Pirfenidone nanoparticles improve corneal wound healing and prevent scarring following alkali burn.
OBJECTIVE
To evaluate the effects of pirfenidone nanoparticles on corneal re-epithelialization and scarring, major clinical challenges after alkali burn.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Department of Laboratory Animal Facility, CSIR-Indian Institute of Chemical Biology, Kolkata, India.
Sushovan
S
Chowdhury
Rajdeep
R
Guha
Ruchit
R
Trivedi
Uday B
UB
Kompella
Aditya
A
Konar
Sarbani
S
Hazra
D000818:Animals; D003315:Cornea / Q000378:metabolism; D065306:Corneal Injuries; D005126:Eye Burns / Q000188:drug therapy* / Q000378:metabolism; D005260:Female; D007150:Immunohistochemistry; D008297:Male; D053758:Nanoparticles / Q000737:chemistry*; D011728:Pyridones / Q000737:chemistry / Q000627:therapeutic use*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014945:Wound Healing / Q000187:drug effects*
D011728:Pyridones; C093844:pirfenidone