Establishment of a Robust and Simple Corneal Organ Culture Model to Monitor Wound Healing

Schumann, Sandra; Dietrich, Eva; Kruse, Charli; Grisanti, Salvatore; Ranjbar, Mahdy

doi:10.3390/jcm10163486

Cited by 7 publications

(10 citation statements)

References 28 publications

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“…Over the last two decades, mammalian animal models have supported a detailed understanding of the mechanisms involved in corneal wound healing [ 11 , 12 , 13 , 14 , 15 ]. According to the published literature, acid- or alkali-based corneal injuries have been reported using rabbit and rodent models [ 16 , 17 , 18 , 19 ].…”

Section: Introductionmentioning

confidence: 99%

Effect of Polydeoxyribonucleotide (PDRN) Treatment on Corneal Wound Healing in Zebrafish (Danio rerio)

Edirisinghe

Nikapitiya

Dananjaya

et al. 2022

IJMS

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This study aimed to develop a corneal epithelial injury model in zebrafish (Danio rerio) and investigate the effectiveness of polydeoxyribonucleotide (PDRN) treatment on in vivo corneal epithelial regeneration and wound healing. Chemical injury to zebrafish cornea was produced by placing a small cotton swab containing 3% acetic acid solution. PDRN treatment was performed by immersing corneal-injured zebrafish in water containing PDRN (2 mg/mL) for 10 min at 0, 24, 48, and 72 h post-injury (hpi). The level of corneal healing was evaluated by fluorescein staining, histological examination, transcriptional profiling, and immunoblotting techniques. Fluorescein staining results demonstrate that PDRN treatment significantly (p < 0.05) reduced the wounded area of the zebrafish eye at 48 and 72 hpi, suggesting that PDRN may accelerate the corneal re-epithelialization. Histopathological evaluation revealed that injured corneal epithelial cells were re-organized at 72 hpi upon PDRN treatment with increased goblet cell density and size. Moreover, transcriptional analysis results demonstrate that PDRN treatment induced the mRNA expression of adora2ab (6.3-fold), pax6a (7.8-fold), pax6b (29.3-fold), klf4 (7.3-fold), and muc2.1 (5.0-fold) after the first treatment. Besides, tnf-α (2.0-fold) and heat-shock proteins (hsp70; 2.8-fold and hsp90ab1; 1.6-fold) have modulated the gene expression following the PDRN treatment. Immunoblotting results convincingly confirmed the modulation of Mmp-9, Hsp70, and Tnf-α expression levels upon PDRN treatment. Overall, our corneal injury model in zebrafish allows for understanding the morphological and molecular events of corneal epithelial healing, and ophthalmic responses for PDRN treatment following acid injury in zebrafish.

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Section: Introductionmentioning

confidence: 99%

Effect of Polydeoxyribonucleotide (PDRN) Treatment on Corneal Wound Healing in Zebrafish (Danio rerio)

Edirisinghe

Nikapitiya

Dananjaya

et al. 2022

IJMS

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“…Finally, we evaluated the host–scaffold response using an ex vivo model of lamellar keratectomy. Organ culture systems are an intermediate model between cell culture and animal studies and have been shown to be a reproducible method to study corneal wound healing. − Corneas that received the PhotoCol-RB gel resulted in faster corneal epithelial wound healing compared to the corneas that received the physical collagen gel. Rabbit corneal epithelial cells migrated over the PhotoCol-RB gel, formed a multilayer, and maintained normal epithelial cell phenotype, which was confirmed by the expression of ZO-1.…”

Section: Discussionmentioning

confidence: 99%

Collagen Gels Crosslinked by Photoactivation of Riboflavin for the Repair and Regeneration of Corneal Defects

Fernandes-Cunha

Brunel

Arboleda

et al. 2023

ACS Appl. Bio Mater.

