Corneal chemical burns (CCBs) frequently result in corneal fibrosis or haze, an opacity of the cornea that obstructs vision and induces corneal blindness. Diverse strategies have been employed to prevent or reduce CCB-related corneal haze. In this study, we evaluated the physicochemical characteristics and biologic effects of a topical pirfenidone (PFD)-loaded liposomal formulation (PL) on a corneal alkali burn mice model. We found that PL was appropriate for ocular application due to its physiologic tear pH, osmolarity and viscosity suitable for topical ophthalmic use. Regarding its therapeutic activity, PL-treated mice had significantly reduced haze size and density, corneal edema, corneal thickness, and corneal inflammatory infiltration, in contrast to PFD in aqueous solution (p < 0.01). Importantly, the antifibrotic activity of PL (reduction of corneal haze) was associated with modulation of transforming growth factor (TGF)-β and Interleukin (IL)-1β genes. PL suppressed TGF-β expression and restored normal IL-1β expression in corneal tissue more efficiently in contrast to PFD in aqueous solution. In conclusion, PFD showed essential anti-inflammatory and anti-fibrotic effects in the treatment of alkali burns. Noteworthy, a new formulation of PFD-loaded liposomes remarkably improved these effects, standing out as a promising treatment for corneal haze.
The harvesting of corneal endothelial cells (CEC) has received special attention due to its potential as a therapy for corneal blindness. The main challenges are related to the culture media formulation, cellular density at the primary isolation, and the number of passages in which CEC can retain their functional characteristics. To alternate different media formulations to harvest CEC has an impact on the cellular yield and morphology. Therefore, we analyzed four different sequences of growth factor-supplemented Stimulatory (S) and non-supplemented Quiescent (Q) media, upon passages to find the optimal S-Q culture sequence. We assessed cell yield, morphology, procollagen I production, Na + /K + -ATPase function, and the expression of ZO-1 and Na + /K + -ATPase. Our results show SQSQ and SQQQ sequences with a balance between an improved cell yield and hexagonal morphology rate. CEC cultured in the SQQQ sequence produced procollagen I, showed Na + /K + -ATPase function, and expression of ZO-1 and Na + /K + -ATPase. Our study sets a culture approach to guarantee CEC expansion, as well as functionality for their potential use in tissue engineering and in vivo analyses. Thus, the alternation of S and Q media improves CEC culture. SQQQ sequence demonstrated CEC proliferation and lower the cost implied in SQSQ sequences. We discarded the use of pituitary extract and ROCK inhibitors as essential for CEC proliferation.
Background The harvesting of corneal endothelial cells (CECs) has received special attention given its potential as therapy for corneal blindness. The main challenges to overcome for this purpose are related to the culture media formulation, cellular density at the primary isolation, and the number of passages in which CECs can retain their functional characteristics. The alternance of different media formulations to harvest CECs has an impact on the cellular yield and morphology. We herein analyzed eight different sequences of growth factor-supplemented proliferative (P) and non-supplemented resting (R) media upon passages to find the optimal P/R culture media sequence in regards of cell yield, morphology, procollagen I production, ATPase function, and the expression of ZO-1 and ATPase. Results PRPR and PRRR sequences showed the higher cell yield and hexagonal morphology rate. CECs cultured in the PRRR sequence produced procollagen I, showed Na/K-ATPase function, and expression of ZO-1 and Na/K-ATPase by immunocytochemistry. Our study sets a culture approach to guarantee CECs expansion, as well as functionality for their potential use in tissue engineering and in vivo analyses. Conclusions Alternation of P and R culture media improves CECs culture. PRRR sequence demonstrated to be effective and for CECs proliferation lowering the cost implied in PRPR sequences. We discarded the use of pituitary extract and ROCK inhibitors as essential for CECs proliferation.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.