Biomaterials Research Centres Annual Day 2015 "Tissue - - PDF document

Biomaterials Research Centre Nov 10, 2015

Biomaterials Research Centre’s Annual Day 2015

"Tissue Engineering" Presentation of speakers

• Dr. Suchitra Sumitran-Holgersson earned her Doctor of Medical Sciences degree

in Clinical Immunology at the Karolinska Institutet in 1991, and became an associate Professor in 2000. In 2008, she joined the Sahlgrenska hospital at the Gothenburg University as the Professor of Transplantation Biology. Her research interests include understanding mechanisms of allograft rejections, stem cell engraftment and tissue- engineering of organs with stem cells. She has contributed to more than 80 articles in peer-reviewed journals and 15 book chapters and has seven scientific patents to her credit. Raimund Strehl, PhD, Chief Technology Officer for NovaHep, holds a degree in cell biology from the University of Regensburg, Germany. He has an academic background within cell culture development and tissue engineering as well as an industrial background in the field of human pluripotent stem cell development and

• application. Raimund has worked over ten years at Cellartis, heading product

development and manufacturing in the role of Chief Technology Officer, responsible for collaborations with major pharmaceutical and biotech companies.

• Dr. de Peppo received a BSc Degree in Biotechnology at La Sapienza University in

Rome and a MSc degree in Medical Biotechnology at Bicocca University in Milan, with a thesis in biomaterial engineering from Politecnico di Milano. Following an advanced course in Bioinformatics in Valencia, in 2007, he was awarded a Marie Curie fellowship at the Department of Biomaterials at the University of Gothenburg in Sweden, where he received an international PhD in Tissue Engineering. In 2013 he was honored the Best Prize for his doctoral studies and the Argos Hippium Award for his professional achievements abroad. After completing his PhD, Dr. de Peppo was awarded a postdoctoral fellowship at the New York Stem Cell Foundation Research Institute, where is now leading the Bone Engineering and Regeneration group. His major research goal is to engineer patient-specific vascularized bone grafts for large and complex skeletal reconstructions using an osteoinductive scaffold – perfusion bioreactor model of bone development. Other research directions include manufacturing of biomaterial scaffolds, testing of implantable biomaterials using tissue-engineered products, and stem cell therapy using pluripotent stem cells. Aleksandar holds a Senior Research Position from the Swedish Research council directed towards Soft Matter Physics and is also director for Chalmers Area of Advance Materials Science. The research spans from fundamental investigations of glass formation, gelation and colloidal aggregation to applied projects on new electrolytes for Li-batteries and fuel cells.

Suchitra Sumitran- Holgersson MSC, PhD

• Prof. of Transplantation

Biology Laboratory for Transplantation and Regenerative Medicine Sahlgrenska Science Park, Göteborg

Giuseppe Maria de Peppo, BSc, MSc, PhD

Principal Investigator The New York Stem Cell Foundation Research Institute www.nyscf.org

Raimund Strehl

PhD, CTO, NovaHep AB, Gothenburg, Sweden.

Alexandar Matic

• Prof. and Director

Chalmers Area of Advance Materials Science, Chalmers University of Technology, Gothenburg, Sweden.

Biomaterials Research Centre Nov 10, 2015

Biomaterials Research Centre’s Annual Day 2015

"Tissue Engineering" Presentation of speakers

Abdelouahab El Ghalbzouri (1973) studied Biotechnology and Biochemistry (BSc, 1994-1997) in Etten-Leur, the Netherlands and Molecular and Cellular Biology (MSc, 1998) at the Medical University Paris VI in Paris (France). In 1999, he started his PhD at the Department of Dermatology of the Leiden University Medical Center (LUMC). During his PhD, he optimized the generation

