Quantitative atomic force microscopy provides new insight into matrix vesicle mineralization

Author(s):Show less -

Plaut, Justin S. ^{[1, 2]} ; Strzelecka-Kiliszek, Agnieszka ^[3] ; Bozycki, Lukasz ^[3] ; Pikula, Slawomir ^[3] ; Buchet, Rene ^[4] ; Mebarek, Saida ^[4] ; Chadli, Meriem ^[4] ; Bolean, Mayte ^[5] ; Simao, Ana M. S. ^[5] ; Ciancaglini, Pietro ^[5] ; Magrini, Andrea ^{[6, 7]} ; Rosato, Nicola ^{[6, 8]} ; Magne, David ^[5] ; Girard-Egrot, Agnes ^[5] ; Farquharson, Colin ^{[9, 10]} ; Esener, Sadik C. ^{[1, 2]} ; Milian, Jose L. ^[11] ; Bottini, Massimo ^{[11, 6, 8]}

Affiliation:Show less -

^[1] Univ Calif San Diego, Dept Bioengn, La Jolla, CA 92093 - USA

^[2] Oregon Hlth & Sci Univ, Canc Early Detect Adv Res Ctr, Knight Canc Inst, Portland, OR 97201 - USA

^[3] Polish Acad Sci, Nencki Inst Expt Biol, Lab Biochem Lipids, PL-02093 Warsaw - Poland

^[4] Univ Lyon 1, Univ Lyon, Inst Chim & Biochim Mol & Supramol, CNRS, UMR 5246, F-69622 Villeurbanne - France

^[5] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Quim, BR-14040901 Ribeirao Preto - Brazil

^[6] Univ Roma Tor Vergata, Nanosci & Nanotechnol & Innovat Instrumentat NAST, Rome - Italy

^[7] Univ Roma Tor Vergata, Dept Biopathol & Imaging Diagnost, Rome - Italy

^[8] Univ Roma Tor Vergata, Dept Expt Med, Via Montpellier 1, I-00133 Rome - Italy

^[9] Univ Edinburgh, Royal Dick Sch Vet Studies, Edinburgh EH25 9RG, Midlothian - Scotland

^[10] Univ Edinburgh, Div Dev Biol, Roslin Inst, Edinburgh EH25 9RG, Midlothian - Scotland

^[11] Sanford Burnham Prebys Med Discovery Inst, La Jolla, CA 92037 - USA

• Document type: Journal article
• Source: Archives of Biochemistry and Biophysics; v. 667, p. 14-21, MAY 30 2019.
• Web of Science Citations: 1

Abstract

Matrix vesicles (MVs) are a class of extracellular vesicles that initiate mineralization in cartilage, bone, and other vertebrate tissues by accumulating calcium ions (Ca2+) and inorganic phosphate (P-i) within their lumen and forming a nucleation core (NC). After further sequestration of Ca2+ and P-i, the NC transforms into crystalline complexes. Direct evidence of the existence of the NC and its maturation have been provided solely by analyses of dried samples. We isolated MVs from chicken embryo cartilage and used atomic force microscopy peak force quantitative nanomechanical property mapping (AFM-PFQNM) to measure the nanomechanical and morphological properties of individual MVs under both mineralizing (+Ca2+) and non-mineralizing (-Ca2+) fluid conditions. The elastic modulus of MVs significantly increased by 4-fold after incubation in mineralization buffer. From AFM mapping data, we inferred the morphological changes of MVs as mineralization progresses: prior to mineralization, a punctate feature, the NC, is present within MVs and this feature grows and stiffens during mineralization until it occupies most of the MV lumen. Dynamic light scattering showed a significant increase in hydrodynamic diameter and no change in the zeta potential of hydrated MVs after incubation with Ca2+. This validates that crystalline complexes, which are strongly negative relative to MVs, were forming within the lumen of MVs. These data were substantiated by transmission electron microscopy energy dispersive X-ray and Fourier transform infrared spectroscopic analyses of dried MVs, which provide evidence that the complexes increased in size, crystallinity, and Ca/P ratio within MVs during the mineralization process. (AU)

• FAPESP's process: 14/11941-3 - Are collagen and Annexin V responsible for the control in the biomineralization process?
• Grantee: Pietro Ciancaglini
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/21236-0 - Extracellular matrix vesicles (MVs) mimetic systems to study the regulation of the biomineralization process: proteoliposomes containing NPP1 and Annexin V
• Grantee: Pietro Ciancaglini
• Support Opportunities: Regular Research Grants