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JoVE Journal
Immunology and Infection
Establecer un modelo de córnea ex vivo porcina para estudiar tratamientos farmacológicos contra l...
Establecer un modelo de córnea ex vivo porcina para estudiar tratamientos farmacológicos contra l...
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Establishing a Porcine Ex Vivo Cornea Model for Studying Drug Treatments against Bacterial Keratitis

Establecer un modelo de córnea ex vivo porcina para estudiar tratamientos farmacológicos contra la queratitis bacteriana

Full Text
6,248 Views
09:22 min
May 12, 2020

DOI: 10.3791/61156-v

Katarzyna Okurowska1,2, Sanhita Roy3, Praveen Thokala4, Lynda Partridge1,5, Prashant Garg3, Sheila MacNeil1,6, Peter N. Monk1,7, Esther Karunakaran1,2

1Sheffield Collaboratorium for Antimicrobial Resistance and Biofilms (SCARAB),University of Sheffield, 2Department of Chemical and Biological Engineering,University of Sheffield, 3Hyderabad Eye Research Foundation,L V Prasad Eye Institute, 4Health Economics and Decision Science, School of Health and Related Research,University of Sheffield, 5Department of Molecular Biology and Biotechnology,University of Sheffield, 6Department of Materials Science and Engineering,University of Sheffield, 7Department of Infection, Immunity and Cardiovascular Disease,University of Sheffield

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Overview

This article presents a protocol for establishing an ex vivo porcine model of bacterial keratitis using Pseudomonas aeruginosa. This model effectively simulates in vivo infection, allowing for the assessment of bacterial proliferation and corneal tissue damage.

Key Study Components

Area of Science

  • Microbiology
  • Infectious Diseases
  • Ophthalmology

Background

  • Bacterial keratitis is a serious eye infection.
  • Pseudomonas aeruginosa is a common pathogen associated with this condition.
  • Ex vivo models are essential for studying infections and testing treatments.
  • This model provides a more accurate representation of in vivo conditions.

Purpose of Study

  • To develop a reliable ex vivo model for studying bacterial keratitis.
  • To evaluate antimicrobial efficacy in a controlled environment.
  • To facilitate research on novel antimicrobials.

Methods Used

  • Use of sterile forceps for transferring eyeballs.
  • Removal of conjunctiva and muscle tissue with a scalpel.
  • Transfer of prepared eyeballs to sterile PBS solution.
  • Subsequent treatment with povidone iodine in PBS.

Main Results

  • The model successfully mimics bacterial keratitis.
  • Demonstrated the ability to assess antimicrobial treatments effectively.
  • Validated the use of Pseudomonas aeruginosa and other organisms.
  • Provided a framework for future research in ocular infections.

Conclusions

  • The ex vivo porcine model is a valuable tool for studying bacterial keratitis.
  • It allows for accurate testing of antimicrobial efficacy.
  • This model can be adapted for various pathogens beyond Pseudomonas aeruginosa.

Frequently Asked Questions

What is the significance of using a porcine model?
Porcine models closely resemble human ocular anatomy and physiology, making them ideal for studying eye infections.
Can this model be used for other types of infections?
Yes, the model can also be adapted for studying infections caused by other bacteria, fungi, and yeast.
What are the advantages of using an ex vivo model?
Ex vivo models provide a controlled environment that closely mimics in vivo conditions, allowing for precise experimental manipulation.
How does this model help in antimicrobial research?
It allows researchers to evaluate the effectiveness of new antimicrobials before clinical trials.
What are the key steps in preparing the model?
Key steps include transferring the eyeball, removing surrounding tissues, and treating with sterile solutions.
Is Pseudomonas aeruginosa the only organism that can be used?
No, while Pseudomonas aeruginosa is used in this study, the model is compatible with other pathogens.

Este artículo describe un protocolo paso a paso para establecer un modelo ex vivo porcino de queratitis bacteriana. Pseudomonas aeruginosa se utiliza como organismo prototípico. Este modelo innovador imita la infección in vivo, ya que la proliferación bacteriana depende de la capacidad de la bacteria para dañar el tejido corneal.

El modelo de queratitis porcina ex vivo presentado aquí proporciona a los investigadores que desarrollan nuevos antimicrobianos un modelo in vitro representativo para determinar con mayor precisión la eficacia antimicrobiana en las etapas preclínicas. Hemos utilizado Pseudomonas aeruginosa para este experimento, pero el modelo también funciona bien con otras bacterias y organismos como hongos y levaduras. Use pinzas estériles para transferir el globo ocular a una placa de Petri.

Retire la conjuntiva y el tejido muscular alrededor del globo ocular con una hoja de bisturí número 15 y pinzas. Luego, mientras sostiene el nervio óptico con las pinzas, levante suavemente el globo ocular y transfiéralo a un frasco de medio litro lleno de PBS estéril. Una vez que todos los globos oculares estén limpios de tejido circundante, use pinzas estériles para moverlos a otro frasco de medio litro lleno de yodo povidona al 3% en PBS.

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