Comparison of corneal epitheliotrophic capacity among different human blood-derived preparations.
OBJECTIVE
To compare the corneal epitheliotrophic capacity of different human blood-derived preparations, including cord blood serum (CBS), peripheral blood serum (PBS), and fresh frozen plasma (FFP) on bovine corneal epithelial cells.


METHODS
The concentrations of epithelial growth factor, transforming growth factor β1, insulin-like growth factor 1, hyaluronic acid, fibronectin, albumin, glucose, and calcium in different human blood derivatives were evaluated using enzyme-linked immunosorbent assay or biochemical methods. Cultivated bovine corneal epithelial cells were incubated with various blood derivatives, and cell proliferation, migration, and differentiation were evaluated by colorimetric assay, Boyden chamber chemotaxis assay, wounding assay, scanning electron microscopy, and transepithelial electric resistance measurements. Wound closure was assessed using a scratch-induced directional wounding assay.


RESULTS
Of the 3 human blood derivatives evaluated, CBS had the highest concentrations of epithelial growth factor, transforming growth factor β1, and hyaluronic acid (P < 0.05). FFP had the lowest concentration of calcium and the highest concentration of glucose (P < 0.05). CBS demonstrated the highest ability to promote cellular proliferation, followed by PBS and FFP (P < 0.05). CBS was also the best in promoting cellular differentiation because scanning electron microscopy demonstrated coherent monolayers of flattened and polygonal-shaped cells with evenly distributed microvilli. Transepithelial electric resistance was noted to be the highest for cells incubated in CBS, indicating formation of well-differentiated cells with functional tight junction (P < 0.05). Cells cultivated with FFP were the least capable of promoting proliferation, migration, and differentiation.


CONCLUSIONS
Different human blood derivatives may have different concentrations of epitheliotrophic factors. CBS is generally superior to PBS in promoting corneal epithelial proliferation and differentiation.
Department of Ophthalmology, Buddhist Tzu Chi General Hospital, Taipei Branch, Taipei, Taiwan.
Elizabeth P
EP
Shen
Fung-Rong
FR
Hu
Shyh-Chyi
SC
Lo
Yan-Ming
YM
Chen
Yi-Chen
YC
Sun
Chung-Tien
CT
Lin
Wei-Li
WL
Chen
D000328:Adult; D000818:Animals; D002417:Cattle; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D019573:Epithelium, Corneal / Q000166:cytology* / Q000648:ultrastructure; D005260:Female; D005312:Fetal Blood / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000097:blood; D008855:Microscopy, Electron, Scanning; D008871:Microvilli / Q000648:ultrastructure; D009994:Osmolar Concentration; D010949:Plasma / Q000737:chemistry / Q000502:physiology*; D044967:Serum / Q000737:chemistry / Q000502:physiology*; D053773:Transforming Growth Factor beta1 / Q000097:blood
D036341:Intercellular Signaling Peptides and Proteins; D053773:Transforming Growth Factor beta1