In Vitro Expression Analysis of Cytokines and ROS-Related Genes in Human Corneal Fibroblasts and Keratocytes of Healthy and Keratoconus Corneas

ABSTRACT

Purpose

To investigate expression of cytokines and ROS-related genes in stromal cells of healthy and keratoconus (KC) corneas.

Methods

Expression analysis was performed for cytokines including several interleukins (IL), Tumor necrosis factor-α (TNF-α), Transforming growth factor-β1 (TGF-β1), Interferon-γ (IFN-γ) and ROS-related genes such as Catalase, Glutathione peroxidase 1, NADPH oxidase 1, superoxide dismutase 1 in corneal fibroblasts (HCFs/KC-HCFs) or keratocytes (Keratocytes/KC-Keratocytes) by qPCR and ELISA.

Results

Gene and protein expression of most inflammatory markers was decreased in keratocytes compared to fibroblasts, whereas no differences were found between healthy and keratoconus cells for the majority of cytokines measured. TNF-α expression was increased at gene (KC keratocytes) and protein levels (supernatant of Keratocytes/KC-Keratocytes) compared to corneal fibroblasts. No differential expression of ROS-related genes was detected between healthy and diseased cells in both fibroblasts and keratocytes.

Conclusion

Increased expression of several inflammatory markers described as altered in KC was not evident in KC cells in vitro.

KEYWORDS:

• Corneal fibroblasts
• cytokines
• inflammation
• keratoconus
• keratocytes
• reactive oxygen species

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data that support the findings of this study are available from the corresponding author, TB, upon reasonable request.

Additional information

Funding

Dr. Tim Berger’s work is funded by the Homburg Research Grant (HOMFOR2022). The work of Dr. Tanja Stachon, Ning Chai and Prof. Dr. Nóra Szentmáry at the Dr. Rolf M. Schwiete Center was supported by the Dr. Rolf M. Schwiete Foundation.