Final Report Summary - INNOVABONE (Novel biomimetic strategy for bone regeneration)
Executive Summary:
InnovaBone teams successfully created and produced the InnovaBone product with biodegradable 2-photon polymerized scaffolds, an extensive family of innovative state-of-the-art SMART biomimetic, bioinspired, biodegradable, thermosensitive Elastin-like recombiners (ELRs) containing biologically active bone growth factors with controlled amounts of hydroxyapatite (HA) nanoparticle to enhance bone repair and healing. Novel Equipment was developed by an InnovaBone team including a novel compact simple, low cost bioreactor for testing bone repair scaffolds in long-term cell culture under mechanical stimulation and state-of-the art 2-photon polymerization equipment able to produce large scaffolds of 15 x 15 mm. An InnovaBone team developed an innovative and flexible Data Sample Lifecycle Management app for real time tracing and tracking of scientific experiments.
To study the biomaterials, several InnovaBone teams worked together to generate a cohesive fully integrated platform for testing real time in vitro degradation, in vitro and in vivo bone repair and healing, high throughput and a subchronic in vivo models for evaluating immune and inflammatory foreign body responses to biomaterials, primary human cells testing, and innovative imaging technology for evaluating bone repair following biomaterial implantation.
Training, Stakeholder engagement and Dissemination was successfully accomplished by InnovaBone teams with networking and participative methodologies to support innovative discussions and brainstorming amongst researchers and stakeholders in Europe. Furthermore, state of the art regulatory guidance was provided to ensure progress beyond the project of the foreground generated during the life of the project.
The InnovaBone project created a novel strategy using a multicomponent approach for the healing and repair of non-union bone lesions. A broad biomaterials evaluation platform for bone healing and repair was developed along with several additional commercial products. The InnovaBone team was engaged primary stakeholders from the beginning of the project to ensure the interaction of the clinician and scientists for the best possible product for clinical care. Wide dissemination to promote the project findings was done and will continue with over 20 manuscripts in progress for publication and 5 patents published or in the application stage.
Project Context and Objectives:
The importance of developing novel approaches for bone repair is underscored by the heavy burden on health care costs and patient suffering caused most significantly by traumatic bone lesions, osteoporosis and osteolytic bone metastases. The incidence and prevalence of osteoporosis and related fractures increases as the population ages. Approximately 33% of women and 12.5% of men over the age of 50 will suffer an osteoporotic fracture. Over 800,000 bone bank-derived cadaver allografts (the most commonly used bone substitute in Europe) are used annually and direct costs for osteoporotic fractures alone are €36 billion (PMID:15618996), illustrating the tremendous burden on health care (PMID:11712842). There are therapeutics that retard bone loss and others, which repair large lesions and fractures that do not undergo self-repair, but many of the current approach are associated with complications: high risk for transferable infections, immune responses, chronic inflammation and rejection, stress-shielding and loosening due to wear, material debris formation. All these undesirable events lead to increased costs and recovery time. New biomaterial approaches must be sought which involve the use of various unconventional technologies, cell seeding, pertinent growth factors and cell migration factors (PMID:18454418). Despite substantial advances in biomaterials, further R&D is essential to overcome the limitations of current implant materials, such as improper mechanical strength and durability and insufficient healing caused by foreign body reactions and inefficient vascularisation and bone cell function. Thus, new bone regeneration strategies are necessary.
Our main objective was to achieve radical innovations in state-of-the-art bone biomaterials in order to address the serious morbidity and mortality associated with bone lesions both in the EU and Worldwide. To accomplish this we devised a plan, guided by the needs of the clinician and patient alike, to design and produce optimally performing bioinspired materials based on the knowledge gained from the natural physiological processes underlying bone repair and regeneration. This pioneering strategy aimed to accelerate bone healing and reduce adverse side effects with currently employed materials. Our approach was to develop a 2-step biomimetic product consisting of a bioactive scaffold and a bioactive self-setting gel. Step 1 will be to introduce a firm but compressible bio-inspired, heparinised 3D scaffold which would reduce the formation of blood clots around the implant (one of the major problems with biomaterials leading to unsuccessful engraftment) and provide structural stability of the lesion. Step 2 involved injecting a bioactive recombinant gel into the scaffold. This was planned to provide a microenvironment that promotes bone repair due to the bioactive factors grafted and dispersed within the biogel and scaffold structure. The expected impact will be a radical innovation that accelerates the treatment of large bone lesions.
The step 1 3D scaffolds were produced using two-photon laser polymerisation (2PP). These polymer scaffolds were designed to be mechanically stable and easily trimmed and fitted into bone lesions. In the bone lesion site, these scaffolds were to provide stability during the formation of new bone and would degrade within approximately a year during the bone healing process. The bioactive gel was created with functional and genetically engineered recombinant elastin-like recombinamers (ELRs) which are thermosensitive and self-solidifying. These polymers have absolute precisely controlled molecular architecture and possess sequences that contain specific domains for cell attachment and growth factors. Calcium phosphate nanoparticles (CaP-NPs) dispersed in the gel were generated and added to the biogels to attract and activate the cells important for initiating bone repair.
The objectives of the InnovaBone project were to design and produce a novel 2-step smart bio-scaffold and smart self-setting gel, evaluate the biological and physical properties of bioscaffold-biogel biomaterials, upscale the technological processes allowing the manufacture of prototype scaffolds, and perform technical characterisation and design strategies for product exploitation.
Project Results:
The main S&T results/foregrounds are divided into 1) Biomaterials development, 2) Equipment development, 3) App development, 3) Biomaterial evaluation platform, 4) Stakeholder engagement, Training and Dissemination 4) Exploitation of InnovaBone products and 5) Summary . In each section, the results are highlighted per partner.
Biomaterials development
iba:
1) successfully established a novel material platform for bone or cartilage tissue engineering and cancer research based on FDA approved components like Lactide and caprolactone biopolymers
2) developed technology and parameters for the 2-photon polymerization (2PP) of “tailor-made” designer scaffolds based on triple periodic minimal surfaces (e.g. Schwarz P surfaces) with variable stiffness and biodegradability for bone repair
3) has acquired one patent which was published in July 2015 on their 2PP approach and especially for the use of highly efficient and fast photoinitiators
4) engaged with industrial partners for further development and commercialization and the potential for industry to purchase this patent
UVa:
1) created a new extensive family of innovative state-of-the-art SMART biomimetic, bioinspired, biodegradable, thermosensitive Elastin-like recombiners (ELRs) containing biologically active bone growth factors that are at room temperature are injectable liquids that gelify at body temperature and provide the a microenvironment conducive for bone repair and healing
2) produced artificial biogels with tremendous potential for clinical applications such as in the repair and healing of bone, cartilage and skin
3) established a Step-change in scale up technology for ELR production, making it accessible to a wider community including the high industrial potential and a broad range of new applications and clinical implications
UPC:
1) developed technology that allows the introduction of controlled amounts of doping ions (e.g. Carbonate, magnesium) into the hydroxyapatite (HA) nanoparticle (NP) crystal structure
2) by controlling the composition of the HA NPs, they produced biologically active, non-toxic HA NPs for bone repair and healing
3) developed an innovative method for the dispersion of NPs into liquid Elastin-like Recombiners (ELRs) allowing for homogenous distribution which is a crucial step for reducing NP aggregates and thus, maintaining NP integrity, effectiveness and biological function
4) produced HA NPs that were critical for in vivo bone formation and healing
Equipment development
CSEM:
1) produced a novel compact simple, low cost bioreactor for testing bone repair scaffolds in long-term cell culture under mechanical stimulation
2) has demonstrated that the mechanical properties of the scaffold can be monitored in real time during culture in the bioreactor.
