Biotextiles for ligament repair using biofunctionalisation to aid regeneration
Lead Research Organisation:
University of Manchester
Department Name: Materials
Abstract
The project will be integrated into the UKRMP Acellular approaches for therapeutic delivery Hub, a collaboration between Manchester, Nottingham, Imperial, Southampton, Keele and others.
http://www.ukrmp.org.uk/hubs/
Anterior cruciate ligament (ACL) ruptures are a common problem and occur mostly as a result of sports injuries. Around 200,000 ACL ruptures per year occur in the United States and it is estimated that more than 100,000 ACL repairs are performed annually in the United States.
Such ruptures can subsequently lead to other, serious degenerative joint diseases and for this reason it is important to treat and tackle ACL injuries efficiently and effectively. The current gold standard for ACL repair involves the use of patellar tendon autografts; this treatment has many limitations including: donor site morbidity and more importantly the inability of the graft to fully integrate with the bone tunnel site (area often referred to as the enthesis). High failure rates often occur particularly at this enthesis site subsequently leading to patient revisits for further improved surgeries. Hence, an intervention is required which could be coupled with the current gold standard method to act as a dual technology, which could provide more robust graft integration.
Some research has been performed in developing alternative grafts fabricated from synthetic FDA approved materials including Poly-e-Caprolactone (PCL) and PET with the hope to manufacture tissue-engineered grafts that would provide the required mechanical properties. However, limitations associated with such constructs include their lack of bio-interaction, lack of functional neo-matrix lay down and remodeling, leading to failure at the enthesis site. Therefore, the problem of graft failure (autograft or synthetic) at the enthesis site is yet to be fully resolved. For this reason this study aims to improve graft integration at the enthesis site through improved healing rate.
In this case, we aim to develop a synthetic graft, which is able to precisely mimic the nano-and micro- architecture of native tissue ACL extracellular matrix along with the appropriate mechanical properties and degradation rate. Working in collaboration with Professor Eithne Comerford (University of Liverpool), we aim to identify key extracellular matrix (ECM) protein fragments within the different regions of the ACL (particularly the enthesis) and functionalise these onto the synthetic graft. We have identified fibrillin-1 as a starting point and preliminary results have shown significantly more rapid attachment and alignment along the electrospun fibres with fibrillin present. Further molecules have been identified and will be explored along with signalling pathways.
Cell behavior will be assessed in terms of cell viability, morphology, proliferation, differentiation, intra- and extracellular signalling pathways and neo-matrix secretion. These will be validated and identified using via immunocytochemistry, ELISA, RT-PCR and histology.
Additionally the electrospun system could be used as a platform technology for other ligaments and also tendon repair.
http://www.ukrmp.org.uk/hubs/
Anterior cruciate ligament (ACL) ruptures are a common problem and occur mostly as a result of sports injuries. Around 200,000 ACL ruptures per year occur in the United States and it is estimated that more than 100,000 ACL repairs are performed annually in the United States.
Such ruptures can subsequently lead to other, serious degenerative joint diseases and for this reason it is important to treat and tackle ACL injuries efficiently and effectively. The current gold standard for ACL repair involves the use of patellar tendon autografts; this treatment has many limitations including: donor site morbidity and more importantly the inability of the graft to fully integrate with the bone tunnel site (area often referred to as the enthesis). High failure rates often occur particularly at this enthesis site subsequently leading to patient revisits for further improved surgeries. Hence, an intervention is required which could be coupled with the current gold standard method to act as a dual technology, which could provide more robust graft integration.
Some research has been performed in developing alternative grafts fabricated from synthetic FDA approved materials including Poly-e-Caprolactone (PCL) and PET with the hope to manufacture tissue-engineered grafts that would provide the required mechanical properties. However, limitations associated with such constructs include their lack of bio-interaction, lack of functional neo-matrix lay down and remodeling, leading to failure at the enthesis site. Therefore, the problem of graft failure (autograft or synthetic) at the enthesis site is yet to be fully resolved. For this reason this study aims to improve graft integration at the enthesis site through improved healing rate.
