Effect of glucocorticoid (triamcinolone acetonide) pretreatment in a murine penetrating keratoplasty and suture model

Abstract

Purpose: To evaluate the effect of glucocorticoid (triamcinolone acetonide injectable suspension) pretreatment on corneal neovascularization, lymphangiogenesis, and inflammation in a murine penetrating keratoplasty (PK) and corneal suture model.

Methods: For the PK model, BALB/c mice were used as recipients and C57BL/6 mice were used as donors. A group pretreated with subconjunctival glucocorticoid and a combination of post-subconjunctival and topical glucocorticoids (group I) was compared with two groups that did not receive glucocorticoid pretreatment [one group received a combination of subconjunctival and topical glucocorticoids postoperatively (group II) and the other group received only topical glucocorticoid treatment postoperatively (group III)]. All groups were treated with subconjunctival glucocorticoid on the day of surgery. For the corneal suture model, BALB/c mice were used. A group receiving only pre-suture glucocorticoid treatment (group A) and a group receiving only post-suture glucocorticoid treatment (group C) were compared with a control group that did not receive glucocorticoid therapy (group B). The degree of neovascularization, lymphangiogenesis, and inflammatory infiltration was compared in each of these models.

Results: In the PK model, the group receiving glucocorticoid pretreatment (group I) showed less neovascularization compared with the posttreatment-only groups (group II, P=0.043; group III, P=0.020) and less lymphangiogenesis compared with group III (P=0.005). In the corneal suture model, the glucocorticoid pretreatment group showed a similar level of neovascularization, lymphangiogenesis, and inflammatory infiltration as the posttreatment-only groups (P>0.05).

Conclusions: Glucocorticoid pretreatment before PK decreases neovascularization and lymphangiogenesis compared with posttransplant glucocorticoid treatment alone.

Figure 1

A. Schedule of glucocorticoid application in the penetrating keratoplasty model. B. Schedule of glucocorticoid application in the corneal suture model.

Figure 2

(A) Kaplan-Meier survival graph of Groups I, II, and III in the PK model. Group I showed improved graft survival compared to Groups II and III, but this did not reach statistical significance (p=0.408, p=0.560). (B) Representative pictures of Group I, II, and III.

Figure 3

(A) & (B) - In the PK model, Group I showed significantly less total neovascularization compared to Groups II and III (p=0.043, p=0.020, respectively) and less total lymphangiogenesis (p=0.005) and less graft lymphangiogenesis (p=0.020) compared to Group III. (C) Representative pictures of Group I, II, and III. Upper row: CD31staining, Lower row: LYVE-1 staining

Figure 4

(A) & (B) In the corneal suture model, Groups A and C showed significantly less neovascularization and lymphangiogenesis compared to Group B (p<0.05). There was no significant difference of neovascularization and lymphangiogenesis between Groups A and C (p>0.05). (C) Representative pictures of Group A, B, and C. Upper row: Fluorescent microscopic picture of CD31 staining (green) in flat mount of whole cornea. Middle row: Fluorescent microscopic picture of LYVE-1 staining (red) in flat mount of whole cornea. Bottom row: Confocal microscopic picture of CD11b staining cells (green) around suture area.