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Experimental Study of the Anti-proliferative Effects of AGS-CsA Membrane on the Glaucoma Filtering Surgery

Author: LiuMaoXiong
Tutor: ZhangXiaoGuang
School: Jilin University
Course: Ophthalmology
Keywords: Sodium alginate delay release system Biocompatibility cyclosporine A Glaucoma filtration surgery Transforming growth factor-β1 Reverse transcription polymerase chain reaction (RT -PCR) electron microscopy
CLC: R779.6
Type: PhD thesis
Year: 2006
Downloads: 111
Quote: 1
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Abstract


Objective To investigate the prospect of sodium alginate as a drugdelivering vehicle in the local application to treat some eye diseases. weobserved the histopathologic changes of sodium alginate membrane onglaucoma filtration surgery. The gene expression and the protein expression ofTGF-β1 in the filtering site tissue were demonstrated by RT-PCR and Westernblot. Alginate sodium-cyclosporine A membrane (AGS-CsA membrane) weremade, Vivo and vitro test was use to analysis the release law of AGS-CsAmembrane. Its toxicitys and the effects of suppresses the proliferation offibroblasts were investigated on the glaucoma filtering surgery.Methods We have established filtration surgery animal model with orwithout sodium alginate membrane implanted under the sclera flap of rabbit andobserved its postoperative inflammatory reaction , histopathologic changes by litlamp microscope and light microscope;RT-PCR was carried out for mRNA halfquantitative and Western blot experiment was use to fix quantify of the proteinexpresses of the TGF-β1 in the filtering site tissue. The spectrophotometer wasuse to measured the medicine content of AGS-CsA membrane. Vitro test wasuse to analysis the release law of AGS-CsA membrane by the spectrophotometer.We performed filtration surgery with AGS-CsA membrane implanted under thesclera flap of rabbit and observed medicine dynamics characteristic of themembrane by HPLC;At the same time, we observed its toxicity effect by slitlamp microscope and ultrastructure of ciliary epithelium by transmissionelectron microscope. AGS-CsA membrane used to glaucoma filtration surgeryand cyclosporine A only once used to glaucoma filtration surgery and simplicityglaucoma filtration surgery were compared with. The postoperative IOP、bleb、corneal endothelium were observed by slit lamp microscope,corneal specularphotomicrographs. At 7、14、28 days after surgery, we observed the histologicalchanges and the expression of proliferation cell nuclear antigen(PCNA) in thefiltering area by Immunohistochemistry methods. B ultrasound of thesclerectomy site was performed 28 and 60 days after surgery.Results No significant conjunctival reaction and anterior chamberreaction were seen after surgery. No sign of damage were observed in cornealendothelium , lens. Conjunctiva wound healing was good at 14 days afterfiltration surgery and not observed graft rejection. The histopathologicalexamination showed that there were not significant statistics differences ofneutrophils ,fibroblasts counting in the filtering site tissue between the trialgroups and the control groups. Western blot experiment showed that the proteinexpresses of the TGF-β1 in the filtering site tissue increase after operative;RT-PCR test shows that operation make mRNA of the TGF-β1 in the filteringsite tissue express obvious increased, but there were not significant statisticsdifferences between the trial groups and the control groups . The weight ofAGS-CsA membrane we manufacture is 5±0.14 mg;Medicine contain quantityof CsA is 99±1.47 ug/mg ,150±2.53 ug/mg ,200±1.84 ug/mg. Vitro test showthat CsA in the membrane is release evenly in 7 days.In Vivo test , The medicinedynamics research of membrane showed that medicine is being even in 2weeksreleasing .No poisonous side effect were observed, The densitys of cornealendothelium were no abvious difference, No significant ultrastructure alterationswere observed in epithelial cells of ciliary body by transmission electronmicroscopy. The anti-proliferative test show: At 60 days after surgery ,The IOPdecreased from a mean preoperative value of 2.84±0.261mmHg to a meanpostoperative value of 1.77±0.175mmHg (P< 0.05) ,The IOP was significantstatistically difference ( P <0. 05 ) between the three groups;The functioningblebs in the trial groups were more than that of the control 1,2 groups. Thecomplications such as flat cornea cloudy , retina haemorrhage and effusion,opticnerve edema were not observed among the trial groups ,the control groups andnatural groups. At 7days after surgery ,The number of positive cells(PCNA) infiltering area was no significant statistically difference between the trial groupswith the control 1 groups ( P >0. 05 ),but there were significant statisticallydifference with the control 2 groups( P <0. 05 ). At 14days after surgery , Thenumber of positive cells in filtering area was significant statistically differencebetween the three groups ( P <0. 05 ),The number of positive cells in the trialgroup was less than that of other groups. At 30days after surgery , The numberof positive cells in filtering area was significant statistically difference betweenthe trial groups with the two control groups (P <0. 05 ),but there were nosignificant statistically difference between the two control groups(P >0. 05) ,At60days after surgery, B ultrasound examination shows that there were moresubconjunctival functioning filtrate blebs in the trial groups, and more scartissue in the control 1,2 groups.Conclusion Sodium alginate has a better biocompatibility, It was suit to adrug delivering vehicle in the eye. The more formation of the bleb scarring andthe more expression of TGF-β1of tissue in the filtering site were not excited bysodium alginate. Vivo and vitro test show that AGS-CsA membrane can delayCsA release uniformity;AGS-CsA medicine release system have not anypoisonous side effect on eye ball. AGS-CsA membrane is better suppressed theproliferation of fibroblasts in the filtering site on anti glaucoma filtering surgeryin the long. and its side-effect is less.

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CLC: > Medicine, health > Ophthalmology > Eye surgery and surgery
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