Citrate Stabilizes Hydroxylapatite Precursors: Implications for Bone Mineralization

Ruiz-Agudo, Encarnacion; Ruiz-Agudo, Cristina; Di Lorenzo, Fulvio; Alvarez-Lloret, Pedro; Ibañez-Velasco, Aurelia; Rodriguez-Navarro, Carlos (2021). Citrate Stabilizes Hydroxylapatite Precursors: Implications for Bone Mineralization. ACS biomaterials science & engineering, 7(6), pp. 2346-2357. American Chemical Society 10.1021/acsbiomaterials.1c00196

acsbiomaterials.1c00196.pdf - Published Version
Available under License Creative Commons: Attribution (CC-BY).
© 2021 American Chemical Society

Download (8MB) | Preview

Mineralization of hydroxylapatite (HAp), the main inorganic phase in bone, follows nonclassical crystallization routes involving metastable precursors and is strongly influenced by organic macromolecules. However, the effect of small organic molecules such as citrate on the formation of HAp is not well constrained. Using potentiometric titration experiments and titration calorimetry, in combination with a multianalytical approach, we show that citrate stabilizes prenucleation species as well as a liquid-like calcium phosphate precursor formed before any solid phase nucleates in the system. The stabilization of a liquid-like precursor phase could facilitate infiltration into the cavities of the collagen fibrils during bone mineralization, explaining the enhancement of collagen-mediated mineralization by citrate reported in previous studies. Hence, citrate can influence bone mineralization way before any solid phase (amorphous or crystalline) is formed. We also show that HAp formation after amorphous calcium phosphate (ACP) in the absence and presence of citrate results in nanoplates of about 5-12 nm thick, elongated along the c axis. Such nanoplates are made up of HAp nanocrystallites with a preferred c axis orientation and with interspersed ACP. The nanoplatelet morphology, size, and preferred crystallographic orientation, remarkably similar to those of bone HAp nanocrystals, appear to be an intrinsic feature of HAp formed from an amorphous precursor. Our results challenge current models for HAp mineralization in bone and the role of citrate, offering new clues to help answer the long-standing question as to why natural evolution favored HAp as the mineral phase in bone.

Item Type:

Journal Article (Original Article)


08 Faculty of Science > Institute of Geological Sciences

UniBE Contributor:

Di Lorenzo, Fulvio


500 Science > 550 Earth sciences & geology




American Chemical Society




Fulvio Di Lorenzo

Date Deposited:

11 Apr 2022 15:04

Last Modified:

11 Apr 2022 15:04

Publisher DOI:


PubMed ID:





Actions (login required)

Edit item Edit item
Provide Feedback