3) applied for a patent based on the new assembly method for the bioreactor
4) engaged industrial partners for further development and commercialization of the bioreactor
5) developed a novel concept for sensing the culture conditions inside scaffold materials and has proof of concept
6) produced new rapid and sensitive oxygen and pH sensing materials for cell and tissue culture applications
Tetra:
1) went beyond the state-of-the art building the first ever 2-photon polymerization equipment able to produce LCM scaffolds of 15 x 15 mm
2) applied for a patent application
3) commercialized the scaled up 2PP equipment is currently on the market and one unit was sold and negotiations are in progress for the sale of 5 more
4) produced new equipment with potential applications in Microfluidics- e.g. cavities, valves, microreactors, tubes with <1µm diameter, mixer structures, connectors, filters; Photonics- e.g. photonic crystals, optical interconnects, waveguides, DOE, lens, refractive elements; Life Science- e.g. cell cultivating electronics, wearable, flexible micro rapid prototyping of Smart Mechanics, micro tools
App development
Promoscience:
1) created an integrative platform for research data management with flexibility and quality control
2) developed an innovative and flexible Data Sample Lifecycle Management app for real time tracing and tracking of scientific experiments.
3) has made the SLM app now available commercially for customers such as Universities, Research Institutes and manufacturing industry
Biomaterial evaluation platform
UNIVIE:
1) established innovative in vitro and in vivo biomaterials platforms for testing bone repair and healing with human and mouse cells and with mouse models of bone repair and healing
2) determined that the InnovaBone products, scaffolds, ELRs and HA NPs contribute to enhance bone healing
UCAM:
1) made a groundbreaking development in the evaluation of novel biomaterials with primary human cells that is predictive of clinical outcome
2) has advanced knowledge in the field in establishing and comparing cell lines assays with primary human cells for the evaluation of novel biomaterials
3) established a novel innovative microCT imaging approach to evaluate interior scaffold deformation
UMG-GOE:
1) used innovative imaging technology for evaluating bone repair following biomaterial implantation
2) correlated optical imaging results with in vivo micro computed tomography (CT) in the mouse calvarial defect model for the first time
3) developed inflammation monitoring with Perkin Elmer’s ProSense probe for in vivo non-invasive imaging following the inflammatory response to biomaterials and bone healing with potential use in the future in clinical settings
4) used non-invasive, longitudinal imaging technology to comply with the 3Rs, Reduction, Replacement and Refinement of experimental animal use
MUW:
1) established high throughput models and a subchronic mouse model for evaluating immune and inflammatory foreign body responses to biomaterials
2) discovered that immune models are predictive of the inflammatory response during bone healing
3) created an unprecedented gene expression profile dataset using Illumina microarrays for elucidating the mechanisms underlying bone repair and healing in the context of biomaterials
UNOTT:
1) generated exciting work linking real time in vitro degradation with faster practical methods and
demonstrated predicted correlation with in vivo performance.
2) developed unique in vitro testing and screening technology for small but complex biodegradable
biomimetic materials with wide ranging physical properties.
3) established predictive values of in vitro degradation to in vivo degradation
4) developed new techniques in the area of physical testing of biomaterials
5) amassed knowledge for characterisation and validation methodologies whilst restricted to small geometries and numbers(n)
6) submitted paper on Accelerated degradation of scaffolds
Stakeholder engagement, Training and Dissemination
Moverim:
1) created a community of stakeholders to support the innovation process of InnovaBone to enhance and expedite the innovation process
2) was a front runner in bringing together researchers form a large community working on different EU funded projects (Marie Curie Actions, R&D inHealth, Nanoscience and advanced materials) and establishing a multidisciplinary platform for creating solid new relationships and to encourage further collaborations
3) established a training platform with on site and webinar training sessions
4) established networking and participative methodologies to support innovative discussions and brainstorming amongst researchers and stakeholders in Europe
5) successfully implemented initiatives for a dialogue between stakeholders and researchers to enhance the impact of InnovaBone for patients, health professionals and industry and fill the gap with civil society
6) boosted internal training (MOOCs) and clustering actions with other projects to build a common vision, exchange information and good practices on bone regeneration and biomaterials
7) enhanced its competence in raising researchers skills, in building up participative networking methodologies to keep research groups active beyond the EU project financing and by starting to actively collaborate with policy makers
Promoscience:
1) provided InnovaBone with excellent branding with an interactive website, a logo registered as trademark and several multimedia productions
Exploitation of InnovaBone products
Qserve:
1) provided essential and state of the art regulatory guidance
2) established a systematic risk-based design process adhering to industrial principles
3) established procedures and plans for a practical application of design controls and risk assessment
4) provided a dedicated series of webinars for a successful implementation of the design process
5) enhanced the valorisation potential of the research outcomes
The InnovaBone project:
1) created a novel strategy using a multicomponent approach for the healing and repair of non-union bone lesions
2) created a broad biomaterials evaluation platform for bone healing and repair
3) engaged primary stakeholders from the beginning of the project to ensure the interaction of the clinician and scientists for the best possible product for clinical care
4) disseminated widely to promote the project and project’s findings
5) combined innovation in several disciplines to arrive at novel concepts, experimental methods and products
6) developed several commercial products
7) has over 20 manuscripts in progress for publication
8) has 5 patents published or in the application stage
9) generated numerous collaborations between partners and with new partners from the clustering event
Potential Impact:
Impact on health and society
InnovaBone addressed the repair of non-union bone lesions. The projected results from this large-scale effort will add an innovative bioinspired product to the market. Non-union fractures constitute an enormous burden on the health system in the EU and globally, especially with an increasingly aged population and our new product could alleviate this huge social and economic strain of this clinical problem in the EU. The majority of treatments currently available do not take into account the myriad of physiologic factors involved in the repair and regenerative process. The ability to offer patients breakthrough efficacy by targetting the clinically relevant issues such as blood clotting, sufficient migration and optimum cellular activity within in the biomaterial within the lesion will be a significant innovation in the field of bone repair and regeneration.
The incidence for bone fractures is staggering. One third of women and 1/8th of men over 50 years of age will experience an osteoporotic fracture costing about €36 billion annually (PMID:15618996). The social and economic impact of a curative treatment would be numerous and influence EU competitiveness and health and well-being in Europe. Non-union bone lesions occur in both men and women. There are, however, differences in the outcomes and impacts of our research between women and men. Osteoporosis afflicts more women then men, but the bone fractures are often more severe in men. Bone lesions resulting from metastatic disease will occur in both genders. The impact of novel products that stabilise and repair bone lesions in all patients regardless of gender, will be high.
