A sequential, multiple-treatment, targeted approach to reduce wound healing and failure of glaucoma filtration surgery in a rabbit model (an American Ophthalmological Society thesis).
OBJECTIVE
The purpose of this study was to evaluate the concept of targeting mediators of the scarring process at multiple points across the course of bleb failure, in order to prolong bleb survival.


METHODS
There were three linked parts to the experiment. In the first part, a cannula glaucoma filtration surgery (GFS) was performed on 32 New Zealand White (NZW) rabbits, and bleb survival was assessed for six different regimens plus controls by grading bleb height and width. For the second part of the study, the same GFS surgery was performed on an additional 10 NZW rabbits. Two additional filtering blebs were treated with balanced saline solution (BSS), two received mitomycin-C (MMC) (0.4 mg/mL), and for the remaining six, a sequential regimen was given consisting of 200 mmol/L mannose-6-phosphate (M-6-P) solution at the time of surgery, followed by subconjunctival injections of antibody to connective tissue growth factor at days 2 and 4, and Ilomastat, a broad-spectrum matrix metalloproteinase inhibitor, at days 7, 12, and 20 postoperatively. Bleb survival was again assessed. In the final part of the experiment, blebs treated with either BSS, MMC, or the above sequential multitreatment regimen were examined histologically at 14 days postoperatively in three additional NZW rabbits.


RESULTS
All six individual therapies selected resulted in some improvement of bleb survival compared to BSS control. Blebs treated with the new sequential, multitreatment protocol survived an average of 29 days (regression slope, P < .0001 compared to control), those receiving BSS an average of 17 days, and those treated with MMC (0.4 mg/mL) an average of 36 days. The sequential, multitreatment regimen was significantly superior to any of the six monotherapies for time to zero analysis (flattening) of the bleb (P < .002). Histologic examination of the bleb tissues showed a markedly less epithelial thinning, subepithelial collagen thinning, and goblet cell loss in the multitreatment group, when compared with the MMC blebs.


CONCLUSIONS
In a rabbit model of GFS, a sequential, targeted, multitreatment approach prolonged bleb survival compared to BSS controls and decreased bleb tissue morphological changes when compared to those treated with MMC. It is not known whether these findings can be reproduced in humans, and further work is needed to determine an optimum regimen and timing of therapeutic delivery.
Sherwood
Mark Brian
MB
Department of Ophthalmology, College of Medicine, University of Florida, Gainesville, Florida, USA.
D000477:Alkylating Agents / Q000008:administration & dosage; D000818:Animals; D000963:Antimetabolites / Q000008:administration & dosage*; D001768:Blister / Q000188:drug therapy* / Q000473:pathology; D003228:Conjunctiva / Q000187:drug effects / Q000473:pathology; D003238:Connective Tissue / Q000187:drug effects / Q000473:pathology; D055513:Connective Tissue Growth Factor; D004195:Disease Models, Animal; D016503:Drug Delivery Systems; D018463:Filtering Surgery; D005901:Glaucoma / Q000601:surgery*; D006877:Hydroxamic Acids; D017874:Immediate-Early Proteins / Q000235:genetics / Q000276:immunology; D007211:Indoles / Q000008:administration & dosage; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology; D007429:Intraocular Pressure; D008297:Male; D008360:Mannosephosphates / Q000008:administration & dosage; D016685:Mitomycin / Q000008:administration & dosage; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage; D011817:Rabbits; D053781:Transforming Growth Factor beta2 / Q000235:genetics / Q000276:immunology; D017211:Treatment Failure; D014945:Wound Healing / Q000187:drug effects*
D000477:Alkylating Agents; D000963:Antimetabolites; C526317:CCN2 protein, human; D006877:Hydroxamic Acids; D017874:Immediate-Early Proteins; D007211:Indoles; D036341:Intercellular Signaling Peptides and Proteins; D008360:Mannosephosphates; D020319:Oligodeoxyribonucleotides, Antisense; D053781:Transforming Growth Factor beta2; D055513:Connective Tissue Growth Factor; C027693:mannose-6-phosphate; D016685:Mitomycin; C119203:ilomastat