Association of tissue engineering techniques for mineralized tissue regeneration under degenerative inflammatory stimulus: analysis on 3D-culture perfusion bioreactor and animal inflammatory models

• Grant number: 16/15674-5
• Support Opportunities: Research Grants - Young Investigators Grants
• Start date: April 01, 2017
• End date: November 30, 2022
• Field of knowledge: Health Sciences - Dentistry - Dental Materials
• Principal Investigator: Diana Gabriela Soares dos Passos
• Grantee: Diana Gabriela Soares dos Passos
• Host Institution: Faculdade de Odontologia de Bauru (FOB). Universidade de São Paulo (USP). Bauru , SP, Brazil
• Associated researchers: Carlos Alberto de Souza Costa ; Gelson Luis Adabo ; Josimeri Hebling Costa
• Associated scholarship(s): 20/15971-5 - Bioprinted multifuncional hydrogels as a strategy for Regenerative Dentistry, BP.PD; 21/11223-7 - Indirect three-dimensional printing as a strategy to modulate the interconnectivity of chitosan-calcium scaffolds, BP.IC; 21/11098-8 - Biological characterization of macro-porous chitosan scaffolds associated to different mineral phases for bone regeneration, BP.IC; + associated scholarships 21/11090-7 - Injectable systems with controlled release of simvastatin: development strategy for dentin regeneration, BP.IC; 20/10369-5 - Development of porous scaffolds with oriented macro-channels network: association of bubbling-effect and three-dimensional printing techniques, BP.DD; 21/00357-2 - Applying tissue engineering techniques for realistic 3D culture models development, BP.TT; 20/01086-0 - Applying tissue engineering techiniques for the development of realistic 3D cell culture models, BP.TT; 19/00020-8 - Development of macro-porous chitosan-calcium scaffolds with controlled release of simvastatin: evaluation of the regenerative potential at inflamed areas, BP.PD; 19/06884-4 - Development and analysis of bioactive scaffolds for dentin regeneration, BP.MS; 19/05157-1 - Applying tissue engineering techniques for the development of realistic 3D culture models, BP.TT; 18/26091-6 - Dose response and time-course assays of statins for human pulp cells biostimulation, BP.IC; 18/16688-5 - Applying tissue engineering techniques for the development of realistic 3D culture models, BP.TT; 18/09378-0 - Application of tissue engineering for pulp-dentin complex regeneration: evaluation of a chitosan-calcium scaffold on in vitro simulated pulp exposure model, BP.IC; 18/09323-0 - Porosity modulation of chisotan scaffolds: effect on adhesion, spread and proliferation of mesenchymal stem cells, BP.IC; 17/24198-5 - Chitosan-calcium scaffolds for regeneration of pulp-dentin complex: chemotactic potential analysis in realist model - in vitro study, BP.IC; 17/20181-0 - Tissue Engineering and Biotechnology applyed to the development of calcium-containing macroporous scaffolds with surface nano-topography to modulate pulp-dentin complex regeneration, BP.DR; 17/05578-1 - Association of tissue engineering techniques for mineralized tissue regeneration under degenerative inflammatory stimulus: analysis on 3D-culture perfusion bioreactor and animal inflammatory models, BP.JP - associated scholarships

Abstract

The regeneration of mineralized tissues under degenerative inflammatory stimulus is a challenging situation, since the reparative potential of resident stem cells may be negatively influenced in the presence of high dosage of pro-inflammatory mediators. In this research project, an association of tissue engineering techniques will be evaluated in order to develop macro-porous chitosan scaffolds with architecture and composition similar with those found on mineralized tissue extracellular matrix, associated with release of pleiotropic drugs (statins) in order to modulate inflammation, angiogenesis and neo-tissue genesis by resident cells. Tissue engineering techniques will also be applied for the development of a realistic 3D-cell culture model, to evaluate the scaffolds at specific application sites in vitro. Initially, an in vitro degenerative inflammatory model capable of reduce in around 50% of mineralized tissue deposition by bone marrow (BMSC) and dental pulp (DPSC) stem cells will be stablished, by means of co-culture with LPS-stimulated monocytes (Phase 1). Then, the anti-inflammatory, pro-angiogenic and bioactive dosage of statins (simvastatin, atorvastatin and rouvastatin) will be selected. The effect of these dosage associations on the regenerative potential of BMSCs and DPSCs under degenerative inflammatory stimulus will be assessed (Phase 2). After that, several parameters will be tested to develop a macro-porous nano-fibrous chitosan scaffold with an interconnected pore network. This scaffold will be biomineralized with hidroxiapatite, in order to simulate mineralized matrix chemical composition. A drug release system based on chitosan microspheres loaded with the anti-inflammatory and bioactive dosage of statin will be added to the scaffold (Phase 3). The biologic effect of the scaffold will be assessed on BMSCs and DPSCs under the inflammatory stimulus, as well as the pro-angiogenic potential with endothelial cells (Phase 4). Finally, two in vitro cell culture models will be developed to evaluate the regenerative potential of the scaffolds at specific application sites (Phase 5). Bovine cancellous bone and dental pulp tissue samples will be collected and subjected to the decellularization process followed by BMSC or DPSC seeding, in co-culture or not with endothelial cells, on perfusion bioreactor. The scaffolds will be positioned on specific sites created on the tissue constructs, and this set will be cultivated on the perfusion bioreactor system. Finally, the effect of the scaffolds at specific application sites on animals will also be assessed (Rattus novergiccus) by means of experimental models of chronic inflammation. (AU)

