Investigation of the extracellular vesicles (VEs) role in the initiation, propagation, regeneration, and modeling of biological mineralization

• Grant number: 19/08568-2
• Support Opportunities: Research Projects - Thematic Grants
• Start date: November 01, 2020
• End date: October 31, 2026
• Field of knowledge: Biological Sciences - Biochemistry - Chemistry of Macromolecules
• Principal Investigator: Pietro Ciancaglini
• Grantee: Pietro Ciancaglini
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Pesquisadores principais: Sandra Yasuyo Fukada Alves
• Associated researchers: Adalberto Luiz Rosa ; Ana Paula Ramos ; Márcio Mateus Beloti ; Maytê Bolean Correia
• Associated research grant(s): 24/19254-7 - Extracellular vesicles in diabetes: Implications for bone health and beyond (ExtraVesiBone), AP.R SPRINT; 24/02130-3 - Development of bio-scaffolds containing extracellular matrix vesicles by 3D printing: an innovative approach in bone regeneration, AP.R; 21/03697-9 - Spectrophotometer with UV-RAMAN Optical Fiber - tematic project 2019/08568-2, AP.EMU
• Associated scholarship(s): 24/22322-4 - Bioactive hybrid coatings for titanium dental implants: evaluation of osteogenic activity and osseointegration potential, BP.DR; 24/22479-0 - Effect of mesenchymal stem cell-derived extracellular vesicles on osteoblastic differentiation and bone repair, BP.DD; 24/17743-0 - Elucidation of the composition and function of the protein corona in matrix vesicles (MVs): a new mechanism for controlling biomineralization?, BP.PD; + associated scholarships 24/09087-6 - Scaffolds based on collagen, carrageenan and doxycycline in a subantimicrobial dose for in vivo treatment of periodontal disease, BP.PD; 24/15867-4 - Role of exosomes isolated from osteosarcoma cell lines in the formation and development of osteoclasts, BP.IC; 24/04140-6 - Effects of Ionizing Radiation on Vesicle-Guided Bone Mineralization: Therapeutic Perspectives, BP.PD; 23/16012-0 - EFFECT OF THE APPLICATION OF MAGNETIC NANOPARTICLES AND HYPERTHERMIA IN OSTEOSARCOMA, BP.MS; 24/02382-2 - THE STING PATHWAY ON OSTEOCLASTOGENESIS AND DEVELOPMENT OF EXPERIMENTAL PERIAPICAL LESION, BP.MS; 23/18097-2 - The role of succinate (SUCNR1/GPR91) in the periodontitis formation and bone loss in an experimental model of apical lesion, BP.MS; 23/16079-7 - Effect of quercetin and quercetin Sr2+ on osteoclastogenesis, BP.IC; 23/17960-9 - Training in the isolation of extracellular vesicles (EVs) and MVs modeling biological mineralization, BP.TT; 23/12550-7 - Functional modification of mitochondria during osteoclast differentiation, BP.IC; 23/07580-4 - Preparation of hydrogels containing type II collagen and carrageenans and their potencial use for cartilage regeneration., BP.IC; 23/07244-4 - Development of cells co-culture models to assess the effect of new strontium and flavonoids complexes with application in bone and cartilage regeneration, BP.PD; 23/10404-3 - Investigation of the extracellular vesicles (VEs) role in the initiation, propagation, regeneration, and modeling of biological mineralization, BP.TT; 23/01644-0 - Mimetic models of matrix vesicles to study the action of sphingomyelinase, BP.IC; 23/03640-2 - Effect of the O-GlcNacylation pathway on osteosarcoma invasion and migration, BP.IC; 23/01756-3 - Langmuir monolayers as models of matrix vesicles and their role in bone biomineralization, BP.DD; 22/11231-2 - Investigation of the extracellular vesicles (VEs) role in the initiation, propagation, regeneration, and modeling of biological mineralization, BP.TT; 22/04885-6 - Investigation of the role of Extracellular Vesicles (EVs) and Matrix Vesicles (MVs) in the initiation, propagation, regeneration and control of biological mineralization, BP.PD; 22/05026-7 - Training in the isolation of extracellular vesicles (EVs) and MVs modeling biological mineralization, BP.TT; 21/13140-1 - Study of PHOSPHO1 and nSMase2 interaction with model membranes: a possible correlation in matrix vesicle secretion, BP.DR; 21/13934-8 - Role of glycolysis in osteoclasts function, BP.IC; 21/08558-7 - Study of the effect of gel formulations containing Sr2+ on dentin and bone mineralization in vivo, BP.DR; 21/12121-3 - Effect of morina-Sr2+ complexes on experimental bone loss, BP.IC; 21/12983-5 - Effect of VEs released by osteoblasts on osteoclastogenesis and bone loss in vivo, BP.PD; 21/01499-5 - Effect of extracellular vesicles on osteoblast differentiation and bone regeneration induced by mesenchymal stem cells, BP.DR; 21/02768-0 - Study of the interaction properties of PHOSPHO1 and Sphingomyelinase with model membrane systems, BP.IC - associated scholarships

