Abstract
Purpose: :
To evaluate the autocrine protein expression of cells from the ocular surface after treatment with autologous PRGF-Endoret.
Methods: :
Blood from healthy donors was collected, centrifuged and Plasma Rich in Growth Factors (PRGF-Endoret) was prepared avoiding the buffy coat. Human cultured cells including keratocytes (HCK), conjunctival fibroblasts (HconF) and immortalized corneal epithelial cells (HCE) were treated 48h or 72h with PRGF-Endoret. Cell culture supernatants (conditioned media) were collected at those times. Enzyme-Linked ImmunoSorbent Assay (ELISAs) method was used to measured in conditioned media several factors involved in different regeneration processes like proliferation, migration, angiogenesis or extracellular matrix remodeling.
Results: :
PRGF-Endoret treatment of all types cells induced an increase of anti-angiogenesis factors like trombospondin-1, and a reduction of some angiogenic factors like VEGF; showing a possible regulatory effect of corneal angiogenesis. Keratocytes and conjunctival fibroblasts showed an increased expression of metallopeptidase inhibitor 1 (TIMP-1), hyaluronic acid and procollagen after PRGF-Endoret treatment, indicating a possible regulatory effect of extracellular matrix remodeling. The increased expression of HGF by HConF and HCK cells after PRGF-Endoret stimulation reveals a potential paracrine stimulation for corneal re-epithelialization.
Conclusions: :
The PRGF-Endoret stimulates significantly the autocrine expression of several factors involved in ocular surface tissue regeneration.
Keywords: wound healing • growth factors/growth factor receptors • cornea: stroma and keratocytes