Methods for In Vitro Analysis of Antimicrobial Activity and Toxicity of Anti-keratitis Peptides: Bacterial Viability in Tears, MTT, and TNF-α Release Assays

Cappiello, Floriana, Casciaro, Bruno, Kolar, Satya Sree, Baidouri, Hasna, McDermott, Alison and Mangoni, Maria Luisa (2016) Methods for In Vitro Analysis of Antimicrobial Activity and Toxicity of Anti-keratitis Peptides: Bacterial Viability in Tears, MTT, and TNF-α Release Assays. In: Antimicrobial Peptides. Springer, London, pp. 395-409. ISBN 978-1-4939-6735-3

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Abstract

Ease of access to the cornea makes antimicrobial peptides (AMPs) ideal candidates for topical drug application. However, before bringing them to the clinic, it is fundamental to evaluate in vitro: (1) the ability of AMPs to kill bacteria in the presence of human tears, by counting the number of surviving bacteria on agar plates; (2) the potential cytotoxicity of AMPs to mammalian cells by a colorimetric method based on the production of a colored formazan crystals by metabolically active cells; and (3) the ability of AMPs to neutralize the toxic effect of the bacterial cell wall component, lipopolysaccharide (LPS), by measuring the level of the pro-inflammatory cytokine, TNF-α, released from LPS-activated macrophages, using a sandwich enzyme-linked immunosorbent assay.

Item Type:	Book Section
Uncontrolled Keywords:	Colony counts, Tear collection, Colorimetric MTT assay, Sandwich immunosorbent assay, LPS detoxification
Subjects:	B500 Ophthalmics
Department:	Faculties > Health and Life Sciences > Applied Sciences
Depositing User:	Becky Skoyles
Date Deposited:	19 May 2017 14:43
Last Modified:	12 Oct 2019 18:31
URI:	http://nrl.northumbria.ac.uk/id/eprint/30774

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