Na Hee Kang1, Shina Lee2, Roo Min Jun3,4. 1. BE Woorieyes Clinic, Wonju, Republic of Korea. 2. Department of Internal Medicine, Division of Nephrology, School of Medicine, Ewha Womans University, Seoul, Republic of Korea. 3. Department of Ophthalmology, School of Medicine, Ewha Womans University, Seoul, Republic of Korea. jrmoph@ewha.ac.kr. 4. Department of ophthalmology, Ewha Womans University Mok-dong Hospital, 1071, Anyangcheon-ro, Yangcheon-gu, Seoul, Korea, 158-710. jrmoph@ewha.ac.kr.
Abstract
PURPOSE: To investigate the composition and biological activity of serum epitheliotrophic factors from chronic renal failure (CRF) patients vs. healthy controls. METHODS: Twenty, 50 and 100 % autologous serum eyedrops (ASEs) were prepared from 16 CRF patients and 16 normal subjects. Serum epithelial growth factor (EGF), platelet-derived growth factor-AB (PDGF-AB), transforming growth factor-β1 (TGF-β1) and fibronectin levels were quantified using enzyme-linked immunosorbent assay kits. A 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide (MTT) assay was used to compare the proliferative effects of serum from CRF patients and healthy donors in a human corneal epithelial cell (HCEC) culture model. Migration assays were conducted via manual scraping of HCECs for migratory potential of ASEs. Morphologic changes were studied with transmission electron microscopy (TEM). RESULTS: EGF, PDGF-AB and TGF-β1 levels in ASEs from healthy donors were significantly higher than in serum of CRF patients. Cellular proliferation was similar in the CRF patient and normal control groups. ASEs from the normal group had a significantly higher effect on cell migration. ASEs in both groups facilitated better proliferation and migration than the negative control. Furthermore, we observed an enhancement after incubation with diluted serum vs. undiluted serum. In TEM analysis, HCECs incubated with CRF patients' 50 % ASEs showed some loss of microvilli without alterations of cytoplasmic organelles. CONCLUSIONS: Epitheliotrophic factors concentrations and biologic activities from CRF patient sera differed from healthy controls. ASEs in CRF patients are also helpful in the corneal healing process, especially when applied at a 50 % concentration.
PURPOSE: To investigate the composition and biological activity of serum epitheliotrophic factors from chronic renal failure (CRF) patients vs. healthy controls. METHODS: Twenty, 50 and 100 % autologous serum eyedrops (ASEs) were prepared from 16 CRF patients and 16 normal subjects. Serum epithelial growth factor (EGF), platelet-derived growth factor-AB (PDGF-AB), transforming growth factor-β1 (TGF-β1) and fibronectin levels were quantified using enzyme-linked immunosorbent assay kits. A 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazoliumbromide (MTT) assay was used to compare the proliferative effects of serum from CRF patients and healthy donors in a human corneal epithelial cell (HCEC) culture model. Migration assays were conducted via manual scraping of HCECs for migratory potential of ASEs. Morphologic changes were studied with transmission electron microscopy (TEM). RESULTS: EGF, PDGF-AB and TGF-β1 levels in ASEs from healthy donors were significantly higher than in serum of CRF patients. Cellular proliferation was similar in the CRF patient and normal control groups. ASEs from the normal group had a significantly higher effect on cell migration. ASEs in both groups facilitated better proliferation and migration than the negative control. Furthermore, we observed an enhancement after incubation with diluted serum vs. undiluted serum. In TEM analysis, HCECs incubated with CRF patients' 50 % ASEs showed some loss of microvilli without alterations of cytoplasmic organelles. CONCLUSIONS: Epitheliotrophic factors concentrations and biologic activities from CRF patient sera differed from healthy controls. ASEs in CRF patients are also helpful in the corneal healing process, especially when applied at a 50 % concentration.