Abstract

Extracellular Vesicles (EVs) have been related to a wide kind of function like developments in immunology, angiogenesis, and stem-cells biology, as well as several pathologies. EVs are bioactive organelles that can carry genetic information, lipids, proteins, and nucleic acids between cells, inducing many molecular functions associated with signalization and regulation of gene expression in the target cells. There is a special group of EVs called Matrix Vesicles (MVs). These vesicles are protagonists in one of the most accepted theories regarding the first hydroxyapatite crystals formation and mineral propagation during the bone formation in vertebrates. However, there are still gaps in these theories that need to be investigated, for example: What is the effective role of vesicles in the process? What is the role of proteins/enzymes present in the MVs? Which interactions between collagen and proteins/enzymes present in the MVs can be responsible for the nucleation and consequently propagation of mineralization? How these interactions regulate the mineralization process mediated by MVs? How pH and phosphosubstrates influence in the formation and propagation of hydroxyapatite crystals mediated by MVs? Through such questions, the project focuses on EVs obtention with enough quality and quantity to understand the interactions source between native vesicles and osteogenic proteins. Proteoliposomes and monolayers of Langmuir as model of membrane enriched with the main proteins of the mineralization process will be used to determine in details the mechanism of apatite formation and proteins function during the process. The project is innovative and interdisciplinary having the proponent and research collaborators from biochemistry, physical chemistry, pharmacy and odontology areas. Thus, this project will comprehend several methodology fields such as the state of biophysical art (rheology, fluidity), tool methodologies (AFM-TERS, TEM microscopies, BAM, microanalysis of X-ray energy dispersive spectroscopy, RAMAN e SPR), biochemical methodologies (isolations and characterization of native EVs, production of recombinant proteins), cellular biology (primary chondrocytes, osteoblasts and osteoclasts), and functions and applications of EVs in vitro and in vivo using biomimetic proteoliposomes for bone regeneration. Highly resolution details regarding the source of interactions between native EVs or proteoliposomes with main proteins of the biomineralization process can be obtained. The research proposal will provide detailed information about the mineralization ability of EVs, specifying the nucleation ability, propagation, regeneration, and degradation. Another strong feature of the proposal is the development of methodological approaches that can be generalized for EVs investigation. This project becomes very innovative because very few laboratories over the world focus their research on the discovery of the EVs functions. Native and model of vesicles present a great biotechnological potential and can also be used for inhibitors selection in order to prevent the pathological calcification, as well as to provide the bone regeneration/reabsorption. (AU)

Scientific publications

(18)

(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)