• f reconstructed human skin models and extensively studied their use for tissue

engineering, research, and clinical purposes. Abdoel continued his work on human skin models as a post-doc (2004-2006) and became senior scientist (universitair docent, UD) at the Department of Dermatology (LUMC). Currently, he is leading the research group ‘disease skin models’, in which various skin diseases (e.g. eczema, squamous cell carcinoma) and skin conditions (e.g. wound healing, skin aging) are mimicked in vitro to better understand the physiopathology of the skin. In 2008, this research group was awarded the ‘Alternative to animal testing award’ from the Dutch Society for Animal Protection and the Netherlands Centre for Alternatives to Animal Use. In 2009, the group received the ‘Pearl project award’ for mimicking skin cancer in vitro from the Netherlands Organization for Health Research and Development (ZonMw). In 2010 he participated in the NGI venture challenge where he presented his spin-off company Biomimiq. Biomimiq offers services with its unique and customizable in vitro human skin models representing both healthy and diseased human skin, ascertaining the company as one of the leaders in a large and rapidly growing market. In January 2013, Biomimiq was incorporated as a separate unit within the Tissue Engineering company Aeon Astron Europe B.V. (Leiden, The Netherlands), where it continued its services on skin models. As founder, he has a scientific advisory role within Biomimiq-AAE Dr Kristina Fant is a research scientist at SP Technical Research Institute of Sweden since 2011. She has a PhD in Physical Chemistry from Chalmers University of Technology. Her area of expertise is in vitro safety testing and the development of alternative test methods, i.e. methods not involving animal

• experiments. She is developing and implementing test methods to be run in

compliance with Good Laboratory Practice principles. The SP cell culture facilities are currently the only Swedish lab to offer GLP approved safety testing to external customers, and Sweden’s representative in the EU-NETVAL network

• f qualified laboratories that can participate in method validation.

Anna Finne Wistrand has a background in polymer synthesis. Her research interests focus today on tissue engineering and the translational, interdisciplinary field of material science and biology. This is an area which requires an enhanced understanding

• f

polymer synthesis, physico-chemical characteristics, characterization and structure-property relationships at both molecular and nanoscale level. A general aim for the research is to design and fabricate complex three-dimensional constructs that attract multiple cell types in a predetermined manner and permit development of extracellular matrices in a well defined way. Anna Finne Wistrand finished her PhD 2003 at KTH Royal Institute of Technology, Fibre and polymer Technology. She has after that spent some time as visiting scientist in the group of (1) Prof. Virgil Percec, Department of Chemistry, University of Pennsylvania (2) Prof. Y. Ito, Nanomedical Engineering, RIKEN. In addition, she has had positions at Akzo Nobel (Casco Adhesive), PP Polymer and she is right now active at Novus Scientific.

Abdelouahab El Ghalbzouri,

PhD, Ass. Prof. LUMC skin research lab, Leiden, The Netherlands.

Anna Finne Wistrand

Ass.Prof. Dep.of Fibre and Polymer Technology, School of Chemical Science and Engineering, KTH Royal Institute of Technology, Sweden

Kristina Fant

Research scientist, SP Technical Research Institute

• f Sweden, Borås,

Sweden

Biomaterials Research Centre Nov 10, 2015

Biomaterials Research Centre’s Annual Day 2015

"Tissue Engineering" Presentation of speakers

Kamal Mustafa is a professor at the University of Bergen, Norway and leader of Tissue Engineering Group. He is also an active member of Bergen Stem Cell

• Consortium. His research activity has been tremendously increased during the last

3 years focusing on the development of a tool box for bone regeneration according to the concept of tissue engineering. The research group is producing innovative research in the field of biomaterials, cell biology, regenerative medicine and translation research. The main research area is addressed to develop translational approaches for regenerative therapies of different skeletal defects. The group involves a vital collaboration between Cell Biologists, Material Scientists, Engineers and Clinicians aiming to develop an appropriate scaffold for bone tissue

