In vitro model for corneal wound healing; organ-cultured human corneas.
Healing of linear, non-perforating thermal burns was studied in 56 human corneas in an air/liquid organ culture system in serum free medium or in media supplemented with 10% fetal bovine serum, 10% human serum or 10% human plasma. The extent of epithelial wound healing was determined by measuring epithelial growth into the wound using digitized computer scanning of light micrographs of 1 micron sections. The cross-sectional area of this epithelial growth entering the wound was significantly greater for corneas incubated with either human serum (16,350 +/- 12,088 microns 2/day; p < 0.0001) or human plasma (20,571 +/- 12,276 microns 2/day; p = 0.0004) than for those incubated in serum free (1,784 +/- 1,957 microns 2/day) medium. There was no significant difference between epithelial growth in the serum free and fetal bovine serum supplemented (3,779 +/- 2,580 microns 2/day) media or between that in human serum and human plasma supplemented media. The thickness of the epithelium adjacent to the wound was greater in corneas cultured in fetal bovine serum supplemented media than in corneas cultured in the presence of human serum. Similarly, the build-up of epithelium at the wound edge for corneas in either serum free or fetal bovine serum supplemented media was significantly greater than for either human serum or human plasma supplemented media. The percentage of basal epithelial nuclei which incorporated bromodeoxyuridine (BrdU) increased during the first three days of culture when it reached a plateau. Comparison of paired wounded and unwounded corneas showed that wounding stimulated an increase in DNA synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)
Collin
H B
HB
Ophthalmology Research Laboratory, National Vision Research Institute, San Diego, CA, USA.
Anderson
J A
JA
Richard
N R
NR
Binder
P S
PS
D001485:Basement Membrane / Q000737:chemistry; D001973:Bromodeoxyuridine; D002465:Cell Movement; D003094:Collagen / Q000032:analysis; D003315:Cornea / Q000473:pathology / Q000502:physiology*; D065306:Corneal Injuries; D016895:Culture Media, Serum-Free; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication / Q000502:physiology; D004848:Epithelium / Q000473:pathology / Q000502:physiology; D005109:Extracellular Matrix / Q000737:chemistry; D005126:Eye Burns / Q000473:pathology / Q000503:physiopathology; D005353:Fibronectins / Q000032:analysis; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007124:Immunoenzyme Techniques; D009924:Organ Culture Techniques; D012038:Regeneration / Q000502:physiology*; D014945:Wound Healing / Q000502:physiology*
D016895:Culture Media, Serum-Free; D005353:Fibronectins; D003094:Collagen; D004247:DNA; D001973:Bromodeoxyuridine