MARCOS A.E. CRUZ; CLAUDIO R. FERREIRA; LUCAS F. B. NOGUEIRA; LUIZ H.S. ANDRILLI; ANA LARA N. DOS SANTOS; JEFERSON M. M. MACEDO; EKEVELINY A. VESCHI; BRUNO Z. FAVARIN; HEITOR G. SEBINELLI; MAYTE BOLEAN; et al. ASPECTOS QUÍMICOS DA BIOMINERALIZAÇÃO ÓSSEA. Química Nova, v. 45, n. 5, p. 578-593, 2022-07-22. (17/20846-2, 19/25054-2, 19/08568-2)

PASQUARELLI, ALBERTO; SILVA ANDRILLI, LUIZ HENRIQUE; BOLEAN, MAYTE; FERREIRA, CLAUDIO REIS; EUFRASIO CRUZ, MARCOS ANTONIO; DE OLIVEIRA, FLAVIA AMADEU; RAMOS, ANA PAULA; MILLAN, JOSE LUIS; BOTTINI, MASSIMO; CIANCAGLINI, PIETRO. Ultrasensitive Diamond Microelectrode Application in the Detection of Ca2+ Transport by AnnexinA5-Containing Nanostructured Liposomes. BIOSENSORS-BASEL, v. 12, n. 7, p. 15-pg., 2022-07-01. (21/13140-1, 19/08568-2, 17/25475-2)

SILVA, LUCAS SANTOS; DE MELO, MARYANNE TRAFANI; SPONCHIADO, PEDRO AUGUSTO INVERNIZZI; BARBOSA JUNIOR, FERNANDO; TAPIA-BLACIDO, DELIA RITA; CIANCAGLINI, PIETRO; RAMOS, ANA PAULA; MANIGLIA, BIANCA CHIEREGATO. Synthesis of composite corn starch/hydroxyapatite nanoparticle biomembranes and their effect on mineralization by osteoblasts. Journal of Applied Polymer Science, v. 140, n. 43, p. 14-pg., 2023-08-18. (19/08568-2, 21/08359-4, 20/08727-0, 19/25054-2, 21/05947-2)

VESCHI, EKEVELINY AMABILE; BOLEAN, MAYTE; DA SILVA ANDRILLI, LUIZ HENRIQUE; SEBINELLI, HEITOR GOBBI; STRZELECKA-KILISZEK, AGNIESZKA; BANDOROWICZ-PIKULA, JOANNA; PIKULA, SLAWOMIR; GRANJON, THIERRY; MEBAREK, SAIDA; MAGNE, DAVID; et al. Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 23, n. 16, p. 15-pg., 2022-08-01. (19/25054-2, 21/13140-1, 17/20846-2, 19/08568-2, 22/05026-7)

ANDRILLI, LUIZ H. S.; SEBINELLI, HEITOR G.; COMINAL, JUCARA G.; BOLEAN, MAYTE; HAYANN, LARWSK; MILLAN, JOSE LUIS; RAMOS, ANA P.; CIANCAGLINI, PIETRO. Differential effects of the lipidic and ionic microenvironment on NPP1's phosphohydrolase and phosphodiesterase activities. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES, v. 1866, n. 4, p. 13-pg., 2024-02-13. (21/03697-9, 22/04885-6, 21/13140-1, 19/25054-2, 19/08568-2)

FAVARIN, BRUNO ZOCCARATTO; RAMOS, ANA PAULA; COSTA FILHO, ANTONIO; CIANCAGLINI, PIETRO. Comparative study of the native and mutant TNAP in in vitro biomineralization. BIOPHYSICAL REVIEWS, v. 13, n. 6, p. 1-pg., 2021-12-01. (18/12092-0, 19/08568-2, 16/21236-0)

