Experimental Study Of Cyclosporine A-containing Cubosomes On Corneal Rejection In The Rat Penetrating Keratoplasty Model

Objectives:1.To observe irritant reaction of different cyclosporine A eye drops on rabbit eyes.2.To establish a rat model of penetrating keratoplasty;To observe the immunosuppressive effect of cyclosporine A eye drops on immunological rejection after corneal transplantation in rats,and evaluate the effect of cyclosporine A-containing cubosomes on corneal allograft rejection.Methods:1.Draize Eye Irritation Test:Pre-selected healthy adult New Zealand white rabbits,animals in each group are 4.Drugs test were dropped in the right conjunctival sac of each animal,while the left conjunctival sac of each eye were dropped control drugs.Drugs were administrated 50μl per eye,every 15min for 2h.At 1h,3h,6h,24h,48h,72h after last administration,evaluate irritant reaction of rabbit conjunctiva,cornea,iris with slit-lamp and score.2.Experimental study of different cyclosporine A eye drops on corneal rejection in the rat penetrating keratoplasty model:Penetrating keratoplasty was performed with Lewis rats as recipients and Wistar rats as donors.55 Lewis rats as recipients were randomly divided into five groups:control group,cubosomes without drug treated group,l%CsA-containing cubosomes treated group,0.5%CsA-containing cubosomes treated group and 1%CsA-containing olive oil treated group.The drugs were administrated 20μl each time and 5 times per day after operation for ten days.The corneal grafts were inspected with slit-lamp and scored every day.At the 10th day postoperatively,the rats of each group were killed respectively.The corneal grafts were examined with HE staining.Results:1.The groups of nanoparticles and cubosomes did not have irritant reaction, only the individual had short-term conjunctival hyperemia and restored after about 1h. The group of olive oil had a little more irritant reaction.There was slight conjunctival hyperemia in about 6h,and iridal hyperemia in one eye.These phenomenons disappeared after 24h.At the same time they blinked frequently.There was more obvious conjunctival hyperemia in two prescription of microemulsion-gel group,and iridal hyperemia in partial eyes.But corneal opacity wasn't observed.At the same time they blinked more frequently.2.Compared to control group and cubosomes without drug treated group,two prescription of CsA-containing cubosomes could extend the survival time of corneal grafts and reduce rejection,and the differences had Statistical significance. Pathological examination revealed that lymphocytic infiltration and neovascularization was less.Conclusion:1.Microemulsion gel and olive oil had mild irritation,while there is almost no irritant in nanoparticles and cubosomes group,which is in line with the requirements of ophthalmic preparations.2.0.5%and 1%CsA-containing cubosomes can reduce the rate of rejection of the rat penetrating keratoplasty model,and extend the survival time of corneal grafts.