Molecular biology of pharmacologic vitreolysis.
OBJECTIVE
Pharmacologic vitreolysis is a promising new therapy to improve vitreoretinal surgery and, ultimately, prevent disease by mitigating the contribution of vitreous to retinopathy. The mechanism of action of the agents being developed for pharmacologic vitreolysis remains unclear. The experiments in this thesis test the hypothesis that pharmacologic vitreolysis agents break down vitreous macromolecules into smaller particles.


METHODS
Microplasmin, hyaluronidase, and collagenase were tested in solutions of hyaluronan (n = 15) and collagen (n = 15), explants of bovine vitreous (n = 15), dissected porcine vitreous (n = 9), and intact porcine eyes (n = 18). There were also 21 controls, totaling 93 specimens. Vitreous macromolecule sizes were determined with dynamic light scattering (DLS), performed at intervals from 10 minutes to 24 hours following injections.


RESULTS
Studies of DLS reproducibility showed a coefficient of variance of less than 3.3% in all but one specimen. Microplasmin decreased porcine vitreous macromolecule size in a dose-dependent manner (correlation coefficient r = 0.93), with an 85% reduction after a 30-minute exposure to the maximum dose. Hyaluronidase decreased vitreous macromolecule size in hyaluronan solutions by 50% at high (1,000 IU/mL, P < .001) doses and in bovine vitreous by 20%. Collagenase decreased macromolecule size in collagen solutions by 20% at both low (1 mg/mL, P < .001) and high (10 mg/mL, P < .001) doses, but not at all in bovine vitreous.


CONCLUSIONS
Pharmacologic vitreolysis can induce a significant decrease in vitreous macromolecule sizes, depending upon the pharmacologic agents and the experimental model. Broad-spectrum agents were more effective than substrate-specific enzymes. Defining the molecular biology of pharmacologic vitreolysis has implications for surgical developments and may impact upon the design of clinical trials to induce prophylactic posterior vitreous detachment.
Sebag
J
J
VMR Institute, Huntington Beach, California, USA.
D000818:Animals; D002417:Cattle; D003094:Collagen; D017364:Collagenases / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D005341:Fibrinolysin / Q000008:administration & dosage / Q000494:pharmacology*; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase / Q000008:administration & dosage / Q000494:pharmacology*; D008027:Light; D046911:Macromolecular Substances / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D008967:Molecular Biology; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012996:Solutions; D013552:Swine; D013997:Time Factors; D014822:Vitreous Body / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology
D046911:Macromolecular Substances; D012996:Solutions; D006820:Hyaluronic Acid; D003094:Collagen; D006821:Hyaluronoglucosaminidase; D005341:Fibrinolysin; D017364:Collagenases