• engineering. Currently, he is sponsoring and leading a maxillofacial clinical trial

which is among the few in Europe with Advanced Therapy Medicinal Products using MSC and biomaterials. Kamal Mustafa received his PhD from Karolinska Institute in 2001. Then, he had a postdoc period at three different institutes; Karolinska Institute, University of Oslo and University of Bergen. He worked also as visiting scientist at The Nordic Institute for Dental Materials Oslo (NIOM), University of North Carolina, Chapel Hill, USA and University of Malmö, Sweden. Annika Enejder is Professor in Molecular Microscopy, Department of Biology and Biological Engineering, Chalmers University of Technology. She started her academic career as a PhD student in the Division of Atomic Physics, Lund University, where she studied the fundamentals of laser light interaction with cells and biomolecules (Prof. Sune Svanberg). After a few years in the industry she returned to academia through a VR postdoc scholarship and spent four years at the Spectroscopy Lab, MIT, USA and the Ludwig-Maximilian University, Munich (Raman and CARS microscopy). She has devoted all her scientific career to interdisciplinary research, integrating advanced physical technologies to the benefit

• f the biosciences. She is currently a part-time Marie Curie Fellow at Stanford

University in Prof. Sarah Heilshorn’s Biomaterials Group and has transferred her unique technologies of recombinant protein engineering of ECM-mimicking proteins for tissue-mimicking environments. The Marie Curie Fellowship has also offered insights in the latest technologies in Biopolymer- and Neuro-sciences, Stem cell and cancer biology as well as neurodegenerative diseases. On the European arena, Annika has been the chair of the European Network for nonlinear Raman microscopy since 2006 involving ~50 EU groups and is the coordinator of a FP7 Marie Curie Innovative Training Network FINON, for the development of novel technology for nano-scale nonlinear microscopy. Patric Wallin is a PhD candidate in Bioscience with a specialization in Educational Science. He is working in the Division of Biological Physics at Chalmers University of Technology, and is associated with the Division of Engineering Education Research (EER). He has tutored projects in a master level course on Tissue Engineering at Chalmers for several years, and has studied how students learn about Tissue engineering with different educational research approaches. Patric’s research interests in the education research field are focused around undergraduate research experiences, communities of practice and situated cognition. He is particular interested in the progress and development processes students experience in these learning situations.

Presentation of speakers

Annika Enejder,

• Prof. Dep. of Biology

and Biological Engineering, Chalmers University

• f Technology,

Gothenburg, Sweden.

Patric Wallin,

PhD candidate Applied Physics and Engineering Education Research, Chalmers University of Technology

Kamal Mustafa

• Prof. Tissue Engineering

Research Group - Leader

• Dep. of Clinical Dentistry-

Center for Clinical Dental Research University of Bergen, Norway

Biomaterials Research Centre´s Annual Day 2015-11-10

“Tissue Engineering”

Abstracts

“Engineering tissues and organs: The next frontier in regenerative medicine” Suchitra Holgersson, Prof. Dep. of Surgery, Sahlgrenska University Hospital, Gothenburg, Sweden

Different pathological end-stage diseases have few effective therapeutic options apart from a whole organ transplant that, however, often meets with a serious shortage of donor organs. To solve these clinical issues we are using a novel tissue engineering (TE) approach in which a whole donor organ is decellularized to

• btain a complex 3D-biomatrix-scaffold maintaining the intrinsic vascular network, that is subsequently

recellularized with recipient's autologous organ-specific differentiated cells or/and stem cells, to build a potentially functional biological substitute.

“NovaHep – Engineering Individualized Blood Vessels” Raimund Strehl, PhD, CTO, NovaHep AB, Gothenburg, Sweden

NovaHep focuses on the commercialization of a unique technology to engineer individualized blood vessels using a decellularized tissue scaffold and the patient's own stem cells. The engineered blood vessels can be used to replace non-functioning blood vessels in patients with different types of vascular disease.

