liverpool 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact The talk resulted in discussion with lecturers and students about significance of the findings, future work, and potential collaborations

Scientists in Liverpool planned new studies and approaches based on our discussion
Year(s) Of Engagement Activity 2013
 
Description Invited seminar at University of Pennsylvania 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Participants in your research or patient groups
Results and Impact We presented our findings in a seminar to the dermatology and orthopaedic departments within the School of Medicine at the University of Pennsylvania.

no actual impacts realised to date
Year(s) Of Engagement Activity 2012
 
Description Invited seminar at University of Southampton 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact We presented the micro-patterned model at an invited seminar at the University of Southampton on November 9th, 2012. The audience consisted of researchers and academics in the Engineering Department and School of Medicine.

no actual impacts realised to date
Year(s) Of Engagement Activity 2012
 
Description Pint of Science 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Public/other audiences
Results and Impact 30 members of the public attended an evening lecture in which I discussed how micro-fabrication technologies are currently being used in biomedical research for creating model 'organs on a chip'. The lecture was followed by a question and answer and discussion session with the audience.
Year(s) Of Engagement Activity 2016