TISSUE ENGINEERED BIOCOMPOSITES FOR BONE REGENERATION

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1 CONTACT CURRICULUM VITAE CHRISTIANE LARANJO SALGADO Born on March 31, 1978, in Ipatinga, Brasil. Biocomposite Group INEB - Instituto de Engenharia Biomédica, University of Porto Rua do Campo Alegre, 823, Porto, Portugal Phone: ; Fax: csalgado@ineb.up.pt FEUP Faculdade de Engenharia da Universidade do Porto Rua Dr. Roberto Frias, s/n Porto PORTUGAL Phone: ; Fax: chris@fe.up.pt ACADEMIC QUALIFICATIONS PhD in Mechanical Engineering. Faculty of Mechanical Engineering of the State University of Campinas (UNICAMP), July Master in Science. Faculty of Sciences of the Federal University of Minas Gerais, September, Dentistry. Faculty of Dentistry of the Federal University of Minas Gerais, July PRESENT POSITIONS Investigator at the Associate Laboratory IBMC-INEB (Instituto de Biologia Molecular e Celular-Instituto de Engenharia Biomédica), University of Porto, since Investigator at FEUP, since AREA OF SCIENTIFIC ACTIVITY TISSUE ENGINEERED BIOCOMPOSITES FOR BONE REGENERATION Over the past decades, bone tissue engineering has undergone a considerable development and now demonstrates a great potential for improved treatment or replacement of damaged bone in comparison with conventional therapies. Engineering bone typically uses a scaffold, osteoblasts or cells that can become osteoblasts (i.e. adult mesenchymal stem cells) and regulating factors that promote cell adhesion, differentiation and bone formation. Therefore highly porous scaffolds have a critical role in cell seeding, proliferation and new tissue formation in three dimensions. Scaffolds are 3D substrates for cells and their main goal is to serve as templates for tissue regeneration. The ideal scaffold must be biocompatible, non-immunogenic, have an interconnected porous network to allow cell penetration and transfer of nutrients, oxygen and waste products. Scaffolds must be biocompatible, osteoconductive, osteointegrative, and have enough mechanical strength to provide structural support during bone growth and remodelling. In addition, scaffolds can serve as delivery vehicles for cytokines such as bone morphogenetic proteins (BMPs), transforming growth factors (TGFs) that can transform recruited precursor cells from the host into bone matrix producing cells, thus providing osteoconduction. Christiane L. Salgado 1/7

2 Parameters such as macroporosity, microporosity and pore interconnectivity (and a biocompatible and bioresorbable substrate with controllable degradation rate), are crucial. The scaffolds should mimic cancellous bone morphology, structure and function in order to optimize integration in the surrounding tissue. Collagen is the main structural protein present in the human body. The good biological properties of collagen, as well as its biocompatibility, biodegradability and easy processing have resulted its frequent use for biomedical applications, being widely used as gels, powders or films. Hydroxyapatite can be easily combined with other materials in order to improve its properties. Among other examples is hydroxyapatite combined with natural polymers, namelly collagen. For bone tissue engineering to be promising it requires the use of scaffolds combined with the appropriate cells to allow for new tissue formation and Angiogenesis. This is a crucial multifactor process, regulated by a number of factors including the action of the vascular endothelial growth factor (VEGF). The development of bioceramics for bone tissue engineering includes chemical and physical modifications such nanostructured and nanocrystalline