The University of Southampton
University of Southampton Institutional Repository

Nanoanalytical electron microscopy reveals a sequential mineralization process involving carbonate-containing amorphous precursors

Nanoanalytical electron microscopy reveals a sequential mineralization process involving carbonate-containing amorphous precursors
Nanoanalytical electron microscopy reveals a sequential mineralization process involving carbonate-containing amorphous precursors
A direct observation and an in-depth characterization of the steps by which bone mineral nucleates and grows in the extracellular matrix during the earliest stages of maturation, using relevant biomineralization models as they grow into mature bone mineral, is an important research goal. To better understand the process of bone mineralization in the extracellular matrix, we used nano-analytical electron microscopy techniques to examine an in vitro model of bone formation. This study demonstrates the presence of three dominant CaP structures in the mineralizing osteoblast cultures: <80 nm dense granules with a low calcium to phosphate ratio (Ca/P) and crystalline domains; calcium phosphate needles emanating from a foci: "needle-like globules" (100-300 nm in diameter); and mature mineral, both with statistically higher Ca/P compared to that of the dense granules. Many of the submicron granules and globules were interspersed around fibrillar structures containing nitrogen, which are most likely the signature of the organic phase. With high spatial resolution electron energy loss spectroscopy (EELS) mapping, spatially resolved maps were acquired showing the distribution of carbonate within each mineral structure. The carbonate was located in the middle of the granules, which suggested the nucleation of the younger mineral starts with a carbonate-containing precursor and that this precursor may act as seed for growth into larger, submicron-sized, needle-like globules of hydroxyapatite with a different stoichiometry. Application of analytical electron microscopy has important implications in deciphering both how normal bone forms and in understanding pathological mineralization.
1936-0851
6826-6835
Nitiputri, Kharissa
d0fb2935-1fc4-4853-a6eb-b9266d1e9508
Ramasse, Quentin M.
30aed351-dfed-4eea-9c87-375b75e24b79
Autefage, Helene
5be3e99a-6cc2-4135-a1fe-a2cc488da694
McGilvery, Catriona M.
d9fcad51-d2ae-463d-9876-3f4e73423f95
Boonrungsiman, Suwimon
265be035-5f1c-478d-af1b-19a01a9bc069
Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Stevens, Molly M.
f0c6dded-e31c-4e40-bfc4-2a70fad118a6
Porter, Alexandra E.
04291b8a-4af2-4132-b847-20bd1d1f48fe
Nitiputri, Kharissa
d0fb2935-1fc4-4853-a6eb-b9266d1e9508
Ramasse, Quentin M.
30aed351-dfed-4eea-9c87-375b75e24b79
Autefage, Helene
5be3e99a-6cc2-4135-a1fe-a2cc488da694
McGilvery, Catriona M.
d9fcad51-d2ae-463d-9876-3f4e73423f95
Boonrungsiman, Suwimon
265be035-5f1c-478d-af1b-19a01a9bc069
Evans, Nicholas D.
06a05c97-bfed-4abb-9244-34ec9f4b4b95
Stevens, Molly M.
f0c6dded-e31c-4e40-bfc4-2a70fad118a6
Porter, Alexandra E.
04291b8a-4af2-4132-b847-20bd1d1f48fe

Nitiputri, Kharissa, Ramasse, Quentin M., Autefage, Helene, McGilvery, Catriona M., Boonrungsiman, Suwimon, Evans, Nicholas D., Stevens, Molly M. and Porter, Alexandra E. (2016) Nanoanalytical electron microscopy reveals a sequential mineralization process involving carbonate-containing amorphous precursors. ACS Nano, 10 (7), 6826-6835. (doi:10.1021/acsnano.6b02443). (PMID:27383526)

Record type: Article

Abstract

A direct observation and an in-depth characterization of the steps by which bone mineral nucleates and grows in the extracellular matrix during the earliest stages of maturation, using relevant biomineralization models as they grow into mature bone mineral, is an important research goal. To better understand the process of bone mineralization in the extracellular matrix, we used nano-analytical electron microscopy techniques to examine an in vitro model of bone formation. This study demonstrates the presence of three dominant CaP structures in the mineralizing osteoblast cultures: <80 nm dense granules with a low calcium to phosphate ratio (Ca/P) and crystalline domains; calcium phosphate needles emanating from a foci: "needle-like globules" (100-300 nm in diameter); and mature mineral, both with statistically higher Ca/P compared to that of the dense granules. Many of the submicron granules and globules were interspersed around fibrillar structures containing nitrogen, which are most likely the signature of the organic phase. With high spatial resolution electron energy loss spectroscopy (EELS) mapping, spatially resolved maps were acquired showing the distribution of carbonate within each mineral structure. The carbonate was located in the middle of the granules, which suggested the nucleation of the younger mineral starts with a carbonate-containing precursor and that this precursor may act as seed for growth into larger, submicron-sized, needle-like globules of hydroxyapatite with a different stoichiometry. Application of analytical electron microscopy has important implications in deciphering both how normal bone forms and in understanding pathological mineralization.

Text
Nitiputri_Nanoanalytical Electron Microscopy Reveals A Sequential Mineralization Process Involving Carbonate-Containing Amorphous Precursors_2016.pdf - Accepted Manuscript
Download (1MB)

More information

Accepted/In Press date: 6 June 2016
e-pub ahead of print date: 6 July 2016
Published date: 26 July 2016
Organisations: Centre for Human Development, Stem Cells and Regeneration, Institute of Developmental Sciences, Bioengineering Group, Human Development & Health

Identifiers

Local EPrints ID: 397987
URI: http://eprints.soton.ac.uk/id/eprint/397987
ISSN: 1936-0851
PURE UUID: 8e5e74cf-2d3b-4cab-8e9e-21fd3007295e

Catalogue record

Date deposited: 14 Jul 2016 08:17
Last modified: 07 Oct 2020 05:12

Export record

Altmetrics

Download statistics

Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.

View more statistics

Atom RSS 1.0 RSS 2.0

Contact ePrints Soton: eprints@soton.ac.uk

ePrints Soton supports OAI 2.0 with a base URL of http://eprints.soton.ac.uk/cgi/oai2

This repository has been built using EPrints software, developed at the University of Southampton, but available to everyone to use.

We use cookies to ensure that we give you the best experience on our website. If you continue without changing your settings, we will assume that you are happy to receive cookies on the University of Southampton website.

×