Mineralization Profile of Annexin A6-Harbouring Proteoliposomes: Shedding Light on the Role of Annexin A6 on Matrix Vesicle-Mediated Mineralization

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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 ; Millan, Jose Luis ; Ramos, Ana Paula ; Buchet, Rene ; Bottini, Massimo ; Ciancaglini, Pietro

• Tipo de documento: Artigo Científico
• Fonte: INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES; v. 23, n. 16, p. 15-pg., 2022-08-01.

Resumo

The biochemical machinery involved in matrix vesicles-mediated bone mineralization involves a specific set of lipids, enzymes, and proteins. Annexins, among their many functions, have been described as responsible for the formation and stabilization of the matrix vesicles ' nucleational core. However, the specific role of each member of the annexin family, especially in the presence of type-I collagen, remains to be clarified. To address this issue, in vitro mineralization was carried out using AnxA6 (in solution or associated to the proteoliposomes) in the presence or in the absence of type-I collagen, incubated with either amorphous calcium phosphate (ACP) or a phosphatidylserine-calcium phosphate complex (PS-CPLX) as nucleators. Proteoliposomes were composed of 1,2-dipalmitoylphosphatidylcholine (DPPC), 1,2-dipalmitoylphosphatidylcholine: 1,2-dipalmitoylphosphatidylserine (DPPC:DPPS), and DPPC:Cholesterol:DPPS to mimic the outer and the inner leaflet of the matrix vesicles membrane as well as to investigate the effect of the membrane fluidity. Kinetic parameters of mineralization were calculated from time-dependent turbidity curves of free Annexin A6 (AnxA6) and AnxA6-containing proteoliposomes dispersed in synthetic cartilage lymph. The chemical composition of the minerals formed was investigated by Fourier transform infrared spectroscopy (FTIR). Free AnxA6 and AnxA6-proteoliposomes in the presence of ACP were not able to propagate mineralization; however, poorly crystalline calcium phosphates were formed in the presence of PS-CPLX, supporting the role of annexin-calcium-phosphatidylserine complex in the formation and stabilization of the matrix vesicles' nucleational core. We found that AnxA6 lacks nucleation propagation capacity when incorporated into liposomes in the presence of PS-CPLX and type-I collagen. This suggests that AnxA6 may interact either with phospholipids, forming a nucleational core, or with type-I collagen, albeit less efficiently, to induce the nucleation process. (AU)

• Processo FAPESP: 19/25054-2 - Nanopartículas contendo estrôncio e sua versatilidade na fabricação de biomateriais: implicações e aplicações em biomineralização
• Beneficiário: Ana Paula Ramos
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 21/13140-1 - Estudo da interação de PHOSPHO1 e nSMase2 com membranas modelo: uma possível correlação na secreção de vesículas da matriz
• Beneficiário: Luiz Henrique da Silva Andrilli
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 17/20846-2 - Explorando o papel da matriz lipídico-proteica-mineral na biomineralização óssea: uma abordagem biofísica empregando-se filmes automontados
• Beneficiário: Marcos Antonio Eufrásio Cruz
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 19/08568-2 - Investigação do papel de vesículas extracelulares (VEs) na iniciação, propagação, regeneração e modelação da mineralização biológica
• Beneficiário: Pietro Ciancaglini
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 22/05026-7 - Treinamento no isolamento de vesículas extracelulares (VEs) e MVs modelação da mineralização biológica
• Beneficiário: Heitor Gobbi Sebinelli
• Modalidade de apoio: Bolsas no Brasil - Programa Capacitação - Treinamento Técnico