HA materials and surface grafting using growth factors, living bone cells or proteins. Adhesive protein, like Fibronectin, contribute to the structural stability of extracellular matrix (ECM) playing an important role in cell attachment to biomaterials surfaces, playing a role in adhesion, growth, differentiation or migration. In bone, they are involved in the early stages of osteogenesis and might be able to nucleate mineralization. Another class of glycoproteins, the matricellular class of secreted glycoproteins that includes osteonectin, thrombospondins, tenascins and osteopontin, exhibit counteradhesive effects. Indeed these can lead to changes in cell shape resulting in the disruption of cell-matrix interactions and cell mobility often require on tissue renewal, tissue remodeling and embryonic development. Osteonectin has a strong affinity for hydroxyapatite. Bone osteonectin strongly binds to type I collagen and synthetic HA and can mediate the in vitro mineralization of the type I collagen. Although these two proteins are both important in bone mineralization or cell matrix adhesion, their role in such events is still far from being well understood. Osteopontin(OPN), osteocalcin(oc), sialoprotein (SP) or phospho-serine (SIBLING) are proteins of the extracellular bone matrix that very efficiently stimulate bone cell activities and bone healing. The modification of collagen/nanoha with these proteins seems to be a promising approach to mimic the situation in bone. The functionalization of the scaffolds should enhance cell adhesion and proliferation; also it would be favorable for mesenquimal stem cells differentiation in bone tissue. DOMAIN OF SPECIALIZATION Research work has been dedicated to developing biocomposites for bone regeneration. Surface modification with different bone proteins will be investigated. Implantation studies revealed the biocompatibility and osteoconductive properties of collagen and nanohydroxyapatite composites. Bone cells and mesenchymal stem cells have been studied on different biomateriais based on collagen conjugated with nanohydroxyapatite. PRESENT RESEARCH INTERESTS New biomaterials development to be applied on porous scaffolds production. These materials would be used for bone tissue regeneration through tissue engineering Christiane L. Salgado 2/7

3 strategies. Macroporous biocomposite materials based on a Collagen type I and III matrix and nanoha/ nanotcp aggregates distributed throughout the surface. The materials will be addressed to enable binding and delivery of growth factors to induce bone tissue regeneration and vascularization. This could involve growth factors encapsulation in the scaffolds. The studies will include in vitro cell cultures (bone cells, endothelial cells and / or stem cells). In vivo experimentation in small animal model (rat and mouse) will be carried out, aiming at foreseeing the behaviour of the scaffolds in regenerating bone at critical size defects or to analyze inflammation induction with subcutaneous implants. Keywords: Regenerative Medicine, Tissue Engineering, Biomaterials Bone regeneration Biocomposites Collagen Artificial extracellular matrix Cell immobilization (3D cell culture) Nanohydroxyapatite Stem cells (substrate-mediated guidance of stem cells fate) COLLABORATIONS At INEB, several colleagues are involved in this activity, namely Fernando Jorge Monteiro, Maria Pia Ferraz, Suzana Sousa, Pedro Quelhas and Maria João Oliveira. HONORS AND AWARDS 2005: Honor, Federação de Sociedades de Biologia Experimental. 