ANDRILLI, LUIZ H. S.; SEBINELLI, HEITOR G.; FAVARIN, BRUNO Z.; CRUZ, MARCOS A. E.; RAMOS, ANA PAULA; BOLEAN, MAYTE; MILLAN, JOSE LUIS; BOTTINI, MASSIMO; CIANCAGLINI, PIETRO. NPP1 and TNAP hydrolyze ATP synergistically during biomineralization. PURINERGIC SIGNALLING, v. N/A, p. 14-pg., 2022-07-23. (19/25054-2, 19/08568-2, 21/13140-1, 17/20846-2)

GUILHERME NETO, JOAO LUIZ; VENTURINI, LUCAS GABRIEL RODRIGUES; SCHNEIDER, AYDA HENRIQUES; TAIRA, THAISE MAYUMI; RODRIGUES, LETICIA FERNANDA DUF FLES; VERAS, FAVIO PROTASIO; OLIVEIRA, SICILIA REZENDE; DA SILVA, TARCILIA APARECIDA; CUNHA, FERNANDO QUEIROZ; FUKADA, SANDRA YASUYO. Neutrophil Extracellular Traps Aggravate Apical Periodontitis by Stimulating Osteoclast Formation. JOURNAL OF ENDODONTICS, v. 49, n. 11, p. 8-pg., 2023-10-28. (19/08568-2, 13/08216-2, 17/24073-8)

FERREIRA, CLAUDIO R.; CRUZ, MARCOS ANTONIO E.; BOLEAN, MAYTE; ANDRILLI, LUIZ HENRIQUE DA S.; MILLAN, JOSE LUIS; RAMOS, ANA PAULA; BOTTINI, MASSIMO; CIANCAGLINI, PIETRO. Annexin A5 stabilizes matrix vesicle-biomimetic lipid membranes: unravelling a new role of annexins in calcification. EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS, v. 52, n. 8, p. 13-pg., 2023-11-08. (19/08568-2, 21/13140-1, 19/25054-2)

SEBINELLI, HEITOR GOBBI; ANDRILLI, LUIZ HENRIQUE SILVA; FAVARIN, BRUNO ZOCCARATTO; CRUZ, MARCOS AANTONIO EUFRASIO; BOLEAN, MAYTE; FIORE, MICHELE; CHIEFFO, CAROLINA; MAGNE, DAVID; MAGRINI, ANDREA; RAMOS, ANA PAULA; et al. Shedding Light on the Role of Na,K-ATPase as a Phosphatase during Matrix-Vesicle-Mediated Mineralization. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, v. 23, n. 23, p. 19-pg., 2022-12-01. (19/25054-2, 21/13140-1, 17/20846-2, 19/08568-2, 22/05026-7)

NOGUEIRA, LUCAS FABRICIO BAHIA; MANIGLIA, BIANCA C.; BUCHET, RENE; MILLAN, JOSE LUIS; CIANCAGLINI, PIETRO; BOTTINI, MASSIMO; RAMOS, ANA PAULA. hree-dimensional cell-laden collagen scaffolds: From biochemistry to bone bioengineerin. JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS, v. 110, n. 4, NOV 2021. (19/08568-2, 20/08727-0, 19/25054-2, 18/25871-8)

MACEDO, JEFERSON MATEUS MOUSSA; KAZMIRCZAK, PAULA CONSUL; HAYANN, LARWSK; DOTTA, TATIANE CRISTINA; CASSIANO, ANA FLAVIA BALESTRERO; PALMA-DIBB, REGINA GUENKA; CIANCAGLINI, PIETRO; BLASIDA, ALMA; ELIZAUR, CONCEPCION; CATIRSE, BENITEZ; et al. Strontium-substituted hydroxyapatite nanoparticles associated with type I-collagen enhances the obliteration of dentinal tubules and the mineralization by human dental pulp stem cells. EMERGENT MATERIALS, v. N/A, p. 16-pg., 2025-01-30. (19/25054-2, 19/08568-2, 23/07244-4)

