Ocular Irritation Evaluation by Reconstructed Human Corneal Epithelium (RhCE) Model

Creative Biolabs utilizes the RhCE model to provide evaluation service of ocular irritation as a validated non-animal alternative to reliably classify mixtures and formulations to be evaluated based on their irritation properties.

RhCE Model Construction

The RhCE model provides a powerful platform to study corneal physiology and evaluate new ophthalmic drugs. This innovative model offers a more cost-effective way to delve into corneal structure, function, and responses, making it a pivotal asset in advancing ocular health and pharmaceutical development. The construction strategy of RhCE model is outlined below:

1. Acquisition of corneal epithelial cells
   Corneal epithelial cells are derived from stable commercially available samples and cultured in appropriate media that maintained stable cellular function and morphological characteristics.
2. Preparation of the RhCE model
   Cellular material obtained by serum-free in vitro culture on appropriate support material to construct multilayer RhCE models. It harbors specific markers such as keratins, CD44, and hemidesmosomes.
3. Recognizing the cell type and structure of the model
   Performed immunohistochemical staining and microscopic observation to confirm the consistency of the major cell types and organization in the constructed multilayered corneal epithelial model with that of the original human corneal epithelium.
4. Validation
   Validate the accuracy and reliability of the constructed RhCE model for known ocular irritants and new chemical substances.

Fig. 1 Observation of the RhCE model by histology and immunohistology.1

RhCE Model Applied to Ocular Irritation Assessment

The following process of drug assessment emphasizes the pivotal role of RhCE model in advancing ocular irritation assessment.

RhCE model adaptation

Equilibrate the RhCE model overnight in the maintenance medium and transfer it to a test culture environment.

Preparation of test conditions

Negative control: PBS without calcium and magnesium ions.
Positive control: methyl acetate.
Object to be measured: PBS without calcium and magnesium ions and substance to be measured.

Collection of stimulus evaluation data

The object to be evaluated (e.g., formulation, drug) is applied to the constructed RhCE model and acted for a given time and concentration (allowing multiple time points and concentration gradients).

Assessment of cellular responses and toxicity

Assess the ocular irritation of the substance to be evaluated on the RhCE model using appropriate cell viability, cell morphology, immunohistochemistry, and genotoxicity. Compare the differences between the treated and control groups.

Data analysis and interpretation

To determine the ocular irritation characteristics of the substances to be evaluated using the mean tissue viability as a predictor. Statistical methods are used to verify the reliability of the results.

Fig. 2 Histological images of apoptotic DNA fragments of SDS-treated RhCE models.2

Strengths

1. Alternative to animal testing
   Reduce reliance on animals such as rabbits and mice in traditional experiments and avoid unnecessary experimental manipulation and injury.
2. Human relevance
   Human corneal epithelial cells are used as a source of model material to incubate the model for stronger human relevance and assessment accuracy.
3. Repeatability and standardization
   The process of constructing and using the RhCE model is relatively standardized, and the reproducibility and comparability of experimental results are ensured by establishing a unified experimental process and standard operation.
4. Controllability
   The use of the RhCE model allows for precise control of experimental conditions, such as concentration of stimulus, time, and mode of exposure. This allows more detailed and reliable data to be obtained.
5. Research efficiency and cost
   RhCE models can be constructed relatively quickly, allowing multiple experiments to be performed and different stimulus substances to be tested simultaneously. It also reduces the time, cost, and resources involved in animal experiments.

Extensive 3D Biology Based Ocular Toxicity Evaluation Services

• Retinal Toxicity Evaluation by Retinal Organoid
• Ocular Toxicity Evaluation by Human Retinal Pigment Epithelial (RPE) Cells
• Ocular Irritation Evaluation by Human 3D Cornea Epithelial Model

At Creative Biolabs, the RhCE model-based ocular irritation evaluation service provides a powerful tool for ocular research with great expertise. Please contact us to get a quote.

Related Sections:

• 3D Biology Based Ocular Toxicity Evaluation Services
• 3D Biology Based Skin Toxicity Evaluation Services
• 3D Biology Based Cardiotoxicity Evaluation Services
• 3D Biology Based Hepatotoxicity Evaluation Services
• 3D Biology-Based Neurotoxicity Evaluation Services
• 3D Biology-Based Nephrotoxicity Evaluation Services
• 3D Biology-Based Gastrointestinal Toxicity Evaluation Services
• 3D Biology Based Phototoxicity & Photoallergy Evaluation Services
• 3D Biology Based Pancreatic Endocrine Toxicity Evaluation Service
• 3D Biology Based Inhalation Toxicity Evaluation Services
• 3D Biology Based Oral Irritation Evaluation Services