2013: FCT Post-doc Grant (SFRH/BPD/84443/2012). ACADEMIC AND PROFESSIONAL EXPERIENCE : Investigator in Faculty of Chemistry Engineering at State University of Campinas (UNICAMP), Brazil today: Investigator in Faculty of Engineering at University of Porto (FEUP), Portugal. EDUCATIONAL ACTIVITIES TEACHING IN UNDERGRADUATE AND POST-GRADUATE PROGRAMMES 2006: Lecturer at Mechanical Engineering in Materials Characterization, State University of Campinas. 2006: Lecturer at Mechanical Engineering in Polymer Process, State University of Campinas. 2012: Invited teacher at Master in Biomedical Engineering Program, University of Porto. LECTURER IN POST-GRADUATE PROGRAMMES 2005: Master Program in Sciences (Physiology and Pharmacology). Faculty of Christiane L. Salgado 3/7

4 Sciences, Federal University of Minas Gerais 2006: Doctoral Program Bioengineering, University of São Paulo. COMMITTEES, BOARDS AND OTHER RELEVANT SCIENTIFIC ACTIVITIES REFEREE BioMatter (2011) Tissue Engineering: part A (2011) PARTICIPATION IN SCIENTIFIC PROJECTS AS RESEARCHER NANOFORBONE - Development of scaffolds based on collagen and nanohydroxyapatyte for bone regeneration. NANOBIOFILM - Development of scaffolds based on nanohydroxyapatyte for bone application with drug delivery for bacterial infection. SUPERVISION OF SCIENTIFIC WORKS MSc STUDENTS Sandra Rodrigues. Preparation of Collagen-nanoHA Biocomposite Cryogels for Bone Tissue Engineering Applications (FEUP - co-supervisor) ( ). Eugênia Teixeira. Adsorption of osteopontin and cell behavior on nanohydroxyapatite/collagen surface (FEUP - co-supervisor) (2012-today). ORGANIZATION OF SCIENTIFIC EVENTS ORGANIZING COMMITTEE XI and XII Summer Course in Physiology and Pharmacology Faculty of Sciences at Federal University of Minas Gerais. Belo Horizonte (2004 and 2005). LANGUAGES English: Advanced Spanish and French: Average PUBLICATIONS AND COMMUNICATIONS PUBLICATIONS (2006-present) THESES Salgado, CL. Development of three-dimensional Polymer Matrixes for Bone Tissue Engineering. PhD Thesis. Faculty of Mechanical Engineering at State University do of Campinas (UNICAMP), Campinas, Salgado, CL. Avaliação da Biocompatibilidade de Biovidro Ativo Poroso - BG60S. Master thesis. Faculty of Sciences at Federal University of Minas Gerais (UFMG), Belo Horizonte, Christiane L. Salgado 4/7

5 BOOK CHAPTERS Salgado, CL; Sanchez, E. M. S., Oliveira, M. F., Maia, I. A., Silva, J. V. L., Zavaglia, C.A.C. Virtual and Rapid Manufacturing: Advanced Research in Virtual and Rapid Prototyping. Londres : Taylor and Francis, 2007, v.1. p.860. PAPERS IN INTERNATIONAL REFEREED JOURNALS Salgado, C. L., Solomao, Z., Silva, P. B., Sanchez, E. M. S., Zavaglia, C. A. C. Biocompatibility And Osteo-Differentiation Study Of Poly(E-Caprolactone) And ß- Tricalcium Phosphate Composite Membranes. Key Engineering Materials., v.396, p , C. L. Salgado, Francischi, J. N., Pereira, M. M., Turchetti-Maia, R. M. M., Salas, C. E., Lopes, M. T. P. Evaluation of Biocompatibility of Bioactive Glass (BG60S). Materials Keys Engineering., v.309, p , C.L.Salgado, E.M.S. Sanchez, J.F. Mano, A.M.Moraes. Characterization Of Chitosan and Polycaprolactone Membranes Designed for Wound Repair Application. Journal of Material Science, v.47(2), p , C.L.Salgado, E.M.S. Sanchez, C.A.C. Zavaglia, A.B.A. Almeida, P.L. Granja. Injectable Biodegradable Polycaprolactone-Sebacic Acid Gels For Bone Tissue Engineering. Tissue Engineering: part A, 18(1-2):137-46, C.L.Salgado, E.M.S. Sanchez, C.A.C. Zavaglia, P.L. Granja. Biocompatibility and Biodegradation of Polycaprolactone-Sebacic Acid Blended Gels. Journal Biomedical Materials Research: Part A, 100(1):243-51, C.L. Salgado, M. Oliveira, J.F. Mano. Combinatorial cell 3D