In this case, we aim to develop a synthetic graft, which is able to precisely mimic the nano-and micro- architecture of native tissue ACL extracellular matrix along with the appropriate mechanical properties and degradation rate. Working in collaboration with Professor Eithne Comerford (University of Liverpool), we aim to identify key extracellular matrix (ECM) protein fragments within the different regions of the ACL (particularly the enthesis) and functionalise these onto the synthetic graft. We have identified fibrillin-1 as a starting point and preliminary results have shown significantly more rapid attachment and alignment along the electrospun fibres with fibrillin present. Further molecules have been identified and will be explored along with signalling pathways.
Cell behavior will be assessed in terms of cell viability, morphology, proliferation, differentiation, intra- and extracellular signalling pathways and neo-matrix secretion. These will be validated and identified using via immunocytochemistry, ELISA, RT-PCR and histology.
Additionally the electrospun system could be used as a platform technology for other ligaments and also tendon repair.
People |
ORCID iD |
| Julie Gough (Primary Supervisor) | |
| Zara Smith (Student) |
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| EP/N509565/1 | 01/10/2016 | 30/09/2021 | |||
| 1735234 | Studentship | EP/N509565/1 | 01/10/2016 | 31/03/2021 | Zara Smith |
| Description | This project investigated the effect of biofunctionalisation of specific fragments of fibrillin-1 on the regenerative potential of anterior cruciate ligament cells on aligned fibres, with cells derived from both canines and humans for use in both medical and veterinary applications. Aligned fibre platforms, produced using an electrospinning technique were found to show high degrees directional alignment. Fibres were capable of undergoing surface treatment in order to make them more hydrophilic, and therefore, more suitable for cell attachment. They also demonstrated the ability to adsorb the protein fragments to their surface, with hydrophilic surfaces displaying the most protein adsorption, when compared to their unmodified controls. Testing for these fragments however, was a challenge due to their small size and low concentrations - indirect ELISA was determined to be the most accurate option for quantitation. It was determined that as little as one microgram of fragment per scaffold is enough to illicit a response in both human and canine cells, with canine cells showing a more prominent response, and appearing to preferentially use mechanically responsive integrins to attach to the fibronectin controls. It was determined that fibrillin-1 modified scaffolds could support initial extracellular matrix (collagen types I and III, fibrillin and fibronectin) deposition on aligned fibre scaffolds, but after long periods (28 days), elastin production was not on observed in either canine or human cell cultures. As a small side project, multiple extracellular matrix components, and their morphology/orientations were displayed in osteoarthritic human ACL tissue, which could further aid in the understanding of the effect of osteoarthritic pathologies on extracellular matrix composition of the ACL. |
| Exploitation Route | Outcomes of this award may highlight the potential of fibrillin-1 as a biofunctionalisation agent in ligament engineering grafts for enhanced repair of the ACL. |
| Sectors | Healthcare,Pharmaceuticals and Medical Biotechnology |
| URL | https://www.ecmconferences.org/abstracts/2018/Collection4/c4.html |
| Description | As the work performed for this project has demonstrated the potential of fibrillin-1 to enhance regeneration potential of ACL cells of both human and canine origin. Though this work was mainly focused on tissue engineering, it has highlighted and added to the evidence of how specific protein fragments could be used in healthcare as a functionalisation method for pre-existing grafts to aid in the repair and recovery of damaged ACLs. The work performed on characterisation of extracellular matrix proteins in osteoarthritic knees has added to current knowledge by determining the morphology, structural positioning and presence of specific matrix proteins are affected, for added understanding of how osteoarthritis affects the surrounding tissues. |
| First Year Of Impact | 2021 |
| Sector | Healthcare,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Societal |
| Description | Henry Royce Institute Research Grant |
| Amount | £5,663 (GBP) |
| Organisation | Henry Royce Institute |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 08/2019 |
| End | 09/2019 |
| Description | President's Doctoral Scholars Award |
| Amount | £3,000 (GBP) |
| Organisation | University of Manchester |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 09/2016 |