Large bone fractures and chronic pain and disability resulting from osteoporosis are difficult to treat and are a heavy burden on the health system in the EU, especially with an increasing aged population. Osteoporosis is a disease in which the bones become porous and break easily and is one of the most common, debilitating and costly chronic diseases in Europe. It is a serious public health concern with an estimated 200 million sufferers worldwide.PMID:9425497 Approximately 30% of postmenopausal women have osteoporosis in Europe and up to 70% of these individualsPMID:16983459, PMID:8695963 will sustain one or more fractures. Statistical data on osteoporosis is available on the Osteoporosis Foundation website: http://www.iofbonehealth.org/facts-and-statistics.html#factsheet-category-22 There is no cure for osteoporosis, but it can be managed through exercise and lifestyle and many drugs like bisphosphonates, selective estrogen receptor modulators, PTH, strontium ranelate, calcium and vitamin D supplements which increase bone density and maintain bone quality. However, some of these therapies have risks such as hormones which have been shown to increase the risk of certain cancers and stroke and bisphosphonates which were associated with osteonecrosis of the jaw and esophageal cancer.PMID:19118315
The socio-economic impact of InnovaBone relates to an aging population and a rising incidence and prevalence of bone diseases. Not only will this project develop new options for cure of bone lesions, but will also provide new mechanistic insights that will impact on therapy of bone disease involving bone loss. An available, commercial source of bone biomaterial with high efficiency and low side effects would alleviate the social and economic strain of these medical problems in the EU.
Impact on the substitution of bank bone or autologous bone by complete industrial production of new biomimetic device is expected. Banked Bone-related problems include the shortage of materials, complicated logistics, immune adverse effects, disease transmission risk and for autologous bone, donor site morbidity, limited availability and additional surgery costs. The advantages to the end user for our proposed InnovaBone biomaterial substitute are simple logistics, no disease transmission, unlimited availability, advantage over (Infuse® Bone Graft), as our product mimics the local bone environment better, through a more physiological composition and dosage and controlled relase of BMPs. The scaffold resembles cancellous bone thereby stabilising the lesion, the scaffolds containing heparin will minimise the blood clotting risk. Additionally, the high doses of BMPs in other therapeutics can be controlled in the ELR gels in this proposal. The impact will be to improve the osteoinductive and osteoconductive phases of bone repair and will additionally biodegrade over time, leading to new bone formed in the time comparable to the speed of natural bone repair.
Development of new, rational design criteria for advanced biomaterials/implants, whereby the specific nano/micro-scale properties, as well as the presentation of signalling molecules, are specifically targeted for a defined clinical use - Novel 3D-scaffolds and biogel with NPs will be developed to meet the specific clinical need of critical sized bone lesions
The develop novel imaging strategies for monitoring bone regeneration in patients with osteoporosis and other bone lesions. This has the potential to improve health care of the European citizens experiencing these serious health complications.
Impact on competitiveness
Enhanced competitiveness of the biomaterials and biomedical industries in the EU - The biomaterials produced in this project will impact a significant clinical need and its associated burden on health care costs. Increased competitiveness from this product will be in the post project production of the scaffold, biogels and Ca-NPs by industry and the combining them to form the 2-step final product
Success of the project will allow additional industrial partners involved in the project to enhance the biotech profile in the EU. contribute favourably to European excellence and EU competitiveness by the resulting publications and patentable biomaterials.
A higher profile of companies: training in EU proposals and projects will increase the probability of receiving more funding from private and public sources, thereby increasing the company profile and infusing the company with adequate funding for further growth and development.
Major R&D investments trends in the field carried out with a preliminary competitive intelligence analysis based on patent search indicates that the field encompasses a wide variety of technologies and thus, requires a general search approach using simple keywords (bone in title, abstract & key content) in combination with the International Patent Classification (IPC) code A61L27/40 Composite materials, i.e. containing one material dispersed in a matrix of the same or different material. New growth in 2008 and 2009 indicates a significant growing trend for R&D investments in this field and realistic expectation for substantial innovations. The USA is the leader in the market, but Europe maintains a good competitiveness at global level, positioning itself ahead of Japan. Australia and Canada are increasing their level of presence, while Germany keeps the best position among European countries and Austria is included in the top10 countries at international level. The top 50 assignees that have issued the greatest number of patents related to bone grafting techniques: the American Depuy corporate, part of the Johnson & Johnson family of companies, holds the first position, followed by Massachusetts Institute for Technology MIT which is the first non industrial actor among the top assignees. Biomet Deutschland Gmbh is the first European company among the top 50 assignees. This analysis, on the one side, confirms the interest towards this technology by some of the most influential industries in the pharmaceutical, biotech and medical devices sectors. On the other hand, it shows a lack of European private investment in such technologies and a related weakness in transforming R&D into application. Several products from InnovaBone will be contribute to the growth of biotech industry in Europe due to high prospects for future marketable innovations in the medium- to long-term.
According to Biomaterial Technology Map by Strategic Business Insights, one of the main factors that constrain the expansion of the biomaterials market is the high cost of development and achieving regulatory approval, due to complexity of procedures and also to wide regulatory variation worldwide. However, the niche markets for biomaterials are high value and medical needs continue to provide opportunities for new products. Taking into consideration these constraints, the InnovaBone consortium defined the proper design control activities on the most promising project’s results. A regulatory strategy plan identifying the needs for the technical dossiers for the countries to market was done.
Impact: Innovation
Biomaterials are a promising technology in terms of business opportunities. For the foreseeable future, biomaterials will have an important place in medical technologies: they have no shortage of potential applications in implants, medical devices and drug-delivery systems. Biomaterials for bone repair and replacement, in particular, will further contributions to the orthopaedics market, driven by the increase in the elderly population and their desire to remain active. According to the Bone and Joint Decade (an international steering committee for musculoskeletal health), approximately 10 million Americans have osteoporosis and 18 million more suffer from low bone mass. In the USA annually, surgeons perform more than 600,000 total joint replacements and 500,000 bone grafts to fill fractures that are unable to heal naturally. Currently, approximately 1.5 million fractures occur in the USA per year as a result of the bone-weakening effects of osteoporosis. Moreover, the USA Census Bureau predicts that more than 54 million people will be over the age of 65 by 2020 in the USA. The mortality rate of USA citizens older than age 50 is 24% after a major bone fracture. As the USA population ages, the number of fractures is likely to rise to 3 million, costing $25.3 billion by 2025. Although a wide variety of biomaterials are already in use in orthopaedics, the research approach chosen by the InnovaBone Consortium highlights significant opportunities for potential market expansion, thanks to improvements in current materials.
The use of novel biomaterials that efficaciously cause bone healing is timely and relevant for the EU. It is estimated that the annual market for treatments for osteoporosis is approximately €10 billion (http://www.bioportfolio.com/) and €560 form bone biomaterials doi:10.1089/tea.2007.0267. Thus, there is clearly a
large market stake for products like the ones in this proposal.
