Abstract
(1) Objective: To study the anti-fibrotic effects of Lycium barbarum polysaccharides (LBP) on corneal stromal fibroblasts and assess LBP’s effect on cell viability. (2) Methods: Primary human corneal keratocytes of passage 3 to 6 were used for all experiments. Cells are pretreated with LBP solution for 24 h and then transforming growth factor beta 1 (TGFβ1) for 48 h and collected for experiments. Fibrotic protein analysis was performed using immunofluorescence and Western blot. The effect of LBP on cell viability was assessed using the MTS assay. (3) Results: LBP significantly reduced the expression of fibrotic proteins, including α-SMA and extracellular matrix proteins (collagen type I and III). LBP significantly decreased the viability of myofibroblasts but not the fibroblasts. Conclusions: In this study, LBP was effective in the prevention of fibrosis gene expression. Further studies to assess the underlying mechanism and pharmacological properties will facilitate the formation of a topical LBP solution for in vivo studies.
Highlights
Corneal scarring is the second most common cause of blindness worldwide [1]
Once cells reached 30% confluence, cells were pre-treated with Lycium barbarum polysaccharides (LBP) at 0.1 mg/mL, 0.5 mg/mL, 1 mg/mL, 1.5 mg/mL, 2.0 mg/mL, 2.5 mg/mL or 3.0 mg/mL treated for 24 h and with transforming growth factor-β1 (TGFβ1) (Millipore, GF439, Burlington, MA, USA) at 10 ng/mL for 48 h to differentiate the cells into myofibroblasts
Phase-contrast microscopy demonstrated bipolar-elongated fibroblast-like morphology of the cells (Figure 1)
Summary
Corneal scarring is the second most common cause of blindness worldwide [1]. Definitive treatment of corneal scarring requires corneal transplantation which is limited by low organ donation rates, especially in the developing world. TGFβ1 is a key cytokine in promoting myofibroblast differentiation with increased production of disorganized stromal matrix proteins, including collagen, fibronectin, tenascin, proteoglycans, thrombospondin and tissue inhibitor of metalloprotease-1 (TIMP-1) This can lead to increased light scattering due to development of corneal haze and scarring [7]. TGFβ2 plays a major role in corneal development the epithelial-mesenchymal interactions, corneal homeostasis and repair [8] Current treatments such as topical corticosteroids are associated with potentially blinding complications, such as cataract, glaucoma and cornea-scleral melting [2]. There were no significant adverse effects detected, including raised intraocular pressure and cornea epithelial toxicity [13] Due to their current availability, good safety profiles and relatively low costs, systemic vitamin C and topical rosiglitazone may serve as more viable forms of therapy, compared to gene therapy, in the near future. We conducted a proof-of-concept experiment using primary corneal fibroblasts
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
