Corneal expression levels of the proinflammatory and pro(lymph)angiogenic cytokines IL-1β, TNFα, VEGF-A, VEGF-C, and VEGF-D were analyzed by real-time PCR after suture placement and treatment with serum eyedrops, bevacizumab eyedrops, or a combination of both. After 2 days of treatment, only bevacizumab affected IL-1β expression (serum mean of 95.6%, n.s.; bevacizumab mean of 74.3%,
P < 0.01; and serum and bevacizumab mean of 96.4%, n.s.), whereas after 7 days of treatment, all treatment groups had significantly reduced IL-1β expression (serum mean of 48.4%,
P < 0.001; bevacizumab mean of 58.9%,
P < 0.001; and serum and bevacizumab mean of 53.2%,
P < 0.001) (
Fig. 5, top left). After 2 days of treatment, TNFα expression was significantly impaired only by bevacizumab (serum mean of 106.1%, n.s.; bevacizumab mean of 68.4%,
P < 0.05; and serum and bevacizumab mean of 93.0%, n.s.), while after 7 days of treatment, the combination of serum and bevacizumab also showed considerable inhibition of TNFα expression (serum mean of 107.5%, n.s.; bevacizumab mean of 86.6%,
P < 0.01; and serum and bevacizumab mean of 84.3%,
P < 0.01) (
Fig. 5, top middle). VEGF-A expression was significantly inhibited by bevacizumab both after 2 days and after 7 days of treatment (bevacizumab after 2 days mean of 77.3%,
P < 0.01; and bevacizumab after 7 days mean of 81.9%,
P < 0.01). While exhibiting no effect after 2 days, serum treatment significantly upregulated VEGF-A expression after 7 days of treatment (serum after 2 days mean of 96.3%, n.s.; and serum after 7 days mean of 120.1%,
P < 0.05). The combination of serum and bevacizumab showed no alteration in VEGF-A expression (serum and bevacizumab after 2 days mean of 100.7%, n.s.; and serum and bevacizumab after 7 days mean of 102.8%, n.s.) (
Fig. 5, bottom left). VEGF-C expression was not significantly altered after 2 days of treatment with serum, bevacizumab, or a combination of both, although bevacizumab showed an apparent tendency to lower expression levels (serum mean of 90.5%, n.s.; bevacizumab mean of 82.8%,
P = 0.06; and serum and bevacizumab mean of 95.4%, n.s.). After 7 days, serum had no effect, whereas bevacizumab and the combination of serum and bevacizumab inhibited VEGF-C expression (serum mean of 89.0%, n.s.; bevacizumab mean of 66.2%,
P < 0.01; and serum and bevacizumab mean of 77.1%,
P < 0.01) (
Fig. 5, bottom middle). VEGF-D expression was reduced by bevacizumab both after 2 days and after 7 days (bevacizumab after 2 days mean of 76.3%,
P < 0.01; and bevacizumab after 7 days mean of 82.3%,
P < 0.05). Serum and the combination of serum and bevacizumab did not modify VEGF-D expression after 2 days or after 7 days of treatment (serum after 2 days mean of 95.2%, n.s.; serum and bevacizumab after 2 days mean of 99.8%, n.s.; serum after 7 days mean of 104.7%, n.s.; and serum and bevacizumab after 7 days mean of 96.0%, n.s.) (
Fig. 5, bottom right).