“Testing Implant Materials using Patient-specific Engineered Bone” Giuseppe Maria de Peppo, BSc, MSc, PhD. Principal Investigator, The New York Stem Cell Foundation Research Institute, New York, USA.

Millions of people worldwide require orthopedic treatments to reconstruct defects caused by traumatic accidents and disease. Metal implants (such as titanium and its alloys) can form a stable bond with bone, an interaction that highly depends on the surface characteristics of the implant material. Osseointegration is traditionally studied using experimental assays and animal studies. However, available in vitro methods fail to provide insights regarding the complex tissue response to the materials in a 3D setting under conditions mimicking the native physiological environment. On the other hand, in vivo studies are time and resource intensive, and often irrelevant due to interspecies differences in tissue quality and metabolism. In order to bridge the existing gap in implantology research, we have engineered patient-specific bone grafts from human induced pluripotent stem cells, and used these as research platforms to study the cellular response to the implant material, the strength of interaction of the implant with the engineered bone, and the quality of the bone-implant interface. In addition to allow development and screening of implant surfaces promoting enhanced osseointegration, this platform could represent the first important step towards the generation of personalized treatments without the need for animal testing.

Biomaterials Research Centre´s Annual Day 2015-11-10

“Tissue Engineering”

Abstracts

“Healthy and disease skin models for research and screening purposes”

• Dr. A. El Ghalbzouri , PhD, Ass. Prof. LUMC skin research lab, Leiden, The Netherlands

Tissue engineering involves the use of living cells to develop biological substitutes for tissue replacements. Next to the treatment of acute and chronic wounds, tissue engineering offers new opportunities as a tool for the industry. For example, reconstructed human skin equivalents (HSEs) are representative models of human skin and widely used for screening purposes, clinical applications and research purposes. Since in vitro HSEs recapitulate most of the in vivo characteristics of human skin, they contribute to the replacement of animal

• experimentation. Human epidermal skin models are mostly used to predict the safety of ingredients used in

various industries, while full thickness HSEs are more often applied for research and tissue engineering

• purposes. These HSEs are all generated with skin tissue obtained from healthy donors. In dermatology, there

is unmet clinical need to develop therapies for a large number of skin diseases, including skin cancer, psoriasis and eczema. Currently there are no such off-the shelf in vitro models available that harbour specific diseased characteristics to screen and validate novel targets, and test the effects of potential new therapies in

• vitro. Therefore, a highly active research area is emerging by combining tissue engineering principles and

knowledge from developmental biology to establish complex three-dimensional in vitro disease model systems representing various skin conditions and diseases, including recessive epidermolysis bullosa simplex, cutaneous squamous cell carcinoma and melanoma, wound healing and skin aging. These HSEs are an excellent tool to gain more insight into the mechanisms of various pathological conditions and contribute to the development of appropriate therapies and novel therapeutics.

“Regulatory use of in vitro skin equivalents” Kristina Fant, Research scientist, SP Technical Research Institute of Sweden, Borås, Sweden

Progress in the development of test systems based on in vitro tissue engineering in the last decades have contributed to a significant decrease in animal use for industrial risk and safety assessment of products and

• ingredients. This is also reflecting extensive research efforts sparked e.g. by stricter legislation foremost

within the EU, where the REACH Regulation stipulates that animal testing should only be undertaken as the last resort when no other methods are available, and the Cosmetics Regulation forbids all animal testing on finished cosmetic products and their ingredients since 2013 regardless of the availability of alternative

• methods. Also for medical devices, recent changes in the ISO 10993 series put an increasing emphasis on the

use of cell-based methods before animal tests both for ethical reasons and to provide improved predictivity and reproducibility. Reconstruction of human epidermis was first described in the scientific literature over thirty years ago and has since then reached a stage where the model is organotypic, i.e. it reflects the histological appearance and the lipid composition of normal epidermis. Thanks to dedicated research efforts in industry and in EU-funded projects, already from the start aiming specifically for possible future validation of methods and regulatory acceptance, RhE technology has finally been adopted as stand-alone methods in OECD test guidelines: OECD TG 431 for skin corrosion (2004) and OECD TG 439 for skin irritation testing of chemicals (2009). However, the methods are not yet validated to also cover testing of medical devices where the irritating potential is usually much lower. An ISO 10993/TC194-organized validation study is underway and some of the considerations in this study will be discussed.

