Exploring the role of lipid-protein-mineral matrix on bone biomineralization: a biophysical approach using self-assembled films

• Grant number: 17/20846-2
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: February 01, 2018
• End date: March 31, 2023
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Physical-Chemistry
• Principal Investigator: Ana Paula Ramos
• Grantee: Marcos Antonio Eufrásio Cruz
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated scholarship(s): 19/26059-8 - Unraveling the mechanism of matrix vesicles biomineralization: a cryo-electron microscopy approach, BE.EP.DR

Abstract

Bone biomineralization is an exquisite process mediated and controlled by cells that culminate in the building of a highly hierarchical hybrid matrix. The building block of this matrix is the mineralized collagen fibrils, where hydroxyapatite (HAp) crystals are infiltrated in the fibril. The origin of these HAp crystals is regarded to the action of Matrix Vesicles (MV). These lipid vesicles are excreted by osteogenic cells and harbor the biochemical machinery required to produce and transport Ca2+ and PO43- ions for the HAp precipitation. Inside the MV, the lipid matrix act as a nucleation core to initiate the HAp crystallization. HAp crystals are them released to the extracellular medium and propagate the mineralization of collagen fibrils. Besides of large knowledge about the MV`s role as the start of bone mineralization and how interactions between collagen and HAp dictates the bone structure, little is known about the interface of MV and collagen mineralization. Our hypothesis is that after the MV`s rupture, lipid-mineral complexes are exposed to the extracellular medium and interact with collagen fibrils to propagate the intrafibrillar mineralization. To access this hypothesis, we will use state-of-art biophysical techniques to characterize the structure of complexes formed between lipids and calcium phosphates and we will explore the ability of these lipid-mineral complexes in propagating the collagen mineralization. We will use Langmuir monolayers composed of lipid found in the MV (phosphatidylserine, sphingomyelin, and cholesterol) to investigate the formation of lipid-mineral complexes. The composition and structure of the organic matrixes will be interconnected with the ability in induce mineral crystallization. We hope to identify with resolution at the molecular level the pathways induced by the lipid matrix in the HAp crystallization. The thermodynamic and kinetic behavior of lipid-induced HAp crystallization will be determined. Once we have identified the nature and ability of lipid-mineral complexes in inducing and control the HAp crystallization, we will evaluate the role of lipid-mineral complexes in propagating the collagen mineralization. For this, we will use collagen hierarchical matrixes mimetizing the structure found in the bone tissue. The creation of hierarchical collagen structures will be done in collaboration with Professor Seung-Wuk Lee (University of California at Berkeley, USA). Our aim with this project is to propose a new vision for bone mineralization, with applications emerging from clinics to the design of bio-inspired synthetic materials. (AU)

Scientific publications

(5)

(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, 19/08568-2, 19/25054-2)

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)

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)

CRUZ, MARCOS A. E.; FERREIRA, CLAUDIO R.; TOVANI, CAMILA B.; DE OLIVEIRA, FLAVIA A.; BOLEAN, MAYTE; CASELI, LUCIANO; MEBAREK, SAIDA; MILLAN, JOSE LUIS; BUCHET, RENE; BOTTINI, MASSIMO; et al. Phosphatidylserine controls calcium phosphate nucleation and growth on lipid monolayers: A physicochemical understanding of matrix vesicle-driven biomineralization. Journal of Structural Biology, v. 212, n. 2, NOV 1 2020. (14/24249-0, 17/08892-9, 17/20846-2)

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)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

CRUZ, Marcos Antonio Eufrásio. Studies on the role of matrix vesicles in bone mineralization: observations from micro to nanoscale. 2023. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PCARP/BC) Ribeirão Preto.