Rapid Bone Graft Synthesis Through Dual Piezoelectric/Nanomechaniocal Stimulation

Lead Research Organisation: University of Glasgow
Department Name: College of Medical, Veterinary &Life Sci

Abstract

Bone graft is regularly used in surgery (plastics, maxillofacial surgery and orthopaedics); bone is actually the second most grafted tissue after blood. Ideally the surgeon wishes to take bone from one area (donor site) to another area (recipient site) to support the operation they are performing or to replace damaged, infected or cancerous tissue . However, a patient's own donor bone is in short supply and its removal can lead to complications at the donor site. This means the surgeon will often recourse to allograft - decellularised (and thus biologically inferior) - bone from cadavers or other people. A third, and growing, option is synthetic graft. Synthetic graft can be made from biologically active materials, but is not viable at the time of implantation and thus not yet as good as living bone, as cells must be made to infiltrate the material prior to regeneration.
It has been known for many decades that bone tissue responds to mechanical loading and commonly, mechanical loading via a bioreactor device is employed to enhanced bone formation in vitro. Recently, it has been also discovered that cells respond to nanoscale mechanical stimulation, and in particular, that nanoscale vibrations induce rapid bone formation from cells cultured in vitro. Interestingly, bone tissue produces minute electrical fields when loaded, a phenomenon which is hypothesised to help with the bone remodelling process during regeneration. However, if this electrical stimulation is important for the rapid generation of bone graft is not yet known and is a focus of this project.
Our bioreactor, that supplies nanoscale 'kicks' to cells in culture can be used to convert mesenchymal stem cells (the stem cells of the bone, simple to isolate from a patient's iliac crest or fat tissue) to bone forming osteoblasts. We will optimise this by designing new 3D architectures for the cells to grow in that work in synergy with the bioreactor. The scaffolds will emit physiological electrical signals that the body uses as cues to heal bone. Further, we will look at the biological building blocks, metabolites, that the cells use to form bone and incorporate these into delivery systems that also work with the bioreactor. The electricity produced by the scaffolds will trigger release of the metabolites, further driving bone formation.
This will ultimately allow us to improve the quality of living bone graft derived from a patient's own cells while reducing the lab time we need to make it.

Technical Summary

We have developed a new bioreactor, the Nanokick, that supplies nanoscale high frequency-low amplitude mechanical cues to MSCs in 2D and 3D to induce osteogenesis. We aim to develop consumables that partner the Nanokick and further stimulate the MSCs. Our goal is to induce large numbers of MSCs to enter osteogenesis as quickly as possible within a 3D environment to provide tissue engineered bone graft and to study this process as a function of electromechanical stimulation.
To achieve this will make full use of the mechanical cues provided by the Nanokick and use piezoactive materials to provide electrical stimulation to the cells in order to potentiate osteogenesis. These will be both in disposable (coverslip that sits in the bottom of the well plate and can be discarded after use) and scaffold format (to enable future tissue engineering as in vivo toxicology is addressed). Cells will be seeded over the coverslips or scaffolds in synthetic hydrogels. We will tailor the hydrogels with adhesive and degradable motifs to optimise the 3D environment; the move to synthetic materials is important as it will facilitate xeno-free protocols as we develop the graft further.
Further, we will use our metabolomics workflows to identify bioactive metabolites that interact with biochemical pathways that we will identify. The metabolites will be incorporated into electrostimulated drug delivery vehicles and incorporated into the piezoactive coverslips or scaffolds. This will enable drug stability and delivery only as the nanokicking process is commenced.
Finally, we will investigate mechanotransductive pathways with particular focus on transmembrane mechanoresponsive ion channels such as piezo 1 and 2.
The work is important primarily as tissue engineered bone graft is required to support the aging population. However, it will also deliver mechanoresponsive-cell stimulating scaffolds an drug delivery systems containing novel metabolite drugs with known biochemical pathway.

Planned Impact

Scientists and Healthcare Professionals.
We will use open access publishing to disseminate as broadly as possible. This will be of direct interest to the biomaterials, tissue engineering, stem cells, pharma and medical communities.