Advantages of the InnovaBone product include the elimination of risk associated with infections (HIV, etc.) due to bone allografts and bank sourced bone, the possibility of shaping the implant for the patient in clinic which offers tailored patient-specific implants and, most importantly, an acellular 2-component implant with a smart biomimetic biogel. Thus, we will avoid the technical challenges and regulatory issues implicit in selling a product containing living cells.
Exploitation of the new technologies and equipment in addition to pre-clinical data from this project will result in new economic opportunities for EU-based ventures. Novel potential therapeutics from these studies will be the entrance into the next level of pre-clinical studies for the involved partners. The production of 2 photon polymerization equipment will have applications in many fields including potential applications in Microfluidics- e.g. cavities, valves, microreactors, tubes with <1µm diameter, mixer structures, connectors, filters; Photonics- e.g. photonic crystals, optical interconnects, waveguides, DOE, lens, refractive elements; Life Science- e.g. cell cultivating electronics, wearable, flexible micro rapid prototyping of Smart Mechanics, micro tools. The project made great strides for commercial products. The development of a mini-bioreactor will have impact in the biological and biomaterials fields.
Impact on European science
InnovaBone will have an impact on elucidating cellular and molecular pathways involved in bone cell response to biomaterials. The basic research component may shed light on the mechanisms underlying bone healing and bone repair, which could lead to new targets. The generation of reliable novel biomaterials and mechanistic insights will have an impact on European competitiveness within the scientific community as well as on the competitiveness of the European-based biotech industry.
In addition, finding new mechanistic insights that would be published in high impact journals would enhance EU scientific excellence. Moreover, new mechanistic insights may lead to new targets that may influence the balance between bone rebuilding and bone resorption in a positive direction. Significant findings in the field as well as high impact publications will also help the trainees in the project.
Reduction of our reliance on complex and costly in vivo experiments to predict the performance of bioactive materials - The use of BioMEMS which closely mimics in vivo conditions and real-time non-invasive imaging will not only reduce the numbers of animals needed for experiments but data will be compared to in vivo studies as a means for future reductions on experimental animal needs
Conclusive remarks
Innovation results or potential achieved concerning the exploitation of the results including the main exploitables from the project and TRL:
• 2PP equipment has reached TRL 4 with its upgraded scanner unit brings it to TRL 5 and the production of scaffolds is at TRL 4
• A 2PP prototype system for rapid scaffold production during the project is considered TRL7. Within the next months a new system called MBZ-2PP-14 will reach TRL8. This new equipment has lead to LCM3 scaffold production in large sizes and amounts which brings it to TRL4
• A bioreactor created at CSEM is at TRL 7
• Calcium phosphate nanoparticles is at present at TRL4 (technology validated in lab), but with the in vivo experiments that are being performed, we expect to soon reach TRL5
• Bacterial growing and elastin-like recombinamer production is carried out in a 20 L fermenter that corresponds to a TRL4 (technology validated in laboratory scale)
• Comprehensive approach to evaluate foreign body reactions to biomaterials in vivo in mice is TRL 3
• Application development for sample lifecycle analysis for further exploitation
Summary of the research results obtained which could lead in the future to tangible innovation
1. Fully developed and optimized gene construction and endotoxin-free BMP2+BMP7 biogel bioproduction
Use of biogels in larger animal pre-clinical studies and eventually in clinical studies
2.Best candidate lactide-caprolactone-methacrylate 3 polymer is the optimal material for bone healing; LCM3 is non toxic and eco-toxicological response is normal; Upscaled LCM3 material by TETRA’s invention for the production of large scafolds
Use of large, non-toxic LCM3 scaffolds in larger animal pre-clinical studies and eventually in clinical studies
3. Hydroxyapatite nanoparticles accentuate bone growth in a pre-clinical mouse calvarial defect model
Use of HA NPs in larger animal pre-clinical studies and eventually in clinical studies to enhance bone healing
4. Gene expression profiling platform of bone healing in the pre-clinical mouse calvarial defect model
New approach to follow bone healing in pre-clinical studies for any biomaterials and for new insights in pathophysiology
5. Several modules derived from co-expression analysis illustrating gene groups in pathways of bone ossification, immune responses and other pathways not yet described in bone healing and repair
New genes to follow bone healing in pre-clinical studies for any biomaterials and for new insights in pathophysiology
6. ProSense is an ideal fluorescence imaging agent for bone inflammation
An innovative approach for screening novel biomaterials for bone healing and the inflammation that occurs during the healing process
7. European Patent Application to be filed by CSEM. Title: A method for the assembly and sealing of bioreactors, authors M. Favre, M. Giazzon, R. Ischer, M.Liley; patent no. EP151965597.
A new bioreactor for evaluating biomaterials in 3D in vivo mimicked conditions
8. German patent application filed by IBA: Zwei-Photonenpolymerisations-Initiatoren für die Anwendung in organischen Lösungsmitteln, Authors: Michael Gottschaldt, Cornelia Bader, Leander Poocza, Felix Siedenbiedel, Gerhard Hildebrand, Klaus Liefeith, Ulrich S. Schubert, Patent subscribed to “Deutsches Patent- und Markenamt” on 18.06.2014 patent accepted und published on 02.07.2015 under patent number: DE 10 2014 008 994 B3 2015.07.02
The use of 2PP for the preparation of scaffolds for tissue regeneration and healing and for endless applications where fine microstructure is required
List of Websites:
www.InnovaBone.eu
Participants
Universitaet Wien / UNIVIE
Oskar.Hoffmann@univie.ac.at
Austria
Institut für Bioprozess- und Analysenmesstechnik e.V. / iba
Klaus.Liefeith@iba-heiligenstadt.de
Germany
University of Cambridge/ UCAM
Serena Best Smb51@cam.ac.uk
United Kingdom
University of Nottingham / UNOTT
David.Grant@nottingham.ac.uk
United Kingdom
Qserve Consultancy BV/ Qserve
Willibrord.driessen@qservegroup.com
The Netherlands
Baxter Innovations GHBH / Baxter
John_Barry@baxter.com
Austria
TETRA Gesellschaft für Sensorik, Robotik und Automation GmbH / TETRA
Olaf Mollenhauer om@tetra-ilmenau.com
Germany
Centre Suisse d'Electronique et de Microtechnique sa / CSEM
Martha.Liley@csem.ch
Switzerland
Medizinische Universität Wien / MUW
Michelle.Epstein@meduniwien.ac.at
Austria
Universidad de Valladolid / UVa
J.Carlos Rodríguez-Cabello roca@bioforge.uva.es
Spain
Universitat Politècnica de Catalunya / UPC
Maria.Pau.Ginebra@upc.edu
Spain
Universitätsmedizin Göttingen / UMG-GOE
Frauke Alves falves@gwdg.de
Germany
Moverim Consulting sprl. / Moverim
Laura Vivani vivani@moverim.eu
Belgium
Promoscience srl. / Promoscience
Riccardo Brancaleon brancaleon@promoscience.com
Italy
InnovaBone teams successfully created and produced the InnovaBone product with biodegradable 2-photon polymerized scaffolds, an extensive family of innovative state-of-the-art SMART biomimetic, bioinspired, biodegradable, thermosensitive Elastin-like recombiners (ELRs) containing biologically active bone growth factors with controlled amounts of hydroxyapatite (HA) nanoparticle to enhance bone repair and healing. Novel Equipment was developed by an InnovaBone team including a novel compact simple, low cost bioreactor for testing bone repair scaffolds in long-term cell culture under mechanical stimulation and state-of-the art 2-photon polymerization equipment able to produce large scaffolds of 15 x 15 mm. An InnovaBone team developed an innovative and flexible Data Sample Lifecycle Management app for real time tracing and tracking of scientific experiments.
