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

doi:10.1007/978-1-4939-6737-7_29

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

Applied Sciences Department

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

5 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 language	English
Title of host publication	Antimicrobial Peptides
Editors	Paul Hansen
Place of Publication	London
Publisher	Springer
Pages	395-409
Volume	1548
ISBN (Print)	978-1-4939-6735-3
DOIs	https://doi.org/10.1007/978-1-4939-6737-7_29
Publication status	Published - 25 Dec 2016

Keywords

Colony counts
Tear collection
Colorimetric MTT assay
Sandwich immunosorbent assay
LPS detoxification

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10.1007/978-1-4939-6737-7_29

Cite this

Cappiello, F., Casciaro, B., Kolar, S. S., Baidouri, H., McDermott, A., & Mangoni, M. L. (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 P. Hansen (Ed.), Antimicrobial Peptides (Vol. 1548, pp. 395-409). Springer. https://doi.org/10.1007/978-1-4939-6737-7_29

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

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.",

keywords = "Colony counts, Tear collection, Colorimetric MTT assay, Sandwich immunosorbent assay, LPS detoxification",

author = "Floriana Cappiello and Bruno Casciaro and Kolar, {Satya Sree} and Hasna Baidouri and Alison McDermott and Mangoni, {Maria Luisa}",

year = "2016",

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doi = "10.1007/978-1-4939-6737-7_29",

language = "English",

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volume = "1548",

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Cappiello, F, Casciaro, B, Kolar, SS, Baidouri, H, McDermott, A & Mangoni, ML 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 P Hansen (ed.), Antimicrobial Peptides. vol. 1548, Springer, London, pp. 395-409. https://doi.org/10.1007/978-1-4939-6737-7_29

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 et al.
Antimicrobial Peptides. ed. / Paul Hansen. Vol. 1548 London: Springer, 2016. p. 395-409.

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

TY - CHAP

T1 - Methods for In Vitro Analysis of Antimicrobial Activity and Toxicity of Anti-keratitis Peptides: Bacterial Viability in Tears, MTT, and TNF-α Release Assays

AU - Cappiello, Floriana

AU - Casciaro, Bruno

AU - Kolar, Satya Sree

AU - Baidouri, Hasna

AU - McDermott, Alison

AU - Mangoni, Maria Luisa

PY - 2016/12/25

Y1 - 2016/12/25

N2 - 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.

AB - 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.

KW - Colony counts

KW - Tear collection

KW - Colorimetric MTT assay

KW - Sandwich immunosorbent assay

KW - LPS detoxification

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DO - 10.1007/978-1-4939-6737-7_29

M3 - Chapter

SN - 978-1-4939-6735-3

VL - 1548

SP - 395

EP - 409

BT - Antimicrobial Peptides

A2 - Hansen, Paul

PB - Springer

CY - London

ER -

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

Abstract

Keywords

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