A novel rat contact lens model for Fusarium keratitis

Mohamed Abou Shousha, Andrea Rachelle C Santos, Rafael A. Oechsler, Alfonso Iovieno, Jorge Maestre-Mesa, Marco Ruggeri, Jose J. Echegaray, Sander Dubovy, Victor L Perez Quinones, Darlene Miller, Eduardo C Alfonso, M. Livia Bajenaru

Research output: Contribution to journalArticle

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Abstract

Purpose: The aim of this study was to develop and characterize a new contact lens-associated fungal keratitis rat model and to assess the ability of non-invasive spectral-domain optical coherence tomography (SD-OCT) to detect pathological changes in vivo in fungal keratitis. Methods: We used SD-OCT to image and measure the cornea of Sprague Dawley rats. Fusarium infection was initiated in the rat eye by fitting Fusarium solani-soaked contact lenses on the experimental eye, while the control animals received contact lenses soaked in sterile saline. The fungal infection was monitored with periodic slit-lamp examination and in vivo SD-OCT imaging of the rat eye, and confirmed by histology, counting of viable fungi in the infected rat cornea, and PCR with specific primers for Fusarium sp. Results: We imaged and measured the rat cornea with SD-OCT. Custom-made contact lenses were developed based on the OCT measurements. Incubation of contact lenses in a F. solani suspension resulted in biofilm formation. We induced contact lens-associated Fusarium keratitis by fitting the rat eyes for 4 h with the Fusarium-contaminated contact lenses. The SD-OCT images of the cornea correlated well with the slit-lamp and histopathological results and clearly defined clinical signs of infection, namely, increased corneal thickening, loss of epithelial continuity, hyper-reflective areas representing infiltrates, and endothelial plaques characteristic of fungal infection. Moreover, in three cases, SD-OCT detected the infection without any clear findings on slit-lamp examination. Infection was confirmed with histological fungal staining, PCR, and microbiological culture positivity. Conclusions: We developed a highly reproducible rat contact lens model and successfully induced contact lens-associated Fusarium keratitis in this model. The clinical presentation of contact lens-associated Fusarium keratitis in the rat model is similar to the human condition. SD-OCT is a valuable tool that non-invasively revealed characteristic signs of the fungal infection and could provide sensitive, objective monitoring in fungal keratitis.

Original languageEnglish
Pages (from-to)2596-2605
Number of pages10
JournalMolecular Vision
Volume19
StatePublished - Dec 27 2013

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Keratitis
Contact Lenses
Fusarium
Optical Coherence Tomography
Cornea
Mycoses
Infection
Fusariosis
Polymerase Chain Reaction
Biofilms
Sprague Dawley Rats
Suspensions
Histology
Fungi
Staining and Labeling

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Abou Shousha, M., Santos, A. R. C., Oechsler, R. A., Iovieno, A., Maestre-Mesa, J., Ruggeri, M., ... Livia Bajenaru, M. (2013). A novel rat contact lens model for Fusarium keratitis. Molecular Vision, 19, 2596-2605.

A novel rat contact lens model for Fusarium keratitis. / Abou Shousha, Mohamed; Santos, Andrea Rachelle C; Oechsler, Rafael A.; Iovieno, Alfonso; Maestre-Mesa, Jorge; Ruggeri, Marco; Echegaray, Jose J.; Dubovy, Sander; Perez Quinones, Victor L; Miller, Darlene; Alfonso, Eduardo C; Livia Bajenaru, M.

In: Molecular Vision, Vol. 19, 27.12.2013, p. 2596-2605.

Research output: Contribution to journalArticle

Abou Shousha, M, Santos, ARC, Oechsler, RA, Iovieno, A, Maestre-Mesa, J, Ruggeri, M, Echegaray, JJ, Dubovy, S, Perez Quinones, VL, Miller, D, Alfonso, EC & Livia Bajenaru, M 2013, 'A novel rat contact lens model for Fusarium keratitis', Molecular Vision, vol. 19, pp. 2596-2605.
Abou Shousha M, Santos ARC, Oechsler RA, Iovieno A, Maestre-Mesa J, Ruggeri M et al. A novel rat contact lens model for Fusarium keratitis. Molecular Vision. 2013 Dec 27;19:2596-2605.
Abou Shousha, Mohamed ; Santos, Andrea Rachelle C ; Oechsler, Rafael A. ; Iovieno, Alfonso ; Maestre-Mesa, Jorge ; Ruggeri, Marco ; Echegaray, Jose J. ; Dubovy, Sander ; Perez Quinones, Victor L ; Miller, Darlene ; Alfonso, Eduardo C ; Livia Bajenaru, M. / A novel rat contact lens model for Fusarium keratitis. In: Molecular Vision. 2013 ; Vol. 19. pp. 2596-2605.
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abstract = "Purpose: The aim of this study was to develop and characterize a new contact lens-associated fungal keratitis rat model and to assess the ability of non-invasive spectral-domain optical coherence tomography (SD-OCT) to detect pathological changes in vivo in fungal keratitis. Methods: We used SD-OCT to image and measure the cornea of Sprague Dawley rats. Fusarium infection was initiated in the rat eye by fitting Fusarium solani-soaked contact lenses on the experimental eye, while the control animals received contact lenses soaked in sterile saline. The fungal infection was monitored with periodic slit-lamp examination and in vivo SD-OCT imaging of the rat eye, and confirmed by histology, counting of viable fungi in the infected rat cornea, and PCR with specific primers for Fusarium sp. Results: We imaged and measured the rat cornea with SD-OCT. Custom-made contact lenses were developed based on the OCT measurements. Incubation of contact lenses in a F. solani suspension resulted in biofilm formation. We induced contact lens-associated Fusarium keratitis by fitting the rat eyes for 4 h with the Fusarium-contaminated contact lenses. The SD-OCT images of the cornea correlated well with the slit-lamp and histopathological results and clearly defined clinical signs of infection, namely, increased corneal thickening, loss of epithelial continuity, hyper-reflective areas representing infiltrates, and endothelial plaques characteristic of fungal infection. Moreover, in three cases, SD-OCT detected the infection without any clear findings on slit-lamp examination. Infection was confirmed with histological fungal staining, PCR, and microbiological culture positivity. Conclusions: We developed a highly reproducible rat contact lens model and successfully induced contact lens-associated Fusarium keratitis in this model. The clinical presentation of contact lens-associated Fusarium keratitis in the rat model is similar to the human condition. SD-OCT is a valuable tool that non-invasively revealed characteristic signs of the fungal infection and could provide sensitive, objective monitoring in fungal keratitis.",
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AU - Abou Shousha, Mohamed

