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Renata Ruoco Loureiro, Priscila Cristovam, Joyce Covre, Jose Alvaro Pereira Gomes; Effect of conditioned medium on corneal epithelial cells wound healing in vitro. Invest. Ophthalmol. Vis. Sci. 2017;58(8):150.
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© ARVO (1962-2015); The Authors (2016-present)
To compare the effectiveness of different conditioned media in the corneal epithelium healing in vitro
The conditioned medium (CM) was collected from: A-) limbal epithelial cells (LEC) grown on the culture plate with 2% FBS; B-) LEC grown on the plate with 5% FBS; C-) LEC cultivated on amniotic membrane (LEC-MA); and d-) limbal fibroblasts (LF) cultured on the culture plate. For the wound healing analysis, corneal epithelial cells were cultured and upon reaching confluence, an injury was caused in the cell layer (wound healing) and treated with the previously collected CM. Fresh medium was used as a control group. The epithelial migration was observed during 72 hours. Also, keratinocyte growth factor (KGF) was quantified in the CM using conventional ELISA assay.
The complete healing was observed between 60 – 72 hours in the groups treated with CM from LEC-MA and LF, followed by CM from LEC 5% FBS with a complete healing after 72 hours of treatment. We could not observe an effective epithelial wound healing in the group treated with CM from LEC 2% FBS even after 72 hours of incubation. The presence of KGF was detected only in the CM from LEC-MA and LF. Using this assay, it was not possible to quantify the KGF in the CM from LEC 2% and in the control group (fresh medium).
These results suggest that CM from LEC-MA and LF seem to be more effective in the corneal epithelial healing in vitro. In the ELISA analysis, KGF was also higher in the groups that have the faster growing. More studies are under way to identify which growth factors are involved in epithelial cell migration and to evaluate the effect of CM to stimulate the epithelial cell healing process of the cornea.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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