Abstract
Purpose: :
Human amniotic fluid (HAF), the liquid surrounding the fetus, is in continuous contact to the child and the amniotic membrane (AM), which has been used in ocular defect surgery. In literature corneal wound healing and anti-inflammatory effect of HAF has been described in an in vivo animal model. Serum eye drops (SED) have been established in clinical trials. Serum supports proliferation and migration of corneal epithelial cells. The positive effect is thought to be due to growth factors, vitamins and other nutrient factors. Both fluids were tested in an in vitro cell culture model for their cell growth support, migration and their content of growth factors.
Methods: :
HAF from ten women (mean age = 36±5 years old) was collected during amniocentesis for prenatal diagnosis in mean at the week 14±0.7 of gestation. Serum was prepared from ten volunteers (mean age = 28±6 years) according to a standardised protocol. Serum and HAF were diluted with balanced salt solution (BSS) to final concentrations of 100, 50, 25, 12.5, 6.25 %. SV-40 immortalised human corneal epithelial cells were cultured in 96-well plates at 37°C, 5% CO2. Cell growth was quantified by means of a luminescence-based ATP-assay at incubation times of 6, 30 and 54 hrs. Cell migration was assessed in a colony dispersion assay with incubation times of 24, 48 and 96 hrs. Growth factors (EGF, TGF-β, fibronectin) were quantified by routine ELISA technology.
Results: :
Serum was found to be significant better in cell growth support during all incubation times. For both serum and HAF cell growth was decreasing by increasing dilution. Colony dispersion assay showed a higher migration potency of serum. All three measured cytokines were found at significant higher quantity in serum.
Conclusions: :
Serum had superior growth supporting and migration effects on corneal epithelial cells than HAF. Wound healing of corneal epithelial defects is among other factors a reason of cell proliferation and cell migration. The results of our experiments show that serum seems to be the better solution. To elucidate the potency of HAF furthermore extensive data should be collected and in vivo trials should be organised.
Keywords: cornea: epithelium • cornea: tears/tear film/dry eye • wound healing