To study the biomaterials, several InnovaBone teams worked together to generate a cohesive fully integrated platform for testing real time in vitro degradation, in vitro and in vivo bone repair and healing, high throughput and a subchronic in vivo models for evaluating immune and inflammatory foreign body responses to biomaterials, primary human cells testing, and innovative imaging technology for evaluating bone repair following biomaterial implantation.
Training, Stakeholder engagement and Dissemination was successfully accomplished by InnovaBone teams with networking and participative methodologies to support innovative discussions and brainstorming amongst researchers and stakeholders in Europe. Furthermore, state of the art regulatory guidance was provided to ensure progress beyond the project of the foreground generated during the life of the project.
The InnovaBone project created a novel strategy using a multicomponent approach for the healing and repair of non-union bone lesions. A broad biomaterials evaluation platform for bone healing and repair was developed along with several additional commercial products. The InnovaBone team was engaged primary stakeholders from the beginning of the project to ensure the interaction of the clinician and scientists for the best possible product for clinical care. Wide dissemination to promote the project findings was done and will continue with over 20 manuscripts in progress for publication and 5 patents published or in the application stage.
Project Context and Objectives:
The importance of developing novel approaches for bone repair is underscored by the heavy burden on health care costs and patient suffering caused most significantly by traumatic bone lesions, osteoporosis and osteolytic bone metastases. The incidence and prevalence of osteoporosis and related fractures increases as the population ages. Approximately 33% of women and 12.5% of men over the age of 50 will suffer an osteoporotic fracture. Over 800,000 bone bank-derived cadaver allografts (the most commonly used bone substitute in Europe) are used annually and direct costs for osteoporotic fractures alone are €36 billion (PMID:15618996), illustrating the tremendous burden on health care (PMID:11712842). There are therapeutics that retard bone loss and others, which repair large lesions and fractures that do not undergo self-repair, but many of the current approach are associated with complications: high risk for transferable infections, immune responses, chronic inflammation and rejection, stress-shielding and loosening due to wear, material debris formation. All these undesirable events lead to increased costs and recovery time. New biomaterial approaches must be sought which involve the use of various unconventional technologies, cell seeding, pertinent growth factors and cell migration factors (PMID:18454418). Despite substantial advances in biomaterials, further R&D is essential to overcome the limitations of current implant materials, such as improper mechanical strength and durability and insufficient healing caused by foreign body reactions and inefficient vascularisation and bone cell function. Thus, new bone regeneration strategies are necessary.
Our main objective was to achieve radical innovations in state-of-the-art bone biomaterials in order to address the serious morbidity and mortality associated with bone lesions both in the EU and Worldwide. To accomplish this we devised a plan, guided by the needs of the clinician and patient alike, to design and produce optimally performing bioinspired materials based on the knowledge gained from the natural physiological processes underlying bone repair and regeneration. This pioneering strategy aimed to accelerate bone healing and reduce adverse side effects with currently employed materials. Our approach was to develop a 2-step biomimetic product consisting of a bioactive scaffold and a bioactive self-setting gel. Step 1 will be to introduce a firm but compressible bio-inspired, heparinised 3D scaffold which would reduce the formation of blood clots around the implant (one of the major problems with biomaterials leading to unsuccessful engraftment) and provide structural stability of the lesion. Step 2 involved injecting a bioactive recombinant gel into the scaffold. This was planned to provide a microenvironment that promotes bone repair due to the bioactive factors grafted and dispersed within the biogel and scaffold structure. The expected impact will be a radical innovation that accelerates the treatment of large bone lesions.
The step 1 3D scaffolds were produced using two-photon laser polymerisation (2PP). These polymer scaffolds were designed to be mechanically stable and easily trimmed and fitted into bone lesions. In the bone lesion site, these scaffolds were to provide stability during the formation of new bone and would degrade within approximately a year during the bone healing process. The bioactive gel was created with functional and genetically engineered recombinant elastin-like recombinamers (ELRs) which are thermosensitive and self-solidifying. These polymers have absolute precisely controlled molecular architecture and possess sequences that contain specific domains for cell attachment and growth factors. Calcium phosphate nanoparticles (CaP-NPs) dispersed in the gel were generated and added to the biogels to attract and activate the cells important for initiating bone repair.
The objectives of the InnovaBone project were to design and produce a novel 2-step smart bio-scaffold and smart self-setting gel, evaluate the biological and physical properties of bioscaffold-biogel biomaterials, upscale the technological processes allowing the manufacture of prototype scaffolds, and perform technical characterisation and design strategies for product exploitation.
Project Results:
The main S&T results/foregrounds are divided into 1) Biomaterials development, 2) Equipment development, 3) App development, 3) Biomaterial evaluation platform, 4) Stakeholder engagement, Training and Dissemination 4) Exploitation of InnovaBone products and 5) Summary . In each section, the results are highlighted per partner.
Biomaterials development
iba:
1) successfully established a novel material platform for bone or cartilage tissue engineering and cancer research based on FDA approved components like Lactide and caprolactone biopolymers
2) developed technology and parameters for the 2-photon polymerization (2PP) of “tailor-made” designer scaffolds based on triple periodic minimal surfaces (e.g. Schwarz P surfaces) with variable stiffness and biodegradability for bone repair
3) has acquired one patent which was published in July 2015 on their 2PP approach and especially for the use of highly efficient and fast photoinitiators
4) engaged with industrial partners for further development and commercialization and the potential for industry to purchase this patent
UVa:
1) created a new extensive family of innovative state-of-the-art SMART biomimetic, bioinspired, biodegradable, thermosensitive Elastin-like recombiners (ELRs) containing biologically active bone growth factors that are at room temperature are injectable liquids that gelify at body temperature and provide the a microenvironment conducive for bone repair and healing
2) produced artificial biogels with tremendous potential for clinical applications such as in the repair and healing of bone, cartilage and skin
3) established a Step-change in scale up technology for ELR production, making it accessible to a wider community including the high industrial potential and a broad range of new applications and clinical implications
UPC:
1) developed technology that allows the introduction of controlled amounts of doping ions (e.g. Carbonate, magnesium) into the hydroxyapatite (HA) nanoparticle (NP) crystal structure
2) by controlling the composition of the HA NPs, they produced biologically active, non-toxic HA NPs for bone repair and healing
3) developed an innovative method for the dispersion of NPs into liquid Elastin-like Recombiners (ELRs) allowing for homogenous distribution which is a crucial step for reducing NP aggregates and thus, maintaining NP integrity, effectiveness and biological function
4) produced HA NPs that were critical for in vivo bone formation and healing
Equipment development
CSEM:
1) produced a novel compact simple, low cost bioreactor for testing bone repair scaffolds in long-term cell culture under mechanical stimulation
2) has demonstrated that the mechanical properties of the scaffold can be monitored in real time during culture in the bioreactor.
