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

Floriana Cappiello, Bruno Casciaro, Satya Sree Kolar, Hasna Baidouri, Alison McDermott, Maria Luisa Mangoni

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

2 Citations (Scopus)

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.
Original languageEnglish
Title of host publicationAntimicrobial Peptides
EditorsPaul Hansen
Place of PublicationLondon
PublisherSpringer
Pages395-409
Volume1548
ISBN (Print)978-1-4939-6735-3
DOIs
Publication statusPublished - 25 Dec 2016

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