Biomaterials Research Centre´s Annual Day 2015-11-10

“Tissue Engineering”

Abstracts

“Material and Clinical Working Scientists -”Hand in Hand”- for Development of Scaffolds Stimulating Stem Cell Growth and Bone Regeneration” A Finne-Wistrand1, K. Mustafa2

1 Department of Fibre and Polymer Technology, School of Chemical Science and Engineering, KTH Royal Institute of Technology,

• Sweden. 2Department of Clinical dentistry, Center for Clinical Dental Research, University of Bergen, Norway

In tissue engineering, the regeneration process is affected by the scaffold microstructure as well as its initial biological and biomechanical properties. There are in general two ways to influence the cell-material interactions, optimize the polymer microstructure and optimize the scaffold design. Our group has strategically evaluated the cell-material interactions and has today a broad knowledge of the influence from functionality, hydrophilicity and crystallinity as well as the scaffold design, porosity, pore structure and mechanical properties. The different strategies presented here are from projects where we are working with scaffold-based therapies aimed at successful engineering of bone constructs for clinical use. The copolymer design is often used for tuning cell-material interactions. We know that even small differences in polymer microstructure influence the material properties and thereby also the cell-material interaction. As an example

• f this, poly(LLA-co-DXO) scaffolds enhance the attachment and differentiation of human osteoblast-like cells in vitro

compared to poly(LLA-co-CL).1 In these copolymer scaffolds pore sizes of >90 was shown to induce better cellular and tissue responses where blood vessels can be recruited. It was also found that endothelial microvascular networks can be generated through the polymer scaffolds and maintained in vivo influencing the osteogenic potential of the tissue engineered construct. Another way to tune the mechanical properties and the hydrophilicity is to add surfactants. In vitro and in vivo data generated recently, indicated that tuning hydrophilicity of copolymer scaffolds with 3% of Tween 80 promotes the proliferation of bone marrow derived stem cells (MSC) and induce bone formation (Un published data). Furthermore, cellular responses was shown to be improved by modifying the surface of the poly(LLA-co-CL) scaffolds with nano- diamond particles (n-DP). n-DP modification significantly increased cell attachment and differentiation of MSC on poly(LLA-co-CL) scaffolds in vitro and enhanced bone formation in vivo.2,3 Another strategy that we are using to tune material-cell interactions is to functionalize the polymers and immobilize growth factors. We have, for example, synthesized epoxy-functionalized polyesters4 and covalently immobilized heparin and we have compared covalently attached and physisorbed BMP2 to the scaffold.5 The physisorption of BMP2

• nto nDP modified copolymer scaffolds appears to hold great promise compared to growth factors adsorbed solely onto

a polymer. A low dose of BMP2 physisorbed onto scaffolds modified with nanodiamond particles was shown to be bioactive for bone regeneration. Obtaining bone after 4 weeks in vivo as demonstrated by histology and µCT analyses suggests accelerated bone regeneration in relation to the modified copolymer scaffolds. The concept of using a combination of a biomaterial and stem cells for bone regeneration has been tested recently in

• human. This phase I study was performed in Bergen employing 11 patients required reconstruction of alveolar bone. It

is among few clinical studies in Europe with Advanced Therapy Medicinal Products. The study indicate that the use of biomaterials combined with MSC in the applied protocol for augmentation of the atrophied mandibular ridge have results comparable to the gold standard; autologous bone transplantation. A close collaboration between material scientists, cell biologists and clinicians is important for finding optimal parameters and this talk will describe how we systematically have worked with this during the years.

