Regulation of corneal repair by particle-mediated gene transfer of opioid growth factor receptor complementary DNA.
OBJECTIVE
To determine whether molecular manipulation of the opioid growth factor receptor (OGFr) alters corneal reepithelialization following central corneal abrasion in rats.


METHODS
The plasmid pcDNA3.1 + OGFr, carrying the rat OGFr complementary DNA in both the sense and antisense orientations, and empty vector (EV), were delivered by gene gun to the rat cornea. After 24 hours, corneas were abraded and reepithelialization was documented by fluorescein photography. Twenty-four hours after wounding, DNA synthesis (with bromodeoxyuridine) was examined.


RESULTS
Eyes transfected with sense constructs of OGFr had corneal defects that were 24%, 52%, and 50% larger than the EV group at 16, 24, and 28 hours, respectively. Conversely, corneas transfected with antisense constructs of OGFr had corneal defects that were 56% and 48% smaller than the EV group at 16 and 24 hours, respectively. Bromodeoxyuridine labeling in the basal and suprabasal layers of the antisense group were increased 3.3- and 3.7-fold, respectively, in DNA synthesis from corresponding EV layers; DNA synthesis was comparable in the sense and EV groups.


CONCLUSIONS
Excess OGFr delays reepithelialization, whereas attenuation of OGFr accelerates repair of the corneal surface. Clinical Relevance Inhibition of opioid growth factor action using gene therapy could be important in the treatment of corneal diseases such as nonhealing and recurrent erosions, diabetic keratopathy, and neurotrophic keratitis.
Zagon
Ian S
IS
Department of Neural and Behavioral Sciences, The Milton S. Hershey Medical Center, Pennsylvania State University, Hershey, PA 17033, USA. isz1@psu.edu
Sassani
Joseph W
JW
Malefyt
Kristin J
KJ
McLaughlin
Patricia J
PJ
D000818:Animals; D019470:Biolistics; D065306:Corneal Injuries; D004247:DNA / Q000096:biosynthesis; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D005131:Eye Injuries / Q000235:genetics / Q000628:therapy*; D019084:Fluorescent Antibody Technique, Indirect; D015590:Fluorophotometry; D005786:Gene Expression Regulation / Q000502:physiology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D008297:Male; D010957:Plasmids; D051381:Rats; D011957:Receptors, Opioid / Q000235:genetics* / Q000378:metabolism; D014945:Wound Healing; D014949:Wounds, Nonpenetrating / Q000378:metabolism / Q000628:therapy*
D016373:DNA, Antisense; D018076:DNA, Complementary; D011957:Receptors, Opioid; C033593:methionine-enkephalin receptor; D004247:DNA