The University of Southampton
University of Southampton Institutional Repository

Live-imaging of bioengineered cartilage tissue using multimodal non-linear molecular imaging

Live-imaging of bioengineered cartilage tissue using multimodal non-linear molecular imaging
Live-imaging of bioengineered cartilage tissue using multimodal non-linear molecular imaging
Coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) are non-linear techniques that allow label-free, non-destructive and non-invasive imaging for cellular and tissue analysis. Although live-imaging studies have been performed previously, concerns that they do not cause any changes at the molecular level in sensitive biological samples have not been addressed. This is important especially for stem cell differentiation and tissue engineering, if CARS/SHG microscopy is to be used as a non-invasive, label-free tool for assessment of the developing neo-tissue. In this work, we monitored the differentiation of human fetal-femur derived skeletal cells into cartilage in three-dimensional cultures using CARS and SHG microscopy and demonstrate the live imaging of the same developing neo-tissue over time. Our work conclusively establishes that non-linear label-free imaging does not alter the phenotype or the gene expression at the different stages of differentiation and has no adverse effect on human skeletal cell growth and behaviour. Additionally, we show that CARS microscopy allows imaging of different molecules of interest, including lipids, proteins and glycosaminoglycans, in the bioengineered neo-cartilage. These studies demonstrate the label-free and truly non-invasive nature of live CARS and SHG imaging and their value and translation potential in skeletal research, regeneration medicine and tissue engineering.
2045-2322
1-9
Moura, Catarina
fdd85e38-f46f-47d0-a33a-6e7328398e90
Bourdakos, Konstantinos
83f6fc3a-db12-476b-9a78-4aad8756f82f
Tare, Rahul
587c9db4-e409-4e7c-a02a-677547ab724a
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9
Moura, Catarina
fdd85e38-f46f-47d0-a33a-6e7328398e90
Bourdakos, Konstantinos
83f6fc3a-db12-476b-9a78-4aad8756f82f
Tare, Rahul
587c9db4-e409-4e7c-a02a-677547ab724a
Oreffo, Richard
ff9fff72-6855-4d0f-bfb2-311d0e8f3778
Mahajan, Sumeet
b131f40a-479e-4432-b662-19d60d4069e9

Moura, Catarina, Bourdakos, Konstantinos, Tare, Rahul, Oreffo, Richard and Mahajan, Sumeet (2019) Live-imaging of bioengineered cartilage tissue using multimodal non-linear molecular imaging. Scientific Reports, 9 (1), 1-9, [5561]. (doi:10.1038/s41598-019-41466-w).

Record type: Article

Abstract

Coherent anti-Stokes Raman scattering (CARS) and second harmonic generation (SHG) are non-linear techniques that allow label-free, non-destructive and non-invasive imaging for cellular and tissue analysis. Although live-imaging studies have been performed previously, concerns that they do not cause any changes at the molecular level in sensitive biological samples have not been addressed. This is important especially for stem cell differentiation and tissue engineering, if CARS/SHG microscopy is to be used as a non-invasive, label-free tool for assessment of the developing neo-tissue. In this work, we monitored the differentiation of human fetal-femur derived skeletal cells into cartilage in three-dimensional cultures using CARS and SHG microscopy and demonstrate the live imaging of the same developing neo-tissue over time. Our work conclusively establishes that non-linear label-free imaging does not alter the phenotype or the gene expression at the different stages of differentiation and has no adverse effect on human skeletal cell growth and behaviour. Additionally, we show that CARS microscopy allows imaging of different molecules of interest, including lipids, proteins and glycosaminoglycans, in the bioengineered neo-cartilage. These studies demonstrate the label-free and truly non-invasive nature of live CARS and SHG imaging and their value and translation potential in skeletal research, regeneration medicine and tissue engineering.

Text
Sci Reports_revised 2-non-highlighted - Accepted Manuscript
Download (5MB)
Text
251218_1 supplement - Accepted Manuscript
Download (527kB)
Text
s41598-019-41466-w - Version of Record
Available under License Creative Commons Attribution.
Download (2MB)

More information

Accepted/In Press date: 4 March 2019
e-pub ahead of print date: 3 April 2019
Published date: 2019

Identifiers

Local EPrints ID: 428678
URI: http://eprints.soton.ac.uk/id/eprint/428678
ISSN: 2045-2322
PURE UUID: 2015c96d-b897-4b03-9624-52d2f8467426
ORCID for Rahul Tare: ORCID iD orcid.org/0000-0001-8274-8837
ORCID for Richard Oreffo: ORCID iD orcid.org/0000-0001-5995-6726
ORCID for Sumeet Mahajan: ORCID iD orcid.org/0000-0001-8923-6666

Catalogue record

Date deposited: 06 Mar 2019 17:30
Last modified: 18 Feb 2021 17:09

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.

×