3) applied for a patent based on the new assembly method for the bioreactor
4) engaged industrial partners for further development and commercialization of the bioreactor
5) developed a novel concept for sensing the culture conditions inside scaffold materials and has proof of concept
6) produced new rapid and sensitive oxygen and pH sensing materials for cell and tissue culture applications
Tetra:
1) went beyond the state-of-the art building the first ever 2-photon polymerization equipment able to produce LCM scaffolds of 15 x 15 mm
2) applied for a patent application
3) commercialized the scaled up 2PP equipment is currently on the market and one unit was sold and negotiations are in progress for the sale of 5 more
4) produced new equipment with potential applications in Microfluidics- e.g. cavities, valves, microreactors, tubes with <1µm diameter, mixer structures, connectors, filters; Photonics- e.g. photonic crystals, optical interconnects, waveguides, DOE, lens, refractive elements; Life Science- e.g. cell cultivating electronics, wearable, flexible micro rapid prototyping of Smart Mechanics, micro tools
App development
Promoscience:
1) created an integrative platform for research data management with flexibility and quality control
2) developed an innovative and flexible Data Sample Lifecycle Management app for real time tracing and tracking of scientific experiments.
3) has made the SLM app now available commercially for customers such as Universities, Research Institutes and manufacturing industry
Biomaterial evaluation platform
UNIVIE:
1) established innovative in vitro and in vivo biomaterials platforms for testing bone repair and healing with human and mouse cells and with mouse models of bone repair and healing
2) determined that the InnovaBone products, scaffolds, ELRs and HA NPs contribute to enhance bone healing
UCAM:
1) made a groundbreaking development in the evaluation of novel biomaterials with primary human cells that is predictive of clinical outcome
2) has advanced knowledge in the field in establishing and comparing cell lines assays with primary human cells for the evaluation of novel biomaterials
3) established a novel innovative microCT imaging approach to evaluate interior scaffold deformation
UMG-GOE:
1) used innovative imaging technology for evaluating bone repair following biomaterial implantation
2) correlated optical imaging results with in vivo micro computed tomography (CT) in the mouse calvarial defect model for the first time
3) developed inflammation monitoring with Perkin Elmer’s ProSense probe for in vivo non-invasive imaging following the inflammatory response to biomaterials and bone healing with potential use in the future in clinical settings
4) used non-invasive, longitudinal imaging technology to comply with the 3Rs, Reduction, Replacement and Refinement of experimental animal use
MUW:
1) established high throughput models and a subchronic mouse model for evaluating immune and inflammatory foreign body responses to biomaterials
2) discovered that immune models are predictive of the inflammatory response during bone healing
3) created an unprecedented gene expression profile dataset using Illumina microarrays for elucidating the mechanisms underlying bone repair and healing in the context of biomaterials
UNOTT:
1) generated exciting work linking real time in vitro degradation with faster practical methods and
demonstrated predicted correlation with in vivo performance.
2) developed unique in vitro testing and screening technology for small but complex biodegradable
biomimetic materials with wide ranging physical properties.
3) established predictive values of in vitro degradation to in vivo degradation
4) developed new techniques in the area of physical testing of biomaterials
5) amassed knowledge for characterisation and validation methodologies whilst restricted to small geometries and numbers(n)
6) submitted paper on Accelerated degradation of scaffolds
Stakeholder engagement, Training and Dissemination
Moverim:
1) created a community of stakeholders to support the innovation process of InnovaBone to enhance and expedite the innovation process
2) was a front runner in bringing together researchers form a large community working on different EU funded projects (Marie Curie Actions, R&D inHealth, Nanoscience and advanced materials) and establishing a multidisciplinary platform for creating solid new relationships and to encourage further collaborations
3) established a training platform with on site and webinar training sessions
4) established networking and participative methodologies to support innovative discussions and brainstorming amongst researchers and stakeholders in Europe
5) successfully implemented initiatives for a dialogue between stakeholders and researchers to enhance the impact of InnovaBone for patients, health professionals and industry and fill the gap with civil society
6) boosted internal training (MOOCs) and clustering actions with other projects to build a common vision, exchange information and good practices on bone regeneration and biomaterials
7) enhanced its competence in raising researchers skills, in building up participative networking methodologies to keep research groups active beyond the EU project financing and by starting to actively collaborate with policy makers
Promoscience:
1) provided InnovaBone with excellent branding with an interactive website, a logo registered as trademark and several multimedia productions
Exploitation of InnovaBone products
Qserve:
1) provided essential and state of the art regulatory guidance
2) established a systematic risk-based design process adhering to industrial principles
3) established procedures and plans for a practical application of design controls and risk assessment
4) provided a dedicated series of webinars for a successful implementation of the design process
5) enhanced the valorisation potential of the research outcomes
The InnovaBone project:
1) created a novel strategy using a multicomponent approach for the healing and repair of non-union bone lesions
2) created a broad biomaterials evaluation platform for bone healing and repair
3) engaged primary stakeholders from the beginning of the project to ensure the interaction of the clinician and scientists for the best possible product for clinical care
4) disseminated widely to promote the project and project’s findings
5) combined innovation in several disciplines to arrive at novel concepts, experimental methods and products
6) developed several commercial products
7) has over 20 manuscripts in progress for publication
8) has 5 patents published or in the application stage
9) generated numerous collaborations between partners and with new partners from the clustering event
Potential Impact:
Impact on health and society
InnovaBone addressed the repair of non-union bone lesions. The projected results from this large-scale effort will add an innovative bioinspired product to the market. Non-union fractures constitute an enormous burden on the health system in the EU and globally, especially with an increasingly aged population and our new product could alleviate this huge social and economic strain of this clinical problem in the EU. The majority of treatments currently available do not take into account the myriad of physiologic factors involved in the repair and regenerative process. The ability to offer patients breakthrough efficacy by targetting the clinically relevant issues such as blood clotting, sufficient migration and optimum cellular activity within in the biomaterial within the lesion will be a significant innovation in the field of bone repair and regeneration.
The incidence for bone fractures is staggering. One third of women and 1/8th of men over 50 years of age will experience an osteoporotic fracture costing about €36 billion annually (PMID:15618996). The social and economic impact of a curative treatment would be numerous and influence EU competitiveness and health and well-being in Europe. Non-union bone lesions occur in both men and women. There are, however, differences in the outcomes and impacts of our research between women and men. Osteoporosis afflicts more women then men, but the bone fractures are often more severe in men. Bone lesions resulting from metastatic disease will occur in both genders. The impact of novel products that stabilise and repair bone lesions in all patients regardless of gender, will be high.
