Anti-fibrotic effects of rosmarinic acid on Tenon’s capsule fibroblasts stimulated with TGF-β: therapeutic potential in ocular surgery

Author(s)

Carolina Maria Módulo ^[1] ; Larissa Domenegueti Ferreira ^[2] ; Lilian Eslaine Costa Mendes da Silva ^[3] ; Marco Andrey Cipriani Frade ^[4] ; Peter Sol Reinach ^[5] ; Eduardo Melani Rocha ^[6] ; Jayter Silva Paula ^[7]

Affiliation

^[1] Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Otorhinolaryngology and Head and Neck Surgery - Brasil

^[2] Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Otorhinolaryngology and Head and Neck Surgery - Brasil

^[3] Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Otorhinolaryngology and Head and Neck Surgery - Brasil

^[4] Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Department of Clinical Medicine - Brasil

^[5] Wenzhou Medical University. Department of Ophthalmology and Optometry

^[6] Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Otorhinolaryngology and Head and Neck Surgery - Brasil

^[7] Universidade de São Paulo. Faculdade de Medicina de Ribeirão Preto. Otorhinolaryngology and Head and Neck Surgery - Brasil

• Document type: Journal article
• Source: Arquivos Brasileiros de Oftalmologia; v. 83, n. 4, p. 305-311, 2020-07-29.

Abstract

ABSTRACT Purpose: Collagen deposition and myofibroblast differentiation are critical factors related to excessive scarring in ocular surgeries. This study evaluated the anti-fibrotic activity of rosmarinic acid on rabbit Tenon’s capsule fibroblasts stimulated with transforming growth factor- β2. Methods: Primary cultures of rabbit Tenon’s capsule fibroblasts were treated with various concentrations of rosmarinic acid for 12 h, in the presence and absence of transforming growth factor-β2. After 48 h, the proliferation index of rabbit Tenon’s capsule fibroblasts and the differentiation of myofibroblasts were investigated through immunofluorescence staining for proliferating cell nuclear antigen and alpha smooth muscle actin. An automated cell counter and colorimetric metabolic activity assay were used to evaluate cell number and viability. Collagen expression and production were determined by quantitative real-time polymerase chain reaction and hydroxyproline assay, respectively. Results: Unstimulated rabbit Tenon’s capsule fibroblasts treated with any concentration of rosmarinic acid exhibited diminished collagen expression (p<0.01) but showed no differences in proliferation index. Transforming growth factor-β2 exposure induced myofibroblast differentiation and increased collagen production. Exposure to rosmarinic acid at 1.0 and 3.0 µM concentrations reduced the proliferation index (p<0.02), as well as the collagen expression and hydroxyproline content (p<0.05). Exposure to 3.0 µM rosmarinic acid reduced viability (p=0.035) in unstimulated rabbit Tenon’s capsule fibroblasts and cell numbers (p=0.001) in both stimulated and unstimulated rabbit Tenon’s capsule fibroblast cultures. Conclusions: Exposure to 1.0 µM rosmarinic acid was noncytotoxic and led to reduced collagen expression and proliferation of stimulated rabbit Tenon’s capsule fibroblasts. These findings suggest that rosmarinic acid is a relatively non-injurious anti-fibrotic compound to rabbit Tenon’s capsule fibroblasts, with potential application as an adjunctive agent in ocular procedures, particularly in glaucoma surgeries. (AU)

• FAPESP's process: 12/00622-9 - Study of potential modulators of the healing process on the proliferation of rabbits tenon's fibroblasts
• Grantee: Jayter Silva de Paula
• Support Opportunities: Regular Research Grants