Use of tissue-engineered skin to study in vitro biofilm development

Carlos A. Charles, Carlos A. Ricotti, Stephen C Davis, Patricia M. Mertz, Robert Kirsner

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

BACKGROUND Biofilms are aggregations of microorganisms that have been identified as potential pathogens in the chronicity of nonhealing wounds. OBJECTIVE To develop an in vitro wound model to study biofilms using Graftskin, a tissue-engineered skin equivalent. MATERIALS AND METHODS Graftskin constructs were divided into sections, and wounds were created on each section. Bacterial suspensions with a concentration of 106 CFU/mL were prepared from cultures of pathogenic isolates of Pseudomonas aeruginosa and Staphylococcus aureus. A 25-μL aliquot of each suspension was deposited in the center of wounds created on the Graftskin. Sections were incubated at various time points, and a biopsy was then taken from the wounded and inoculated area. Sections were visualized with light (hematoxylin and eosin) and epifluorescent microscopy (calcofluor white and ethidium bromide). RESULTS Biofilm was observed on the wound model. Biofilm formation was dependent on time of Graftskin exposure to the bacteria. Biofilm was visualized in the S. aureus group at an earlier time point than in the P. aeruginosa group. CONCLUSIONS We demonstrated biofilm formation in vitro using a wound model. This model may provide a basis on which future studies may explore therapeutic modalities to prevent and eradicate pathogenic bacterial biofilm.

Original languageEnglish
Pages (from-to)1334-1341
Number of pages8
JournalDermatologic Surgery
Volume35
Issue number9
DOIs
StatePublished - Sep 1 2009

Fingerprint

Biofilms
Skin
Wounds and Injuries
Pseudomonas aeruginosa
Staphylococcus aureus
Suspensions
Ethidium
Hematoxylin
Eosine Yellowish-(YS)
In Vitro Techniques
Microscopy
Bacteria
Biopsy
Light

ASJC Scopus subject areas

  • Dermatology
  • Surgery

Cite this

Use of tissue-engineered skin to study in vitro biofilm development. / Charles, Carlos A.; Ricotti, Carlos A.; Davis, Stephen C; Mertz, Patricia M.; Kirsner, Robert.

In: Dermatologic Surgery, Vol. 35, No. 9, 01.09.2009, p. 1334-1341.

Research output: Contribution to journalArticle

Charles, Carlos A. ; Ricotti, Carlos A. ; Davis, Stephen C ; Mertz, Patricia M. ; Kirsner, Robert. / Use of tissue-engineered skin to study in vitro biofilm development. In: Dermatologic Surgery. 2009 ; Vol. 35, No. 9. pp. 1334-1341.
@article{bab0baf259de4ad9846a48715c469143,
title = "Use of tissue-engineered skin to study in vitro biofilm development",
abstract = "BACKGROUND Biofilms are aggregations of microorganisms that have been identified as potential pathogens in the chronicity of nonhealing wounds. OBJECTIVE To develop an in vitro wound model to study biofilms using Graftskin, a tissue-engineered skin equivalent. MATERIALS AND METHODS Graftskin constructs were divided into sections, and wounds were created on each section. Bacterial suspensions with a concentration of 106 CFU/mL were prepared from cultures of pathogenic isolates of Pseudomonas aeruginosa and Staphylococcus aureus. A 25-μL aliquot of each suspension was deposited in the center of wounds created on the Graftskin. Sections were incubated at various time points, and a biopsy was then taken from the wounded and inoculated area. Sections were visualized with light (hematoxylin and eosin) and epifluorescent microscopy (calcofluor white and ethidium bromide). RESULTS Biofilm was observed on the wound model. Biofilm formation was dependent on time of Graftskin exposure to the bacteria. Biofilm was visualized in the S. aureus group at an earlier time point than in the P. aeruginosa group. CONCLUSIONS We demonstrated biofilm formation in vitro using a wound model. This model may provide a basis on which future studies may explore therapeutic modalities to prevent and eradicate pathogenic bacterial biofilm.",
author = "Charles, {Carlos A.} and Ricotti, {Carlos A.} and Davis, {Stephen C} and Mertz, {Patricia M.} and Robert Kirsner",
year = "2009",
month = "9",
day = "1",
doi = "10.1111/j.1524-4725.2009.01238.x",
language = "English",
volume = "35",
pages = "1334--1341",
journal = "Dermatologic Surgery",
issn = "1076-0512",
publisher = "Wiley-Blackwell",
number = "9",

}

TY - JOUR

T1 - Use of tissue-engineered skin to study in vitro biofilm development

AU - Charles, Carlos A.

AU - Ricotti, Carlos A.

AU - Davis, Stephen C

AU - Mertz, Patricia M.

AU - Kirsner, Robert

PY - 2009/9/1

Y1 - 2009/9/1

N2 - BACKGROUND Biofilms are aggregations of microorganisms that have been identified as potential pathogens in the chronicity of nonhealing wounds. OBJECTIVE To develop an in vitro wound model to study biofilms using Graftskin, a tissue-engineered skin equivalent. MATERIALS AND METHODS Graftskin constructs were divided into sections, and wounds were created on each section. Bacterial suspensions with a concentration of 106 CFU/mL were prepared from cultures of pathogenic isolates of Pseudomonas aeruginosa and Staphylococcus aureus. A 25-μL aliquot of each suspension was deposited in the center of wounds created on the Graftskin. Sections were incubated at various time points, and a biopsy was then taken from the wounded and inoculated area. Sections were visualized with light (hematoxylin and eosin) and epifluorescent microscopy (calcofluor white and ethidium bromide). RESULTS Biofilm was observed on the wound model. Biofilm formation was dependent on time of Graftskin exposure to the bacteria. Biofilm was visualized in the S. aureus group at an earlier time point than in the P. aeruginosa group. CONCLUSIONS We demonstrated biofilm formation in vitro using a wound model. This model may provide a basis on which future studies may explore therapeutic modalities to prevent and eradicate pathogenic bacterial biofilm.

AB - BACKGROUND Biofilms are aggregations of microorganisms that have been identified as potential pathogens in the chronicity of nonhealing wounds. OBJECTIVE To develop an in vitro wound model to study biofilms using Graftskin, a tissue-engineered skin equivalent. MATERIALS AND METHODS Graftskin constructs were divided into sections, and wounds were created on each section. Bacterial suspensions with a concentration of 106 CFU/mL were prepared from cultures of pathogenic isolates of Pseudomonas aeruginosa and Staphylococcus aureus. A 25-μL aliquot of each suspension was deposited in the center of wounds created on the Graftskin. Sections were incubated at various time points, and a biopsy was then taken from the wounded and inoculated area. Sections were visualized with light (hematoxylin and eosin) and epifluorescent microscopy (calcofluor white and ethidium bromide). RESULTS Biofilm was observed on the wound model. Biofilm formation was dependent on time of Graftskin exposure to the bacteria. Biofilm was visualized in the S. aureus group at an earlier time point than in the P. aeruginosa group. CONCLUSIONS We demonstrated biofilm formation in vitro using a wound model. This model may provide a basis on which future studies may explore therapeutic modalities to prevent and eradicate pathogenic bacterial biofilm.

UR - http://www.scopus.com/inward/record.url?scp=69949154805&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=69949154805&partnerID=8YFLogxK

U2 - 10.1111/j.1524-4725.2009.01238.x

DO - 10.1111/j.1524-4725.2009.01238.x

M3 - Article

VL - 35

SP - 1334

EP - 1341

JO - Dermatologic Surgery

JF - Dermatologic Surgery

SN - 1076-0512

IS - 9

ER -