Large bone fractures and chronic pain and disability resulting from osteoporosis are difficult to treat and are a heavy burden on the health system in the EU, especially with an increasing aged population. Osteoporosis is a disease in which the bones become porous and break easily and is one of the most common, debilitating and costly chronic diseases in Europe. It is a serious public health concern with an estimated 200 million sufferers worldwide.PMID:9425497 Approximately 30% of postmenopausal women have osteoporosis in Europe and up to 70% of these individualsPMID:16983459, PMID:8695963 will sustain one or more fractures. Statistical data on osteoporosis is available on the Osteoporosis Foundation website: http://www.iofbonehealth.org/facts-and-statistics.html#factsheet-category-22 There is no cure for osteoporosis, but it can be managed through exercise and lifestyle and many drugs like bisphosphonates, selective estrogen receptor modulators, PTH, strontium ranelate, calcium and vitamin D supplements which increase bone density and maintain bone quality. However, some of these therapies have risks such as hormones which have been shown to increase the risk of certain cancers and stroke and bisphosphonates which were associated with osteonecrosis of the jaw and esophageal cancer.PMID:19118315
The socio-economic impact of InnovaBone relates to an aging population and a rising incidence and prevalence of bone diseases. Not only will this project develop new options for cure of bone lesions, but will also provide new mechanistic insights that will impact on therapy of bone disease involving bone loss. An available, commercial source of bone biomaterial with high efficiency and low side effects would alleviate the social and economic strain of these medical problems in the EU.
Impact on the substitution of bank bone or autologous bone by complete industrial production of new biomimetic device is expected. Banked Bone-related problems include the shortage of materials, complicated logistics, immune adverse effects, disease transmission risk and for autologous bone, donor site morbidity, limited availability and additional surgery costs. The advantages to the end user for our proposed InnovaBone biomaterial substitute are simple logistics, no disease transmission, unlimited availability, advantage over (Infuse® Bone Graft), as our product mimics the local bone environment better, through a more physiological composition and dosage and controlled relase of BMPs. The scaffold resembles cancellous bone thereby stabilising the lesion, the scaffolds containing heparin will minimise the blood clotting risk. Additionally, the high doses of BMPs in other therapeutics can be controlled in the ELR gels in this proposal. The impact will be to improve the osteoinductive and osteoconductive phases of bone repair and will additionally biodegrade over time, leading to new bone formed in the time comparable to the speed of natural bone repair.
Development of new, rational design criteria for advanced biomaterials/implants, whereby the specific nano/micro-scale properties, as well as the presentation of signalling molecules, are specifically targeted for a defined clinical use - Novel 3D-scaffolds and biogel with NPs will be developed to meet the specific clinical need of critical sized bone lesions
The develop novel imaging strategies for monitoring bone regeneration in patients with osteoporosis and other bone lesions. This has the potential to improve health care of the European citizens experiencing these serious health complications.
Impact on competitiveness
Enhanced competitiveness of the biomaterials and biomedical industries in the EU - The biomaterials produced in this project will impact a significant clinical need and its associated burden on health care costs. Increased competitiveness from this product will be in the post project production of the scaffold, biogels and Ca-NPs by industry and the combining them to form the 2-step final product
Success of the project will allow additional industrial partners involved in the project to enhance the biotech profile in the EU. contribute favourably to European excellence and EU competitiveness by the resulting publications and patentable biomaterials.
A higher profile of companies: training in EU proposals and projects will increase the probability of receiving more funding from private and public sources, thereby increasing the company profile and infusing the company with adequate funding for further growth and development.
Major R&D investments trends in the field carried out with a preliminary competitive intelligence analysis based on patent search indicates that the field encompasses a wide variety of technologies and thus, requires a general search approach using simple keywords (bone in title, abstract & key content) in combination with the International Patent Classification (IPC) code A61L27/40 Composite materials, i.e. containing one material dispersed in a matrix of the same or different material. New growth in 2008 and 2009 indicates a significant growing trend for R&D investments in this field and realistic expectation for substantial innovations. The USA is the leader in the market, but Europe maintains a good competitiveness at global level, positioning itself ahead of Japan. Australia and Canada are increasing their level of presence, while Germany keeps the best position among European countries and Austria is included in the top10 countries at international level. The top 50 assignees that have issued the greatest number of patents related to bone grafting techniques: the American Depuy corporate, part of the Johnson & Johnson family of companies, holds the first position, followed by Massachusetts Institute for Technology MIT which is the first non industrial actor among the top assignees. Biomet Deutschland Gmbh is the first European company among the top 50 assignees. This analysis, on the one side, confirms the interest towards this technology by some of the most influential industries in the pharmaceutical, biotech and medical devices sectors. On the other hand, it shows a lack of European private investment in such technologies and a related weakness in transforming R&D into application. Several products from InnovaBone will be contribute to the growth of biotech industry in Europe due to high prospects for future marketable innovations in the medium- to long-term.
According to Biomaterial Technology Map by Strategic Business Insights, one of the main factors that constrain the expansion of the biomaterials market is the high cost of development and achieving regulatory approval, due to complexity of procedures and also to wide regulatory variation worldwide. However, the niche markets for biomaterials are high value and medical needs continue to provide opportunities for new products. Taking into consideration these constraints, the InnovaBone consortium defined the proper design control activities on the most promising project’s results. A regulatory strategy plan identifying the needs for the technical dossiers for the countries to market was done.
Impact: Innovation
Biomaterials are a promising technology in terms of business opportunities. For the foreseeable future, biomaterials will have an important place in medical technologies: they have no shortage of potential applications in implants, medical devices and drug-delivery systems. Biomaterials for bone repair and replacement, in particular, will further contributions to the orthopaedics market, driven by the increase in the elderly population and their desire to remain active. According to the Bone and Joint Decade (an international steering committee for musculoskeletal health), approximately 10 million Americans have osteoporosis and 18 million more suffer from low bone mass. In the USA annually, surgeons perform more than 600,000 total joint replacements and 500,000 bone grafts to fill fractures that are unable to heal naturally. Currently, approximately 1.5 million fractures occur in the USA per year as a result of the bone-weakening effects of osteoporosis. Moreover, the USA Census Bureau predicts that more than 54 million people will be over the age of 65 by 2020 in the USA. The mortality rate of USA citizens older than age 50 is 24% after a major bone fracture. As the USA population ages, the number of fractures is likely to rise to 3 million, costing $25.3 billion by 2025. Although a wide variety of biomaterials are already in use in orthopaedics, the research approach chosen by the InnovaBone Consortium highlights significant opportunities for potential market expansion, thanks to improvements in current materials.
The use of novel biomaterials that efficaciously cause bone healing is timely and relevant for the EU. It is estimated that the annual market for treatments for osteoporosis is approximately €10 billion (http://www.bioportfolio.com/) and €560 form bone biomaterials doi:10.1089/tea.2007.0267. Thus, there is clearly a
large market stake for products like the ones in this proposal.