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Bioengineered corneal tissue is a promising therapeutic modality for the treatment of corneal blindness as a substitute for cadaveric graft tissue. In this study, we fabricated a collagen gel using ultraviolet-A (UV-A) light and riboflavin as a photosensitizer (PhotoCol-RB) as an in situ-forming matrix to fill corneal wounds and create a cohesive interface between the crosslinked gel and adjacent collagen. The PhotoCol-RB gels supported corneal epithelialization and exhibited higher transparency compared to physically crosslinked collagen. We showed that different riboflavin concentrations yielded gels with different mechanical and biological properties. In vitro experiments using human corneal epithelial cells (hCECs) showed that hCECs are able to proliferate on the gel and express corneal cell markers such as cytokeratin 12 (CK12) and tight junctions (ZO-1). Using an ex vivo burst assay, we also showed that the PhotoCol-RB gels are able to seal corneal perforations. Ex vivo organ culture of the gels filling lamellar keratectomy wounds showed that the epithelium that regenerated over the PhotoCol-RB gels formed a multilayer compared to just a double layer for those that grew over physically cross-linked collagen. These gels can be formed either in situ directly on the wound site to conform to the geometry of a defect, or can be preformed and then applied to the corneal wound. Our results indicate that PhotoCol-RB gels merit further investigation as a way to stabilize and repair deep and perforating corneal wounds.

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“…Typically, tears supply growth factors to aid the healing process (Klenkler et al, 2007) and this is a limitation of my ex vivo cornea model. Studies on ex vivo porcine corneas showed that corneal stratified squamous epithelium could regenerate within 72 hours (Schumann et al, 2021) to six days (Castro et al, 2019). In ex vivo human corneas, complete reepithelialization was observed after seven days, resulting in five to six layers of the stratified squamous epithelium.…”

Section: Discussionmentioning

confidence: 99%

The effect of ciprofloxacin and gentamicin on wound healing inex vivosheep cornea model

Okurowska,

Suarez,

Karunakaran

2023

Preprint

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PurposeOur objective was to assess the efficacy of anex vivosheep corneal model as an alternative for live animal testing in screening drug cytotoxicity. In pursuit of this goal, we investigated the impact of two commonly used topical antibiotics, ciprofloxacin and gentamicin, on wound healing. Furthermore, we examined different antibiotic dosages and dosing regimens to understand their effects comprehensively.MethodsThe epithelium onex vivo sheep corneas was removed with a scalpel, and the area was treated with ciprofloxacin (0.1, 0.3, and 1 mg mL-1), gentamicin (0.25, 1, and 3 mg mL-1), or phosphate-buffered saline (control). The corneas were exposed to treatments continuously or twice daily for ten minutes. Wound closure was observed by fluorescein retention and histological staining.ResultsUntreated corneas healed within 41 hours. Continuous exposure to both ciprofloxacin and gentamicin significantly reduced the corneal healing ability in a time- and concentration-dependent manner. Overall, ciprofloxacin was found to be more toxic than gentamycin. However, this model showed that the corneal epithelium could heal effectively when both antibiotics were administered intermittently.ConclusionCiprofloxacin demonstrated greater inhibition of wound healing compared to gentamicin, aligning within vivostudies. The administration of drops several times daily mitigated the toxic effects of antibiotics. Theex vivosheep wound healing model holds promise as an alternative approach toin vivotoxicity testing, enabling the swift evaluation of novel antimicrobial treatments and eye drop additives.

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Establishment of a Robust and Simple Corneal Organ Culture Model to Monitor Wound Healing

Cited by 7 publications

References 28 publications

Effect of Polydeoxyribonucleotide (PDRN) Treatment on Corneal Wound Healing in Zebrafish (Danio rerio)

Effect of Polydeoxyribonucleotide (PDRN) Treatment on Corneal Wound Healing in Zebrafish (Danio rerio)

Collagen Gels Crosslinked by Photoactivation of Riboflavin for the Repair and Regeneration of Corneal Defects

The effect of ciprofloxacin and gentamicin on wound healing inex vivosheep cornea model

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