Pirfenidone nanoparticles improve corneal wound healing and prevent scarring following alkali burn

• Published in: PloS one Vol. 8; no. 8; p. e70528
• Main Authors: Chowdhury, Sushovan, Guha, Rajdeep, Trivedi, Ruchit, Kompella, Uday B, Konar, Aditya, Hazra, Sarbani
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 05.08.2013; Public Library of Science (PLoS)
• Subjects: Actin; Animals; Biology; Collagen; Collagen (type I); Cornea; Cornea - metabolism; Corneal Injuries; Cytokines; Eye Burns - drug therapy; Eye Burns - metabolism; Eye diseases; Female; Fibroblasts; Fibrosis; Fishery sciences; Fluorescence; Fluorescence microscopy; Glaucoma; Growth factors; Immunoblotting; Immunocytochemistry; Immunohistochemistry; Laboratory animals; Male; Materials Science; Medicine; Microscopy; Morphology; Muscle proteins; Muscles; Nanoparticles; Nanoparticles - chemistry; Pharmaceutical sciences; Polylactide-co-glycolide; Polyvinyl alcohol; Pulmonary fibrosis; Pyridones - chemistry; Pyridones - therapeutic use; Rats; Rats, Sprague-Dawley; Scars; Smooth muscle; Surgery; Synthesis; Transforming growth factors; Wound healing; Wound Healing - drug effects; United States > US; Colorado; India
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0070528

To evaluate the effects of pirfenidone nanoparticles on corneal re-epithelialization and scarring, major clinical challenges after alkali burn. Effect of pirfenidone on collagen I and α-smooth muscle actin (α-SMA) synthesis by TGFβ induced primary corneal fibroblast cells was evaluated by immunoblotting and immunocytochemistry. Pirfenidone loaded poly (lactide-co-glycolide) (PLGA) nanoparticles were prepared, characterized and their cellular entry was examined in primary corneal fibroblast cells by fluorescence microscopy. Alkali burn was induced in one eye of Sprague Dawley rats followed by daily topical treatment with free pirfenidone, pirfenidone nanoparticles or vehicle. Corneal re-epithelialization was assessed daily by flourescein dye test; absence of stained area indicated complete re-epithelialization and the time for complete re-epithelialization was determined. Corneal haze was assessed daily for 7 days under slit lamp microscope and graded using a standard method. After 7 days, collagen I deposition in the superficial layer of cornea was examined by immunohistochemistry. Pirfenidone prevented (P<0.05) increase in TGF β induced collagen I and α-SMA synthesis by corneal fibroblasts in a dose dependent manner. Pirfenidone could be loaded successfully within PLGA nanoparticles, which entered the corneal fibroblasts within 5 minutes. Pirfenidone nanoparticles but not free pirfenidone significantly (P<0.05) reduced collagen I level, corneal haze and the time for corneal re-epithelialization following alkali burn. Pirfenidone decreases collagen synthesis and prevents myofibroblast formation. Pirfenidone nanoparticles improve corneal wound healing and prevent fibrosis. Pirfenidone nanoparticles are of potential value in treating corneal chemical burns and other corneal fibrotic diseases.