AU - Santos, Andrea Rachelle C

AU - Oechsler, Rafael A.

AU - Iovieno, Alfonso

AU - Maestre-Mesa, Jorge

AU - Ruggeri, Marco

AU - Echegaray, Jose J.

AU - Dubovy, Sander

AU - Perez Quinones, Victor L

AU - Miller, Darlene

AU - Alfonso, Eduardo C

AU - Livia Bajenaru, M.

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N2 - Purpose: The aim of this study was to develop and characterize a new contact lens-associated fungal keratitis rat model and to assess the ability of non-invasive spectral-domain optical coherence tomography (SD-OCT) to detect pathological changes in vivo in fungal keratitis. Methods: We used SD-OCT to image and measure the cornea of Sprague Dawley rats. Fusarium infection was initiated in the rat eye by fitting Fusarium solani-soaked contact lenses on the experimental eye, while the control animals received contact lenses soaked in sterile saline. The fungal infection was monitored with periodic slit-lamp examination and in vivo SD-OCT imaging of the rat eye, and confirmed by histology, counting of viable fungi in the infected rat cornea, and PCR with specific primers for Fusarium sp. Results: We imaged and measured the rat cornea with SD-OCT. Custom-made contact lenses were developed based on the OCT measurements. Incubation of contact lenses in a F. solani suspension resulted in biofilm formation. We induced contact lens-associated Fusarium keratitis by fitting the rat eyes for 4 h with the Fusarium-contaminated contact lenses. The SD-OCT images of the cornea correlated well with the slit-lamp and histopathological results and clearly defined clinical signs of infection, namely, increased corneal thickening, loss of epithelial continuity, hyper-reflective areas representing infiltrates, and endothelial plaques characteristic of fungal infection. Moreover, in three cases, SD-OCT detected the infection without any clear findings on slit-lamp examination. Infection was confirmed with histological fungal staining, PCR, and microbiological culture positivity. Conclusions: We developed a highly reproducible rat contact lens model and successfully induced contact lens-associated Fusarium keratitis in this model. The clinical presentation of contact lens-associated Fusarium keratitis in the rat model is similar to the human condition. SD-OCT is a valuable tool that non-invasively revealed characteristic signs of the fungal infection and could provide sensitive, objective monitoring in fungal keratitis.

AB - Purpose: The aim of this study was to develop and characterize a new contact lens-associated fungal keratitis rat model and to assess the ability of non-invasive spectral-domain optical coherence tomography (SD-OCT) to detect pathological changes in vivo in fungal keratitis. Methods: We used SD-OCT to image and measure the cornea of Sprague Dawley rats. Fusarium infection was initiated in the rat eye by fitting Fusarium solani-soaked contact lenses on the experimental eye, while the control animals received contact lenses soaked in sterile saline. The fungal infection was monitored with periodic slit-lamp examination and in vivo SD-OCT imaging of the rat eye, and confirmed by histology, counting of viable fungi in the infected rat cornea, and PCR with specific primers for Fusarium sp. Results: We imaged and measured the rat cornea with SD-OCT. Custom-made contact lenses were developed based on the OCT measurements. Incubation of contact lenses in a F. solani suspension resulted in biofilm formation. We induced contact lens-associated Fusarium keratitis by fitting the rat eyes for 4 h with the Fusarium-contaminated contact lenses. The SD-OCT images of the cornea correlated well with the slit-lamp and histopathological results and clearly defined clinical signs of infection, namely, increased corneal thickening, loss of epithelial continuity, hyper-reflective areas representing infiltrates, and endothelial plaques characteristic of fungal infection. Moreover, in three cases, SD-OCT detected the infection without any clear findings on slit-lamp examination. Infection was confirmed with histological fungal staining, PCR, and microbiological culture positivity. Conclusions: We developed a highly reproducible rat contact lens model and successfully induced contact lens-associated Fusarium keratitis in this model. The clinical presentation of contact lens-associated Fusarium keratitis in the rat model is similar to the human condition. SD-OCT is a valuable tool that non-invasively revealed characteristic signs of the fungal infection and could provide sensitive, objective monitoring in fungal keratitis.

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