MICA: Open-access biomaterials microfabrication and non-invasive imaging facilities for Regenerative Medicine.

We propose a transformative instrumentation bid to catalyse collaborative regenerative medicine research. We are requesting to purchase capital equipment which will allow the regenerative medicine community in Sheffield (who are all members of the University Centre for Biomaterials and Tissue Engineering) and the UK to keep at the forefront of internationally competitive research. Between them the applicants have experience of tackling clinical problems, which are not currently amenable to conventional treatment - MacNeil has developed tissue engineered skin for patients with extensive skin loss due to burns injuries and commercialised this as a project (MySkin). She has also developed tissue engineered buccal mucosa and translated this into early stage clinical evaluation for patients with scarring of the urethra. She is now working with colleagues in India to develop a simpler and safer alternative to the amniotic membrane, which is currently used to culture corneal stem cells for transfer to the clinic for patients with scarring of the cornea. This work on microfabricating a rapidly degrading scaffold (with Claeyssens) has also led to a new methodology for scaling up and speeding up the projection of electrospun scaffolds which contain 3D structures to protect these fragile cells.

Haycock has made significant breakthroughs in how to culture stem cells for peripheral nerve repair and working with Claeyssens has developed nerve tissue guides into which these cells are placed. This work is showing significant promise for nerve regeneration in animal models of nerve transection, with plans for clinical translation. The development of these tissue-engineering technologies is very much underpinned by current instrumentation and expertise within the laboratory, but has been strengthened in recent years with the appointment of Matcher who has international expertise in biophotonics, and this is being applied to imaging 3D tissue constructs. The purchase of new equipment will allow this work to progress into the clinic to image cells and constructs post implantation in the cornea, skin and nerve. Claeyssens brings expertise in microfabrication which has already led to significant advances in ongoing programmes of cornea and nerve repair and has the potential to advance to the production of patient specific constructs and to scale up the production of constructs for clinical trials. Rehman brings a wealth of expertise on FTIR and Raman spectroscopy of biological tissues, including bone, cell, tissues and histopathologically fixed biopsies, and these techniques are currently being applied to investigating 3D constructs of skin and analysing surface properties of scaffolds. In addition, his expertise in developing bioactive-guided regenerative membranes for various applications, such as periodontal treatment and restorative materials adds considerably to the biomaterials manufacturing portfolio of this centre. The latter technique is information rich while imaging cells with optical coherence tomography has relatively poor resolution but is fast and convenient and can be translated to the clinic. We anticipate synergy between having both technologies available.

Thus, the applicants have between them a range of applications and instrumentation technologies to address problems in regenerative medicine. The equipment requested will not only support these projects but will benefit other members of the UK Regenerative Medicine community both locally and across the UK research community. This will be achieved by running the technologies as open access facilities for the UK research community, as it is unlikely that all such Centres in the UK will have access to microfabrication and state-of the art non-invasive imaging facilities. These facilities underpinned by the research expertise available in Sheffield will lead to new collaborations and will benefit UK Regenerative Medicine.

Biomaterials fabrication: The biomaterials fabrication suite will house a femtosecond laser-based microstructuring facility, able to structure macroscopic 3D tissue engineering scaffolds on a high speed and with sub-micrometer resolution. This set-up will be complemented with a fused-deposition modelling set-up to produce scaffolds from thermoplastic polymers and composite materials and for precise protein patterning and cell printing. Additionally, a commercial electrospinning rig will be used to allow for reproducible manufacture of electrospun scaffolds for clinical applications.
Non-invasive imaging:
Light Sheet Fluorescence Microscopy provides enhanced sectioning capabilities by exploiting thin laser light sheet for imaging samples perpendicularly to the direction of observation. The advantage of this equipment is its application to living samples as it causes less photo-damage and stress, as only the observed section is illuminated, leading to higher contrast imaging.
A Raman Spectrometer including 455, 532, 633, 780 nm excitation lasers and a FTIR spectrometer equipped with a microscope, Attenuated Total Reflectance (ATR), Diffuse Internal Reflectance (DRIFT) and Photo-Acoustic Sampling (PAS). Both experiments will provide micro-spectroscopic information of a wide range of natural tissues and scaffold materials. The FTIR set-up will also enable time- and phase resolved spectroscopy. In addition, these set-ups will allow monitoring of single cell, cell interactions with the scaffolds and cell proliferation.
Advanced clinical biophotonic imaging based on a clinical OCT and multi-photon imaging facility: (i) an in-vivo multi-beam OCT will enable monitoring in-vivo tissue regeneration and angiogenesis. (ii) a commercial in-vivo second harmonic and multi-photon platform will be used to study In-vivo collagen formation. This system also provides label-free imaging of lipids, to study barrier layer formation and damage mechanisms in-vivo.

UK Academia:
We will be pro-active in promoting new collaborations with other academics for access to this equipment and the underlying expertise behind it. Thus the equipment will have immediate impact on the on-going research of the applicant and co-applicants but will also contribute to the on-going research of other members of the Centre for Biomaterials and Tissue Engineering and the Centre for Stem Cell Research in Sheffield and to any other interested parties in the UK.
NHS Clinicians:
The applicants MacNeil, Haycock and Claeyssens are working with clinical colleagues in the UK and overseas - MacNeil on the cornea (UK and India) and Haycock on tissue engineering of nerve (UK and Sweden) on research which we intend to translate into first in man clinical studies within the next 18 months. The availability of the biomaterials microfabrication equipment will strongly aid achieving reproducible production of biodegradable constructs for combination with cells for implantation into patients. The OCT equipment we request is suitable for use in man and hence will help us evaluate materials and cells post implantation in a non-invasive manner. There is a particular need for equipment that will allow the monitoring of scaffolds and cells into patients - this is in itself a major challenge and the OCT equipment we request will be only part of the solution for this for the UK.
UK Industry:
The impact for industry is that these open access equipment facilities will provide an attractive solution for manufacturers to send prospective customers to visit equipment that is in use and in particular to learn about the performance of the equipment from other academic researchers rather than directly from a company alone. This model for using equipment as a demonstrator site for industry is a successful one and can be very much to the advantage of equipment manufacturers.
On the basis of this manufacturers have already offered significant discounts in the quotations for the equipment we propose to buy. Further through the Centre for Biomaterials and Tissue Engineering we will invite Industry to our annual/bi-annual meetings where work is communicated openly. The Centre also runs small themed workshops on particular clinical problems or technologies and we would anticipate running such workshops to showcase the biomaterials microfabrication suite of equipment and a separate workshop to showcase the new equipment for non-invasive imaging. From experience demonstrating data obtained with new equipment is the best way to engage the interest of researchers and often leads to new collaborations.
General public and Schools:
The applicants are involved in communicating science to the general public in a number of ways. The Kroto Research Institute every year has a visit from Professor Sir Harry and Lady Margaret Kroto and we have a school outreach programme associated with the visit. Many of the staff in the KRI also engage in outreach programmes in schools describing research particularly in regenerative medicine. The translational stories that we have to tell - of developing skin cells to benefit burns patients and developing better ways to treat nerve injuries are stories that are easily understood in terms of their application and attractive to school children as examples of how regenerative medicine can contribute to society and provide an interesting career.