Laboratories.
There is a range of bioreactors available to academic laboratories including mechanical (eg BOSE, EBERS), perfusion (eg BOSE, Minucell) and rotary (eg Synthecon). All have drawbacks for bone production ranging from affordability, sterility, usability and functional outcome. The advantage of our system is that it uses standard cell culture consumables and thus standard protocols. This will clearly open up 3D bone research to many more labs.

Industry.
In the first instance our work will be of interest to industrialists working in the mechanotransduction (bioreactor) and material science (synthetic bioactive graft) areas. We will develop a bioreactor, consumables/scaffolds for it and try to move to use synthetic environments for our 3D bone graft. Companies working in this area include those with interest in bioreactors (e.g. BOSE, EBERS, Minuth, Synthecon etc) and providers of innovative tissue engineering scaffolds (e.g. BiogelX, Stem Cell Technologies, Baxter Healthcare, Medtronic etc). These companies would generate economic benefit from co-developing, licensing and exploiting our bioreactors and consumables for them. Secondly, we will appeal to Pharma with our novel mass spectrometry approaches to drug design (e.g. GSK). Thirdly, we will appeal to companies with an interest in advanced technologies, such as Medtronic, Genzyme and Advanced Tissue Solutions, who will be interested in tissue engineering approaches to bone graft.

Patients.
Bone graft is the second most transplanted tissue after blood. Autograft harvest from e.g. the iliac crest has a range of associated complications including herniation, instability, nerve damage, infection, deep hematoma requiring surgical intervention and pain [1]. Synthetic bone graft is increasing in use but is biologically inferior and tends to be used with BMP (e.g. Medtronic Infuse)[2]. BMP has to be used in high doses [3] and this has led to serious respiratory, neurological and inflammatory complications and the issuing of a public health warning by the FDA [4]. Subsequently, reports of the carcinogenic potential of BMP have also been noted. The development of "clean" tissue engineered bone graft with good volume (cms3) would be a major advantage. All surgeons we have surveyed strongly agree there is urgent unmet need.

Health Service Providers.
- Delivery of tissue engineered bone graft would reduce the need for two surgeries. This would represent a saving on surgical time and could be competitively placed compared to e.g. BMP treatment, which can cost up to £3k per patient in non-union fixation [4].
- Delivery of novel drugs and delivery systems. We will identify bioactive metabolites to enhance graft engineering. Our model could also be used to test 3rd party drugs and to advise personal therapies.
Public
Our work will raise interest in science, increasing awareness of the scientific challenges of supporting an aging population. We will engage with the next generation of scientists to inspire them to overcome the burden and limitations of bone grafting and bone regeneration.

1. Arrington 1996. Clin Orthop Relat Res. 329, 300-9.
2. Epstein 2013. Surg Neurol. 4, S343-S352.
3. Garrison et al 2007. NIHR Health Technology Assessment programme.
4. FDA Public Health Notification.
5. Dahabreh 2007. JBJ Editorial.

Publications

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Donnelly H (2018) Designing stem cell niches for differentiation and self-renewal. in Journal of the Royal Society, Interface

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Robertson SN (2018) Control of cell behaviour through nanovibrational stimulation: nanokicking. in Philosophical transactions. Series A, Mathematical, physical, and engineering sciences

 
Description We have adapted our nanovibrational bioreactor to work with piezo active scaffold materials to deliver small electrical potentials to the cell sin the bioreactor.
Exploitation Route We are lookign to develop novel tissue engineering scaffolds that can allow cells to be preconditioned in the lab.
Sectors Healthcare

 
Description training of practioners and researchers
Geographic Reach Multiple continents/international 
Policy Influence Type Influenced training of practitioners or researchers
 
Description Early diagnosis and intervention of osteoporosis using nanovibrational stimulation
Amount £480,000 (GBP)
Funding ID ST/S000852/1 
Organisation Science and Technologies Facilities Council (STFC) 
Sector Academic/University
Country United Kingdom
Start 11/2018 
End 10/2020
 
Description Engineering growth factor microenvironments - a new therapeutic paradigm for regenerative medicine
Amount £3,661,144 (GBP)
Funding ID EP/P001114/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Academic/University
Country United Kingdom
Start 10/2016 
End 09/2021
 
Description Excellence with Impact - Outreach and Stakeholder engagement at the Royal Society Summer Science Exhibition
Amount £3,000 (GBP)
Organisation University of Glasgow 
Sector Academic/University
Country United Kingdom
Start 06/2018 
End 05/2019
 