REFERENCES:

1 S. Dånmark, A. Finne-Wistrand, M. Wendel, et al (2010) J Bioact Compat Polym 25:207-223.2 Z. Xing, T. O. Pedersen, X. Wu, et al (2013) Tissue

engineering 19:1783-1791. 3 Y. Sun, A. Finne-Wistrand, T. Waag, et al (2015) Macromol. Mater. Eng. 300: 436–447 4J. Undin, A. Finne-Wistrand, A.-C. Albertsson et al (2013) Biomacromolecules 14: 2095-2102. 5 S. Suliman, Z. Xing, XJ. Wu, et al (2015) J. Control Release, 197, 148-157. ACKNOWLEDGEMENTS: The authors acknowledge the European Union 7th Frame Program, VascuBone (project number 242175), The Swedish Research Council (Dnr 621- 2013-3764), The Norwegian Research Council (17734/V50, 180383/V4

Biomaterials Research Centre´s Annual Day 2015-11-10

“Tissue Engineering”

Abstracts

“Biomaterials in the year of light – Emerging microscopy techniques for 3D imaging of tissue-mimicking milieus” Annika Enejder, Prof. Dep. of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.

We develop an emerging category of laser-based microscopy techniques for 3D visualization of biopolymers (native and bioengineered fibrous proteins, carbohydrates, membrane lipids etc.) in living cells and extra-cellular mimicking matrices/surfaces. This allows us to monitor the dynamic and dual interaction between cells and ECM components/different surface chemistries under close to native conditions without any sample preparations. Instead of attaching a bulky fluorophore to the target molecules, their inherent vibrations are resonantly enhanced by frequency- matched electromagnetic waves from pico-second short laser pulses and then detected as blue-shifted scattered light; Coherent Anti-Stokes Raman Scattering (CARS) and/or Second Harmonic Generation (SHG) signals. The group hosts

• ne of the most technically well-equipped laboratories for non-linear

microscopy in Europe and the single one in Scandinavia, allowing for simultaneous CARS, Raman, 2-photon fluorescence, Second and Third Harmonic Generation (SHG & THG) microscopy. Unique images of adipocyte- derived stem cells in tissue-mimicking recombinant protein-engineered scaffolds (collaboration with the Heilshorn group, Stanford University) and fibroblast cells on lipid coated surfaces (Julie Gold, Chalmers) will be shown without any sample preparation, sectioning or labeling.

CARS microscopy image at the carbon-hydrogen vibration 2930 cm-1 characteristic for proteins, showing the distribution of fibers in an electrospun scaffold of elastin-like proteins synthesized by recombinant protein-engineering by the Heilshorn group, Stanford University. The image covers 35x35x10 µm.

“How to educate the next generation of tissue engineering researchers” Patric Wallin, PhD cand. Applied Physics and Engineering Education Research, Chalmers University of Technology, Gothenburg, Sweden.

Tissue engineering is very research intensive both in academia and industry, and graduates that are going to work in the field are required to have the ability to advance the scientific frontier. Therefore, tissue engineering education needs to reflect the nature of the discipline and establish a strong link between research and teaching. In this talk, I will present a framework that can be used to establish different approaches to link research and teaching. In the second part, I will focus on my own case study to describe how teaching and research is linked in a master’s course on tissue engineering. A central component of the course is an authentic research project that the students carry

• ut in smaller groups and in collaboration with faculty. I will discuss how the students experience learning in this kind
• f discovery-oriented environment based on data from reflective writing and interviews. From the data three themes

related to the students’ learning experiences were identified: learning to navigate the field, learning to do real research and learning to work with others. Overall, the students strongly valued learning in a discovery-oriented environment and three aspects of the course contributed to much of its success: taking a holistic approach to linking teaching and research, engaging students in the whole inquiry process, and situating authentic problems in an authentic physical and social context.