Advantages of the InnovaBone product include the elimination of risk associated with infections (HIV, etc.) due to bone allografts and bank sourced bone, the possibility of shaping the implant for the patient in clinic which offers tailored patient-specific implants and, most importantly, an acellular 2-component implant with a smart biomimetic biogel. Thus, we will avoid the technical challenges and regulatory issues implicit in selling a product containing living cells.
Exploitation of the new technologies and equipment in addition to pre-clinical data from this project will result in new economic opportunities for EU-based ventures. Novel potential therapeutics from these studies will be the entrance into the next level of pre-clinical studies for the involved partners. The production of 2 photon polymerization equipment will have applications in many fields including potential applications in Microfluidics- e.g. cavities, valves, microreactors, tubes with <1µm diameter, mixer structures, connectors, filters; Photonics- e.g. photonic crystals, optical interconnects, waveguides, DOE, lens, refractive elements; Life Science- e.g. cell cultivating electronics, wearable, flexible micro rapid prototyping of Smart Mechanics, micro tools. The project made great strides for commercial products. The development of a mini-bioreactor will have impact in the biological and biomaterials fields.
Impact on European science
InnovaBone will have an impact on elucidating cellular and molecular pathways involved in bone cell response to biomaterials. The basic research component may shed light on the mechanisms underlying bone healing and bone repair, which could lead to new targets. The generation of reliable novel biomaterials and mechanistic insights will have an impact on European competitiveness within the scientific community as well as on the competitiveness of the European-based biotech industry.
In addition, finding new mechanistic insights that would be published in high impact journals would enhance EU scientific excellence. Moreover, new mechanistic insights may lead to new targets that may influence the balance between bone rebuilding and bone resorption in a positive direction. Significant findings in the field as well as high impact publications will also help the trainees in the project.
Reduction of our reliance on complex and costly in vivo experiments to predict the performance of bioactive materials - The use of BioMEMS which closely mimics in vivo conditions and real-time non-invasive imaging will not only reduce the numbers of animals needed for experiments but data will be compared to in vivo studies as a means for future reductions on experimental animal needs
Conclusive remarks
Innovation results or potential achieved concerning the exploitation of the results including the main exploitables from the project and TRL:
• 2PP equipment has reached TRL 4 with its upgraded scanner unit brings it to TRL 5 and the production of scaffolds is at TRL 4
• A 2PP prototype system for rapid scaffold production during the project is considered TRL7. Within the next months a new system called MBZ-2PP-14 will reach TRL8. This new equipment has lead to LCM3 scaffold production in large sizes and amounts which brings it to TRL4
• A bioreactor created at CSEM is at TRL 7
• Calcium phosphate nanoparticles is at present at TRL4 (technology validated in lab), but with the in vivo experiments that are being performed, we expect to soon reach TRL5
• Bacterial growing and elastin-like recombinamer production is carried out in a 20 L fermenter that corresponds to a TRL4 (technology validated in laboratory scale)
• Comprehensive approach to evaluate foreign body reactions to biomaterials in vivo in mice is TRL 3
• Application development for sample lifecycle analysis for further exploitation
Summary of the research results obtained which could lead in the future to tangible innovation
1. Fully developed and optimized gene construction and endotoxin-free BMP2+BMP7 biogel bioproduction
Use of biogels in larger animal pre-clinical studies and eventually in clinical studies
2.Best candidate lactide-caprolactone-methacrylate 3 polymer is the optimal material for bone healing; LCM3 is non toxic and eco-toxicological response is normal; Upscaled LCM3 material by TETRA’s invention for the production of large scafolds
Use of large, non-toxic LCM3 scaffolds in larger animal pre-clinical studies and eventually in clinical studies
3. Hydroxyapatite nanoparticles accentuate bone growth in a pre-clinical mouse calvarial defect model
Use of HA NPs in larger animal pre-clinical studies and eventually in clinical studies to enhance bone healing
4. Gene expression profiling platform of bone healing in the pre-clinical mouse calvarial defect model
New approach to follow bone healing in pre-clinical studies for any biomaterials and for new insights in pathophysiology
5. Several modules derived from co-expression analysis illustrating gene groups in pathways of bone ossification, immune responses and other pathways not yet described in bone healing and repair
New genes to follow bone healing in pre-clinical studies for any biomaterials and for new insights in pathophysiology
6. ProSense is an ideal fluorescence imaging agent for bone inflammation
An innovative approach for screening novel biomaterials for bone healing and the inflammation that occurs during the healing process
7. European Patent Application to be filed by CSEM. Title: A method for the assembly and sealing of bioreactors, authors M. Favre, M. Giazzon, R. Ischer, M.Liley; patent no. EP151965597.
A new bioreactor for evaluating biomaterials in 3D in vivo mimicked conditions
8. German patent application filed by IBA: Zwei-Photonenpolymerisations-Initiatoren für die Anwendung in organischen Lösungsmitteln, Authors: Michael Gottschaldt, Cornelia Bader, Leander Poocza, Felix Siedenbiedel, Gerhard Hildebrand, Klaus Liefeith, Ulrich S. Schubert, Patent subscribed to “Deutsches Patent- und Markenamt” on 18.06.2014 patent accepted und published on 02.07.2015 under patent number: DE 10 2014 008 994 B3 2015.07.02
The use of 2PP for the preparation of scaffolds for tissue regeneration and healing and for endless applications where fine microstructure is required
List of Websites:
www.InnovaBone.eu
Participants
Universitaet Wien / UNIVIE
Oskar.Hoffmann@univie.ac.at
Austria
Institut für Bioprozess- und Analysenmesstechnik e.V. / iba
Klaus.Liefeith@iba-heiligenstadt.de
Germany
University of Cambridge/ UCAM
Serena Best Smb51@cam.ac.uk
United Kingdom
University of Nottingham / UNOTT
David.Grant@nottingham.ac.uk
United Kingdom
Qserve Consultancy BV/ Qserve
Willibrord.driessen@qservegroup.com
The Netherlands
Baxter Innovations GHBH / Baxter
John_Barry@baxter.com
Austria
TETRA Gesellschaft für Sensorik, Robotik und Automation GmbH / TETRA
Olaf Mollenhauer om@tetra-ilmenau.com
Germany
Centre Suisse d'Electronique et de Microtechnique sa / CSEM
Martha.Liley@csem.ch
Switzerland
Medizinische Universität Wien / MUW
Michelle.Epstein@meduniwien.ac.at
Austria
Universidad de Valladolid / UVa
J.Carlos Rodríguez-Cabello roca@bioforge.uva.es
Spain
Universitat Politècnica de Catalunya / UPC
Maria.Pau.Ginebra@upc.edu
Spain
Universitätsmedizin Göttingen / UMG-GOE
Frauke Alves falves@gwdg.de
Germany
Moverim Consulting sprl. / Moverim
Laura Vivani vivani@moverim.eu
Belgium
Promoscience srl. / Promoscience
Riccardo Brancaleon brancaleon@promoscience.com
Italy