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Serum from plasma rich in growth factors regenerates rabbit corneas by promoting cell proliferation, migration, differentiation, adhesion and limbal stemness
Author(s) -
Etxebarria Jaime,
SanzLázaro Sara,
HernáezMoya Raquel,
Freire Vanesa,
Durán Juan A.,
Morales María–Celia,
Andollo Noelia
Publication year - 2017
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/aos.13371
Subject(s) - wound healing , in vivo , stromal cell , cell growth , in vitro , biology , microbiology and biotechnology , cell migration , fetal bovine serum , immunohistochemistry , stem cell , andrology , pathology , immunology , cancer research , medicine , biochemistry
Purpose To evaluate the regenerating potential and the mechanisms through which the autologous serum derived from plasma rich in growth factors (s‐ PRGF ) favours corneal wound healing in vitro and in vivo . Methods We compared the effect of various concentrations of s‐ PRGF versus fetal bovine serum ( FBS ) and control treatment in rabbit primary corneal epithelial and stromal cells and wounded rabbit corneas. Cell proliferation was measured using an enzymatic colorimetric assay. In vitro and in vivo wound‐healing progression was assessed by image‐analysis software. Migration and invasion were evaluated using transfilter assays. Histological structure was analysed in stained sections. Protein expression was evaluated by immunohistochemistry. Results s‐ PRGF promoted the robust proliferation of epithelial cultures at any concentration, similar to FBS . Likewise, s‐ PRGF and FBS produced similar re‐epithelialization rates in in vitro wound‐healing assays. In vivo , s‐ PRGF treatment accelerated corneal wound healing in comparison with control treatment. This difference was significant only for 100% s‐ PRGF treatment in our healthy rabbit model. Histological analysis confirmed normal epithelialization in all cases. Immunohistochemistry showed a higher expression of cytokeratins 3/76 and 15, zonula occludens ‐1 and alpha‐smooth muscle actin proteins as a function of s‐ PRGF concentration. Notably, keratocyte density in the anterior third of the stroma increased with increase in s‐ PRGF concentration, suggesting an in vivo chemotactic effect of s‐ PRGF on keratocytes that was further confirmed in vitro . Conclusion s‐ PRGF promotes proliferation and migration and influences limbal stemness, adhesion and fibrosis during corneal healing.