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A bioengineered three-dimensional cell culture platform integrated with microfluidics to address antimicrobial resistance in tuberculosis

A bioengineered three-dimensional cell culture platform integrated with microfluidics to address antimicrobial resistance in tuberculosis
A bioengineered three-dimensional cell culture platform integrated with microfluidics to address antimicrobial resistance in tuberculosis
Antimicrobial resistance presents one of the most significant threats to human health, with the emergence of totally drug-resistant organisms. We have combined bioengineering, genetically modified bacteria, longitudinal readouts, and fluidics to develop a transformative platform to address the drug development bottleneck, utilizing Mycobacterium tuberculosis as the model organism. We generated microspheres incorporating virulent reporter bacilli, primary human cells, and an extracellular matrix by using bioelectrospray methodology. Granulomas form within the three-dimensional matrix, and mycobacterial stress genes are upregulated. Pyrazinamide, a vital first-line antibiotic for treating human tuberculosis, kills M. tuberculosis in a three-dimensional culture but not in a standard two-dimensional culture or Middlebrook 7H9 broth, demonstrating that antibiotic sensitivity within microspheres reflects conditions in patients. We then performed pharmacokinetic modeling by combining the microsphere system with a microfluidic plate and demonstrated that we can model the effect of dynamic antibiotic concentrations on mycobacterial killing. The microsphere system is highly tractable, permitting variation of cell content, the extracellular matrix, sphere size, the infectious dose, and the surrounding medium with the potential to address a wide array of human infections and the threat of antimicrobial resistance.
2150-7511
1-14
Bielecka, M.K.
90391ea3-aa1f-4104-a893-568c138718a2
Tezera, L.B.
c5598dbf-23a8-4934-96a4-7c783bf9e776
Zmijan, R.
6461a3ba-03b6-43c0-89dd-8e221457b45d
Drobniewski, F.
1b3af90f-5f68-4eb5-bf1f-cc5b89b8f7f5
Zhang, Xunli
d7cf1181-3276-4da1-9150-e212b333abb1
Jayasinghe, S.
7f141099-bb9a-4862-8cfa-7b5dd85d798d
Elkington, P.
60828c7c-3d32-47c9-9fcc-6c4c54c35a15
Bielecka, M.K.
90391ea3-aa1f-4104-a893-568c138718a2
Tezera, L.B.
c5598dbf-23a8-4934-96a4-7c783bf9e776
Zmijan, R.
6461a3ba-03b6-43c0-89dd-8e221457b45d
Drobniewski, F.
1b3af90f-5f68-4eb5-bf1f-cc5b89b8f7f5
Zhang, Xunli
d7cf1181-3276-4da1-9150-e212b333abb1
Jayasinghe, S.
7f141099-bb9a-4862-8cfa-7b5dd85d798d
Elkington, P.
60828c7c-3d32-47c9-9fcc-6c4c54c35a15

Bielecka, M.K., Tezera, L.B., Zmijan, R., Drobniewski, F., Zhang, Xunli, Jayasinghe, S. and Elkington, P. (2017) A bioengineered three-dimensional cell culture platform integrated with microfluidics to address antimicrobial resistance in tuberculosis. mBio, 8 (1), 1-14. (doi:10.1128/mBio.02073-16).

Record type: Article

Abstract

Antimicrobial resistance presents one of the most significant threats to human health, with the emergence of totally drug-resistant organisms. We have combined bioengineering, genetically modified bacteria, longitudinal readouts, and fluidics to develop a transformative platform to address the drug development bottleneck, utilizing Mycobacterium tuberculosis as the model organism. We generated microspheres incorporating virulent reporter bacilli, primary human cells, and an extracellular matrix by using bioelectrospray methodology. Granulomas form within the three-dimensional matrix, and mycobacterial stress genes are upregulated. Pyrazinamide, a vital first-line antibiotic for treating human tuberculosis, kills M. tuberculosis in a three-dimensional culture but not in a standard two-dimensional culture or Middlebrook 7H9 broth, demonstrating that antibiotic sensitivity within microspheres reflects conditions in patients. We then performed pharmacokinetic modeling by combining the microsphere system with a microfluidic plate and demonstrated that we can model the effect of dynamic antibiotic concentrations on mycobacterial killing. The microsphere system is highly tractable, permitting variation of cell content, the extracellular matrix, sphere size, the infectious dose, and the surrounding medium with the potential to address a wide array of human infections and the threat of antimicrobial resistance.

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More information

Accepted/In Press date: 12 January 2017
e-pub ahead of print date: 7 February 2017
Published date: 7 February 2017
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 404834
URI: https://eprints.soton.ac.uk/id/eprint/404834
ISSN: 2150-7511
PURE UUID: 8112503e-7926-4fd6-a786-6335c52102b4
ORCID for L.B. Tezera: ORCID iD orcid.org/0000-0002-7898-6709
ORCID for P. Elkington: ORCID iD orcid.org/0000-0003-0390-0613

Catalogue record

Date deposited: 24 Jan 2017 10:01
Last modified: 06 Jun 2018 12:28

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