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IN VITRO EFFECTS OF THREE BLOOD DERIVATIVES ON HUMAN CORNEAL EPITHELIAL CELLS.
Purpose:
To compare the effects of three blood derivatives (Autologous Serum, AS; Platelet-Rich Plasma, PRP and serum derived from Plasma Rich in Growth Factors, PRGF) on a human corneal epithelial (HCE) cell line, in order to evaluate their potential as an effective treatment for corneal epithelial disorders.
Methods:
The concentrations of EGF, FGF, VEGF, HGF, PDGF and fibronectin were quantified by ELISA. The proliferation and viability of HCE cells were measured by an MTT colorimetric assay. Cell morphology was assessed by phase-contrast microscopy. The patterns of expression of several CONNEXIN, INVOLUCRIN and INTEGRIN α6 genes were analyzed by real-time RT-PCR.
Results:
We have found significantly higher levels of EGF in PRGF compared to AS and PRP. However, both AS and PRGF induced robust proliferation of HCE cells. In addition, PRGF cultured cells grew as heterogeneous colonies, exhibiting differentiated and non-differentiated cell phenotypes, whereas AS- and PRP-treated cultures exhibited quite homogeneous colonies. Finally, PRGF up-regulated the expression of several genes associated with communication and cell differentiation, in comparison to AS or PRP.
Conclusions:
PRGF promotes biological processes required for corneal epithelialization, such as proliferation and differentiation. Since PRGF effects are similar to those associated with routinely used blood derivatives, the present findings warrant further research on PRGF as a novel alternative treatment for ocular surface diseases.
To compare the effects of three blood derivatives (Autologous Serum, AS; Platelet-Rich Plasma, PRP and serum derived from Plasma Rich in Growth Factors, PRGF) on a human corneal epithelial (HCE) cell line, in order to evaluate their potential as an effective treatment for corneal epithelial disorders.
Methods:
The concentrations of EGF, FGF, VEGF, HGF, PDGF and fibronectin were quantified by ELISA. The proliferation and viability of HCE cells were measured by an MTT colorimetric assay. Cell morphology was assessed by phase-contrast microscopy. The patterns of expression of several CONNEXIN, INVOLUCRIN and INTEGRIN α6 genes were analyzed by real-time RT-PCR.
Results:
We have found significantly higher levels of EGF in PRGF compared to AS and PRP. However, both AS and PRGF induced robust proliferation of HCE cells. In addition, PRGF cultured cells grew as heterogeneous colonies, exhibiting differentiated and non-differentiated cell phenotypes, whereas AS- and PRP-treated cultures exhibited quite homogeneous colonies. Finally, PRGF up-regulated the expression of several genes associated with communication and cell differentiation, in comparison to AS or PRP.
Conclusions:
PRGF promotes biological processes required for corneal epithelialization, such as proliferation and differentiation. Since PRGF effects are similar to those associated with routinely used blood derivatives, the present findings warrant further research on PRGF as a novel alternative treatment for ocular surface diseases.
Freire V, Andollo N, Etxebarria J, Duran JA, Morales MC.
Source
R&D&I Department, Instituto Clinico-Quirurgico de Oftalmologia, Bilbao, Spain.