Scientific publications

(13)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

FERNANDA BALESTRERO CASSIANO; DIANA GABRIELA SOARES; ESTER ALVES FERREIRA BORDINI; GIOVANA ANOVAZZI; JOSIMERI HEBLING; CARLOS ALBERTO DE SOUZA COSTA. Simvastatin-Enriched Macro-Porous Chitosan-Calcium-Aluminate Scaffold for Mineralized Tissue Regeneration. Brazilian Dental Journal, v. 31, n. 4, p. 385-391, 2020-09-04. (17/20181-0, 16/06704-8, 16/15674-5)

BORDINI, ESTER ALVES FERREIRA; CASSIANO, FERNANDA BALESTRERO; SILVA, ISABELA SANCHES POMPEO; USBERTI, FELIPE ROCHELLE; ANOVAZZI, GIOVANA; PACHECO, LEANDRO EDGAR; PANSANI, TAISA NOGUEIRA; LEITE, MARIA LUISA; HEBLING, JOSIMERI; COSTA, CARLOS ALBERTO DE SOUZA; et al. Synergistic potential of 1 alpha,25-dihydroxyvitamin D3 and calcium-aluminate-chitosan scaffolds with dental pulp cells. CLINICAL ORAL INVESTIGATIONS, v. 24, n. 2, p. 663-674, FEB 2020. (17/20181-0, 16/15674-5)

SUELEN ALINE DE LIMA BARROS; DIANA GABRIELA SOARES; MARIA LUÍSA LEITE; FERNANDA GONÇALVES BASSO; CARLOS ALBERTO DE SOUZA COSTA; GELSON LUÍS ADABO. Influence of Zirconia-Coated Bioactive Glass on Gingival Fibroblast Behavior. Brazilian Dental Journal, v. 30, n. 4, p. 333-341, 2019-07-22. (16/15674-5, 15/19034-8)

ORTECHO-ZUTA, U.; DE OLIVEIRA DUQUE, C. C.; LEITE, M. L.; BORDINI, E. A. F.; BASSO, F. G.; HEBLING, J.; DE SOUZA COSTA, C. A.; SOARES, D. G.. Effects of Enzymatic Activation of Bleaching Gels on Hydrogen Peroxide Degradation Rates, Bleaching Effectiveness, and Cytotoxicity. Operative Dentistry, v. 44, n. 4, p. 414-423, JUL-AUG 2019. (16/15674-5, 15/21770-4)

SOARES, DIANA GABRIELA; BORDINI, ESTER ALVES FERREIRA; CASSIANO, FERNANDA BALESTRERO; BRONZE-UHLE, ERIKA SOARES; PACHECO, LEANDRO EDGAR; ZABEO, GIOVANA; HEBLING, JOSIMERI; LISBOA-FILHO, PAULO NORONHA; BOTTINO, MARCO CICERO; DE SOUZA COSTA, CARLOS ALBERTO. Characterization of novel calcium hydroxide-mediated highly porous chitosan-calcium scaffolds for potential application in dentin tissue engineering. JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS, v. 108, n. 6, FEB 2020. (16/15674-5)