| End | 02/2020 |
| Description | President's Doctoral Scholars Travel Award |
| Amount | £500 (GBP) |
| Organisation | University of Manchester |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 09/2018 |
| End | 09/2018 |
| Description | Collaboration with Liverpool Musculoskeletal Biobank - human ACL cells |
| Organisation | University of Liverpool |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We are using the tissue to digest and remove the cells for experimentation relevant to the biobank and associated academic's interests. They will have access to all our work and data and we will be providing them with regular updates. |
| Collaborator Contribution | In turn, the Liverpool Musculoskeletal Biobank (LMB) will be providing us with human anterior cruciate ligament tissue and advice for the continuing experimentation. |
| Impact | No outputs or outcomes as of yet, though one paper is being prepared for publication. |
| Start Year | 2019 |
| Description | Collaboration with Liverpool University - Canine CL cells |
| Organisation | University of Liverpool |
| Department | Institute of Ageing and Chronic Disease |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Though we have not had any contribution in terms of training, we will be including the research team at Liverpool University as authors in any papers we publish and we will be furthering work which is of interest to their research focus as a whole. |
| Collaborator Contribution | They have provided primary canine cells, which have been used in the first two parts of the project, I have worked with them at Liverpool University, performing experiments with post doctoral staff. |
| Impact | Presentations at two conferences, one European and one national. Posters presented at two conferences, one European and one national. We are also currently writing a paper on the data gathered thus far and intend to publish in the very near future. |
| Start Year | 2016 |
| Description | Lecture for a girl's STEM visit |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Approximately 50 female students attended a lecture where I discussed research and the area in which I perform my research. I have discussed parts of my research with the students and explained to them the importance of ongoing research in my immediate area. |
| Year(s) Of Engagement Activity | 2018 |
| URL | http://www.mub.eps.manchester.ac.uk/science-engineering/2019/02/14/dragonfly-day-women-can-do-anythi... |
| Description | Postgraduate summer research showcase |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | A poster fair regarding postgraduate student's research projects. Posters are hung and discussed and general public, staff and students are welcomed to discuss research going on within the university. Visitors reported fascination with the work and interest in the potential long-term development of the research. |
| Year(s) Of Engagement Activity | 2018 |
| URL | http://www.psrs.manchester.ac.uk/ |
| Description | STEM Fair Stand at Manchester Cricket Grounds |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | A stand was made for the interest of 4 years groups of 11 - 15 year old students at the Old Trafford cricket grounds centre in Manchester. Students could take part in small stand-suitable activities and learn about biomaterials and talk about the research concerning this area. Teachers reported extreme interest in the areas discussed and have requested further events for the purpose of introducing students to new and fascinating STEM areas. |
| Year(s) Of Engagement Activity | 2018 |
| Description | STEM for girls workshops held at the University of Manchester |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | 45 female pupils from secondary schools in the greater Manchester area attended outreach events for widening participation. We discussed and performed experiments relevant to the area I am conducting my project work. The school reported that they very much enjoyed the workshop and that it had sparked a lot of interest from the attending pupils. |
| Year(s) Of Engagement Activity | 2018,2019 |
| URL | http://www.mub.eps.manchester.ac.uk/science-engineering/2019/02/14/dragonfly-day-women-can-do-anythi... |
| Description | School visit to talk about STEM subjects |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Visit made to a school in the greater Manchester area for 5 workshops spread across the day. In these workshop sessions, I interacted with boy male and female pupils of ages between 12 and 15 and introduced them to my research area and the work we do. I also introduced them to my research and explained why it was important for furthering orthopedic medicine. We performed a large practical experiment where the students discovered how to make simple biomaterials of their own. |
| Year(s) Of Engagement Activity | 2018 |