Development of a curved, stratified, in vitro model to assess ocular biocompatibility

Autor: Ann M. Wright, Sara Williams, Maud Gorbet, Robert Pintwala, Denise Hileeto, Cameron K Postnikoff
Jazyk: angličtina
Rok vydání: 2014
Předmět:
Integrins
Pathology
Silicones
Gene Expression
lcsh:Medicine
Toxicology
Epithelium
Cornea
Spectrum Analysis Techniques
Tissue engineering
Molecular Cell Biology
Materials Testing
Biological Systems Engineering
Medicine and Health Sciences
lcsh:Science
Corneal epithelium
Confluency
Multidisciplinary
Cell Death
medicine.diagnostic_test
Chemistry
Hydrogels
Flow Cytometry
Cell biology
medicine.anatomical_structure
Cell Processes
Spectrophotometry
Engineering and Technology
Cytophotometry
Anatomy
Cellular Types
Benzalkonium Compounds
Research Article
Biotechnology
medicine.medical_specialty
Biocompatibility
Cell Survival
Predictive Toxicology
Bioengineering
In Vitro Techniques
Research and Analysis Methods
Cell Growth
Flow cytometry
Biomaterials
Cell Adhesion
Genetics
medicine
Humans
Viability assay
Tissue Engineering
lcsh:R
Biology and Life Sciences
Epithelial Cells
Cell Biology
Ophthalmology
Biological Tissue
Medical Devices and Equipment
lcsh:Q
sense organs
Cytometry
Zdroj: PLoS ONE, Vol 9, Iss 5, p e96448 (2014)
PLoS ONE
ISSN: 1932-6203
Popis: Purpose To further improve in vitro models of the cornea, this study focused on the creation of a three-dimensional, stratified, curved epithelium; and the subsequent characterization and evaluation of its suitability as a model for biocompatibility testing. Methods Immortalized human corneal epithelial cells were grown to confluency on curved cellulose filters for seven days, and were then differentiated and stratified using an air-liquid interface for seven days before testing. Varying concentrations of a commercial ophthalmic solution containing benzalkonium chloride (BAK), a known cytotoxic agent, and two relevant ocular surfactants were tested on the model. A whole balafilcon A lens soaked in phosphate buffered saline (BA PBS) was also used to assess biocompatibility and verify the validity of the model. Viability assays as well as flow cytometry were performed on the cells to investigate changes in cell death and integrin expression. Results The reconstructed curved corneal epithelium was composed of 3–5 layers of cells. Increasing concentrations of BAK showed dose-dependent decreased cell viability and increased integrin expression and cell death. No significant change in viability was observed in the presence of the surfactants. As expected, the BA PBS combination appeared to be very biocompatible with no adverse change in cell viability or integrin expression. Conclusions The stratified, curved, epithelial model proved to be sensitive to distinct changes in cytotoxicity and is suitable for continued assessment for biocompatibility testing of contact lenses. Our results showed that flow cytometry can provide a quantitative measure of the cell response to biomaterials or cytotoxic compounds for both the supernatant and adherent cell populations. As a specifically designed in vitro model of the corneal epithelium, this quantitative model for biocompatibility at the ocular surface may help improve our understanding of cell-material interactions and reduce the use of animal testing.
Databáze: OpenAIRE
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