Description Wellcome Trust ISSF Early Career Researcher Fund
Amount £18,350 (GBP)
Organisation Wellcome Trust 
Department Wellcome Trust Institutional Strategic Support Fund
Sector Charity/Non Profit
Country United Kingdom
Start 03/2018 
End 12/2018
 
Title Polymer surfaces to control growth factor presentation. 
Description We have made a polymeric surface that drives organisation of fibronectin into nanonetworks that bind growth factors at ultra-low dose but with high biological efficacy. 
Type Of Material Technology assay or reagent 
Year Produced 2016 
Provided To Others? Yes  
Impact We have performed a first veterinal trial with the technology that has been very successful. We are now progressing towards human use. 
 
Title Use of NanoKick Bioreactor for chondrogenic differentiation of MSCs 
Description Modification of cell culture method to induce chondrogenic differentiation of MSCs when using the NanoKick Bioreactor. 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2017 
Provided To Others? No  
Impact Preliminary data obtained further funding from a Wellcome Trust ISSF Early Career Catalyst Fund (~£18300). 
 
Title Control of cell behaviour through nanovibrational stimulation: nanokicking 
Description  
Type Of Material Database/Collection of data 
Year Produced 2018 
Provided To Others? Yes  
 
Description Collaboration for Synthesis of Osteogenic Metabolites and Chemical Derivatives 
Organisation University of Glasgow
Department School of Chemistry
Country United Kingdom 
Sector Academic/University 
PI Contribution Identification of interesting metabolites in osteogenesis and in vitro testing of osteogenic potential
Collaborator Contribution Chemical synthesis of metabolite molecules
Impact We will shortly publish results from this collaboration.
Start Year 2017
 
Description Collaboration with Dr. Manus Biggs in National University of Ireland, Galway to creat Dual Piezoelectric/Nanovibration system for rapid bone regeneration 
Organisation National University of Ireland, Galway
Country Ireland 
Sector Academic/University 
PI Contribution Combining piezoelectric scaffold provided by Dr Manus group with our nanovibrator for MSCs osteogenic stimulation.
Collaborator Contribution Synthesizing piezoelectric scaffold.
Impact The collaboration just started recently. We will publish a paper in near future.
Start Year 2017
 
Description Collaboration with Geistlich 
Organisation Geistlich
Country Switzerland 
Sector Private 
PI Contribution Testing of in vitro cell culture on Geistlich implant materials
Collaborator Contribution Contribution of implant materials from Geistlich
Impact Disciplines involved - Dentistry, cell engineering, materials
Start Year 2018
 
Description Collaboration with Prof Stuart Reid, University of Strathclyde 
Organisation University of Strathclyde
Country United Kingdom 
Sector Academic/University 
PI Contribution Together, we have designed a new bioreactor and consumables for it.
Collaborator Contribution They design and build the bioreactors.
Impact THis is a long standing collaboration in which we designed and build the nanovibrational bioreactor (originally Nanoforce, now Nanokick). We have worked togther, produced patents and publications as well as securing further funding.
Start Year 2011
 
Description Collaboration with Prof. Salmeron in school of engineering of University of Glasgow for functional and controlable bone scaffold. 
Organisation University of Glasgow
Country United Kingdom 
Sector Academic/University 
PI Contribution Using our noval nanovibrator conbining 3D cell culture system to assess PEG-gel based functional and controllable bone scaffold.
Collaborator Contribution Helping us to synthesize the PEG-Gel based functional and controllable bone scaffold.
Impact No outcomes yet, as the collaboration has been recently started.
Start Year 2018
 
Description Souhtampton Collaboration 
Organisation University of Southampton
Department Bone and Joint Research Group
Country United Kingdom 
Sector Academic/University 
PI Contribution This is a long term collaboration where cells, materials and expertise are shared to drive research
Collaborator Contribution This is a long term collaboration where cells, materials and expertise are shared to drive research
Impact A large number of papers, grants and IP.
 