CHAVES FILHO, GILDACIO PEREIRA; TAVARES, PEDRO DE ANDRADE; DE JESUS, ANANDA FERNANDA; CIANCAGLINI, PIETRO; SEGUNDO, JOSE EDUARDO SANTAREM; RAMOS, ANA PAULA. Building a digital library on research into mineralizing vesicles: a systematic review-based approach. BIOPHYSICAL REVIEWS, v. N/A, p. 25-pg., 2025-02-13. (19/25054-2, 19/08568-2, 21/03349-0, 23/07244-4)

RAMOS, ANA PAULA; BOLEAN, MAYTE; CRUZ, MARCOS A. E.; ANDRILLI, LUIZ H. S.; NOGUEIRA, LUCAS F. B.; SEBINELLI, HEITOR G.; DOS SANTOS, ANA LARA N.; FAVARIN, BRUNO Z.; MACEDO, JEFERSON M. M.; VESCHI, EKEVELINY A.; et al. Langmuir monolayers and proteoliposomes as models of matrix vesicles involved in biomineralization. BIOPHYSICAL REVIEWS, v. 13, n. 6, p. 3-pg., 2021-12-01. (18/25871-8, 17/20846-2, 19/25054-2, 19/08568-2)

PIMENTA, ISRAEL B.; CHAVES FILHO, GILDACIO; SILVA, ELIAS G. B.; NOGUEIRA, LUCAS F. B.; DE MENDONCA, TOMAZ SANTANA; FURTADO, TAISSA C. S.; FERREIRA GASPARNETO, PAULO CESAR; DIAS, LUIS GUSTAVO; FUKADA, SANDRA YASUYO; CIANCAGLINI, PIETRO; et al. Synthesis and Characterization of a Strontium-Quercetin Complex and Its In Vitro and In Vivo Potential for Application in Bone Regeneration. ACS OMEGA, v. 10, n. 5, p. 11-pg., 2025-01-30. (19/25054-2, 19/08568-2, 23/07244-4)

RIBEIRO, MARIANA S. P.; VENTURINI, LUCAS G. R.; SPECK-HERNANDEZ, CESAR A.; ALABARSE, PAULO V. G.; XAVIER, THAIS; TAIRA, THAISE M.; CUNHA, FERNANDO Q.; FUKADA, SANDRA Y.. AMPKα1 negatively regulates osteoclastogenesis and mitigates pathological bone loss. Journal of Biological Chemistry, v. 299, n. 12, p. 8-pg., 2023-12-01. (19/08568-2, 13/08216-2, 17/24073-8)

NOGUEIRA, LUCAS FABRICIO BAHIA; CRUZ, MARCOS ANTONIO EUFRASIO; TOVANI, CAMILA BUSSOLA; LOPES, HELENA BACHA; BELOTI, MARCIO MATEUS; CIANCAGLINI, PIETRO; BOTTINI, MASSIMO; RAMOS, ANA PAULA. Curcumin-loaded carrageenan nanoparticles: Fabrication, characterization, and assessment of the effects on osteoblasts mineralization. COLLOIDS AND SURFACES B-BIOINTERFACES, v. 217, p. 9-pg., 2022-09-01. (19/25054-2, 19/08568-2, 18/25871-8)

NOGUEIRA, LUCAS FABRICIO BAHIA; CRUZ, MARCOS ANTONIO EUFRASIO; MELO, MARYANNE TRAFANI DE; MANIGLIA, BIANCA CHIEREGATO; CAROLEO, FABRIZIO; PAOLESSE, ROBERTO; LOPES, HELENA BACHA; BELOTI, MARCIO M.; CIANCAGLINI, PIETRO; RAMOS, ANA PAULA; et al. Collagen/Kappa-Carrageenan-Based Scaffolds as Biomimetic Constructs for In Vitro Bone Mineralization Studies. Biomacromolecules, v. 24, n. 3, p. 9-pg., 2023-02-14. (19/08568-2, 20/08727-0, 22/01219-5, 19/25054-2, 16/25955-1, 18/25871-8, 21/05947-2)