MARJORIE DE OLIVEIRA GALLINARI; ESTER ALVES FERREIRA BORDINI; VITOR DE TOLEDO STUANI; FERNANDA BALESTRERO CASSIANO; CAMILA CORREA DA SILVA BRAGA DE MELO; JULIANO MILANEZI DE ALMEIDA; LUCIANO TAVARES ÂNGELO CINTRA; CARLOS ALBERTO DE SOUZA COSTA; DIANA GABRIELA SOARES. Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells. Brazilian Oral Research, v. 37, 2023-02-13. (19/00020-8, 16/15674-5)

SOARES, DIANA G.; BORDINI, ESTER A. F.; SWANSON, W. BENTON; DE SOUZA COSTA, CARLOS A.; BOTTINO, MARCO C.. Platform technologies for regenerative endodontics from multifunctional biomaterials to tooth-on-a-chip strategies. CLINICAL ORAL INVESTIGATIONS, JUN 2021. (16/15674-5)

SOARES, DIANA G.; BORDINI, ESTER A. F.; SWANSON, W. BENTON; DE SOUZA COSTA, CARLOS A.; BOTTINO, MARCO C.. Platform technologies for regenerative endodontics from multifunctional biomaterials to tooth-on-a-chip strategies. CLINICAL ORAL INVESTIGATIONS, v. 25, n. 8, p. 4749-4779, AUG 2021. (16/15674-5)

BORDINI, ESTER A. F.; FERREIRA, JESSICA A.; DUBEY, NILESHKUMAR; RIBEIRO, JULIANA S.; DE SOUZA COSTA, CARLOS A.; SOARES, DIANA G.; BOTTINO, MARCO C.. Injectable Multifunctional Drug Delivery System for Hard Tissue Regeneration under Inflammatory Microenvironments. ACS APPLIED BIO MATERIALS, v. 4, n. 9, p. 6993-7006, SEP 20 2021. (18/14257-7, 16/15674-5)

SOARES, DIANA G.; ANOVAZZI, GIOVANNA; BORDINI, ESTER ALVES F.; ZUTA, UXUA O.; SILVA LEITE, MARIA LUISA A.; BASSO, FERNANDA G.; HEBLING, JOSIMERI; DE SOUZA COSTA, CARLOS A.. Biological Analysis of Simvastatin-releasing Chitosan Scaffold as a Cell-free System for Pulp-dentin Regeneration. JOURNAL OF ENDODONTICS, v. 44, n. 6, p. 971-976, JUN 2018. (13/23520-0, 16/15674-5)

FERNANDES, PAULO HENRIQUE MARTINS; BORDINI, ESTER ALVES FERREIRA; CASSIANO, FERNANDA BALESTRERO; DE AZEVEDO-SILVA, LUCAS JOSE; FERRAIRO, BRUNNA MOTA; LISBOA-FILHO, PAULO NORONHA; FORTULAN, CARLOS ALBERTO; DOS PASSOS, DIANA GABRIELA SOARES; BORGES, ANA FLAVIA SANCHES. TiO2 nanoparticles added to dense bovine hydroxyapatite bioceramics increase human osteoblast mineralization activity. Dental Materials, v. 38, n. 11, p. 9-pg., 2022-10-14. (18/23639-0, 13/07296-2, 16/15674-5)

GALLINARI, MARJORIE DE OLIVEIRA; FERREIRA BORDINI, ESTER ALVES; STUANI, VITOR DE TOLEDO; CASSIANO, FERNANDA BALESTRERO; DA SILVA BRAGA DE MELO, CAMILA CORREA; DE ALMEIDA, JULIANO MILANEZI; ANGELO CINTRA, LUCIANO TAVARES; DE SOUZA COSTA, CARLOS ALBERTO; SOARES, DIANA GABRIELA. Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells. BRAZILIAN ORAL RESEARCH, v. 37, p. 15-pg., 2023-01-01. (16/15674-5, 19/00020-8)

DA SILVA BRAGA MELO, CAMILA CORREA; CASSIANO, FERNANDA BALESTRERO; BRONZE-UHLE, ERIKA SOARES; STUANI, VITOR DE TOLEDO; FERREIRA BORDINI, ESTER ALVES; GALLINARI, MARJORIE DE OLIVEIRA; SOUZA COSTA, CARLOS ALBERTO; SOARES, DIANA GABRIELA. Mineral-induced bubbling effect and biomineralization as strategies to create highly porous and bioactive scaffolds for dentin tissue engineering. JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS, v. 110, n. 8, p. 14-pg., 2022-02-09. (16/15674-5, 19/06884-4)