In vitro Effects of Doxorubicin and Mitomycin C on Human Tenon’s Capsule Fibroblasts

Abstract

Doxorubicin is an antiproliferative agent, that also inhibits prolyl 4-hydroxylase in vitro. We evaluated the effects of doxorubicin and mitomycin C (MMC) on the proliferation of human Tenon's capsule fibroblasts and the secretion of type I collagen in these cells to explore the potential use of doxorubicin as an antifibrotic agent after glaucoma filtering surgery. Standard immunoassays were used to determine the concentrations of type I procollagen COOH-terminal peptide (PIP), laminin and vitronectin receptor in conditioned media and cell lysates in the presence or absence of doxorubicin or MMC. The mitotic activity and viability of these cells were also determined. Cellular ultrastructure was evaluated by transmission electron microscopy. Doxorubicin and MMC decreased the cellular viability, the mitotic activity, and the production of the peptides measured. The inhibition of PIP secretion into the culture medium was higher in doxorubicin-treated cultures than in MMC-treated cultures at the tested concentrations (5-100 microM). The decrease in PIP levels in cell lysates was less in doxorubicin-treated culture (25 microM) than in MMC-treated culture (25 microM), suggesting that procollagen synthesis and secretion might be attenuated by doxorubicin. Ultrastructural studies revealed increased numbers of lysosomes in the cytoplasm of doxorubicin-treated cells relative to MMC-treated cells. Doxorubicin and MMC reduced cell viability and inhibited the proliferation and synthesis of PIP, laminin, and vitronectin receptor peptide. Inhibitory effects of doxorubicin on PIP secretion were more potent than those of MMC. Doxorubicin may be useful for inhibiting the fibrotic response at the site of ocular filtering surgery.