biomaterials interactions screening for tissue engineering using bioinspired superhydrophobic substrates. Integrative Biology: Technical Innovation, v.4(3), p , Rodrigues SC, Salgado CL, Sahu A, Monteiro FJ. Preparation of Collagen- Hydroxyapatite Biocomposite Scaffolds by Cryogelation Method for Tissue Engineering Applications. Journal of Biomedical Materials Research Part A, v.101 (4), p , Oliveira MB, Salgado CL, Song W, Mano JF. Combinatorial On-Chip Study of Miniaturized 3D Porous Scaffolds Using a Patterned Superhydrophobic Platform. Small, 11;9(5): , ABSTRACTS IN REFEREED JOURNALS Salgado CL, Duarte ECB, Aguiar APV, Alfenas ER, Santos VR. Microbiological test: acrylic resin bottons disinfected in microwave energy. Journal Of Dental Research, V.81, P.B63, Salgado CL, Oliveira MB, Mano JF. Innovative three-dimensional platform for combinatorial analysis of cell/biomaterials interactions. Tissue Engineering, v.26(s1), p.25, Rodrigues SC, Sahu A, Salgado CL, Monteiro FJ. Preparation Of Collagen- Hydroxyapatite Biocomposite Scaffolds By Cryogelation Method For Tissue Engineering Applications. Tissue Engineering, v.26(s1), p.276, Salgado, C. L.; Oliveira, M. B.; Mano, J. F. Combinatorial Cell-Porous Scaffolds Interactions Study and Scaffold Physicochemical Characterization in an Innovative Bioinspired High-Throughput Platform. International Journal of Artificial Organs: 34 (8), p , Oliveira, M. B.; Salgado, C. L.; Song, W. L.; Mano, J.F. Superhydrophobic patterned chips for the combinatorial and rapid study of 3D biomaterials-cells interactions and protein delivery systems. Journal Of Tissue Engineering And Regenerative Medicine,Volume: 6, Special Issue: SI, Supplement: 2, Oliveira, M. B.; Salgado, C. L.; Mano, J. F. Patterned superhydrophobic surfaces Christiane L. Salgado 5/7

6 for the combinatorial assessment of 3D biomaterials-cells interactions. Journal Of Tissue Engineering And Regenerative Medicine,Volume: 6, Special Issue: SI, Supplement: 1, Pages: , Oliveira, M. B.; Salgado, C. L.; Mano, J. F. Superhydrophobic platforms for the combinatorial analysis of biomaterials-cells interactions using arrays of 3D scaffolds with distinct mechanical and morphological properties. Journal Of Tissue Engineering And Regenerative Medicine, Volume: 6, Special Issue: SI, Supplement: 1, Pages: , PATENT Salgado, C. L., Sanchez, E. M. S., Granja, P. L., Zavaglia, C. A. C. Método De Confecção De Polímero Biodegradável Injetável, 2008 Patente: Privilégio De Inovação N , Método De Confecção De Polímero Biodegradável. 23 de Setembro de 2008 (Depósito). COMMUNICATIONS ORAL COMMUNICATIONS Salgado CL, Oliveira MB, Mano JF. Innovative three-dimensional platform for combinatorial analysis of cell/biomaterials interactions. Termis 2011, Granada, Salgado, C. L., Sanchez, E. M. S., Oliveira, M. F., Silva, J. V. L., Zavaglia, C. A. C. Evaluation Of Degradation Of Bioabsorbable Polycaprolactone Used In Rapid Prototyping For Medical Application In: 3rd International Conference On Advanced Research In Virtual And Rapid Prototyping, Leiria, Salgado, C. L., Salas, C. E., Francischi, J. N., Pereira, M. M., Lopes, M. T. P., Turchetti-Maia, R. M. M. Determinação de citotoxicidade e da resposta inflamatória aguda induzida por biovidro ativo poroso In: III Congresso latinoamericano de Biomateriais e Órgãos Artificiais, Campinas, Salgado, C. L., Solomao, Z., Silva, P. B., Sanchez, E. M. S., Zavaglia, C. A. C. Biocompatibility And Osteo-Differentiation Study Of Poly(E-Caprolactone) And ß- Tricalcium Phosphate Composite Membranes. 