Title Mechanical Bioreactor 
Description Design of a novel nanovibrational bioreactor for bone graft production 
IP Reference WO2017029393A1 
Protection Patent application published
Year Protection Granted 2017
Licensed No
Impact None yet
 
Description FEBS Workshop Biological Surfaces and Interfaces: Interface Dynamics, 2nd-7th July 2017, Sant Feliu de Guixols, Catalonia, Spain 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Mr W. Orapiriyakul had a Poster presentation at FEBS Workshop Biological Surfaces and Interfaces: Interface Dynamics, 2nd-7th July 2017, Sant Feliu de Guixols, Catalonia, Spain
Year(s) Of Engagement Activity 2017
 
Description Glasgow Orthopaedic Research Initiative (GLORI 2018), 23d February 2018, Glasgow, UK. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Mr W. Orapiriyakul had a Poster presentation at Glasgow Orthopaedic Research Initiative (GLORI 2018), 23th Febuary 2018, Glasgow, UK.
Year(s) Of Engagement Activity 2018
 
Description 28th Annual Conference of the European Society for Biomaterials (ESB) 4-8th September 2017, Athens, Greece. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Mr W. Orapiriyakul: Oral presentation at 28th Annual Conference of the European Society for Biomaterials (ESB) 4-8th September 2017, Athens, Greece.
Year(s) Of Engagement Activity 2017
 
Description 29th European Conference on Biomaterials (ESB 2018), 9-13th September 2018, Maastricht, Netherlands. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Mr W. Orapiriyakul had a Poster presentation in 29th European Conference on Biomaterials (ESB 2018), 9-13th September 2018, Maastricht, Netherlands.
Year(s) Of Engagement Activity 2018
 
Description 5th TERMIS World Congress 2018, 4-7th September 2018, Kyoto, Japan. 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact mr W.Orapiriyakul had a Poster presentation in 5th TERMIS World Congress 2018, 4-7th September 2018, Kyoto, Japan.
Year(s) Of Engagement Activity 2018
 
Description BLOG CeMi: NANOKICKING STEM CELLS TO GROW NEW BONE 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Monica P. Tsimbouri wrote a blog about the people, work and challenges involved for the recent paper
Tsimbouri, Childs et al. Stimulation of 3D osteogenesis by mesenchymal stem cells using a nanovibrational bioreactor. Nat. Biomed. Eng. http://doi.org/10.1038/s41551-017-0127-4 (2017).
Year(s) Of Engagement Activity 2017
URL https://www.gla.ac.uk/research/az/thecemi/blog/nanokickingstemcellstogrownewbone/
 
Description Biomaterials seminar 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact Mr W Orapiriyakul gave a talk at the Biomaterials seminars session on 13/11/18
Year(s) Of Engagement Activity 2018
 
Description Conference presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Identification and in vitro screening of osteogenic metabolites through supplement-free nanovibration-driven mesenchymal stem cell differentiation (2018) Tom Hodgkinson, Karl Burgess, David France, Paul Campsie, Shaun Robertson, David Phillips, Peter Childs, Richard Oreffo, Stuart Reid, Manuel Salmeron-Sanchez, Matthew Dalby - Matrix Biology Europe, Manchester, UK.
Year(s) Of Engagement Activity 2018
 
Description Conference presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Conference presentation by Tom Hodgkinson at European Society for Biomaterials, Maastricht
Year(s) Of Engagement Activity 2018
 
Description Conference presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Identification and in vitro screening of osteogenic metabolites through supplement-free nanovibration-driven mesenchymal stem cell differentiation (2018) Tom Hodgkinson, Karl Burgess, David France, Paul Campsie, Shaun Robertson, David Phillips, Peter Childs, Richard Oreffo, Stuart Reid, Manuel Salmeron-Sanchez, Matthew Dalby - Matrix Biology Europe, Manchester, UK.
Year(s) Of Engagement Activity 2018
 
Description Conference presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Identification and in vitro screening of osteogenic metabolites through supplement-free nanovibration-driven mesenchymal stem cell differentiation (2018) Tom Hodgkinson, Karl Burgess, David France, Paul Campsie, Shaun Robertson, David Phillips, Peter Childs, Richard Oreffo, Stuart Reid, Manuel Salmeron-Sanchez, Matthew Dalby - TCES 2018, Keele, UK.
Year(s) Of Engagement Activity 2018
 
