Intra-stromal Injection of Honey-Treated Keratocytes as a Cell-Based Therapy for Experimental Corneal Laceration
Fatemeh SanieJahromi1 , M. Hossein Nowroozzadeh1 *, Zahra Emadi1 , Masoomeh Eghtedari1 , Zohreh Khajehahmadi1
- Poostchi Ophthalmology Research Center, Department of Ophthalmology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Abstract: This study aimed to assess the potential of low concentration honey supplemented medium (HSM) for expansion of human native keratocytes that express the genes involved in the secretion of extracellular matrix needed for corneal transparency. The in-vivo effect of HSM-treated keratocytes was also examined in an experimental model of corneal laceration.
Methods: Human keratocytes were cultured in 1% HSM versus 10% fetal bovine serum (FBS)- supplemented media for 24 h. The effect of HSM on keratocyte proliferation was evaluated using the MTT assay. The relative expression of Lum, Kera, ALDH3A1, and Thy-1 was quantified by real-time PCR as the known markers of native keratocytes and cell proliferation. Finally, we evaluated the safety and efficiency of HSM-treated keratocytes intrastromal injection in a rabbit model of corneal laceration.
Results: MTT assay showed that cell viability was not significantly affected in HSM-treated keratocytes compared to those treated with FBS-supplemented medium (84.71 ± 2.38 vs 100.08 ± 10.92, respectively; P = 0.076). Moreover, the expression of Lum, Kera, and ALDH3A1 was significantly increased in HSM-treated keratocytes compared to cells treated with FBS, while the expression of proliferation biomarkers of Thy-1 was not significantly changed between the two treatments. Intrastromal injection of HSM-treated keratocytes in the laceration animal model was also safe and uneventful and showed less stromal inflammation and neovascularization, and consequently, a better final architecture with less residual haze compared to the group injected with FBS-treated keratocytes.
Conclusion: This study showed a favorable phenotype of HSM-treated keratocytes to be used as a cell-based therapy to improve corneal wound healing. These findings were confirmed in a rabbit model of corneal laceration, in which intrastromal injection of HSM-treated keratocytes was associated with promising clinical outcomes.
