Donor age and gestational age influence on growth factor levels in human amniotic membrane.
OBJECTIVE
Amniotic membrane (AM) is used as a biomaterial for reconstruction in ocular surface surgery. This study investigated the influence of interdonor variations and processing and preservation procedures applied to the AM on growth factors and protein levels.


METHODS
Samples of human AM from thirteen donors were analysed. Collected donor data were age, parity and gestational age. Total protein amount was measured in extracts of intact AM nonpreserved, lyophilized and cryopreserved, at -80°C and in liquid nitrogen. An enzyme-linked immunosorbent assay (ELISA) was used to assay growth factors protein levels for epidermal growth factor, basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), transforming growth factor beta1 (TFG-β(1) ) and nerve growth factor (NGF). Univariate and multivariate statistical analyses were used to study the influence of the preservation method applied and interdonor variations on growth factors levels.


RESULTS
We detected important variations in growth factors and protein concentrations between samples from different donors. Total protein amount, bFGF, HGF, KGF and TGF-β(1) showed lower levels in samples from donors with higher gestational ages and donor ages, for all groups.


CONCLUSIONS
The variability in the biochemical composition of AM from different donors is considerable, and it is related with donor factors as donor age and gestational age. As AM biochemical composition has a role in its therapeutic effects, these variations could affect the clinical results of amniotic membrane transplantation and must be taken into account in donor selection processes.
Department of Ophthalmology, USC University Hospital Complex of Santiago de Compostela, Santiago de Compostela, Spain. lopez.mariajesus@gmail.com
Maria J
MJ
López-Valladares
M
M
Teresa Rodríguez-Ares
Rosario
R
Touriño
Francisco
F
Gude
M
M
Teresa Silva
Jose
J
Couceiro
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000650:Amnion / Q000378:metabolism*; D015925:Cryopreservation; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D051523:Fibroblast Growth Factor 7 / Q000378:metabolism; D005865:Gestational Age; D020073:Gravidity; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D020932:Nerve Growth Factor / Q000378:metabolism; D014019:Tissue Donors; D014021:Tissue Preservation / Q000379:methods; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D055815:Young Adult
D036341:Intercellular Signaling Peptides and Proteins; D053773:Transforming Growth Factor beta1; D016222:Fibroblast Growth Factor 2; D051523:Fibroblast Growth Factor 7; D017228:Hepatocyte Growth Factor; D020932:Nerve Growth Factor