Description Conference presentation 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Third sector organisations
Results and Impact Identification and in vitro screening of osteogenic metabolites through supplement-free nanovibration-driven mesenchymal stem cell differentiation (2018) Tom Hodgkinson, Karl Burgess, David France, Paul Campsie, Shaun Robertson, David Phillips, Peter Childs, Richard Oreffo, Stuart Reid, Manuel Salmeron-Sanchez, Matthew Dalby - GLORI, Glasgow UK.
Year(s) Of Engagement Activity 2018
 
Description GLORI at the CeMi 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Tom Hodgkinson, Talk
"Identification and in vitro screening of osteogenic metabolites through supplement-free
nanovibration-driven mesenchymal stem cell differentiation"
Glasgow Orthopedic Research Initiative 2018
23rd November 2018
Year(s) Of Engagement Activity 2018
 
Description GLORI at the CeMi 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Mr Ian Kennedy presented a talk on
"Nanoscale vibrations to modulate osteogenesis"
Glasgow Orthopedic Research Initiative 2018
23rd November 2018
Year(s) Of Engagement Activity 2018
 
Description GLORI conference organisation 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Planning and Organisation of Dr Monica P Tsimbouri the scientific mentor of the Glasgow Orthopaedic Research Initiative (GLORI). GLORI has been established to encourage collaboration between the basic sciences, applied sciences, engineering and clinic. This has the aim of bringing the latest ideas in basic materials research into use to deliver the next-generation of orthopaedic care. It combines expertise from orthopaedic surgeons, biologists, engineers and chemists not only from Glasgow and Strathclyde Universities but the rest of Scotland too and also with the view to expand to the rest of the UK.
Year(s) Of Engagement Activity 2018
 
Description GLORI-2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact National orthopedic conference in Glasgow where about 60 audience attended.
Year(s) Of Engagement Activity 2018
 
Description Glori2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Submitted a research abstract, stimulating discussion and chaired a conference session on bone regeneration strategies
Year(s) Of Engagement Activity 2018
 
Description IKEA at home with life sciences 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Dr Monica P. Tsimbouri
participated at the
IKEA at Home with Life Sciences
A team of students, researchers and support staff represented the Institute at IKEA on Saturday 24th November 2018 as part of the MVLS at IKEA event.
Year(s) Of Engagement Activity 2018
URL https://www.gla.ac.uk/researchinstitutes/biology/newsevents/headline_581051_en.html?utm_medium=email...
 
Description NEB community: the story behind the paper 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr P.M. Tsimbouri and Dr P. Childs were asked by the NEB community to write a story behind their recent paper
Tsimbouri, Childs et al. Stimulation of 3D osteogenesis by mesenchymal stem cells using a nanovibrational bioreactor. Nat. Biomed. Eng. http://doi.org/10.1038/s41551-017-0127-4 (2017).

Story title:
Growing bone using nanovibration:
A reproducible method for mechanically driving stem-cell osteogenesis in 3D culture.
Year(s) Of Engagement Activity 2017
URL http://go.nature.com/2xVgSNN
 
Description Oral Presentation at Biomaterials Seminar, University of Glasgow 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Presented research on nano vibrational differentiation of mesenchymal stem cells as part of the biomaterials seminar series at the University of Glasgow
Year(s) Of Engagement Activity 2018
 
Description Poster presentation 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Wich Orapiriyakul presented a poster at GLORI2018
Year(s) Of Engagement Activity 2018
 
Description Royal Society Summer Exhibition 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Schools
Results and Impact We presented our growth factor based bone regeneration technology at the Exhibition
Year(s) Of Engagement Activity 2018
 
Description Scottish Gocernment Outreach Event Edinburgh 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Policymakers/politicians
Results and Impact Outreach event to mark the Scottish Governments science policy launch
Year(s) Of Engagement Activity 2018
 
Description Talk at GLORI2018 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Dr Monica P Tsimbouri delivered a talk on behalf of Prof Dalby at GLORI2018
Year(s) Of Engagement Activity 2018
 
Description The University Network ((TUN) 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Dr Monica P. Tsimbouri was invited by the The University Network ((TUN), a global media company for college students available in 108 languages) to write an article about the nanokicking project. (https://www.tun.com/blog/nanokicking-technology-bone-growth-development/)
Year(s) Of Engagement Activity 2017
URL https://www.tun.com/blog/nanokicking-technology-bone-growth-development