20º Bioceramics, Búzios, POSTER COMMUNICATIONS Salgado CL, Oliveira MB, Mano JF. Combinatorial Cell-Porous Scaffolds Interactions Study and Scaffold Physicochemical Characterization in an Innovative Bioinspired High-Throughput Platform. XXXVIII Congress of the European Society for Artificial Organs (ESAO), Porto, Rodrigues SC, Sahu A, Salgado CL, Monteiro FJ. Preparation Of Collagen- Hydroxyapatite Biocomposite Scaffolds By Cryogelation Method For Tissue Engineering Applications. Termis 2011, Granada, Salgado, C. L., Moraes, A. M. Caracterização Da Degradação E Da Biocompatibilidade Das Membranas De Quitosana E Policaprolactona Para Aplicação Em Engenharia Tecidual In: XVIII Congresso Brasileiro De Engenharia Química, Foz Do Iguaçu, Salgado, C. L., Sanchez, E. M. S., Moraes, A. M. Morphology and biocompatibility analyses of chitosan and polycaprolactone membranes for tissue engineering application In: 19 th International Congress of Chemical and Process Engineering, Praga, Salgado, C. L., Sanchez, E. M. S., Zavaglia, C. A. C. Evaluation Of Polycaprolactone And Sebacic Acid Blend Scaffolds For Bone Tissue Engineering. 5 o Congresso De Órgãos Artificiais E Biomateriais, Ouro Preto, Christiane L. Salgado 6/7

7 Salgado, C. L., Sanchez, E. M. S., Zavaglia, C. A. C. Evaluation Of Polycaprolactone And Sebacic Acid Blends Fused For Bone Tissue Engineering In: 8th World Biomaterials Congress, 2008, Amsterdan. 8th World Biomaterials Congress, Salgado, C. L., Sanchez, E. M. S., Zavaglia, C. A. C., Granja, P. L. Injectable Polymeric Matrices for Bone Tissue Engineering In: 8th World Biomaterial Congress, 2008, Amsterdan. 8th World Biomaterial Congress, Sanchez, E. M. S., Aismoto, M. C., Salgado, C. L., Sanchez, C., Zavaglia, C. A. C. Comparação Entre Dois Cimentos Ortopédicos A Base De Poli Metacrilato De Metila (Pmma) Segundo Alguns Requisitos Da Norma ABNT NBR-ISO 5833 In: ABPOL 2007, São Paulo, Salgado, C. L., Sanchez, E. M. S., Zavaglia, C. A. C. Evaluation Of Polycaprolactone And Sebacic Acid Blends Fused For Bone Tissue Engineering In: Euromat 2007, 2007, Nurembergue. Euromat Frankfurt:, Sanchez, E. M. S., Salgado, C. L., Silva, J. V. L., Maia, I. A., Oliveira, M. F., Zavaglia, C. A. C. Mechanical Evaluation Of Polyamides Used In Rapid Prototyping After Accelerated Aging In: 3rd China-Europe Symposium - Processing And Properties Of Reinforced Polymers. Budapeste:, Sanchez, E. M. S., Salgado, C. L., Silva, J. V. L., Maia, I. A., Oliveira, M. F., Zavaglia, C. A. C. Morphological And Mechanical Evaluation Of Polyamides Used In Rapid Prototyping After Accelerated Aging In: Polychar15 - World Forum On Advanced Materials, 2007 Polychar15 - World Forum On Advanced Materials. Búzios, Salgado, C. L., Bossi, R. I., C. Cutrim, Zakia, M. B., Zavaglia, C. A. C. Degradação De Polímero Bioreabsorvível Para Liberação Controlada De Fármacos In: IV Congresso Latino Americano De Órgãos Artificiais E Biomateriais, Caxambu, Sanchez, E. M. S., Salgado, C. L., Silva, J. V. L., Maia, I. A., Oliveira, M. F., Zavaglia, C. A. C. Evaluation Of Mechanical Properties Of Polyamides Used In Rapid Prototyping In: V Sociedade Brasileira De Pesquisa Em Materiais, 2006, Florianópolis. V SBPmat., Salgado, C. L., Bossi, R. I., C. Cutrim, Zakia, M. B., Zavaglia, C. A. C. Degradação de polímero bioreabsorvível para liberação controlada de fármacos In: IV congresso Latino americano de órgãos artificiais e biomateriais, Caxambu, Salgado, C. L., Turchetti-Maia, R. M. M., Pereira, M. M., Francischi, J. N., Salas, C. E., Lopes, M. T. P. Citotoxity and Inflammatory Evaluation of Porous Bioactive Glass Scaffolds In: Sociedade Brasileira de Pesquisa em Materiais, Recife, February 27 th, Christiane L. Salgado 7/7